Compositions and methods of treating lupus nephritis

Abstract

The present disclosure provides, inter alia, methods for depleting kidney-resident B cells in an individual that has lupus nephritis, comprising administering obinutuzumab to the individual in three antibody exposures, as well as uses, compositions for use, and kits related thereto. In some embodiments, the methods result in an average of less than one B cell per mm2 tissue, a 95% or greater reduction in average number of B cells per mm2 tissue, and / or an activity index (Al) of less than or equal to 1 in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure.

Description

Attorney Docket No.14639-20729.40COMPOSITIONS AND METHODS OF TREATING LUPUS NEPHRITISCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority benefit of U.S. Provisional Application No.63 / 699,081, filed on September 25, 2024; U.S. Provisional Application No.63 / 754,441, filed on February 5, 2025; and International Application No. PCT / US2025 / 025954, filed on April 23, 2025, each of which is hereby incorporated by reference in its entirety.REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (146392072940SEQLIST.xml; Size: 41,776 bytes; and Date of Creation: September 18, 2025) are incorporated herein by reference in their entirety.FIELD OF THE INVENTION

[0003] Provided herein are methods for depleting kidney-resident B cells in an individual that has lupus nephritis, comprising administering obinutuzumab to the individual in three antibody exposures, as well as uses, compositions for use, and kits related thereto.BACKGROUND

[0004] Proliferative lupus nephritis is the most common organ-threatening manifestation of systemic lupus erythematosus. Glomerular injury and tubulointerstitial inflammation result in proteinuria, hematuria, and progressive renal impairment. Goals of treatment include reduction in proteinuria, prevention of renal damage, and minimization of toxicities of immunosuppressive therapies. Hahn et al., Arthritis Care and Research , 2012; Fanouriakis et al., Ann. Rheum. Dis.78:736-45, 2019. Even with treatment, many patients have a poor outcome such as the development of end-stage renal disease (ESRD), need for hemodialysis or renal transplantation, or death, and the risk of ESRD has not substantially improved during the last twenty years. Hanly, et al., Rheumatology 55(2):252-62, 2016; Tektonidou et al., Arthritis Rheumatol 68(6):1432-1441, 2016). There are currently no therapies approved for the treatment1MOFO-360876942Attorney Docket No.14639-20729.40of lupus nephritis in the United States. Current non-approved, standard of care treatments are associated with toxicities and low rates of complete response.

[0005] Two anti-CD20 antibodies have been tested in clinical studies for efficacy in treating lupus nephritis. Rituximab, a type I anti-CD20 antibody, depleted peripheral CD19+ B cells in 71 of 72 patients and led to more responders and greater reductions in anti-dsDNA and C3 / C4 levels in a clinical study (LUNAR). Dose regimen for the LUNAR study consisted of administration to patients with class III or class IV lupus nephritis (LN), rituximab (1,000 mg) or placebo on days 1, 15, 168, and 182 (week 0, 2, 24, 26). However, rituximab therapy did not improve clinical outcomes after 1 year of treatment.

[0006] The efficacy and safety of obinutuzumab in lupus nephritis have now been confirmed in the Phase II NOBILITY (NCT02550652) and the Phase III REGENCY (NCT04221477) trials (Furie RA, et al. Ann Rheum Dis.2022;81:100-107; Furie RA, et al. N Engl J Med.2025;392:1471-1483). In the REGENCY trial, the primary endpoint of complete renal response at Week 76 was achieved by 46.4% of patients treated with obinutuzumab plus standard therapy compared with 33.1% of patients treated with placebo plus standard therapy (adjusted difference 13.4%; 95% CI, 2.0 to 24.8; P=0.02; Furie RA, et al. N Engl J Med.2025;392:1471-1483).

[0007] Persistent kidney inflammation, driven by tissue-resident immune cells, remains a significant challenge in lupus nephritis, frequently leading to irreversible damage despite existing therapies. Demonstration of a direct therapeutic impact on kidney tissue-resident B cell populations, which are central to local inflammation and autoantibody production, has remained an unmet need.

[0008] All references cited herein, including patent applications and publications, are incorporated by reference in their entirety.SUMMARY

[0009] In certain aspects, provided herein is a method for depleting kidney-resident B cells in an individual in need thereof (e.g., an individual having lupus, or an individual having lupus nephritis), comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not2MOFO-360876942Attorney Docket No.14639-20729.40provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human; and wherein treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure. In some embodiments, kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure have an average of at least 10, at least 12, at least 13, or at least 15 B cells per mm2tissue. In some embodiments, treatment of the individual with obinutuzumab results in less than one B cell per mm2tissue in a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure. In some embodiments, a kidney biopsy sample obtained from the individual prior to treatment with obinutuzumab has at least 10, at least 15, at least 25, at least 50, at least 100, or at least 200 B cells per mm2tissue.

[0010] In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between3MOFO-360876942Attorney Docket No.14639-20729.40about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human; and wherein treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure. In some embodiments, kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure have an average of at least 10, at least 12, at least 13, or at least 15 B cells per mm2tissue. In some embodiments, treatment of the individual with obinutuzumab results in less than one B cell per mm2tissue in a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure. In some embodiments, a kidney biopsy sample obtained from the individual prior to treatment with obinutuzumab has at least 10, at least 15, at least 25, at least 50, at least 100, or at least 200 B cells per mm2tissue.

[0011] In other aspects, provided herein is a method for depleting kidney-resident B cells in an individual in need thereof (e.g., an individual having lupus, or an individual having lupus nephritis), comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human; and wherein treatment with obinutuzumab in a plurality of individuals results in a 90% or greater, 91% or greater, 92% or greater, 93% or greater, 94% or greater, 95% or greater, 96% or greater, 97% or greater, or 98% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to an average number of B cells per mm2tissue in kidney4MOFO-360876942Attorney Docket No.14639-20729.40biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure. In some embodiments, treatment of the individual with obinutuzumab results in a 90% or greater, 91% or greater, 92% or greater, 93% or greater, 94% or greater, 95% or greater, 96% or greater, 97% or greater, or 98% or greater reduction in number of B cells per mm2tissue in a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to a number of B cells per mm2tissue in a kidney biopsy sample obtained from the individual prior to treatment with obinutuzumab.

[0012] In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human; and wherein treatment with obinutuzumab in a plurality of individuals results in a 90% or greater, 91% or greater, 92% or greater, 93% or greater, 94% or greater, 95% or greater, 96% or greater, 97% or greater, or 98% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to an average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure. In some embodiments, treatment of the individual with obinutuzumab results in a 90% or greater, 91% or greater, 92% or greater, 93% or greater, 94% or greater, 95% or greater, 96% or greater, 97% or greater, or 98% or greater reduction in number of B cells per mm2tissue in a kidney biopsy sample obtained from the individual at about 76 weeks to about 805MOFO-360876942Attorney Docket No.14639-20729.40weeks after administration of the first antibody exposure, as compared to a number of B cells per mm2tissue in a kidney biopsy sample obtained from the individual prior to treatment with obinutuzumab.

[0013] In other aspects, provided herein is a method for depleting and / or reducing activity of kidney-resident B cells in an individual in need thereof (e.g., an individual having lupus, or an individual having lupus nephritis), comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human; and wherein a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1. In some embodiments, a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0. In some embodiments, a kidney biopsy sample obtained from the individual prior to administration of the first antibody exposure exhibits an AI of greater than or equal to 4 or greater than or equal to 6.

[0014] In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the6MOFO-360876942Attorney Docket No.14639-20729.40first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human; and wherein a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1. In some embodiments, a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0. In some embodiments, a kidney biopsy sample obtained from the individual prior to administration of the first antibody exposure exhibits an AI of greater than or equal to 4 or greater than or equal to 6.

[0015] In some embodiments according to any one of the embodiments disclosed herein, the kidney-resident B cells are CD79a+ / CD138- B cells. In some embodiments, prior to administration of the first antibody exposure, the individual has one or more of the following: (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female.

[0016] In certain aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to a type II anti-CD20 antibody and a second antibody exposure to the type II anti-CD20 antibody; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure; wherein the first antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; wherein the second antibody exposure comprises one or two doses of the type II7MOFO-360876942Attorney Docket No.14639-20729.40anti-CD20 antibody, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab and a second antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; and wherein, prior to administration of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g.

[0017] In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering intravenously to the individual a first and a second antibody exposure to a type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.08MOFO-360876942Attorney Docket No.14639-20729.40g / g. In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering intravenously to the individual a first, a second, and a third antibody exposure to a type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises one dose of 1000mg of the type II anti-CD20 antibody on week 52 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering intravenously to the individual a first, a second, and a third antibody exposure to a type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 50 and 52 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In some embodiments, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 3.0 g / g.9MOFO-360876942Attorney Docket No.14639-20729.40

[0018] In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; and wherein the individual has or is one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female.

[0019] In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising intravenously administering to the individual a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab in combination with mycophenolate mofetil (MMF); wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab administered on week 52 of treatment; and wherein the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (b) an estimated10MOFO-360876942Attorney Docket No.14639-20729.40glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; (h) is female.

[0020] In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising intravenously administering to the individual a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab in combination with mycophenolate mofetil (MMF); wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab administered on weeks 50 and 52 of treatment; and wherein the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; (h) is female.

[0021] In other aspects, provided herein is a method for reducing risk of lupus nephritis flare in an individual that has lupus, comprising intravenously administering to the individual a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab in combination with mycophenolate mofetil (MMF) and a glucocorticoid; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab administered on week 52 of treatment; and wherein the administration of obinutuzumab in combination with MMF and a glucocorticoid to a plurality of individuals11MOFO-360876942Attorney Docket No.14639-20729.40results in a reduction in risk of lupus nephritis flare by about 51% to about 61% at week 76 of treatment, as compared to risk of lupus nephritis flare in a plurality of individuals receiving MMF and the glucocorticoid in the absence of obinutuzumab. In other aspects, provided herein is a method for reducing risk of lupus nephritis flare in an individual that has lupus, comprising intravenously administering to the individual a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab in combination with mycophenolate mofetil (MMF) and a glucocorticoid; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab administered on weeks 50 and 52 of treatment; and wherein the administration of obinutuzumab in combination with MMF and a glucocorticoid to a plurality of individuals results in a reduction in risk of lupus nephritis flare by about 51% to about 61% at week 76 of treatment, as compared to risk of lupus nephritis flare in a plurality of individuals receiving MMF and the glucocorticoid in the absence of obinutuzumab. In some embodiments, the administration of obinutuzumab in combination with MMF and a glucocorticoid to a plurality of individuals results in a reduction in risk of lupus nephritis flare by 56% at week 76 of treatment, as compared to risk of lupus nephritis flare in a plurality of individuals receiving MMF and the glucocorticoid in the absence of obinutuzumab. In some embodiments, the lupus nephritis flare is defined as eGFR decrease >20% compared with Week 24 in patients with UPCR >1 g / g and / or cellular casts; UPCR increase (i) to >1 g / g if Week 24 UPCR was <0.2 g / g, (ii) to >2 g / g if Week 24 UPCR was 0.2-1 g / g or (iii) to doubling if Week 24 UPCR was >1 g / g; receipt of rescue therapy, except for corticosteroid-only rescue.

[0022] In other aspects, provided herein is a method for reducing risk of unfavorable kidney outcome in an individual that has lupus, comprising intravenously administering to the individual a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab in combination with mycophenolate mofetil (MMF) and a glucocorticoid; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of12MOFO-360876942Attorney Docket No.14639-20729.40obinutuzumab administered on week 52 of treatment; and wherein the administration of obinutuzumab in combination with MMF and a glucocorticoid to a plurality of individuals results in a reduction in risk of unfavorable kidney outcome by about 58% to about 68% at week 76 of treatment, as compared to risk of unfavorable kidney outcome in a plurality of individuals receiving MMF and the glucocorticoid in the absence of obinutuzumab. In other aspects, provided herein is a method for reducing risk of unfavorable kidney outcome in an individual that has lupus, comprising intravenously administering to the individual a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab in combination with mycophenolate mofetil (MMF) and a glucocorticoid; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab administered on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab administered on weeks 50 and 52 of treatment; and wherein the administration of obinutuzumab in combination with MMF and a glucocorticoid to a plurality of individuals results in a reduction in risk of unfavorable kidney outcome by about 58% to about 68% at week 76 of treatment, as compared to risk of unfavorable kidney outcome in a plurality of individuals receiving MMF and the glucocorticoid in the absence of obinutuzumab. In some embodiments, the administration of obinutuzumab in combination with MMF and a glucocorticoid to a plurality of individuals results in a reduction in risk of unfavorable kidney outcome by 63% at week 76 of treatment, as compared to risk of unfavorable kidney outcome in a plurality of individuals receiving MMF and the glucocorticoid in the absence of obinutuzumab. In some embodiments, the unfavorable kidney outcome is defined as the first of the following events: death, doubling of serum creatinine or treatment failure.

[0023] In some embodiments according to any one of the embodiments disclosed herein, prior to administration of the first antibody exposure, the individual has a UPCR of greater than or equal to about 2.0 g / g. In some embodiments, the individual has a baseline UPCR of greater than or equal to about 2.0 g / g. In some embodiments, prior to administration of the first antibody exposure, the individual has a UPCR of greater than or equal to about 3.0 g / g. In some embodiments, the individual has a baseline UPCR of greater than or equal to about 3.0 g / g. In some embodiments, prior to administration of the first antibody exposure, the individual has an13MOFO-360876942Attorney Docket No.14639-20729.40eGFR that is less than or equal to 90 mL / min / 1.73m2. In some embodiments, the individual has a baseline eGFR that is less than or equal to 90 mL / min / 1.73m2. In some embodiments, after administration of the third antibody exposure, the individual has: an eGFR that is greater than or equal to 85% of an eGFR of the individual prior to administration of the first antibody exposure; and / or a UPCR of less than or equal to 0.5 g / g. In some embodiments, after administration of the third antibody exposure, the individual has: an eGFR that is greater than or equal to 85% of an eGFR of the individual prior to administration of the first antibody exposure; and a UPCR of less than or equal to 0.5 g / g. In some embodiments, at Week 76 of treatment (e.g., at about 76 weeks after administration of a first dose of the first antibody exposure), the individual has: an eGFR that is greater than or equal to 85% of an eGFR of the individual prior to administration of the first antibody exposure; and / or a UPCR of less than or equal to 0.5 g / g. In some embodiments, at Week 76 of treatment (e.g., at about 76 weeks after administration of a first dose of the first antibody exposure), the individual has: an eGFR that is greater than or equal to 85% of an eGFR of the individual prior to administration of the first antibody exposure; and a UPCR of less than or equal to 0.5 g / g. In some embodiments, prior to administration of the first antibody exposure, a serum sample obtained from the individual prior to administration of the first antibody exposure is positive for anti-dsDNA antibodies, e.g., having anti-dsDNA antibodies present at greater than about 120 kU / L. In some embodiments, a baseline serum sample obtained from the individual is positive for anti-dsDNA antibodies, e.g., having anti-dsDNA antibodies present at greater than about 120 kU / L. In some embodiments, prior to administration of the first antibody exposure, a serum sample obtained from the individual prior to administration of the first antibody exposure has a low C3 complement level, e.g., having C3 complement present at less than about 0.9 g / L. In some embodiments, a baseline serum sample obtained from the individual has a low C3 complement level, e.g., having C3 complement present at less than about 0.9 g / L. In some embodiments, prior to administration of the first antibody exposure, a serum sample obtained from the individual prior to administration of the first antibody exposure has a low C4 complement level, e.g., having C4 complement present at less than about 0.1 g / L. In some embodiments, a baseline serum sample obtained from the individual has a low C4 complement level, e.g., having C4 complement present at less than about 0.1 g / L. In some embodiments, prior to treatment, the individual has no prior history or diagnosis of lupus nephritis. In some embodiments, prior to administration of the first antibody exposure, the individual has no prior14MOFO-360876942Attorney Docket No.14639-20729.40history or diagnosis of lupus nephritis. In some embodiments, prior to treatment, the individual has no prior history or diagnosis of lupus (SLE). In some embodiments, prior to administration of the first antibody exposure, the individual has no prior history or diagnosis of lupus (SLE).

[0024] In some embodiments according to any of the embodiments described herein, the methods further comprise (e.g., prior to administration of the first antibody exposure) identifying the individual as having a UPCR of greater than or equal to about 2.0 g / g. In some embodiments, the methods further comprise (e.g., prior to administration of the first antibody exposure) determining that the individual has a UPCR of greater than or equal to about 2.0 g / g. In some embodiments, the identification and / or determination comprise(s) measuring UPCR in a urine sample obtained from the individual. In some embodiments, the methods further comprise obtaining a urine sample from the individual.

[0025] In some embodiments according to any of the embodiments described herein, prior to administration of the first antibody exposure, the individual has a UPCR of greater than or equal to about 3.0 g / g. In some embodiments, the methods further comprise (e.g., prior to administration of the first antibody exposure, such as prior to a first dose of the first antibody exposure) identifying the individual as having a UPCR of greater than or equal to about 3.0 g / g. In some embodiments, the methods further comprise (e.g., prior to administration of the first antibody exposure, such as prior to a first dose of the first antibody exposure) determining that the individual has a UPCR of greater than or equal to about 3.0 g / g. In some embodiments, the identification and / or determination comprises measuring UPCR in a urine sample obtained from the individual. In some embodiments, the methods further comprise obtaining a urine sample from the individual.

[0026] In some embodiments according to any one of the embodiments disclosed herein, after administration of the third antibody exposure, the individual has a UPCR of less than 0.8 g / g. In some embodiments, the individual has a UPCR of less than 0.8 g / g at about 76 weeks after administration of a first dose of the first antibody exposure. In some embodiments, the method results in the individual having a UPCR of less than 0.8 g / g, less than or equal to about 0.7 g / g, or less than or equal to about 0.5 g / g. In some embodiments, the methods result in preventing or delaying progression to end-stage kidney disease, e.g., in the treated individual. In some embodiments, the method results in a complete renal response at week 76 of treatment. In some embodiments, administering such treatment to a plurality of human patients with lupus nephritis15MOFO-360876942Attorney Docket No.14639-20729.40results in an improvement in complete renal response at week 76 of treatment, e.g., as compared to treatment of a plurality of human patients with lupus nephritis with standard-of-care treatment. In some embodiments, administering such treatment to a plurality of human patients with lupus nephritis results in at least 46% of treated individuals (e.g., of the plurality) achieving a complete renal response at week 76 of treatment.

[0027] In some embodiments according to any of the embodiments described herein, after administration of the second antibody exposure, the individual has a UPCR of less than or equal to about 0.7 g / g. In some embodiments, the methods result in a UPCR of less than or equal to about 0.7 g / g in the individual. In some embodiments, the individual has a UPCR of less than or equal to about 0.7 g / g at about 52 weeks, at about 76 weeks, and / or at about 104 weeks after administration of a first dose of the first antibody exposure. In some embodiments, after administration of the second antibody exposure, the individual has a UPCR of less than or equal to about 0.5 g / g. In some embodiments, the methods result in a UPCR of less than or equal to about 0.5 g / g in the individual. In some embodiments, the individual has a UPCR of less than or equal to about 0.5 g / g at about 52 weeks, at about 76 weeks, and / or at about 104 weeks after administration of a first dose of the first antibody exposure. In some embodiments, after administration of the second antibody exposure, the individual has an estimated glomerular filtration rate (eGFR) that is greater than or equal to 80% of an eGFR of the individual prior to administration of the first antibody exposure; and / or an eGFR that is greater than or equal to about 60 mL / minute. In some embodiments, the methods result in an eGFR that is greater than or equal to 80% of an eGFR of the individual prior to administration of the first antibody exposure; and / or an eGFR that is greater than or equal to about 60 mL / minute. In some embodiments, the eGFR of the individual is measured at about 52 weeks, at about 76 weeks, or at about 104 weeks after administration of the first antibody exposure (e.g., after administration of a first dose of the first antibody exposure).

[0028] In some embodiments according to any of the embodiments described herein, the first antibody exposure comprises a first dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody and a second dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody. In some embodiments, the first dose of the first antibody exposure is about 1000mg of the type II anti-CD20 antibody. In some embodiments, the second dose of the first antibody exposure is about 1000mg of the type II anti-CD20 antibody. In some16MOFO-360876942Attorney Docket No.14639-20729.40embodiments, the first antibody exposure comprises a first dose of the type II anti-CD20 antibody and a second dose of the type II anti-CD20 antibody, and the second dose of the first antibody exposure is not provided until from about 1.5 weeks to about 2.5 weeks after the first dose of the first antibody exposure. In some embodiments, the first antibody exposure comprises a first dose of the type II anti-CD20 antibody and a second dose of the type II anti-CD20 antibody, and the second dose of the first antibody exposure is not provided until about 2 weeks after the first dose of the first antibody exposure. In some embodiments, the second antibody exposure comprises a first dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody and a second dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody. In some embodiments, the first dose of the second antibody exposure is about 1000mg of the type II anti-CD20 antibody. In some embodiments, the second dose of the second antibody exposure is about 1000mg of the type II anti-CD20 antibody. In some embodiments, the second antibody exposure comprises a first dose of the type II anti-CD20 antibody and a second dose of the type II anti-CD20 antibody, and the second dose of the second antibody exposure is not provided until from about 1.5 weeks to about 2.5 weeks after the first dose of the second antibody exposure. In some embodiments, the second antibody exposure comprises a first dose of the type II anti-CD20 antibody and a second dose of the type II anti-CD20 antibody, and the second dose of the second antibody exposure is not provided until about 2 weeks after the first dose of the second antibody exposure. In some embodiments, the first antibody exposure comprises a first dose of about 1000mg of obinutuzumab and a second dose of about 1000mg of obinutuzumab. In some embodiments, the first antibody exposure comprises a first dose of obinutuzumab and a second dose of obinutuzumab, and wherein the second dose of the first antibody exposure is not provided until about 2 weeks after the first dose of the first antibody exposure. In some embodiments, the second antibody exposure comprises a first dose of about 1000mg of obinutuzumab and a second dose of about 1000mg of obinutuzumab. In some embodiments, the second antibody exposure comprises a first dose of obinutuzumab and a second dose of obinutuzumab, and wherein the second dose of the second antibody exposure is not provided until about 2 weeks after the first dose of the second antibody exposure.

[0029] In some embodiments according to any of the embodiments described herein, the methods further comprise administering to the individual a third antibody exposure to the type II anti-CD20 antibody, wherein the third antibody exposure comprises one or two doses of the type17MOFO-360876942Attorney Docket No.14639-20729.40II anti-CD20 antibody, and wherein the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure. In some embodiments, the third antibody exposure comprises a single dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody. In some embodiments, the single dose of the third antibody exposure is about 1000mg of the type II anti-CD20 antibody. In some embodiments, the single dose of the third antibody exposure is not provided until about 52 weeks after the first dose of the first antibody exposure or until about 28 weeks after the first dose of the second antibody exposure. In some embodiments, the third antibody exposure comprises two doses of between about 900mg and about 1100mg of the type II anti-CD20 antibody. In some embodiments, each of the two doses of the third antibody exposure is about 1000mg of the type II anti-CD20 antibody. In some embodiments, the first dose of the third antibody exposure is not provided until about 50 weeks after the first dose of the first antibody exposure or until about 26 weeks after the first dose of the second antibody exposure, and the second dose of the third antibody exposure is not provided until about 52 weeks after the first dose of the first antibody exposure or until about 28 weeks after the first dose of the second antibody exposure. In some embodiments, the methods further comprise administering to the individual a third antibody exposure to obinutuzumab, wherein the third antibody exposure comprises one or two doses of obinutuzumab, and wherein the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure. In some embodiments, the third antibody exposure comprises a single dose of about 1000mg of obinutuzumab. In some embodiments, the single dose of the third antibody exposure is not provided until about 52 weeks after the first dose of the first antibody exposure or until about 28 weeks after the first dose of the second antibody exposure. In some embodiments, the third antibody exposure comprises two doses of about 1000mg of obinutuzumab. In some embodiments, the first dose of the third antibody exposure is not provided until about 50 weeks after the first dose of the first antibody exposure or until about 26 weeks after the first dose of the second antibody exposure, and wherein the second dose of the third antibody exposure is not provided until about 52 weeks after the first dose of the first antibody exposure or until about 28 weeks after the first dose of the second antibody exposure. In some embodiments, the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; the second antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; and the third18MOFO-360876942Attorney Docket No.14639-20729.40antibody exposure comprises one dose of 1000 mg of obinutuzumab on week 52 of treatment. In some embodiments, the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab on weeks 50 and 52 of treatment. In some embodiments, the first antibody exposure comprises two doses of 1000mg of obinutuzumab on days 1 and 15 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on days 168 and 182 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab on day 364 of treatment. In some embodiments, the first antibody exposure comprises two doses of 1000mg of obinutuzumab on days 1 and 15 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on days 168 and 182 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab on days 350 and 364 of treatment.

[0030] In some embodiments, after administration of the third antibody exposure (e.g., after the single dose of the third antibody exposure, or after the 2nddose of the third antibody exposure), the individual has a UPCR of less than or equal to about 0.7 g / g. In some embodiments, after administration of the third antibody exposure (e.g., after the single dose of the third antibody exposure, or after the 2nddose of the third antibody exposure), the individual has a UPCR of less than or equal to about 0.5 g / g. In some embodiments, after administration of the third antibody exposure (e.g., after the single dose of the third antibody exposure, or after the 2nddose of the third antibody exposure), the individual has an estimated glomerular filtration rate (eGFR) that is greater than or equal to 80% of an eGFR of the individual prior to administration of the first antibody exposure; and / or an eGFR that is greater than or equal to about 60 mL / minute.

[0031] In some embodiments according to any of the embodiments described herein, the individual has lupus nephritis. In some embodiments, the individual has class III or class IV lupus nephritis. In some embodiments, the individual has class III (C) or class IV (C) lupus nephritis. In some embodiments, the individual has concomitant class V lupus nephritis. In some embodiments, the individual has active lupus nephritis. In some embodiments, prior to administration of the first antibody exposure, the individual has active lupus nephritis. In some19MOFO-360876942Attorney Docket No.14639-20729.40embodiments, prior to treatment, the individual has active lupus nephritis. In some embodiments, the individual is at risk for developing class III or class IV lupus nephritis. In some embodiments, the individual is a human. In some embodiments, the individual is a woman. In some embodiments, the individual is female. In some embodiments, prior to administration of the first antibody exposure, the individual with lupus nephritis has active disease. In some embodiments, prior to treatment, the individual with lupus nephritis has active disease.

[0032] In some embodiments according to any of the embodiments described herein, the methods further comprise administering to the individual an effective amount of a standard of care treatment. In some embodiments, the standard of care treatment comprises treatment with one or more of an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin-receptor blocker, cyclophosphamide, azathioprine, a glucocorticoid or corticosteroid, and mycophenolic acid or a derivative or salt thereof. In some embodiments, the methods further comprise administering to the individual an effective amount of an immunosuppressive agent. In some embodiments, the immunosuppressive agent comprises mycophenolic acid, a derivative thereof, or a salt thereof. In some embodiments, the immunosuppressive agent or standard of care treatment comprises mycophenolate mofetil (MMF). In some embodiments, the methods comprise administering to the individual an effective amount of mycophenolate mofetil (MMF). In some embodiments, MMF is administered to the individual at a target dose of about 2.0g / day to about 2.5g / day. In some embodiments, MMF is administered to the individual at a dose of 1200mg / m2 / day in divided doses with a maximum of 2.5g / day. In some embodiments, the immunosuppressive agent or standard of care treatment comprises an effective amount of a glucocorticoid or corticosteroid. In some embodiments, the methods further comprise administering to the individual an effective amount of a glucocorticoid or corticosteroid. In some embodiments, the glucocorticoid or corticosteroid comprises prednisone (e.g., oral prednisone). In some embodiments, the methods comprise administering to the individual about 0.5mg / kg / day oral prednisone, with a maximum of about 60mg / day and tapered to about 5mg / day beginning at Week 2. In some embodiments, the glucocorticoid or corticosteroid comprises methylprednisolone. In some embodiments, the methods comprise administering to the individual methylprednisolone by intravenous infusion prior to one or more dose(s) (e.g., each dose) of the type II anti-CD20 antibody (e.g., obinutuzumab), e.g., at weeks 0, 2, 24, 26,20MOFO-360876942Attorney Docket No.14639-20729.40and 52 or at weeks 0, 2, 24, 26, 50, and 52 of treatment. In some embodiments, the methods comprise intravenously administering 80mg methylprednisolone to the individual prior to one or more dose(s) of obinutuzumab. In some embodiments, the glucocorticoid or corticosteroid comprises prednisone. In some embodiments, the methods further comprise administering to the individual an effective amount of oral prednisone. In some embodiments, oral prednisone is administered to the individual at a dose of 0.5-2mg / kg / day with a maximum of 60mg / day. In some embodiments, oral prednisone is administered to the individual at a dose of 0.5-2mg / kg / day until week 2, then tapered to a dose of 5mg / day by week 24 of treatment. In some embodiments, oral prednisone is tapered to a dose of less than or equal to 7.5mg / day from Week 64 through Week 76 of treatment. In some embodiments, the methods further comprise administering to the individual an effective amount of a glucocorticoid or corticosteroid prior to one or more dose(s) of obinutuzumab. In some embodiments, the methods further comprise administering to the individual an effective amount of an antihistamine, e.g., prior to one or more dose(s) of obinutuzumab. In some embodiments, the antihistamine comprises diphenhydramine. In some embodiments, the methods comprise administering to the individual about 50mg of diphenhydramine prior to one or more dose(s) of obinutuzumab. In some embodiments, the methods further comprise administering to the individual an effective amount of acetaminophen, e.g., prior to one or more dose(s) of obinutuzumab. In some embodiments, the acetaminophen is administered orally at a dose of 15mg / kg. In some embodiments, the methods comprise administering to the individual about 650mg to about 1000mg of acetaminophen prior to one or more dose(s) of obinutuzumab. In some embodiments, the methods further comprise administering to the individual an effective amount of an antihypertensive agent. In some embodiments, the antihypertensive agent is an angiotensin-converting enzyme (ACE) inhibitor or an angiotensin-receptor blocker. In some embodiments, the first antibody exposure, and / or the second antibody exposure, and / or the third antibody exposure, are administered intravenously.

[0033] In some embodiments according to any of the embodiments described herein, the antibody is humanized. In some embodiments, the antibody is afucosylated. In some embodiments, the heavy chain of the type II anti-CD20 antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7. In some embodiments, the light chain of the type II anti-CD20 antibody comprises a light chain variable region21MOFO-360876942Attorney Docket No.14639-20729.40comprising the amino acid sequence of SEQ ID NO:8. In some embodiments, the heavy chain of the type II anti-CD20 antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:7, and the light chain of the type II anti-CD20 antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:8. In some embodiments, the heavy chain of the type II anti-CD20 antibody comprises the amino acid sequence of SEQ ID NO: 9. In some embodiments, the light chain of the type II anti-CD20 antibody comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the heavy chain of the type II anti-CD20 antibody comprises the amino acid sequence of SEQ ID NO: 9, and the light chain of the type II anti-CD20 antibody comprises the amino acid sequence of SEQ ID NO: 10. In some embodiments, the type II anti-CD20 antibody is obinutuzumab.

[0034] In some embodiments according to any of the embodiments described herein, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on days 1 and 15 of treatment; the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on days 168 and 182 of treatment; and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on days 1 and 15 of treatment; the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on days 168 and 182 of treatment; the method further comprises administering to the individual a third antibody exposure comprising one dose of 1000 mg of the type II anti-CD20 antibody on day 364 of treatment; and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on days 1 and 15 of treatment; the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on days 168 and 182 of treatment; the method further comprises administering to the individual a third antibody exposure comprising two doses of 1000 mg of the type II anti-CD20 antibody on days 350 and 364 of treatment; and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; the second antibody exposure comprises two doses of 1000mg of the22MOFO-360876942Attorney Docket No.14639-20729.40type II anti-CD20 antibody on weeks 24 and 26 of treatment; the method further comprises administering to the individual a third antibody exposure comprising one dose of 1000 mg of the type II anti-CD20 antibody on week 52 of treatment; and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; the method further comprises administering to the individual a third antibody exposure comprising two doses of 1000 mg of the type II anti-CD20 antibody on weeks 50 and 52 of treatment; and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody in the first month of treatment (e.g., on weeks 0 and 2); the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody at 6 months of treatment (e.g., on weeks 24 and 26); and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody in the first month of treatment (e.g., on weeks 0 and 2); the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody at 6 months of treatment (e.g., on weeks 24 and 26); the method further comprises administering to the individual a third antibody exposure comprising one dose of 1000 mg of the type II anti-CD20 antibody at 12 months of treatment (e.g., on week 52); and the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody in the first month of treatment (e.g., on weeks 0 and 2); the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody at 6 months of treatment (e.g., on weeks 24 and 26); the method further comprises administering to the individual a third antibody exposure comprising two doses of 1000 mg of the type II anti-CD20 antibody at 12 months of treatment (e.g., on weeks 50 and 52); and the type II anti-CD20 antibody is obinutuzumab.

[0035] In some embodiments according to any one of the embodiments disclosed herein, the methods further comprise administering to the individual a fourth antibody exposure of 1000 mg of obinutuzumab on week 80 of treatment. In some embodiments, the methods further comprise, after week 80 of treatment, administering to the individual one or more additional exposure(s) of 1000 mg of obinutuzumab every 6 months. In some embodiments, the methods further comprise administering to the individual additional exposures of 1000 mg of obinutuzumab on weeks 80,23MOFO-360876942Attorney Docket No.14639-20729.40106, 132, 158, and 184 of treatment. In some embodiments, the methods further comprise administering to the individual additional exposures of 1000 mg of obinutuzumab on weeks 80, 82, 104, 106, 132, 158, and 184 of treatment. In some embodiments, the methods further comprise, after week 184 of treatment, administering to the individual one or more additional exposure(s) of 1000 mg of obinutuzumab every 6 months.

[0036] In other aspects, provided herein is a method for depleting and / or reducing activity of kidney-resident B cells in an individual in need thereof (e.g., an individual having lupus, or an individual having lupus nephritis), comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment; wherein the individual is a human; and wherein: (a) treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure; (b) treatment with obinutuzumab in a plurality of individuals results in a 95% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure; and / or (c) a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1, optionally a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0. In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the24MOFO-360876942Attorney Docket No.14639-20729.40individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment; wherein the individual is a human; and wherein: (a) treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure; (b) treatment with obinutuzumab in a plurality of individuals results in a 95% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure; and / or (c) a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1, optionally a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0. In other aspects, provided herein is a method for depleting and / or reducing activity of kidney-resident B cells in an individual in need thereof (e.g., an individual having lupus, or an individual having lupus nephritis), comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment; wherein the individual is a human; and wherein: (a) treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure; (b) treatment with obinutuzumab in a plurality of individuals results in a 95% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after25MOFO-360876942Attorney Docket No.14639-20729.40administration of the first antibody exposure, as compared to average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure; and / or (c) a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1, optionally a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0. In other aspects, provided herein is a method for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment; wherein the individual is a human; and wherein: (a) treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure; (b) treatment with obinutuzumab in a plurality of individuals results in a 95% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure; and / or (c) a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1, optionally a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0. In some embodiments, the methods further comprise administering to the individual an effective amount of a standard of care treatment. In some embodiments, the standard of care treatment comprises mycophenolate mofetil (MMF). In some embodiments, MMF is administered to the individual at a target dose of about 2.0g / day to about 2.5g / day. In some embodiments, the methods further comprise26MOFO-360876942Attorney Docket No.14639-20729.40administering to the individual an effective amount of a glucocorticoid or corticosteroid. In some embodiments, the glucocorticoid or corticosteroid comprises prednisone. In some embodiments, the methods comprise administering to the individual about 0.5mg / kg / day oral prednisone, with a maximum of about 60mg / day and tapered to about 5mg / day beginning at Week 2. In some embodiments, the methods further comprise, prior to one or more dose(s) of obinutuzumab, administering to the individual an effective amount of a glucocorticoid or corticosteroid, acetaminophen, and / or an antihistamine. In some embodiments, the methods comprise intravenously administering 80mg methylprednisolone to the individual prior to one or more dose(s) of obinutuzumab; administering to the individual about 650mg to about 1000mg of acetaminophen prior to one or more dose(s) of obinutuzumab; and / or administering to the individual about 50mg of diphenhydramine prior to one or more dose(s) of obinutuzumab.In other aspects, provided herein is a kit or article of manufacture for depleting kidney-resident B cells in an individual in need thereof, comprising: (a) a container comprising obinutuzumab; and (b) a package insert with instructions for depleting and / or reducing activity of kidney-resident B cells in an individual according to any one of the embodiments disclosed herein, wherein the individual is a human. In some embodiments, the instructions indicate that obinutuzumab is to be administered to the individual in a first antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment, a second antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment, and a third antibody exposure that comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment. In some embodiments, the instructions indicate that obinutuzumab is to be administered to the individual in a first antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment, a second antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment, and a third antibody exposure that comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment. In some embodiments, the individual has or has been diagnosed with lupus nephritis. In some embodiments, the kits further comprise a container that comprises: a second medicament, wherein obinutuzumab is a first medicament; and wherein the package insert further comprises instructions for administering the second medicament to the individual.27MOFO-360876942Attorney Docket No.14639-20729.40In some embodiments, the second medicament is a standard-of-care medicament for treating lupus nephritis, e.g.., MMF.

[0037] In other aspects, provided herein is a kit or article of manufacture for treating lupus nephritis in an individual that has lupus, comprising a container comprising a type II anti-CD20 antibody, wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and a package insert with instructions for treating lupus nephritis in an individual according to any one of the methods of the present disclosure. In some embodiments, the type II anti-CD20 antibody is obinutuzumab. In some embodiments, the instructions indicate that the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g prior to treatment. In some embodiments, the instructions indicate that the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 3.0 g / g prior to treatment. In some embodiments, the instructions further indicate that a first and a second antibody exposure to the type II anti-CD20 antibody are administered to the individual, wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure; wherein the first antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; and wherein the second antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody. In some embodiments, the instructions further indicate that a first, a second, and a third antibody exposure to the type II anti-CD20 antibody are administered to the individual, wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure; wherein the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; wherein the second antibody exposure comprises one or two doses of the type II28MOFO-360876942Attorney Docket No.14639-20729.40anti-CD20 antibody, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; and wherein the third antibody exposure comprises one dose of the type II anti-CD20 antibody, the third antibody exposure comprising a total exposure of between about 800mg and about 1200mg of the type II anti-CD20 antibody. In some embodiments, the instructions further indicate that a first, a second, and a third antibody exposure to the type II anti-CD20 antibody are administered to the individual, wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure; wherein the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; wherein the second antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; and wherein the third antibody exposure comprises two doses of the type II anti-CD20 antibody, the third antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody. In some embodiments, the instructions further indicate that a first and a second antibody exposure to obinutuzumab are administered to the individual; wherein the first antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 1 or week 0 of treatment and a second dose of about 1000mg of obinutuzumab on day 15 or week 2 of treatment; and wherein the second antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 168 or week 24 of treatment and a second dose of about 1000mg of obinutuzumab on day 182 or week 26 of treatment. In some embodiments, the instructions further indicate that a first, a second, and a third antibody exposure to obinutuzumab are administered to the individual; wherein the first antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 1 or week 0 of treatment and a second dose of about 1000mg of obinutuzumab on day 15 or week 2 of treatment; wherein the second antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 168 or week 24 of treatment and a second dose of about 1000mg of obinutuzumab on day 182 or week 26 of treatment; and wherein the third antibody exposure comprises a single dose of about 1000mg of obinutuzumab on day 364 or week 52 of treatment. In some embodiments, the instructions29MOFO-360876942Attorney Docket No.14639-20729.40further indicate that a first, a second, and a third antibody exposure to obinutuzumab are administered to the individual; wherein the first antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 1 or week 0 of treatment and a second dose of about 1000mg of obinutuzumab on day 15 or week 2 of treatment; wherein the second antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 168 or week 24 of treatment and a second dose of about 1000mg of obinutuzumab on day 182 or week 26 of treatment; and wherein the third antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 350 or week 50 of treatment and a second dose of about 1000mg of obinutuzumab on day 364 or week 52 of treatment. In some embodiments, the kits further comprise a container that comprises a second medicament, wherein the type II anti-CD20 antibody is a first medicament; and instructions on the package insert for administering the second medicament to the individual. In some embodiments, the second medicament is an immunosuppressive agent, a glucocorticoid, an anti-malarial agent, or a corticosteroid, e.g., as disclosed herein. In some embodiments, the second medicament is a standard-of-care medicament for treating lupus nephritis, e.g., as disclosed herein. In some embodiments, the second medicament is MMF. In some embodiments, the instructions further indicate that the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (i) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (ii) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (iii) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (iv) a serum sample that has a low C3 complement level; (v) a serum sample that has a low C4 complement level; (vi) no prior history or diagnosis of lupus and / or lupus nephritis; (vii) concomitant class V lupus nephritis; and / or (viii) is female. In some embodiments, the instructions indicate that a first, a second, and a third antibody exposure to obinutuzumab are administered to the individual; wherein the first antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 1 or week 0 of treatment and a second dose of about 1000mg of obinutuzumab on day 15 or week 2 of treatment; wherein the second antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 168 or week 24 of treatment and a second dose of about 1000mg of obinutuzumab on day 182 or week 26 of treatment; and wherein the third antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 350 or week 50 of treatment and a second dose of about 1000mg of30MOFO-360876942Attorney Docket No.14639-20729.40obinutuzumab on day 364 or week 52 of treatment, and the instructions further indicate that the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (i) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (ii) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (iii) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (iv) a serum sample that has a low C3 complement level; (v) a serum sample that has a low C4 complement level; (vi) no prior history or diagnosis of lupus and / or lupus nephritis; (vii) concomitant class V lupus nephritis; (viii) is female. In some embodiments, the instructions indicate that a first, a second, and a third antibody exposure to obinutuzumab are administered to the individual; wherein the first antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 1 or week 0 of treatment and a second dose of about 1000mg of obinutuzumab on day 15 or week 2 of treatment; wherein the second antibody exposure comprises a first dose of about 1000mg of obinutuzumab on day 168 or week 24 of treatment and a second dose of about 1000mg of obinutuzumab on day 182 or week 26 of treatment; and wherein the third antibody exposure comprises a single dose of about 1000mg of obinutuzumab on day 364 or week 52 of treatment, and the instructions further indicate that the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (i) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (ii) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (iii) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (iv) a serum sample that has a low C3 complement level; (v) a serum sample that has a low C4 complement level; (vi) no prior history or diagnosis of lupus and / or lupus nephritis; (vii) concomitant class V lupus nephritis; and / or (viii) is female.

[0038] In other aspects, provided herein is obinutuzumab for use in a method of depleting and / or reducing activity of kidney-resident B cells in an individual that has lupus nephritis. In some embodiments, the method comprises administering intravenously to the individual a first, a second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of31MOFO-360876942Attorney Docket No.14639-20729.40obinutuzumab intravenously administered to the individual on week 52 of treatment. In some embodiments, the method comprises administering intravenously to the individual a first, a second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment.

[0039] In other aspects, provided herein is a type II anti-CD20 antibody (e.g., obinutuzumab) for use in any one of the methods disclosed herein. In other aspects, provided herein is a type II anti-CD20 antibody for use in a method for treating lupus nephritis in an individual that has lupus, said method comprising administering intravenously to the individual a first and a second antibody exposure to a type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In other aspects, provided herein is a type II anti-CD20 antibody for use in a method for treating lupus nephritis in an individual that has lupus, said method comprising administering intravenously to the individual a first, a second, and a third antibody exposure to a type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises one dose of 1000mg of the type II anti-CD20 antibody on week 52 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of32MOFO-360876942Attorney Docket No.14639-20729.40SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In other aspects, provided herein is a type II anti-CD20 antibody for use in a method for treating lupus nephritis in an individual that has lupus, said method comprising administering intravenously to the individual a first, a second, and a third antibody exposure to a type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 50 and 52 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In some embodiments, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 3.0 g / g. In some embodiments, the type II anti-CD20 antibody is obinutuzumab. In other aspects, provided herein is a type II anti-CD20 antibody (e.g., obinutuzumab) for use in a method for treating lupus nephritis in an individual that has lupus, said method comprising administering intravenously to the individual a first, a second, and a third antibody exposure to a type II anti-CD20 antibody, wherein the type II anti-CD20 antibody is obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises one dose of 1000mg of obinutuzumab on week 52 of treatment; and wherein the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (i) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g;33MOFO-360876942Attorney Docket No.14639-20729.40(ii) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (iii) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (iv) a serum sample that has a low C3 complement level; (v) a serum sample that has a low C4 complement level; (vi) no prior history or diagnosis of lupus and / or lupus nephritis; (vii) concomitant class V lupus nephritis; (viii) is female. In other aspects, provided herein is a type II anti-CD20 antibody (e.g., obinutuzumab) for use in a method for treating lupus nephritis in an individual that has lupus, said method comprising administering intravenously to the individual a first, a second, and a third antibody exposure to a type II anti-CD20 antibody, wherein the type II anti-CD20 antibody is obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 50 and 52 of treatment; and wherein the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (i) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (ii) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (iii) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (iv) a serum sample that has a low C3 complement level; (v) a serum sample that has a low C4 complement level; (vi) no prior history or diagnosis of lupus and / or lupus nephritis; (vii) concomitant class V lupus nephritis; (viii) is female.

[0040] In other aspects, provided herein is the use of obinutuzumab in the manufacture of a medicament for depleting and / or reducing activity of kidney-resident B cells in an individual that has lupus nephritis. In some embodiments, the obinutuzumab or medicament is to be administered intravenously to the individual in a first, a second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment. In some embodiments, the obinutuzumab or medicament is to be administered intravenously to the individual in a first, a34MOFO-360876942Attorney Docket No.14639-20729.40second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment.

[0041] In other aspects, provided herein is the use of a type II anti-CD20 antibody (e.g., obinutuzumab) in the manufacture of a medicament for treating lupus nephritis in an individual that has lupus, e.g., according to any one of the methods disclosed herein. In other aspects, provided herein is the use of a type II anti-CD20 antibody in the manufacture of a medicament for treating lupus nephritis in an individual that has lupus, wherein the medicament is to be administered intravenously to the individual in a first and a second antibody exposure to the type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In other aspects, provided herein is the use of a type II anti-CD20 antibody in the manufacture of a medicament for treating lupus nephritis in an individual that has lupus, wherein the medicament is to be administered intravenously to the individual in a first, a second, and a third antibody exposure to the type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises one dose of 1000mg of the type II anti-CD20 antibody on week 52 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H135MOFO-360876942Attorney Docket No.14639-20729.40sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In other aspects, provided herein is the use of a type II anti-CD20 antibody in the manufacture of a medicament for treating lupus nephritis in an individual that has lupus, wherein the medicament is to be administered intravenously to the individual in a first, a second, and a third antibody exposure to the type II anti-CD20 antibody; wherein the first antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises two doses of 1000mg of the type II anti-CD20 antibody on weeks 50 and 52 of treatment; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6; and wherein, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g. In some embodiments, prior to administration of the first dose of the first antibody exposure, the individual has a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 3.0 g / g. In some embodiments, the type II anti-CD20 antibody is obinutuzumab. In other aspects, provided herein is the use of a type II anti-CD20 antibody (e.g., obinutuzumab) in the manufacture of a medicament for treating lupus nephritis in an individual that has lupus, wherein the medicament is to be administered intravenously to the individual in a first, a second, and a third antibody exposure to the type II anti-CD20 antibody, wherein the type II anti-CD20 antibody is obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises one dose of 1000mg of obinutuzumab on week 52 of36MOFO-360876942Attorney Docket No.14639-20729.40treatment; and wherein the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (i) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (ii) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (iii) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (iv) a serum sample that has a low C3 complement level; (v) a serum sample that has a low C4 complement level; (vi) no prior history or diagnosis of lupus and / or lupus nephritis; (vii) concomitant class V lupus nephritis; and / or (viii) is female. In other aspects, provided herein is the use of a type II anti-CD20 antibody (e.g., obinutuzumab) in the manufacture of a medicament for treating lupus nephritis in an individual that has lupus, wherein the medicament is to be administered intravenously to the individual in a first, a second, and a third antibody exposure to the type II anti-CD20 antibody, wherein the type II anti-CD20 antibody is obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; wherein the third antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 50 and 52 of treatment; and wherein the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (i) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 2.0 g / g; (ii) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (iii) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (iv) a serum sample that has a low C3 complement level; (v) a serum sample that has a low C4 complement level; (vi) no prior history or diagnosis of lupus and / or lupus nephritis; (vii) concomitant class V lupus nephritis; and / or (viii) is female.

[0042] It is to be understood that one, some, or all of the properties of the various embodiments described herein may be combined to form other embodiments of the present invention. These and other aspects of the invention will become apparent to one of skill in the art. These and other embodiments of the invention are further described by the detailed description that follows.BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG.1 provides a schematic diagram of the randomized, placebo-controlled, Phase II NOBILITY trial (NCT02550652; Furie, R.A. et al. (2022) Ann Rheum Dis.81(1):100-107). CRR, complete renal response; CS, corticosteroids; dsDNA, double-stranded DNA; eGFR,37MOFO-360876942Attorney Docket No.14639-20729.40estimated glomerular filtration rate; HPF, high-power field; ISN, International Society of Nephrology; IV, intravenous; LN, lupus nephritis; MMF, mycophenolate mofetil; MPA, mycophenolic acid; OBI, obinutuzumab; PBO, placebo; PRR, partial renal response; R, randomization; RBC, red blood cell; RPS, Renal Pathology Society; SOC, standard of care; ULN, upper limit of normal; UPCR, urine protein-to-creatinine ratio. *One patient randomly assigned to obinutuzumab did not receive obinutuzumab due to pregnancy. A compositeworsening of baseline serum creatinine by >15% and inactive urinary sediment (<10 RBCs / HPF without RBC casts).

[0044] FIG.2 shows the assessment of different primary endpoint measures using data from the NOBILITY trial. Endpoint measures were defined as follows: REGENCY CRR (estimated -baseline eG -mL CRR, complete renalresponse; eGFR, estimated glomerular filtration rate; PERR, primary efficacy renal response; UPCR, urine protein-to-creatinine ratio.

[0045] FIG.3 shows the achievement of REGENCY CRR by level of baseline proteinuria in patients treated with obinutuzumab with standard-of-care therapy or placebo with standard-of-care therapy in the NOBILITY trial. Modified intention-to-not exactly correspond to the subtraction of the patient percentages due to rounding. P values were obtained using the Cochran-Mantel-Haenszel test stratified for region (US vs non-US) and race (Afro-Caribbean / African American vs others). CRR, complete renal response; UPCR, urine protein-to-creatinine ratio.

[0046] FIG.4 provides a schematic illustrating a Phase III study (NCT04221477) evaluating the efficacy, safety, and pharmacokinetics of obinutuzumab compared with placebo in patients with ISN / RPS Class III or IV lupus nephritis when added onto standard-of-care therapy comprising mycophenolate mofetil (MMF) and corticosteroids.38MOFO-360876942Attorney Docket No.14639-20729.40

[0047] FIG.5 shows a disposition of the Phase III study shown in FIG.4. *one patient randomized to placebo received obinutuzumab; three patients randomized to placebo did not receive study drug.

[0048] FIG.6 shows the proportion of patients achieving a complete renal response (CRR) at Week 76 in the efficacy-evaluable population for the obinutuzumab and placebo groups.

[0049] FIGS.7A & 7B show forest plots of the differences in proportion of patients in CRR at Week 76 by subgroup (as indicated) in efficacy-evaluable patients.

[0050] FIG.8 shows the analysis populations for the Phase III study.

[0051] FIGS.9A & 9B show the demographics of the efficacy-evaluable patients in the obinutuzumab group, placebo group, and all patients.

[0052] FIGS.10A-10C show the baseline characteristics of the efficacy-evaluable patients in the obinutuzumab group, placebo group, and all patients.

[0053] FIGS.11A-11H show supportive analyses of the differences in proportion of patients in CRR at Week 76 by subgroup in efficacy-evaluable patients using different analyses for the Phase III study.

[0054] FIG.12A shows a plot of the time to a lupus nephritis (LN) flare from week 24 through Week 76 in obinutuzumab treated and placebo groups. Obinutuzumab treatment reduced the risk of lupus nephritis flare in patients by 56% at week 76 of treatment. LN flare is defined as eGFR decrease >20% compared with Week 24 in patients with UPCR >1 g / g and / or cellular casts; UPCR increase (i) to >1 g / g if Week 24 UPCR was <0.2 g / g, (ii) to >2 g / g if Week 24 UPCR was 0.2-1 g / g or (iii) to doubling if Week 24 UPCR was >1 g / g; receipt of rescue therapy, except for corticosteroid-only rescue.aPlus standard therapy of mycophenolate mofetil plus glucocorticoids.bStatistical significance cannot be claimed as there was no correction for multiplicity. FIG.12B shows a plot of the time to an unfavourable kidney outcome through Week 76 in obinutuzumab treated and placebo groups. Obinutuzumab treatment reduced the risk of unfavourable kidney outcomes in patients by 63% at week 76 of treatment. Unfavorable kidney outcome is defined as the first of the following events: death, doubling of serum creatinine or treatment failure.aPlus standard therapy of mycophenolate mofetil plus glucocorticoids.bStatistical significance cannot be claimed as there was no correction for multiplicity. CI, confidence interval; eGFR, estimated glomerular filtration rate; HR, hazard ratio; UPCR, urine protein-to-creatinine ratio.39MOFO-360876942Attorney Docket No.14639-20729.40

[0055] FIG.13 shows counts of tissue-resident B cells (CD79a+ / CD138-) at baseline and week 76 from the longitudinal kidney biopsy cohort of the REGENCY trial (NCT04221477). B cell counts are shown by treatment group in the biopsy cohort, with results from obinutuzumab treated patients on the left and placebo treated patients on the right. Timepoints are baseline and week 76 of the study, with B cell counts (per mm2tissue) on the y-axis. In cases of replicates (more than one slide stained for the same participant), a single value was calculated as the sum of B cells divided by the sum of areas. Complete renal response (CRR) was achieved by 7 participants (50%) in the obinutuzumab group and 3 (20%) in the placebo group.

[0056] FIG.14AAI=0 (right) at week 76 among patients treated with obinutuzumab or placebo in the kidney biopsy cohort of the REGENCY trial. Error bars show 95% confidence interval (CI). Patients -only rescue prior to week 76 were considered non-responders.aPlus standard therapy consisting of mycophenolate mofetil plus glucocorticoids.

[0057] FIG.14B shows the respective changes in activity index (AI; left) and chronicity index (right) from baseline biopsy to Week 76 biopsy among patients treated with obinutuzumab or placebo in the kidney biopsy cohort of the REGENCY trial, expressed as mean with standard deviation error bars.aPlus standard therapy consisting of mycophenolate mofetil plus glucocorticoids.DETAILED DESCRIPTION

[0058] The Phase III study (REGENCY; NCT04221477) evaluated the efficacy, safety, and pharmacokinetics of obinutuzumab compared with placebo in patients with ISN / RPS Class III or IV lupus nephritis (LN) when added onto standard-of-care therapy comprising mycophenolate mofetil (MMF) and corticosteroids. The results of this trial indicated that obinutuzumab treatment as described herein was superior to standard therapy alone in helping people with lupus nephritis achieve a complete renal response and greater peripheral B cell depletion, showing a statistically significant and clinically meaningful improvement in treatment of individuals with active LN.

[0059] Persistent kidney inflammation, driven by tissue-resident immune cells, remains a significant challenge in lupus nephritis (LN), frequently leading to irreversible damage despite existing therapies. The present disclosure demonstrates that obinutuzumab potently depleted B 40MOFO-360876942Attorney Docket No.14639-20729.40cells in kidney tissue of individuals with LN, representing the first demonstration of kidney tissue-level B cell depletion by an anti-CD20 therapeutic agent in any glomerular disease.Achievement of histologic remission reduces future LN flare risk to preserve functional kidney mass. In addition, the higher proportion of patients who achieved histologic remission (as compared to the proportion that achieved CRR) suggests that the impact of obinutuzumab in LN is greater than that implied by CRR rate.

[0060] In one aspect, provided herein is a method for depleting and / or reducing activity of kidney-resident B cells in an individual in need thereof (e.g., an individual having lupus, or an individual having lupus nephritis), comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human. In some embodiments, treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure. In some embodiments, treatment with obinutuzumab in a plurality of individuals results in a 90% or greater, 91% or greater, 92% or greater, 93% or greater, 94% or greater, 95% or greater, 96% or greater, 97% or greater, or 98% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to an average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure. In some embodiments, a kidney biopsy sample obtained from the individual at about 76 weeks to about41MOFO-360876942Attorney Docket No.14639-20729.4080 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1, or an AI of 0.I. General Techniques

[0061] The techniques and procedures described or referenced herein are generally well understood and commonly employed using conventional methodology by those skilled in the art, such as, for example, the widely utilized methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 3d edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; Current Protocols in Molecular Biology (F.M. Ausubel, et al. eds., (2003)); the series Methods in Enzymology (Academic Press, Inc.): PCR 2: A Practical Approach (M.J. MacPherson, B.D. Hames and G.R. Taylor eds. (1995)), Harlow and Lane, eds. (1988) Antibodies, A Laboratory Manual, and Animal Cell Culture (R.I. Freshney, ed. (1987));Oligonucleotide Synthesis (M.J. Gait, ed., 1984); Methods in Molecular Biology, Humana Press; Cell Biology: A Laboratory Notebook (J.E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R.I. Freshney), ed., 1987); Introduction to Cell and Tissue Culture (J.P. Mather and P.E. Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J.B. Griffiths, and D.G. Newell, eds., 1993-8) J. Wiley and Sons; Handbook of Experimental Immunology (D.M. Weir and C.C. Blackwell, eds.); Gene Transfer Vectors for Mammalian Cells (J.M. Miller and M.P. Calos, eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (J.E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology (C.A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: A Practical Approach (D. Catty., ed., IRL Press, 1988-1989); Monoclonal Antibodies: A Practical Approach (P. Shepherd and C. Dean, eds., Oxford University Press, 2000); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory Press, 1999); The Antibodies (M. Zanetti and J. D. Capra, eds., Harwood Academic Publishers, 1995); and Cancer: Principles and Practice of Oncology (V.T. DeVita et al., eds., J.B. Lippincott Company, 1993).42MOFO-360876942Attorney Docket No.14639-20729.40II. Definitions

[0062] The term "lupus nephritis (LN)" refers to a manifestation of lupus (e.g., systemic lupus erythematosus, drug-induced lupus, neonatal lupus, or discoid lupus) in the kidney(s).

[0063] The term “antibody” includes monoclonal antibodies (including full length antibodies which have an immunoglobulin Fc region), antibody compositions with polyepitopic specificity, multispecific antibodies (e.g., bispecific antibodies, diabodies, and single-chain molecules, as well as antibody fragments (e.g., Fab, F(ab')2, and Fv). The term “immunoglobulin” (Ig) is used interchangeably with “antibody” herein.

[0064] The basic 4-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light (L) chains and two identical heavy (H) chains. An IgM antibody consists of 5 of the basic heterotetramer units along with an additional polypeptide called a J chain, and contains 10 antigen binding sites, while IgA antibodies comprise from 2-5 of the basic 4-chain units which can polymerize to form polyvalent assemblages in combination with the J chain. In the case of IgGs, the 4-chain unit is generally about 150,000 daltons. Each L chain is linked to an H chain by one covalent disulfide bond, while the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has at the N-terminus, a variable domain (VH) followed by three constant domains (CH) for each of the and chains and four CHdomains for μ and isotypes. Each L chain has at the N-terminus, a variable domain (VL) followed by a constant domain at its other end. The VLis aligned with the VHand the CLis aligned with the first constant domain of the heavy chain (CH1). Particular amino acid residues are believed to form an interface between the light chain and heavy chain variable domains. The pairing of a VHand VLtogether forms a single antigen-binding site. For the structure and properties of the different classes of antibodies, see e.g., Basic and Clinical Immunology, 8th Edition, Daniel P. Sties, Abba I. Terr and Tristram G. Parsolw (eds), Appleton & Lange, Norwalk, CT, 1994, page 71 and Chapter 6. The L chain from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains (CH), immunoglobulins can be assigned to different classes or isotypes. There are five classes of immunoglobulins: IgA, IgD, IgE, IgG and IgM, having heavy chains designated , , , and μ, respectively. The and classes are further divided into subclasses on the basis of relatively 43MOFO-360876942Attorney Docket No.14639-20729.40minor differences in the CH sequence and function, e.g., humans express the following subclasses: IgG1, IgG2A, IgG2B, IgG3, IgG4, IgA1 and IgA2.

[0065] The “variable region” or “variable domain” of an antibody refers to the amino-terminal domains of the heavy or light chain of the antibody. The variable domains of the heavy chain and light chain may be referred to as “VH” and “VL”, respectively. These domains are generally the most variable parts of the antibody (relative to other antibodies of the same class) and contain the antigen binding sites.

[0066] The term "variable" refers to the fact that certain segments of the variable domains differ extensively in sequence among antibodies. The V domain mediates antigen binding and defines the specificity of a particular antibody for its particular antigen. However, the variability is not evenly distributed across the entire span of the variable domains. Instead, it is concentrated in three segments called hypervariable regions (HVRs) both in the light-chain and the heavy chain variable domains. The more highly conserved portions of variable domains are called the framework regions (FR). The variable domains of native heavy and light chains each comprise four FR regions, largely adopting a beta-sheet configuration, connected by three HVRs, which form loops connecting, and in some cases forming part of, the beta-sheet structure. The HVRs in each chain are held together in close proximity by the FR regions and, with the HVRs from the other chain, contribute to the formation of the antigen binding site of antibodies (see Kabat et al., Sequences of Immunological Interest, Fifth Edition, National Institute of Health, Bethesda, MD (1991)). The constant domains are not involved directly in the binding of antibody to an antigen, but exhibit various effector functions, such as participation of the antibody in antibody-dependent cellular toxicity.

[0067] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations and / or post-translation modifications (e.g., isomerizations, amidations) that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. In contrast to polyclonal antibody preparations which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, the monoclonal antibodies are advantageous in that they are synthesized by the hybridoma culture, uncontaminated by other44MOFO-360876942Attorney Docket No.14639-20729.40immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by a variety of techniques, including, for example, the hybridoma method (e.g., Kohler and Milstein., Nature, 256:495-97 (1975); Hongo et al., Hybridoma, 14 (3): 253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nded.1988); Hammerling et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Patent No.4,816,567), phage-display technologies (see, e.g., Clackson et al., Nature, 352: 624-628 (1991); Marks et al., J. Mol. Biol.222: 581-597 (1992); Sidhu et al., J. Mol. Biol.338(2): 299-310 (2004); Lee et al., J. Mol. Biol.340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004), and technologies for producing human or human-like antibodies in animals that have parts or all of the human immunoglobulin loci or genes encoding human immunoglobulin sequences (see, e.g., WO 1998 / 24893; WO 1996 / 34096; WO 1996 / 33735; WO 1991 / 10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA 90: 2551 (1993); Jakobovits et al., Nature 362: 255-258 (1993); Bruggemann et al., Year in Immunol.7:33 (1993); U.S. Patent Nos.5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016; Marks et al., Bio / Technology 10: 779-783 (1992); Lonberg et al., Nature 368: 856-859 (1994);Morrison, Nature 368: 812-813 (1994); Fishwild et al., Nature Biotechnol.14: 845-851 (1996); Neuberger, Nature Biotechnol.14: 826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol.13: 65-93 (1995).

[0068] The term “naked antibody” refers to an antibody that is not conjugated to a cytotoxic moiety or radiolabel.

[0069] The terms “full-length antibody,” “intact antibody” or “whole antibody” are used interchangeably to refer to an antibody in its substantially intact form, as opposed to an antibody fragment. Specifically whole antibodies include those with heavy and light chains including an Fc region. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. In some cases, the intact antibody may have one or more effector functions.45MOFO-360876942Attorney Docket No.14639-20729.40

[0070] An “antibody fragment” comprises a portion of an intact antibody, preferably the antigen binding and / or the variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2and Fv fragments; diabodies; linear antibodies (see U.S. Patent 5,641,870, Example 2; Zapata et al., Protein Eng.8(10): 1057-1062

[1995] ); single-chain antibody molecules and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies produced two identical antigen-binding fragments, called "Fab" fragments, and a residual "Fc" fragment, a designation reflecting the ability to crystallize readily. The Fab fragment consists of an entire L chain along with the variable region domain of the H chain (VH), and the first constant domain of one heavy chain (CH1). Each Fab fragment is monovalent with respect to antigen binding, i.e., it has a single antigen-binding site. Pepsin treatment of an antibody yields a single large F(ab')2fragment which roughly corresponds to two disulfide linked Fab fragments having different antigen-binding activity and is still capable of cross-linking antigen. Fab' fragments differ from Fab fragments by having a few additional residues at the carboxy terminus of the CH1 domain including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine residue(s) of the constant domains bear a free thiol group. F(ab')2antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0071] The Fc fragment comprises the carboxy-terminal portions of both H chains held together by disulfides. The effector functions of antibodies are determined by sequences in the Fc region, the region which is also recognized by Fc receptors (FcR) found on certain types of cells.

[0072] “Fv” is the minimum antibody fragment which contains a complete antigen-recognition and -binding site. This fragment consists of a dimer of one heavy- and one light-chain variable region domain in tight, non-covalent association. From the folding of these two domains emanate six hypervariable loops (3 loops each from the H and L chain) that contribute the amino acid residues for antigen binding and confer antigen binding specificity to the antibody.However, even a single variable domain (or half of an Fv comprising only three HVRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.46MOFO-360876942Attorney Docket No.14639-20729.40

[0073] “Single-chain Fv” also abbreviated as “sFv” or “scFv” are antibody fragments that comprise the VHand VLantibody domains connected into a single polypeptide chain.Preferably, the sFv polypeptide further comprises a polypeptide linker between the VHand VLdomains which enables the sFv to form the desired structure for antigen binding. For a review of the sFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol.113, Rosenburg and Moore eds., Springer-Verlag, New York, pp.269-315 (1994).

[0074] “Functional fragments” of the antibodies of the invention comprise a portion of an intact antibody, generally including the antigen binding or variable region of the intact antibody or the Fc region of an antibody which retains or has modified FcR binding capability. Examples of antibody fragments include linear antibody, single-chain antibody molecules and multispecific antibodies formed from antibody fragments.

[0075] The term “diabodies” refers to small antibody fragments prepared by constructing sFv fragments (see preceding paragraph) with short linkers (about 5-10) residues) between the VHand VLdomains such that inter-chain but not intra-chain pairing of the V domains is achieved, thereby resulting in a bivalent fragment, i.e., a fragment having two antigen-binding sites.Bispecific diabodies are heterodimers of two "crossover" sFv fragments in which the VHand VLdomains of the two antibodies are present on different polypeptide chains. Diabodies are described in greater detail in, for example, EP 404,097; WO 93 / 11161; Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993).

[0076] The monoclonal antibodies herein specifically include "chimeric" antibodies (immunoglobulins) in which a portion of the heavy and / or light chain is identical with or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chain(s) is(are) identical with or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (U.S. Patent No.4,816,567; Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). Chimeric antibodies of interest herein include PRIMATIZED®antibodies wherein the antigen-binding region of the antibody is derived from an antibody produced by, e.g., immunizing macaque monkeys with an antigen of interest. As used herein, “humanized antibody” is used a subset of “chimeric antibodies.”47MOFO-360876942Attorney Docket No.14639-20729.40

[0077] "Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies that contain minimal sequence derived from non-human immunoglobulin. In one embodiment, a humanized antibody is a human immunoglobulin (recipient antibody) in which residues from an HVR (hereinafter defined) of the recipient are replaced by residues from an HVR of a non-human species (donor antibody) such as mouse, rat, rabbit or non-human primate having the desired specificity, affinity, and / or capacity. In some instances, framework (“FR”) residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or in the donor antibody. These modifications may be made to further refine antibody performance, such as binding affinity. In general, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin sequence, and all or substantially all of the FR regions are those of a human immunoglobulin sequence, although the FR regions may include one or more individual FR residue substitutions that improve antibody performance, such as binding affinity, isomerization, immunogenicity, etc. The number of these amino acid substitutions in the FR are typically no more than 6 in the H chain, and in the L chain, no more than 3. The humanized antibody optionally will also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol.2:593-596 (1992). See also, for example, Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol.1:105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech.5:428-433 (1994); and U.S. Pat. Nos.6,982,321 and 7,087,409.

[0078] A “human antibody” is an antibody that possesses an amino-acid sequence corresponding to that of an antibody produced by a human and / or has been made using any of the techniques for making human antibodies as disclosed herein. This definition of a human antibody specifically excludes a humanized antibody comprising non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage-display libraries. Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol., 222:581 (1991). Also available for the preparation of human monoclonal antibodies are methods described in Cole et al., Monoclonal Antibodies and Cancer48MOFO-360876942Attorney Docket No.14639-20729.40Therapy, Alan R. Liss, p.77 (1985); Boerner et al., J. Immunol., 147(1):86-95 (1991). See also van Dijk and van de Winkel, Curr. Opin. Pharmacol., 5: 368-74 (2001). Human antibodies can be prepared by administering the antigen to a transgenic animal that has been modified to produce such antibodies in response to antigenic challenge, but whose endogenous loci have been disabled, e.g., immunized xenomice (see, e.g., U.S. Pat. Nos.6,075,181 and 6,150,584 regarding XENOMOUSETMtechnology). See also, for example, Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006) regarding human antibodies generated via a human B-cell hybridoma technology.

[0079] The term “hypervariable region,” “HVR,” or “HV,” when used herein refers to the regions of an antibody variable domain which are hypervariable in sequence and / or form structurally defined loops. Generally, antibodies comprise six HVRs; three in the VH (H1, H2, H3), and three in the VL (L1, L2, L3). In native antibodies, H3 and L3 display the most diversity of the six HVRs, and H3 in particular is believed to play a unique role in conferring fine specificity to antibodies. See, e.g., Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, in Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, NJ, 2003). Indeed, naturally occurring camelid antibodies consisting of a heavy chain only are functional and stable in the absence of light chain. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol.3:733-736 (1996).

[0080] A number of HVR delineations are in use and are encompassed herein. The Kabat Complementarity Determining Regions (CDRs) are based on sequence variability and are the most commonly used (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)). Chothia refers instead to the location of the structural loops (Chothia and Lesk, J. Mol. Biol.196:901-917 (1987)). The AbM HVRs represent a compromise between the Kabat HVRs and Chothia structural loops, and are used by Oxford Molecular's AbM antibody modeling software. The “contact” HVRs are based on an analysis of the available complex crystal structures. The residues from each of these HVRs are noted below.Loop Kabat AbM Chothia ContactL1 L24-L34 L24-L34 L26-L32 L30-L36L2 L50-L56 L50-L56 L50-L52 L46-L55L3 L89-L97 L89-L97 L91-L96 L89-L9649MOFO-360876942Attorney Docket No.14639-20729.40H1 H31-H35B H26-H35B H26-H32 H30-H35B (Kabat numbering)H1 H31-H35 H26-H35 H26-H32 H30-H35 (Chothia numbering)H2 H50-H65 H50-H58 H53-H55 H47-H58H3 H95-H102 H95-H102 H96-H101 H93-H101

[0081] HVRs may comprise “extended HVRs” as follows: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2) and 89-97 or 89-96 (L3) in the VL and 26-35 (H1), 50-65 or 49-65 (H2) and 93-102, 94-102, or 95-102 (H3) in the VH. The variable domain residues are numbered according to Kabat et al., supra, for each of these definitions.

[0082] The expression “variable-domain residue-numbering as in Kabat” or “amino-acid-position numbering as in Kabat,” and variations thereof, refers to the numbering system used for heavy-chain variable domains or light-chain variable domains of the compilation of antibodies in Kabat et al., supra. Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to a shortening of, or insertion into, a FR or HVR of the variable domain. For example, a heavy-chain variable domain may include a single amino acid insert (residue 52a according to Kabat) after residue 52 of H2 and inserted residues (e.g. residues 82a, 82b, and 82c, etc. according to Kabat) after heavy-chain FR residue 82. The Kabat numbering of residues may be determined for a given antibody by alignment at regions of homology of the sequence of the antibody with a “standard” Kabat numbered sequence.

[0083] "Framework" or "FR" residues are those variable-domain residues other than the HVR residues as herein defined.

[0084] A “human consensus framework” or “acceptor human framework” is a framework that represents the most commonly occurring amino acid residues in a selection of human immunoglobulin VL or VH framework sequences. Generally, the selection of human immunoglobulin VL or VH sequences is from a subgroup of variable domain sequences.Generally, the subgroup of sequences is a subgroup as in Kabat et al., Sequences of Proteins of Immunological Interest, 5thEd. Public Health Service, National Institutes of Health, Bethesda, MD (1991). Examples include for the VL, the subgroup may be subgroup kappa I, kappa II, kappa III or kappa IV as in Kabat et al., supra. Additionally, for the VH, the subgroup may be subgroup I, subgroup II, or subgroup III as in Kabat et al., supra. Alternatively, a human consensus framework can be derived from the above in which particular residues, such as when a human framework residue is selected based on its homology to the donor framework by aligning 50MOFO-360876942Attorney Docket No.14639-20729.40the donor framework sequence with a collection of various human framework sequences. An acceptor human framework “derived from” a human immunoglobulin framework or a human consensus framework may comprise the same amino acid sequence thereof, or it may contain pre-existing amino acid sequence changes. In some embodiments, the number of pre-existing amino acid changes are 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, or 2 or less.

[0085] A “VH subgroup III consensus framework” comprises the consensus sequence obtained from the amino acid sequences in variable heavy subgroup III of Kabat et al., supra. In one embodiment, the VH subgroup III consensus framework amino acid sequence comprises at least a portion or all of each of the following sequences: EVQLVESGGGLVQPGGSLRLSCAAS (HC-FR1)(SEQ ID NO:35), WVRQAPGKGLEWV (HC-FR2), (SEQ ID NO:36), RFTISADTSKNTAYLQMNSLRAEDTAVYYCAR (HC-FR3, SEQ ID NO:37), WGQGTLVTVSA (HC-FR4), (SEQ ID NO:38).

[0086] A “VL kappa I consensus framework” comprises the consensus sequence obtained from the amino acid sequences in variable light kappa subgroup I of Kabat et al., supra. In one embodiment, the VH subgroup I consensus framework amino acid sequence comprises at least a portion or all of each of the following sequences: DIQMTQSPSSLSASVGDRVTITC (LC-FR1) (SEQ ID NO:39), WYQQKPGKAPKLLIY (LC-FR2) (SEQ ID NO:40), GVPSRFSGSGSGTDFTLTISSLQPEDFATYYC (LC-FR3)(SEQ ID NO:41), FGQGTKVEIKR (LC-FR4)(SEQ ID NO:42).

[0087] An “amino-acid modification” at a specified position, e.g. of the Fc region, refers to the substitution or deletion of the specified residue, or the insertion of at least one amino acid residue adjacent the specified residue. Insertion “adjacent” to a specified residue means insertion within one to two residues thereof. The insertion may be N-terminal or C-terminal to the specified residue. The preferred amino acid modification herein is a substitution.

[0088] An “affinity-matured” antibody is one with one or more alterations in one or more HVRs thereof that result in an improvement in the affinity of the antibody for antigen, compared to a parent antibody that does not possess those alteration(s). In one embodiment, an affinity-matured antibody has nanomolar or even picomolar affinities for the target antigen. Affinity-matured antibodies are produced by procedures known in the art. For example, Marks et al., Bio / Technology 10:779-783 (1992) describes affinity maturation by VH- and VL-domain51MOFO-360876942Attorney Docket No.14639-20729.40shuffling. Random mutagenesis of HVR and / or framework residues is described by, for example: Barbas et al. Proc Nat. Acad. Sci. USA 91:3809-3813 (1994); Schier et al. Gene 169:147-155 (1995); Yelton et al. J. Immunol.155:1994-2004 (1995); Jackson et al., J. Immunol.154(7):3310-9 (1995); and Hawkins et al, J. Mol. Biol.226:889-896 (1992).

[0089] As use herein, the term "specifically binds to" or is "specific for" refers to measurable and reproducible interactions such as binding between a target and an antibody, which is determinative of the presence of the target in the presence of a heterogeneous population of molecules including biological molecules. For example, an antibody that specifically binds to a target (which can be an epitope) is an antibody that binds this target with greater affinity, avidity, more readily, and / or with greater duration than it binds to other targets. In one embodiment, the extent of binding of an antibody to an unrelated target is less than about 10% of the binding of the antibody to the target as measured, e.g., by a radioimmunoassay (RIA). In certainspecifically binds to an epitope on a protein that is conserved among the protein from different species. In another embodiment, specific binding can include, but does not require exclusive binding.

[0090] The term “Fc region” herein is used to define a C-terminal region of an immunoglobulin heavy chain, including native-sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an immunoglobulin heavy chain might vary, the human IgG heavy-chain Fc region is usually defined to stretch from an amino acid residue at position Cys226, or from Pro230, to the carboxyl-terminus thereof. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during production or purification of the antibody, or by recombinantly engineering the nucleic acid encoding a heavy chain of the antibody. Accordingly, a composition of intact antibodies may comprise antibody populations with all K447 residues removed, antibody populations with no K447 residues removed, and antibody populations having a mixture of antibodies with and without the K447 residue. Suitable native-sequence Fc regions for use in the antibodies of the invention include human IgG1, IgG2 (IgG2A, IgG2B), IgG3 and IgG4.

[0091] “Fc receptor” or “FcR” describes a receptor that binds to the Fc region of an antibody. The preferred FcR is a native sequence human FcR. Moreover, a preferred FcR is one which52MOFO-360876942Attorney Docket No.14639-20729.40binds an IgG antibody (a gamma receptor) and includes receptors of the Fc RI, Fc RII, andFc RIII subclasses, including allelic variants and alternatively spliced forms of these receptors,Fc RII receptors include Fc RIIA (an "activating receptor") and Fc RIIB (an "inhibitingreceptor"), which have similar amino acid sequences that differ primarily in the cytoplasmicdomains thereof. Activating receptor Fc RIIA contains an immunoreceptor tyrosine-basedactivation motif (ITAM) in its cytoplasmic domain. Inhibiting receptor Fc RIIB contains animmunoreceptor tyrosine-based inhibition motif (ITIM) in its cytoplasmic domain. (see M. Daëron, Annu. Rev. Immunol.15:203-234 (1997). FcRs are reviewed in Ravetch and Kinet, Annu. Rev. Immunol.9: 457-92 (1991); Capel et al., Immunomethods 4: 25-34 (1994); and de Haas et al., J. Lab. Clin. Med.126: 330-41 (1995). Other FcRs, including those to be identified in the future, are encompassed by the term "FcR" herein.

[0092] The term “Fc receptor” or “FcR” also includes the neonatal receptor, FcRn, which is responsible for the transfer of maternal IgGs to the fetus. Guyer et al., J. Immunol.117: 587 (1976) and Kim et al., J. Immunol.24: 249 (1994). Methods of measuring binding to FcRn are known (see, e.g., Ghetie and Ward, Immunol. Today 18: (12): 592-8 (1997); Ghetie et al., Nature Biotechnology 15 (7): 637-40 (1997); Hinton et al., J. Biol. Chem.279 (8): 6213-6 (2004); WO 2004 / 92219 (Hinton et al.). Binding to FcRn in vivo and serum half-life of human FcRn high-affinity binding polypeptides can be assayed, e.g., in transgenic mice or transfected human cell lines expressing human FcRn, or in primates to which the polypeptides having a variant Fc region are administered. WO 2004 / 42072 (Presta) describes antibody variants which improved or diminished binding to FcRs. See also, e.g., Shields et al., J. Biol. Chem.9(2): 6591-6604 (2001).

[0093] The phrase “substantially reduced,” or “substantially different,” as used herein, denotes a sufficiently high degree of difference between two numeric values (generally one associated with a molecule and the other associated with a reference / comparator molecule) such that one of skill in the art would consider the difference between the two values to be of statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values). The difference between said two values is, for example, greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, and / or greater than about 50% as a function of the value for the reference / comparator molecule.53MOFO-360876942Attorney Docket No.14639-20729.40

[0094] The term “substantially similar” or “substantially the same,” as used herein, denotes a sufficiently high degree of similarity between two numeric values (for example, one associated with an antibody of the invention and the other associated with a reference / comparator antibody), such that one of skill in the art would consider the difference between the two values to be of little or no biological and / or statistical significance within the context of the biological characteristic measured by said values (e.g., Kd values). The difference between said two values is, for example, less than about 50%, less than about 40%, less than about 30%, less than about 20%, and / or less than about 10% as a function of the reference / comparator value.

[0095] "Carriers" as used herein include pharmaceutically acceptable carriers, excipients, or stabilizers that are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffered solution. Examples of physiologically acceptable carriers include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as TWEEN™, polyethylene glycol (PEG), and PLURONICS™.

[0096] A “package insert” refers to instructions customarily included in commercial packages of medicaments that contain information about the indications customarily included in commercial packages of medicaments that contain information about the indications, usage, dosage, administration, contraindications, other medicaments to be combined with the packaged product, and / or warnings concerning the use of such medicaments, etc.

[0097] As used herein, the term “treatment” refers to clinical intervention designed to alter the natural course of the individual or cell being treated during the course of clinical pathology. Desirable effects of treatment include, but are not limited to, decreasing the rate of disease progression, ameliorating or palliating the disease state, remission or improved prognosis, and delaying disease progression. For example, an individual is successfully “treated” if one or more symptoms associated with lupus nephritis are mitigated or eliminated, including, but are not limited to, elevated serum creatinine, proteinuria, red cell casts, reduced renal function, nephrotic54MOFO-360876942Attorney Docket No.14639-20729.40syndrome, granular casts, microhematuria, macrohematuria, hypertension, tubular abnormalities, hyperkalemia, rapidly progressive glomerulonephritis (RPGN), and acute renal failure (ARF). Delaying progression of a disease (e.g., lupus nephritis) means to defer, hinder, slow, retard, stabilize, and / or postpone development of the disease. This delay can be of varying lengths of time, depending on the history of the disease and / or individual being treated. As is evident to one skilled in the art, a sufficient or significant delay can, in effect, encompass prevention, in that the individual, e.g., an individual at risk for developing the disease, does not develop the disease. For example, the progression of SLE in an individual before the onset of LN symptoms and / or pathology may be delayed such that the development of LN is postponed or prevented.

[0098] “CD20” as used herein refers to the human B-lymphocyte antigen CD20 (also known as CD20, B-lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5; the sequence is characterized by the SwissProt database entry P11836) is a hydrophobic transmembrane protein with a molecular weight of approximately 35 kD located on pre-B and mature B lymphocytes. (Valentine, M.A., et al., J. Biol. Chem.264(19) (198911282-11287; Tedder, T.F., et al, Proc. Natl. Acad. Sci. U.S.A.85 (1988) 208-12; Stamenkovic, I., et al., J. Exp. Med.167 (1988) 1975-80; Einfeld, D.A., et al., EMBO J.7 (1988) 711-7; Tedder, T.F., et al., J. Immunol.142 (1989) 2560-8). The corresponding human gene is Membrane-spanning 4-domains, subfamily A, member 1, also known as MS4A1. This gene encodes a member of the membrane-spanning 4A gene family. Members of this nascent protein family are characterized by common structural features and similar intron / exon splice boundaries and display unique expression patterns among hematopoietic cells and nonlymphoid tissues. This gene encodes the B-lymphocyte surface molecule which plays a role in the development and differentiation of B-cells into plasma cells. This family member is localized to 11q12, among a cluster of family members. Alternative splicing of this gene results in two transcript variants which encode the same protein.

[0099] The terms "CD20" and "CD20 antigen" are used interchangeably herein, and include any variants, isoforms and species homologs of human CD20 which are naturally expressed by cells or are expressed on cells transfected with the CD20 gene. Binding of an antibody of the invention to the CD20 antigen mediate the killing of cells expressing CD20 (e.g., a tumor cell) by inactivating CD20. The killing of the cells expressing CD20 may occur by one or more of the following mechanisms: Cell death / apoptosis induction, ADCC and CDC.55MOFO-360876942Attorney Docket No.14639-20729.40

[0100] Synonyms of CD20, as recognized in the art, include B-lymphocyte antigen CD20, B-lymphocyte surface antigen B1, Leu-16, Bp35, BM5, and LF5.

[0101] The term “anti-CD20 antibody” according to the invention is an antibody that binds specifically to CD20 antigen. Depending on binding properties and biological activities of anti-CD20 antibodies to the CD20 antigen, two types of anti-CD20 antibodies (type I and type II anti-CD20 antibodies) can be distinguished according to Cragg, M.S., et al., Blood 103 (2004) 2738-2743; and Cragg, M.S., et al., Blood 101 (2003) 1045-1052, see Table 1 below.Table 1. Properties of type I and type II anti-CD20 antibodies

[0102] Examples of type II anti-CD20 antibodies include e.g. humanized B-Ly1 antibody IgG1 (a chimeric humanized IgG1 antibody as disclosed in WO 2005 / 044859), 11B8 IgG1 (as disclosed in WO 2004 / 035607), and AT80 IgG1. Typically type II anti-CD20 antibodies of the IgG1 isotype show characteristic CDC properties. Type II anti-CD20 antibodies have a decreased CDC (if IgG1 isotype) compared to type I antibodies of the IgG1 isotype.

[0103] Examples of type I anti-CD20 antibodies include e.g. rituximab, HI47 IgG3 (ECACC, hybridoma), 2C6 IgG1 (as disclosed in WO 2005 / 103081), 2F2 IgG1 (as disclosed and WO 2004 / 035607 and WO 2005 / 103081) and 2H7 IgG1 (as disclosed in WO 2004 / 056312).

[0104] The afucosylated anti-CD20 antibodies according to the invention are preferably type II anti-CD20 antibodies, more preferably afucosylated humanized B-Ly1 antibodies as described in WO 2005 / 044859 and WO 2007 / 031875.

[0105] The “rituximab” antibody (reference antibody; example of a type I anti-CD20 antibody) is a genetically engineered chimeric human gamma 1 murine constant domain containing monoclonal antibody directed against the human CD20 antigen. However this antibody is not glycoengineered and not afocusylates and thus has an amount of fucose of at least 85 %. This chimeric antibody contains human gamma 1 constant domains and is identified by the name 56MOFO-360876942Attorney Docket No.14639-20729.40"C2B8" in US 5,736,137 (Andersen, et. al.) issued on April 17, 1998, assigned to IDEC Pharmaceuticals Corporation. Rituximab is approved for the treatment of patients with non-Hodgkin's lymphoma (NHL), chronic lymphocytic leukemia (CLL), rheumatoid arthritis, granulomatosis with polyangiitis and microscopic polyangiitis, and pemphigus vulgaris. In vitro mechanism of action studies have shown that rituximab exhibits human complement-dependent cytotoxicity (CDC) (Reff, M.E., et. al, Blood 83(2) (1994) 435-445). Additionally, it exhibits activity in assays that measure antibody-dependent cellular cytotoxicity (ADCC).

[0106] The term “GA101 antibody” as used herein refers to any one of the following antibodies that bind human CD20: (1) an antibody comprising an HVR-H1 comprising the amino acid sequence of SEQ ID NO:1, an HVR-H2 comprising the amino acid sequence of SEQ ID NO:2, an HVR-H3 comprising the amino acid sequence of SEQ ID NO:3, an HVR-L1 comprising the amino acid sequence of SEQ ID NO:4, an HVR-L2 comprising the amino acid sequence of SEQ ID NO:5, and an HVR-L3 comprising the amino acid sequence of SEQ ID NO:6; (2) an antibody comprising a VH domain comprising the amino acid sequence of SEQ ID NO:7 and a VL domain comprising the amino acid sequence of SEQ ID NO:8, (3) an antibody comprising an amino acid sequence of SEQ ID NO:9 and an amino acid sequence of SEQ ID NO: 10; (4) an antibody known as obinutuzumab, or (5) an antibody that comprises an amino acid sequence that has at least 95%, 96%, 97%, 98% or 99% sequence identity with amino acid sequence of SEQ ID NO:9 and that comprises an amino acid sequence that has at least 95%, 96%, 97%, 98% or 99% sequence identity with an amino acid sequence of SEQ ID NO:10. In one embodiment, the GA101 antibody is an IgG1 isotype antibody. In some embodiments, the anti-CD20 antibody is a humanized B-Ly1 antibody.

[0107] The term “humanized B-Ly1 antibody” refers to humanized B-Ly1 antibody as disclosed in WO 2005 / 044859 and WO 2007 / 031875, which were obtained from the murine monoclonal anti-CD20 antibody B-Ly1 (variable region of the murine heavy chain (VH): SEQ ID NO: 11; variable region of the murine light chain (VL): SEQ ID NO: 12- see Poppema, S. and Visser, L., Biotest Bulletin 3 (1987) 131-139) by chimerization with a human constant domain from IgG1 and following humanization (see WO 2005 / 044859 and WO 2007 / 031875). These “humanized B-Ly1 antibodies” are disclosed in detail in WO 2005 / 044859 and WO 2007 / 031875.Variable region of the murine monoclonal anti-CD20 antibody B-Ly1 heavy chain (VH) (SEQ ID NO: 11)57MOFO-360876942Attorney Docket No.14639-20729.40Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys Lys 1 5 10 15Ala Ser Gly Tyr Ala Phe Ser Tyr Ser Trp Met Asn Trp Val Lys Leu 20 25 30 Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly Arg Ile Phe Pro Gly Asp 35 40 45Gly Asp Thr Asp Tyr Asn Gly Lys Phe Lys Gly Lys Ala Thr Leu Thr 50 55 60Ala Asp Lys Ser Ser Asn Thr Ala Tyr Met Gln Leu Thr Ser Leu Thr 65 70 75 80 Ser Val Asp Ser Ala Val Tyr Leu Cys Ala Arg Asn Val Phe Asp Gly 85 90 95Tyr Trp Leu Val Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 100 105 110 Variable region of the murine monoclonal anti-CD20 antibody B-Ly1 light chain (VL) (SEQ ID NO: 12)Asn Pro Val Thr Leu Gly Thr Ser Ala Ser Ile Ser Cys Arg Ser Ser 1 5 10 15Lys Ser Leu Leu His Ser Asn Gly Ile Thr Tyr Leu Tyr Trp Tyr Leu 20 25 30 Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn 35 40 45Leu Val Ser Gly Val Pro Asp Arg Phe Ser Ser Ser Gly Ser Gly Thr 50 55 60Asp Phe Thr Leu Arg Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val 65 70 75 80 Tyr Tyr Cys Ala Gln Asn Leu Glu Leu Pro Tyr Thr Phe Gly Gly Gly 85 90 95Thr Lys Leu Glu Ile Lys Arg100

[0108] In one embodiment, the “humanized B-Ly1 antibody” has variable region of the heavy chain (VH) selected from group of SEQ ID NO:7, 8, and 13 to 33 (corresponding to, inter alia, B-HH2 to B-HH9 and B-HL8 to B-HL17 of WO 2005 / 044859 and WO 2007 / 031875). In one specific embodiment, such variable domain is selected from the group consisting of SEQ ID NOS:14, 15, 7, 19, 25, 27, and 29 (corresponding to B-HH2, BHH-3, B-HH6, B-HH8, B-HL8, B-HL11 and B-HL13 of WO 2005 / 044859 and WO 2007 / 031875). In one specific embodiment, the “humanized B-Ly1 antibody” has variable region of the light chain (VL) of SEQ ID NO:8 (corresponding to B-KV1 of WO 2005 / 044859 and WO 2007 / 031875). In one specific embodiment, the “humanized B-Ly1 antibody” has a variable region of the heavy chain (VH) of SEQ ID NO:7 (corresponding to B-HH6 of WO 2005 / 044859 and WO 2007 / 031875) and a58MOFO-360876942Attorney Docket No.14639-20729.40variable region of the light chain (VL) of SEQ ID NO:8 (corresponding to B-KV1 ofWO 2005 / 044859 and WO 2007 / 031875). Furthermore in one embodiment, the humanized B-Ly1 antibody is an IgG1 antibody. According to the invention such afocusylated humanized B-Ly1 antibodies are glycoengineered (GE) in the Fc region according to the procedures described in WO 2005 / 044859, WO 2004 / 065540, WO 2007 / 031875, Umana, P. et al., Nature Biotechnol.17 (1999) 176-180 and WO 99 / 154342. In one embodiment, the afucosylated glyco-engineered humanized B-Ly1 is B-HH6-B-KV1 GE. In one embodiment, the anti-CD20 antibody is obinutuzumab (recommended INN, WHO Drug Information, Vol.26, No.4, 2012, p.453). As used herein, obinutuzumab is synonymous for GA101 or RO5072759. This replaces all previous versions (e.g. Vol.25, No.1, 2011, p.75-76), and is formerly known as afutuzumab (recommended INN, WHO Drug Information, Vol.23, No.2, 2009, p.176;Vol.22, No.2, 2008, p.124). As used herein, references to obinutuzumab refer to GAZYVA® as well as biosimilar antibodies thereof. In some embodiments, the humanized B-Ly1 antibody is an antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10 or an antigen-binding fragment thereof. In some embodiments, the humanized B-Ly1 antibody comprises a heavy chain variable region comprising the three heavy chain CDRs of SEQ ID NO:9 and a light chain variable region comprising the three light chain CDRs of SEQ ID NO:10.Heavychain (SEQ ID NO:9)QVQLVQSGAE VKKPGSSVKV SCKASGYAFSYSWINWVRQA PGQGLEWMGR50 IFPGDGDTDY NGKFKGRVTI TADKSTSTAYMELSSLRSED TAVYYCARNV100 FDGYWLVYWG QGTLVTVSSA STKGPSVFPLAPSSKSTSGG TAALGCLVKD150 YFPEPVTVSW NSGALTSGVH TFPAVLQSSGLYSLSSVVTV PSSSLGTQTY200 ICNVNHKPSN TKVDKKVEPK SCDKTHTCPPCPAPELLGGP SVFLFPPKPK250 DTLMISRTPE VTCVVVDVSH EDPEVKFNWYVDGVEVHNAK TKPREEQYNS300 TYRVVSVLTV LHQDWLNGKE YKCKVSNKALPAPIEKTISK AKGQPREPQV350 YTLPPSRDEL TKNQVSLTCL VKGFYPSDIAVEWESNGQPE NNYKTTPPVL400 DSDGSFFLYS KLTVDKSRWQ QGNVFSCSVMHEALHNHYTQ KSLSLSPG 449Light chain (SEQ ID NO:10)DIVMTQTPLS LPVTPGEPAS ISCRSSKSLLHSNGITYLYW YLQKPGQSPQ50 LLIYQMSNLV SGVPDRFSGS GSGTDFTLKISRVEAEDVGV YYCAQNLELP100 YTFGGGTKVE IKRTVAAPSV FIFPPSDEQLKSGTASVVCL LNNFYPREAK150 VQWKVDNALQ SGNSQESVTE QDSKDSTYSLSSTLTLSKAD YEKHKVYACE200 VTHQGLSSPV TKSFNRGEC 21959MOFO-360876942Attorney Docket No.14639-20729.40

[0109] In some embodiments, the humanized B-Ly1 antibody is an afucosylated glyco-engineered humanized B-Ly1. Such glycoengineered humanized B-Ly1 antibodies have an altered pattern of glycosylation in the Fc region, preferably having a reduced level of fucose residues. Preferably the amount of fucose is 60 % or less of the total amount of oligosaccharides at Asn297 (in one embodiment the amount of fucose is between 40 % and 60 %, in another embodiment the amount of fucose is 50 % or less, and in still another embodiment the amount of fucose is 30 % or less). Furthermore the oligosaccharides of the Fc region are preferably bisected. These glycoengineered humanized B-Ly1 antibodies have an increased ADCC.

[0110] The “ratio of the binding capacities to CD20 on Raji cells (ATCC-No. CCL-86) of an anti-CD20 antibodies compared to rituximab” is determined by direct immunofluorescence measurement (the mean fluorescence intensities (MFI) is measured) using said anti-CD20 antibody conjugated with Cy5 and rituximab conjugated with Cy5 in a FACSArray (Becton Dickinson) with Raji cells (ATCC-No. CCL-86), as described in Example No.2, and calculated as follows:Ratio of the binding capacities to CD20 on Raji cells (ATCC-No. CCL-86) =MFI (Cy5 -anti- CD20 antibody) Cy5- labeling ratio (Cy5 - rituximab) MFI (Cy5 - rituximab) Cy5- labeling ratio (Cy5 -anti- CD20 antibody)

[0111] MFI is the mean fluorescent intensity. The “Cy5-labeling ratio” as used herein means the number of Cy5-label molecules per molecule antibody.

[0112] Typically said type II anti-CD20 antibody has a ratio of the binding capacities to CD20 on Raji cells (ATCC-No. CCL-86) of said second anti-CD20 antibody compared to rituximab of 0.3 to 0.6, and in one embodiment, 0.35 to 0.55, and in yet another embodiment, 0.4 to 0.5.

[0113] In one embodiment said type II anti-CD20 antibody, e.g., a GA101 antibody, has increased antibody dependent cellular cytotoxicity (ADCC).

[0114] By “antibody having increased antibody dependent cellular cytotoxicity (ADCC)”, it is meant an antibody, as that term is defined herein, having increased ADCC as determined by any suitable method known to those of ordinary skill in the art. One accepted in vitro ADCC assay is as follows:60MOFO-360876942Attorney Docket No.14639-20729.401) the assay uses target cells that are known to express the target antigen recognized by the antigen-binding region of the antibody;2) the assay uses human peripheral blood mononuclear cells (PBMCs), isolated from blood of a randomly chosen healthy donor, as effector cells;3) the assay is carried out according to following protocol:i) the PBMCs are isolated using standard density centrifugation procedures and are suspended at 5 x 106cells / ml in RPMI cell culture medium;ii) the target cells are grown by standard tissue culture methods, harvested from the exponential growth phase with a viability higher than 90%, washed in RPMI cell culture medium, labeled with 100 micro-Curies of51Cr, washed twice with cell culture medium, and resuspended in cell culture medium at a density of 105cells / ml;iii) 100 microliters of the final target cell suspension above are transferred to each well of a 96-well microtiter plate;iv) the antibody is serially-diluted from 4000 ng / ml to 0.04 ng / ml in cell culture medium and 50 microliters of the resulting antibody solutions are added to the target cells in the 96-well microtiter plate, testing in triplicate various antibody concentrations covering the whole concentration range above;v) for the maximum release (MR) controls, 3 additional wells in the plate containing the labeled target cells, receive 50 microliters of a 2% (VN) aqueous solution of non-ionic detergent (Nonidet, Sigma, St. Louis), instead of the antibody solution (point iv above);vi) for the spontaneous release (SR) controls, 3 additional wells in the plate containing the labeled target cells, receive 50 microliters of RPMI cell culture medium instead of the antibody solution (point iv above);vii) the 96-well microtiter plate is then centrifuged at 50 x g for 1 minute and incubated for 1 hour at 4°C;viii) 50 microliters of the PBMC suspension (point i above) are added to each well to yield an effector:target cell ratio of 25:1 and the plates are placed in an incubator under 5% CO2 atmosphere at 37°C for 4 hours;61MOFO-360876942Attorney Docket No.14639-20729.40ix) the cell-free supernatant from each well is harvested and the experimentally released radioactivity (ER) is quantified using a gamma counter;x) the percentage of specific lysis is calculated for each antibody concentration according to the formula (ER-MR) / (MR-SR) x 100, where ER is the average radioactivity quantified (see point ix above) for that antibody concentration, MR is the average radioactivity quantified (see point ix above) for the MR controls (see point V above), and SR is the average radioactivity quantified (see point ix above) for the SR controls (see point vi above);4) "increased ADCC" is defined as either an increase in the maximum percentage of specific lysis observed within the antibody concentration range tested above, and / or a reduction in the concentration of antibody required to achieve one half of the maximum percentage of specific lysis observed within the antibody concentration range tested above. In one embodiment, the increase in ADCC is relative to the ADCC, measured with the above assay, mediated by the same antibody, produced by the same type of host cells, using the same standard production, purification, formulation and storage methods, which are known to those skilled in the art, except that the comparator antibody (lacking increased ADCC) has not been produced by host cells engineered to overexpress GnTIII and / or engineered to have reduced expression from the fucosyltransferase 8 (FUT8) gene (e.g., including, engineered for FUT8 knock out).

[0115] Said "increased ADCC" can be obtained by, for example, mutating and / or glycoengineering of said antibodies. In one embodiment, the antibody is glycoengineered to have a biantennary oligosaccharide attached to the Fc region of the antibody that is bisected by GlcNAc, e.g., in WO 2003 / 011878 (Jean-Mairet et al.); US Patent No.6,602,684 (Umana et al.); US 2005 / 0123546 (Umana et al.), Umana, P., et al., Nature Biotechnol.17 (1999) 176-180). In another embodiment, the antibody is glycoengineered to lack fucose on the carbohydrate attached to the Fc region by expressing the antibody in a host cell that is deficient in protein fucosylation (e.g., Lec13 CHO cells or cells having an alpha-1,6-fucosyltransferase gene (FUT8) deleted or the FUT gene expression knocked down (see, e.g., Yamane-Ohnuki et al. Biotech. Bioeng.87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107). In yet another embodiment, the antibody sequence has been engineered in its Fc region to enhance ADCC (e.g., in one embodiment, such engineered antibody variant62MOFO-360876942Attorney Docket No.14639-20729.40comprises an Fc region with one or more amino acid substitutions at positions 298, 333, and / or 334 of the Fc region (EU numbering of residues)).

[0116] The term "complement-dependent cytotoxicity (CDC)" refers to lysis of o tumor target cells by the antibody according to the invention in the presence of complement. CDC can be measured by the treatment of a preparation of CD20 expressing cells with an anti-CD20 antibody according to the invention in the presence of complement. CDC is found if the antibody induces at a concentration of 100 nM the lysis (cell death) of 20% or more of the tumor cells after 4 hours. In one embodiment, the assay is performed with51Cr or Eu labeled tumor cells and measurement of released51Cr or Eu. Controls include the incubation of the tumor target cells with complement but without the antibody.

[0117] The term "expression of the CD20” antigen is intended to indicate a significant level of expression of the CD20 antigen in a cell, e.g., a T- or B- Cell. In one embodiment, patients to be treated according to the methods of this invention express significant levels of CD20 on a B-cell. CD20 expression on a B-cell can be determined by standard assays known in the art. e.g., CD20 antigen expression is measured using immunohistochemical (IHC) detection, FACS or via PCR-based detection of the corresponding mRNA.

[0118] As used in this specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to “a molecule” optionally includes a combination of two or more such molecules, and the like.

[0119] The term “about” as used herein refers to the usual error range for the respective value readily known to the skilled person in this technical field. Reference to “about” a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se.

[0120] It is understood that aspects and embodiments of the invention described herein include “comprising,” “consisting,” and “consisting essentially of” aspects and embodiments.III. Methods

[0121] In one aspect, provided herein are methods for depleting and / or reducing activity of kidney-resident B cells in an individual in need thereof (e.g., an individual having lupus, or an individual having lupus nephritis), comprising administering to the individual at least a first 63MOFO-360876942Attorney Docket No.14639-20729.40antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human.

[0122] In another aspect, provided herein are methods for treating lupus nephritis in an individual that has lupus, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure; wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab; wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab; wherein the individual is a human.

[0123] In some embodiments, the methods result in depletion of kidney-resident B cells in the individual. In some embodiments, treatment with obinutuzumab in a plurality of individuals according to the methods of the present disclosure results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure (e.g., after administration of the first dose of the first antibody exposure at Week 0 or Day 1). In some embodiments, kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure64MOFO-360876942Attorney Docket No.14639-20729.40(e.g., prior to a first dose of the first antibody exposure) have an average of at least 10, at least 12, at least 13, or at least 15 B cells per mm2tissue. In some embodiments, treatment of an individual with obinutuzumab results in one or no B cells per mm2tissue in a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure (e.g., after administration of the first dose of the first antibody exposure at Week 0 or Day 1). In some embodiments, a kidney biopsy sample obtained from the individual prior to treatment with obinutuzumab according to the methods of the present disclosure (e.g., prior to a first dose of the first antibody exposure) has at least 10, at least 15, at least 25, at least 50, at least 100, or at least 200 B cells per mm2tissue. In some embodiments, treatment with obinutuzumab in a plurality of individuals results in a 90% or greater, 91% or greater, 92% or greater, 93% or greater, 94% or greater, 95% or greater, 96% or greater, 97% or greater, or 98% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure (e.g., after administration of the first dose of the first antibody exposure at Week 0 or Day 1), as compared to an average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure (e.g., prior to a first dose of the first antibody exposure).

[0124] In some embodiments, the methods result in reduced or eliminated activity of kidney-resident B cells in the individual. In some embodiments, a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure (e.g., after administration of the first dose of the first antibody exposure at Week 0 or Day 1) exhibits an activity index (AI) of less than or equal to 1. In some embodiments, a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure (e.g., after administration of the first dose of the first antibody exposure at Week 0 or Day 1) exhibits an AI of 0. In some embodiments, a kidney biopsy sample obtained from the individual prior to administration of the first antibody exposure (e.g., prior to a first dose of the first antibody exposure) exhibits an AI of greater than or equal to 4 or greater than or equal to 6.

[0125] In some embodiments, references to “baseline” and “prior to administration of the first antibody exposure” interchangeably refer to a time prior to administration of a first dose of a first antibody exposure of the present disclosure. In some embodiments, a “baseline” sample is65MOFO-360876942Attorney Docket No.14639-20729.40one that is obtained from an individual prior to treatment as disclosed herein, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure.

[0126] In some embodiments, the individual has one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female. In some embodiments, the individual has one, two, three, four, five, six, seven, or all of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female. In some embodiments, the individual is a human. In some embodiments, the individual is female.

[0127] In some embodiments, the methods comprise administering to the individual at least a first antibody exposure to a type II anti-CD20 antibody and a second antibody exposure to the type II anti-CD20 antibody; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure; wherein the first antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; wherein the second antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody. For example, in some embodiments, the methods comprise administering to the individual two doses of 1000mg66MOFO-360876942Attorney Docket No.14639-20729.40of the type II anti-CD20 antibody on weeks 0 and 2 of treatment and two doses of 1000mg of the type II anti-CD20 antibody on weeks 24 and 26 of treatment.

[0128] In some embodiments, the methods further comprise administering to the individual a third antibody exposure to the type II anti-CD20 antibody, wherein the third antibody exposure comprises one or two doses of the type II anti-CD20 antibody, and wherein the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure. For example, in some embodiments, the methods comprise administering to the individual a single dose of 1000mg of the type II anti-CD20 antibody on week 52 of treatment. In some embodiments, the methods comprise administering to the individual two doses of 1000mg of the type II anti-CD20 antibody on weeks 50 and 52 of treatment.

[0129] In some embodiments, the first antibody exposure comprises a first dose of between about 900mg and about 1100mg (e.g., about 1000mg) of the type II anti-CD20 antibody and a second dose of between about 900mg and about 1100mg (e.g., about 1000mg) of the type II anti-CD20 antibody. In some embodiments, the second dose of the first antibody exposure is not provided until between about 1.5 weeks and about 2.5 weeks (e.g., about 2 weeks) after the first dose of the first antibody exposure. In some embodiments, the second dose of the first antibody exposure is not provided until between about 10 weeks and about 17 days (e.g., about 14 days) after the first dose of the first antibody exposure.

[0130] In some embodiments, the second antibody exposure comprises a first dose of between about 900mg and about 1100mg (e.g., about 1000mg) of the type II anti-CD20 antibody and a second dose of between about 900mg and about 1100mg (e.g., about 1000mg) of the type II anti-CD20 antibody. In some embodiments, the second dose of the second antibody exposure is not provided until between about 1.5 weeks and about 2.5 weeks (e.g., about 2 weeks) after the first dose of the second antibody exposure. In some embodiments, the second dose of the second antibody exposure is not provided until between about 10 days and about 17 days (e.g., about 14 days) after the first dose of the second antibody exposure.

[0131] In some embodiments, the third antibody exposure one or two doses of the type II anti-CD20 antibody. In some embodiments, the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure. In some embodiments, the third exposure comprises a single dose of between about 900mg and about 1100mg (e.g., about 1000mg) of the type II anti-CD20 antibody. In some embodiments, the67MOFO-360876942Attorney Docket No.14639-20729.40single dose of the third antibody exposure is not provided until between about 48 weeks and about 56 weeks (e.g., about 52 weeks) after the first dose of the first antibody exposure or until between about 24 weeks and about 32 weeks (e.g., about 28 weeks) after the first dose of the second antibody exposure. In some embodiments, the single dose of the third antibody exposure is not provided until between about 48 weeks and about 56 weeks (e.g., about 52 weeks) after the first dose of the first antibody exposure or until between about 336 days and about 392 days (e.g., about 365 days) after the first dose of the second antibody exposure. In some embodiments, the third exposure comprises two doses of between about 900mg and about 1100mg (e.g., about 1000mg) of the type II anti-CD20 antibody. In some embodiments, the first dose of the third antibody exposure is not provided until between about 322 days and about 378 days (e.g., about 350 days) after the first dose of the first antibody exposure or until between about 168 days and about 224 days (e.g., about 196 days) after the first dose of the second antibody exposure. In some embodiments, the first dose of the third antibody exposure is not provided until between about 46 weeks and about 54 weeks (e.g., about 50 weeks) after the first dose of the first antibody exposure or until between about 24 weeks and about 32 weeks (e.g., about 28 weeks) after the first dose of the second antibody exposure. In some embodiments, the second dose of the third antibody exposure is not provided until between about 48 weeks and about 56 weeks (e.g., about 52 weeks) after the first dose of the first antibody exposure or until between about 24 weeks and about 32 weeks (e.g., about 28 weeks) after the first dose of the second antibody exposure. In some embodiments, the second dose of the third antibody exposure is not provided until between about 48 weeks and about 56 weeks (e.g., about 52 weeks) after the first dose of the first antibody exposure or until between about 336 days and about 392 days (e.g., about 365 days) after the first dose of the second antibody exposure.

[0132] In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on days 1 and 15 of treatment and a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on days 168 and 182 of treatment. In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on weeks 0 and 2 of treatment and a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on68MOFO-360876942Attorney Docket No.14639-20729.40weeks 24 and 26 of treatment. In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) in the first month of treatment (e.g., on weeks 0 and 2) and a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) at 6 months of treatment (e.g., on weeks 24 and 26).

[0133] In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on days 1 and 15 of treatment, a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on days 168 and 182 of treatment, and a third antibody exposure comprising one dose of 1000 mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on day 364 of treatment. In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on weeks 0 and 2 of treatment, a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on weeks 24 and 26 of treatment, and a third antibody exposure comprising one dose of 1000 mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on week 52 of treatment. In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) in the first month of treatment (e.g., on weeks 0 and 2), a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) at 6 months of treatment (e.g., on weeks 24 and 26), and a third antibody exposure comprising one dose of 1000 mg of the type II anti-CD20 antibody (e.g., obinutuzumab) at 12 months of treatment (e.g., on week 52).

[0134] In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on days 1 and 15 of treatment, a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on days 168 and 182 of treatment, and a third antibody exposure comprising two doses of 1000 mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on days 350 and 364 of treatment. In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on weeks 0 and 269MOFO-360876942Attorney Docket No.14639-20729.40of treatment, a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on weeks 24 and 26 of treatment, and a third antibody exposure comprising two doses of 1000 mg of the type II anti-CD20 antibody (e.g., obinutuzumab) on weeks 50 and 52 of treatment. In some embodiments, the methods comprise administering to the individual a first antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) in the first month of treatment (e.g., on weeks 0 and 2), a second antibody exposure that comprises two doses of 1000mg of the type II anti-CD20 antibody (e.g., obinutuzumab) at 6 months of treatment (e.g., on weeks 24 and 26), and a third antibody exposure comprising two doses of 1000 mg of the type II anti-CD20 antibody (e.g., obinutuzumab) at 12 months of treatment (e.g., on weeks 50 and 52).

[0135] In some embodiments, the methods further comprise (e.g., after a third antibody exposure of the present disclosure) administering to the individual a fourth antibody exposure of 1000 mg of obinutuzumab. In some embodiments, the fourth antibody exposure is administered at week 80 of treatment. In some embodiments, the methods further comprise administering to the individual one or more additional exposure(s) of 1000 mg of obinutuzumab every 6 months, e.g., after week 80 of treatment. In some embodiments, the methods further comprise (e.g., after a third antibody exposure of the present disclosure) administering to the individual one or more doses of 1000 mg of obinutuzumab every 6 months. In some embodiments, the methods further comprise (e.g., after a third antibody exposure of the present disclosure) administering to the individual additional exposures of 1000 mg of obinutuzumab on weeks 80, 106, 132, 158, and 184 of treatment. In some embodiments, the methods further comprise (e.g., after a third antibody exposure of the present disclosure) administering to the individual additional doses of 1000 mg of obinutuzumab on weeks 80, 106, 132, 158, and 184 of treatment. In some embodiments, the methods further comprise (e.g., after week 184 of treatment) administering to the individual one or more doses of 1000 mg of obinutuzumab every 6 months.

[0136] In some embodiments, the methods further comprise (e.g., prior to administration of a first dose of the first antibody exposure) identifying the individual as having a UPCR of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g. In some embodiments, the methods further comprise (e.g., prior to administration of a first dose of the first antibody exposure) determining that the individual has a UPCR of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or70MOFO-360876942Attorney Docket No.14639-20729.40equal to about 3.0 g / g. In some embodiments, the identification and / or determination comprise(s) measuring UPCR in a urine sample obtained from the individual. In some embodiments, the methods further comprise obtaining a urine sample from the individual.

[0137] In some embodiments, the methods further comprise (e.g., prior to administration of a first dose of the first antibody exposure) identifying the individual as having or being one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female. In some embodiments, the individual has or is one, two, three, four, five, six, seven, or all of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female. In some embodiments, the individual has or is one or more of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female. In some embodiments, the individual has or is one, two, three, four, five, six, seven, or all of the following at baseline (e.g., prior to administration of the first antibody exposure): (a) a urine protein-to-creatinine ratio (UPCR) of71MOFO-360876942Attorney Docket No.14639-20729.40greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g; (b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2; (c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level; (e) a serum sample that has a low C4 complement level; (f) no prior history or diagnosis of lupus and / or lupus nephritis; (g) concomitant class V lupus nephritis; and / or (h) is female. Methods for measuring anti-dsDNA antibodies (e.g., in a sample such as a serum sample) are known in the art; see, e.g., Kavanaugh, A.F. et al. (2002) Arthritis Rheum.47(5):546-555. Methods for measuring complement C3 and C4 (e.g., in a sample such as a serum sample) are known in the art; see, e.g., Ayano, M. and Horiuchi, T. (2023) Biomolecules 13(2):367. In some embodiments, prior to administration of the first antibody exposure, the individual with lupus nephritis has active disease. In some embodiments, prior to treatment, the individual with lupus nephritis has active disease. In some embodiments, active disease refers to: (a) active or active / chronic Class III or IV proliferative LN, e.g., as evidenced by renal biopsy performed within 6 months (and optional Class V disease); and / or (b) baseline UPCR of greater than or equal to 1.0 g / g, e.g., on a 24-hour collection.

[0138] In some embodiments, the methods of the present disclosure result in histological remission in a treated individual. For example, as described herein, in some embodiments, the methods result in depletion of kidney-resident B cells and / or reduced or eliminated activity of kidney-resident B cells in the individual.

[0139] In some embodiments, the methods of the present disclosure result in depletion of kidney-resident B cells in an individual. In some embodiments, the kidney-resident B cells are CD79a+ / CD138- B cells. Methods for assessing kidney-resident B cells are known in the art, and exemplary such methods are described herein. In some embodiments, depletion is assessed by counting B cells in a kidney biopsy sample obtained from an individual. For example, number of B cells (e.g., CD79a+ / CD138- B cells) can be quantified using one or more formalin-fixed, paraffin-embedded tissue sections (e.g., from a kidney biopsy sample) by immunofluorescence microscopy. In some embodiments, depletion is assessed by counting kidney-resident B cells in kidney biopsy samples obtained from the same individual before and after treatment, e.g., obtained before treatment at Week 0 and at Week 76 (e.g., between Week 76 and Week 80) of treatment.72MOFO-360876942Attorney Docket No.14639-20729.40

[0140] In some embodiments, the methods of the present disclosure result in reduced or eliminated activity of kidney-resident B cells in an individual. In some embodiments, the kidney-resident B cells are CD79a+ / CD138- B cells. In some embodiments, activity of kidney-resident B cells is assessed using the NIH activity index (AI), which provides a semiquantitative assessment of LN activity in the kidney, to one or more kidney biopsy samples. See Bajema, I.M. et al. Kidney Int.2018;93(4):789-796. The AI provides an overall score that takes into account sub-scores related to endocapillary hypercellularity, neutrophils / karyorrhexis, fibrinoid necrosis, hyaline deposits, cellular / fibrocellular crescents, and interstitial inflammation. In some embodiments, activity of kidney-resident B cells is assessed by measuring AI in kidney biopsy samples obtained from the same individual before and after treatment, e.g., obtained before treatment at Week 0 and at Week 76 (e.g., between Week 76 and Week 80) of treatment.

[0141] In some embodiments, the methods of the present disclosure result in prevention or delaying progression of end-stage kidney disease. In some embodiments, the methods of the present disclosure result in a complete renal response at week 76 of treatment. In some embodiments, the methods of the present disclosure result in a complete renal response at week 76 of treatment. In some embodiments, administering such treatment to a plurality of human patients with lupus nephritis (e.g., according to the methods of the present disclosure) results in an improvement in complete renal response at week 76 of treatment, as compared to treatment of a plurality of human patients with lupus nephritis with standard-of-care treatment.

[0142] In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.8 g / g after treatment as disclosed herein. In some embodiments, the methods of the present disclosure result in a UPCR of less than about 0.8 g / g after treatment as disclosed herein. In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g after treatment as disclosed herein. In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g, e.g., sustained through about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a73MOFO-360876942Attorney Docket No.14639-20729.40first antibody exposure of the present disclosure). In some embodiments, the treated individual has a UPCR of less than or equal to about 0.7 g / g, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, a UPCR of less than or equal to about 0.7 g / g is measured in a sample obtained from the treated individual, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).

[0143] In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.5 g / g after treatment as disclosed herein. In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.5 g / g, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.5 g / g, e.g., sustained through about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the treated individual has a UPCR of less than or equal to about 0.5 g / g, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, a UPCR of less than or equal to about 0.5 g / g is measured in a sample obtained from the treated individual, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).

[0144] Certain aspects of the present disclosure relate to measurement of UPCR. In some embodiments, UPCR is measured on a 24-hour urine collection. Assays for measuring UPCR (e.g., in a urine sample) are known in the art.74MOFO-360876942Attorney Docket No.14639-20729.40

[0145] In some embodiments, the methods of the present disclosure result in an eGFR of the treated individual that is greater than or equal to about 80% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the methods of the present disclosure result in an eGFR of the treated individual that is greater than or equal to 60mL / min or greater than or equal to about 80% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, an eGFR of a treated individual is measured in a sample obtained from the individual at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).

[0146] In some embodiments, the methods of the present disclosure result in an eGFR of the treated individual that is greater than or equal to about 85% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, an eGFR of a treated individual is measured in a sample obtained from the individual at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).

[0147] Certain aspects of the present disclosure relate to measurement of eGFR. Assays for measuring eGFR (e.g., in a urine sample) are known in the art. In some embodiments, eGFR is calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) creatinine equation (see, e.g., Levey, A.S. and Stevens, L.A. (2010) Am J Kidney Dis 55(4):622-627).

[0148] In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.5 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure) in an individual after treatment as disclosed herein. In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.5 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR in an individual, e.g., at75MOFO-360876942Attorney Docket No.14639-20729.40about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.5 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR in an individual, e.g., sustained through about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the treated individual has a UPCR of less than or equal to about 0.5 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, a UPCR of less than or equal to about 0.5 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR are measured in a sample obtained from the treated individual, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the treated individual does not experience treatment failure or require rescue therapy, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).

[0149] In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure) in an individual after treatment as disclosed herein. In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR in an individual, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the methods of the76MOFO-360876942Attorney Docket No.14639-20729.40present disclosure result in a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR in an individual, e.g., sustained through about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the treated individual has a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 85% of a baseline eGFR are measured in a sample obtained from the treated individual, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the treated individual does not experience treatment failure or require rescue therapy, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).

[0150] In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 60mL / min or greater than or equal to about 80% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure) in an individual after treatment as disclosed herein. In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 60mL / min or greater than or equal to about 80% of a baseline eGFR in an individual, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 60mL / min or greater than or equal to about 80% of a baseline eGFR in an individual, e.g.,77MOFO-360876942Attorney Docket No.14639-20729.40sustained through about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the treated individual has a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 60mL / min or greater than or equal to about 80% of a baseline eGFR, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, a UPCR of less than or equal to about 0.7 g / g and an eGFR that is greater than or equal to about 60mL / min or greater than or equal to about 80% of a baseline eGFR are measured in a sample obtained from the treated individual, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the treated individual does not experience treatment failure or require rescue therapy, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).Anti-CD20 antibodies

[0151] Certain aspects of the present disclosure relate to anti-CD20 antibodies, e.g., for use in methods described herein, e.g., for treating or preventing progression of lupus nephritis. In some embodiments, the anti-CD20 antibody is a type II antibody. In some embodiments, the anti-CD20 antibody is human or humanized. In some embodiments, the anti-CD20 antibody is afucosylated. In some embodiments, the anti-CD20 antibody is a GA101 antibody.

[0152] Examples of type II anti-CD20 antibodies include e.g. humanized B-Ly1 antibody IgG1 (a chimeric humanized IgG1 antibody as disclosed in WO 2005 / 044859), 11B8 IgG1 (as disclosed in WO 2004 / 035607), and AT80 IgG1. Typically type II anti-CD20 antibodies of the IgG1 isotype show characteristic CDC properties. Type II anti-CD20 antibodies have a decreased CDC (if IgG1 isotype) compared to type I antibodies of the IgG1 isotype.

[0153] In some embodiments, the anti-CD20 antibody is a GA101 antibody described herein. In some embodiments, the anti-CD20 is any one of the following antibodies that bind human 78MOFO-360876942Attorney Docket No.14639-20729.40CD20: (1) an antibody comprising an HVR-H1 comprising the amino acid sequence of GYAFSY (SEQ ID NO:1), an HVR-H2 comprising the amino acid sequence of FPGDGDTD (SEQ ID NO:2), an HVR-H3 comprising the amino acid sequence of NVFDGYWLVY (SEQ ID NO:3), an HVR-L1 comprising the amino acid sequence of RSSKSLLHSNGITYLY (SEQ ID NO:4), an HVR-L2 comprising the amino acid sequence of QMSNLVS (SEQ ID NO:5), and an HVR-L3 comprising the amino acid sequence of AQNLELPYT (SEQ ID NO:6); (2) an antibody comprising a VH domain comprising the amino acid sequence of SEQ ID NO:7 and a VL domain comprising the amino acid sequence of SEQ ID NO:8, (3) an antibody comprising an amino acid sequence of SEQ ID NO:9 and an amino acid sequence of SEQ ID NO:10; (4) an antibody known as obinutuzumab, or (5) an antibody that comprises an amino acid sequence that has at least 95%, 96%, 97%, 98% or 99% sequence identity with amino acid sequence of SEQ ID NO:9 and that comprises an amino acid sequence that has at least 95%, 96%, 97%, 98% or 99% sequence identity with an amino acid sequence of SEQ ID NO:10. In one embodiment, the GA101 antibody is an IgG1 isotype antibody. In some embodiments, the anti-CD20 antibody comprises an HVR-H1, HVR-H2, HVR-H3, HVR-L1, HVR-L2, and HVR-L3 of any of the antibodies described herein, e.g., 3 HVRs from SEQ ID NO:7 and 3 HVRs from SEQ ID NO:8, 3 HVRs from SEQ ID NO:9 and 3 HVRs from SEQ ID NO:10, or any HVRs of the amino acid sequences provided in Table 2.

[0154] In some embodiments, the anti-CD20 antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:7, and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:8.QVQLVQSGAEVKKPGSSVKVSCKASGYAFSYSWINWVRQAPGQGLEWMGRIFPGDGD TDYNGKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARNVFDGYWLVYWGQGTL VTVSS (SEQ ID NO:7) DIVMTQTPLSLPVTPGEPASISCRSSKSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLV SGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCAQNLELPYTFGGGTKVEIKRTV (SEQ ID NO:8).

[0155] In some embodiments, the anti-CD20 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO:9, and a light chain comprising the amino acid sequence of SEQ ID NO:10.79MOFO-360876942Attorney Docket No.14639-20729.40QVQLVQSGAEVKKPGSSVKVSCKASGYAFSYSWINWVRQAPGQGLEWMGRIFPGDGDTDYNGKF KGRVTITADKSTSTAYMELSSLRSEDTAVYYCARNVFDGYWLVYWGQGTLVTVSSASTKGPSVF PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPS SSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWE SNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO:9) DIVMTQTPLSLPVTPGEPASISCRSSKSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLVSGVP DRFSGSGSGTDFTLKISRVEAEDVGVYYCAQNLELPYTFGGGTKVEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:10)

[0156] In some embodiments, the anti-CD20 antibody is a humanized B-Ly1 antibody. In some embodiments, the humanized B-Ly1 antibody comprises a heavy chain variable region comprising the three heavy chain CDRs of SEQ ID NO:9 and a light chain variable region comprising the three light chain CDRs of SEQ ID NO:10. In some embodiments, the humanized B-Ly1 antibody comprises a heavy chain comprising the sequence of SEQ ID NO:9 and a light chain comprising the sequence of SEQ ID NO:10.

[0157] In some embodiments, the anti-CD20 antibody comprises an amino acid sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a polypeptide sequence listed in Table 2 below.Table 2. Polypeptide sequences.80MOFO-360876942Attorney Docket No.14639-20729.4081MOFO-360876942Attorney Docket No.14639-20729.4082MOFO-360876942Attorney Docket No.14639-20729.40

[0158] In some embodiments, the anti-CD20 antibody (e.g., a type II anti-CD20 antibody) is an afucosylated glyco-engineered antibody. Such glycoengineered antibodies have an altered pattern of glycosylation in the Fc region, preferably having a reduced level of fucose residues. Preferably the amount of fucose is 60 % or less of the total amount of oligosaccharides at Asn297 (in one embodiment the amount of fucose is between 40 % and 60 %, in another embodiment the amount of fucose is 50 % or less, and in still another embodiment the amount of fucose is 30 % or less). Furthermore the oligosaccharides of the Fc region are preferably bisected. In some embodiments, the type II anti-CD20 antibody comprises an Fc region comprising a biantennary oligosaccharide that is bisected by N-acetyl glucosamine (GlcNAc). These glycoengineered humanized anti-CD20 (e.g., B-Ly1) antibodies have an increased ADCC.

[0159] The oligosaccharide component can significantly affect properties relevant to the efficacy of a therapeutic glycoprotein, including physical stability, resistance to protease attack, interactions with the immune system, pharmacokinetics, and specific biological activity. Such properties may depend not only on the presence or absence, but also on the specific structures, of oligosaccharides. Some generalizations between oligosaccharide structure and glycoprotein function can be made. For example, certain oligosaccharide structures mediate rapid clearance of the glycoprotein from the bloodstream through interactions with specific carbohydrate binding proteins, while others can be bound by antibodies and trigger undesired immune reactions.(Jenkins, N., et al., Nature Biotechnol.14 (1996) 975-81).

[0160] Mammalian cells are the preferred hosts for production of therapeutic glycoproteins, due to their capability to glycosylate proteins in the most compatible form for human application. (Cumming, D.A., et al., Glycobiology 1 (1991) 115-30; Jenkins, N., et al., Nature Biotechnol.1483MOFO-360876942Attorney Docket No.14639-20729.40(1996) 975-81). Bacteria very rarely glycosylate proteins, and like other types of common hosts, such as yeasts, filamentous fungi, insect and plant cells, yield glycosylation patterns associated with rapid clearance from the blood stream, undesirable immune interactions, and in some specific cases, reduced biological activity. Among mammalian cells, Chinese hamster ovary (CHO) cells have been most commonly used during the last two decades. In addition to giving suitable glycosylation patterns, these cells allow consistent generation of genetically stable, highly productive clonal cell lines. They can be cultured to high densities in simple bioreactors using serum free media, and permit the development of safe and reproducible bioprocesses. Other commonly used animal cells include baby hamster kidney (BHK) cells, NSO- and SP2 / 0-mouse myeloma cells. More recently, production from transgenic animals has also been tested. (Jenkins, N., et al., Nature Biotechnol.14 (1996) 975-981).

[0161] Antibodies may contain carbohydrate structures at conserved positions in the heavy chain constant regions, with each isotype possessing a distinct array of N-linked carbohydrate structures, which variably affect protein assembly, secretion or functional activity. (Wright, A., and Morrison, S.L., Trends Biotech.15 (1997) 26-32). The structure of the attached N-linked carbohydrate varies considerably, depending on the degree of processing, and can include high-mannose, multiply-branched as well as biantennary complex oligosaccharides. (Wright, A., and Morrison, S.L., Trends Biotech.15 (1997) 26-32). Typically, there is heterogeneous processing of the core oligosaccharide structures attached at a particular glycosylation site such that even monoclonal antibodies exist as multiple glycoforms. Likewise, it has been shown that major differences in antibody glycosylation occur between cell lines, and even minor differences are seen for a given cell line grown under different culture conditions. (Lifely, M.R., et al., Glycobiology 5(8) (1995) 813-22).

[0162] One way to obtain large increases in potency, while maintaining a simple production process and potentially avoiding significant, undesirable side effects, is to enhance the natural, cell-mediated effector functions of monoclonal antibodies by engineering their oligosaccharide component as described in Umana, P., et al., Nature Biotechnol.17 (1999) 176-180 and US 6,602,684. IgG1 type antibodies, the most commonly used antibodies in cancer immunotherapy, are glycoproteins that have a conserved N-linked glycosylation site at Asn297 in each CH2 domain. The two complex biantennary oligosaccharides attached to Asn297 are buried between the CH2 domains, forming extensive contacts with the polypeptide backbone, and their presence84MOFO-360876942Attorney Docket No.14639-20729.40is essential for the antibody to mediate effector functions such as antibody dependent cellular cytotoxicity (ADCC) (Lifely, M.R., et al., Glycobiology 5 (1995) 813-822; Jefferis, R., et al., Immunol. Rev.163 (1998) 59-76; Wright, A., and Morrison, S.L., Trends Biotechnol.15 (1997) 26-32).

[0163] It was previously shown that overexpression in Chinese hamster ovary (CHO) cells of ß(1,4)-N-acetylglucosaminyltransferase I11 ("GnTII17y), a glycosyltransferase catalyzing the formation of bisected oligosaccharides, significantly increases the in vitro ADCC activity of an antineuroblastoma chimeric monoclonal antibody (chCE7) produced by the engineered CHO cells. (See Umana, P., et al., Nature Biotechnol.17 (1999) 176-180; and WO 99 / 154342, the entire contents of which are hereby incorporated by reference). The antibody chCE7 belongs to a large class of unconjugated monoclonal antibodies which have high tumor affinity and specificity, but have too little potency to be clinically useful when produced in standard industrial cell lines lacking the GnTIII enzyme (Umana, P., et al., Nature Biotechnol.17 (1999) 176-180). That study was the first to show that large increases of ADCC activity could be obtained by engineering the antibody producing cells to express GnTIII, which also led to an increase in the proportion of constant region (Fc)-associated, bisected oligosaccharides, including bisected, non-fucosylated oligosaccharides, above the levels found in naturally-occurring antibodies.

[0164] In some embodiments, the anti-CD20 antibody (e.g., a type II anti-CD20 antibody) comprises a human Fc region (e.g., a human IgG1 Fc region). In some embodiments, the Fc region comprises an N-linked oligosaccharide that has been modified. In some embodiments, the N-linked oligosaccharides of the Fc region have reduced fucose residues as compared to an antibody with non-modified N-linked oligosaccharides. In some embodiments, the bisected oligosaccharide is a bisected complex oligosaccharide. In some embodiments, the N-linked oligosaccharides have been modified to have increased bisected, nonfucosylated oligosaccharides. In some embodiments, the bisected, nonfucosylated oligosaccharides are the hybrid type. In some embodiments, the bisected, nonfucosylated oligosaccharides are the complex type. For more detailed description, see, e.g., WO 2003 / 011878 (Jean-Mairet et al.); US Patent No.6,602,684 (Umana et al.); US 2005 / 0123546 (Umana et al.); and U.S. Patent No.8,883,980 (Umana et al.).

[0165] In some embodiments, the type II anti-CD20 antibody is obinutuzumab.85MOFO-360876942Attorney Docket No.14639-20729.40Antibody Preparation

[0166] An antibody according to any of the above embodiments (e.g., a type II anti-CD20 antibody of the present disclosure) may incorporate any of the features, singly or in combination, as described in Sections 1-7 below:1. Antibody Affinity

[0167] In certain embodiments, an antibody provided herein has a dissociation constant (Kd)0.001 nM (e.g. 10-8M or less, e.g. from 10-8M to 10-13M, e.g., from 10-9M to 10-13M).

[0168] In one embodiment, Kd is measured by a radiolabeled antigen binding assay (RIA). In one embodiment, an RIA is performed with the Fab version of an antibody of interest and its antigen. For example, solution binding affinity of Fabs for antigen is measured by equilibrating Fab with a minimal concentration of (125I)-labeled antigen in the presence of a titration series of unlabeled antigen, then capturing bound antigen with an anti-Fab antibody-coated plate (see, e.g., Chen et al., J. Mol. Biol.293:865-881(1999)). To establish conditions for the assay, MICROTITER®multi-capturing anti-Fab antibody (Cappel Labs) in 50 mM sodium carbonate (pH 9.6), and subsequently blocked with 2% (w / v) bovine serum albumin in PBS for two to five hours at room temperature (approximately 23°C). In a non-adsorbent plate (Nunc #269620), 100 pM or 26 pM [125I]-antigen are mixed with serial dilutions of a Fab of interest (e.g., consistent with assessment of the anti-VEGF antibody, Fab-12, in Presta et al., Cancer Res.57:4593-4599 (1997)). The Fab of interest is then incubated overnight; however, the incubation may continue for a longer period (e.g., about 65 hours) to ensure that equilibrium is reached. Thereafter, the mixtures are transferred to the capture plate for incubation at room temperature (e.g., for one hour). The solution is then removed and the plate washed eight times with 0.1% polysorbate 20 (TWEEN-20®-20TM;Packard) is added, and the plates are counted on a TOPCOUNTTMgamma counter (Packard) for ten minutes. Concentrations of each Fab that give less than or equal to 20% of maximal binding are chosen for use in competitive binding assays.86MOFO-360876942Attorney Docket No.14639-20729.40

[0169] According to another embodiment, Kd is measured using a BIACORE®surface plasmon resonance assay. For example, an assay using a BIACORE®-2000 or a BIACORE®-3000 (BIAcore, Inc., Piscataway, NJ) is performed at 25°C with immobilized antigen CM5 chips at ~10 response units (RU). In one embodiment, carboxymethylated dextran biosensor chips (CM5, BIACORE, Inc.) are activated with N-ethyl-N’- (3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS) according to the supplier’s instructions.Following the injection of antigen, 1 M ethanolamine is injected to block unreacted groups. For kinetics measurements, two-fold serial dilutions of Fab (0.78 nM to 500 nM) are injected in PBS with 0.05% polysorbate 20 (TWEEN-20TM) surfactant (PBST) at 25°C at a flow rate oftion rates (kon) and dissociation rates (koff) are calculatedusing a simple one-to-one Langmuir binding model (BIACORE®Evaluation Software version 3.2) by simultaneously fitting the association and dissociation sensorgrams. The equilibriumdissociation constant (Kd) is calculated as the ratio koff / kon.See, e.g., Chen et al., J. Mol. Biol.293:865-881 (1999). If the on-rate exceeds 106 M-1 s-1 by the surface plasmon resonance assay above, then the on-rate can be determined by using a fluorescent quenching technique that measures the increase or decrease in fluorescence emission intensity (excitation = 295 nm; emission = 340 nm, 16 nm band-pass) at 25oC of a 20 nM anti-antigen antibody (Fab form) in PBS, pH 7.2, in the presence of increasing concentrations of antigen as measured in a spectrometer, such as a stop-flow equipped spectrophometer (Aviv Instruments) or a 8000-series SLM-AMINCOTMspectrophotometer (ThermoSpectronic) with a stirred cuvette.2. Antibody Fragments

[0170] In certain embodiments, an antibody provided herein is an antibody fragment.Antibody fragments include, but are not limited to, Fab, Fab’, Fab’-SH, F(ab’)2, Fv, and scFv fragments, and other fragments described below. For a review of certain antibody fragments, see Hudson et al. Nat. Med.9:129-134 (2003). For a review of scFv fragments, see, e.g., Pluckthün, in The Pharmacology of Monoclonal Antibodies, vol.113, Rosenburg and Moore eds., (Springer-Verlag, New York), pp.269-315 (1994); see also WO 93 / 16185; and U.S. Patent Nos.5,571,89487MOFO-360876942Attorney Docket No.14639-20729.40and 5,587,458. For discussion of Fab and F(ab')2fragments comprising salvage receptor binding epitope residues and having increased in vivo half-life, see U.S. Patent No.5,869,046.

[0171] Diabodies are antibody fragments with two antigen-binding sites that may be bivalent or bispecific. See, for example, EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med.9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat. Med.9:129-134 (2003).

[0172] Single-domain antibodies are antibody fragments comprising all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U.S. Patent No.6,248,516 B1).

[0173] Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody as well as production by recombinant host cells (e.g. E. coli or phage), as described herein.3. Chimeric and Humanized Antibodies

[0174] In certain embodiments, an antibody provided herein is a chimeric antibody. Certain chimeric antibodies are described, e.g., in U.S. Patent No.4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate, such as a monkey) and a human constant region. In a further example, a chimeric antibody is a “class switched” antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.

[0175] In certain embodiments, a chimeric antibody is a humanized antibody. Typically, a non-human antibody is humanized to reduce immunogenicity to humans, while retaining the specificity and affinity of the parental non-human antibody. Generally, a humanized antibody comprises one or more variable domains in which HVRs, e.g., CDRs, (or portions thereof) are derived from a non-human antibody, and FRs (or portions thereof) are derived from human antibody sequences. A humanized antibody optionally will also comprise at least a portion of a human constant region. In some embodiments, some FR residues in a humanized antibody are substituted with corresponding residues from a non-human antibody (e.g., the antibody from which the HVR residues are derived), e.g., to restore or improve antibody specificity or affinity.88MOFO-360876942Attorney Docket No.14639-20729.40

[0176] Humanized antibodies and methods of making them are reviewed, e.g., in Almagro and Fransson, Front. Biosci.13:1619-1633 (2008), and are further described, e.g., in Riechmann et al., Nature 332:323-329 (1988); Queen et al., Proc. Nat’l Acad. Sci. USA 86:10029-10033 (1989); US Patent Nos.5, 821,337, 7,527,791, 6,982,321, and 7,087,409; Kashmiri et al., Methods 36:25-34 (2005) (describing specificity determining region (SDR) grafting); Padlan, Mol. Immunol.28:489-498 (1991) (describing “resurfacing”); Dall’Acqua et al., Methods 36:43-60 (2005) (describing “FR shuffling”); and Osbourn et al., Methods 36:61-68 (2005) and Klimka et al., Br. J. Cancer, 83:252-260 (2000) (describing the “guided selection” approach to FR shuffling).

[0177] Human framework regions that may be used for humanization include but are not limited to: framework regions selected using the "best-fit" method (see, e.g., Sims et al. J.Immunol.151:2296 (1993)); framework regions derived from the consensus sequence of human antibodies of a particular subgroup of light or heavy chain variable regions (see, e.g., Carter et al. Proc. Natl. Acad. Sci. USA, 89:4285 (1992); and Presta et al. J. Immunol., 151:2623 (1993)); human mature (somatically mutated) framework regions or human germline framework regions (see, e.g., Almagro and Fransson, Front. Biosci.13:1619-1633 (2008)); and framework regions derived from screening FR libraries (see, e.g., Baca et al., J. Biol. Chem.272:10678-10684 (1997) and Rosok et al., J. Biol. Chem.271:22611-22618 (1996)).4. Human Antibodies

[0178] In certain embodiments, an antibody provided herein is a human antibody. Human antibodies can be produced using various techniques known in the art. Human antibodies are described generally in van Dijk and van de Winkel, Curr. Opin. Pharmacol.5: 368-74 (2001) and Lonberg, Curr. Opin. Immunol.20:450-459 (2008).

[0179] Human antibodies may be prepared by administering an immunogen to a transgenic animal that has been modified to produce intact human antibodies or intact antibodies with human variable regions in response to antigenic challenge. Such animals typically contain all or a portion of the human immunoglobulin loci, which replace the endogenous immunoglobulin loci, or which are present extrachromosomally or integrated randomly into the animal’s chromosomes. In such transgenic mice, the endogenous immunoglobulin loci have generally been inactivated. For review of methods for obtaining human antibodies from transgenic animals, see Lonberg, Nat. Biotech.23:1117-1125 (2005). See also, e.g., U.S. Patent Nos.89MOFO-360876942Attorney Docket No.14639-20729.406,075,181 and 6,150,584 describing XENOMOUSETMtechnology; U.S. Patent No.5,770,429 describing HUMAB® technology; U.S. Patent No.7,041,870 describing K-M MOUSE® technology, and U.S. Patent Application Publication No. US 2007 / 0061900, describing VELOCIMOUSE® technology). Human variable regions from intact antibodies generated by such animals may be further modified, e.g., by combining with a different human constant region.

[0180] Human antibodies can also be made by hybridoma-based methods. Human myeloma and mouse-human heteromyeloma cell lines for the production of human monoclonal antibodies have been described. (See, e.g., Kozbor J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, pp.51-63 (Marcel Dekker, Inc., New York, 1987); and Boerner et al., J. Immunol., 147: 86 (1991).) Human antibodies generated via human B-cell hybridoma technology are also described in Li et al., Proc. Natl. Acad. Sci. USA, 103:3557-3562 (2006). Additional methods include those described, for example, in U.S. Patent No.7,189,826 (describing production of monoclonal human IgM antibodies from hybridoma cell lines) and Ni, Xiandai Mianyixue, 26(4):265-268 (2006) (describing human-human hybridomas). Human hybridoma technology (Trioma technology) is also described in Vollmers and Brandlein, Histology and Histopathology, 20(3):927-937 (2005) and Vollmers and Brandlein, Methods and Findings in Experimental and Clinical Pharmacology, 27(3):185-91 (2005).

[0181] Human antibodies may also be generated by isolating Fv clone variable domain sequences selected from human-derived phage display libraries. Such variable domain sequences may then be combined with a desired human constant domain. Techniques for selecting human antibodies from antibody libraries are described below.5. Library-Derived Antibodies

[0182] Antibodies of the invention may be isolated by screening combinatorial libraries for antibodies with the desired activity or activities. For example, a variety of methods are known in the art for generating phage display libraries and screening such libraries for antibodies possessing the desired binding characteristics. Such methods are reviewed, e.g., in Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O’Brien et al., ed., Human Press, Totowa, NJ, 2001) and further described, e.g., in the McCafferty et al., Nature 348:552-554; Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol.222: 581-597 (1992); Marks and Bradbury, in Methods in Molecular Biology 248:161-175 (Lo, ed., Human Press, Totowa, NJ,90MOFO-360876942Attorney Docket No.14639-20729.402003); Sidhu et al., J. Mol. Biol.338(2): 299-310 (2004); Lee et al., J. Mol. Biol.340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132(2004).

[0183] In certain phage display methods, repertoires of VH and VL genes are separately cloned by polymerase chain reaction (PCR) and recombined randomly in phage libraries, which can then be screened for antigen-binding phage as described in Winter et al., Ann. Rev.Immunol., 12: 433-455 (1994). Phage typically display antibody fragments, either as single-chain Fv (scFv) fragments or as Fab fragments. Libraries from immunized sources provide high-affinity antibodies to the immunogen without the requirement of constructing hybridomas.Alternatively, the naive repertoire can be cloned (e.g., from human) to provide a single source of antibodies to a wide range of non-self and also self antigens without any immunization as described by Griffiths et al., EMBO J, 12: 725-734 (1993). Finally, naive libraries can also be made synthetically by cloning unrearranged V-gene segments from stem cells, and using PCR primers containing random sequence to encode the highly variable CDR3 regions and to accomplish rearrangement in vitro, as described by Hoogenboom and Winter, J. Mol. Biol., 227: 381-388 (1992). Patent publications describing human antibody phage libraries include, for example: US Patent No.5,750,373, and US Patent Publication Nos.2005 / 0079574, 2005 / 0119455, 2005 / 0266000, 2007 / 0117126, 2007 / 0160598, 2007 / 0237764, 2007 / 0292936, and 2009 / 0002360.

[0184] Antibodies or antibody fragments isolated from human antibody libraries are considered human antibodies or human antibody fragments herein.6. Multispecific Antibodies

[0185] In certain embodiments, an antibody provided herein is a multispecific antibody, e.g. a bispecific antibody. Multispecific antibodies are monoclonal antibodies that have binding specificities for at least two different sites. In certain embodiments, one of the binding specificities is for CD20 and the other is for any other antigen. In certain embodiments, bispecific antibodies may bind to two different epitopes of CD20. Bispecific antibodies may also be used to localize cytotoxic agents to cells which express CD20. Bispecific antibodies can be prepared as full length antibodies or antibody fragments.

[0186] Techniques for making multispecific antibodies include, but are not limited to, recombinant co-expression of two immunoglobulin heavy chain-light chain pairs having91MOFO-360876942Attorney Docket No.14639-20729.40different specificities (see Milstein and Cuello, Nature 305: 537 (1983)), WO 93 / 08829, and Traunecker et al., EMBO J.10: 3655 (1991)), and “knob-in-hole” engineering (see, e.g., U.S. Patent No.5,731,168). Multi-specific antibodies may also be made by engineering electrostatic steering effects for making antibody Fc-heterodimeric molecules (WO 2009 / 089004A1); cross-linking two or more antibodies or fragments (see, e.g., US Patent No.4,676,980, and Brennan et al., Science, 229: 81 (1985)); using leucine zippers to produce bi-specific antibodies (see, e.g., Kostelny et al., J. Immunol., 148(5):1547-1553 (1992)); using "diabody" technology for making bispecific antibody fragments (see, e.g., Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993)); and using single-chain Fv (sFv) dimers (see,e.g. Gruber et al., J. Immunol., 152:5368 (1994)); and preparing trispecific antibodies as described, e.g., in Tutt et al. J.Immunol.147: 60 (1991).

[0187] Engineered antibodies with three or more functional antigen binding sites, including “Octopus antibodies,” are also included herein (see, e.g. US 2006 / 0025576A1).

[0188] The antibody or fragment herein also includes a “Dual Acting FAb” or “DAF” comprising an antigen binding site that binds to CD20 as well as another, different antigen (see, US 2008 / 0069820, for example).7. Antibody Variants

[0189] In certain embodiments, amino acid sequence variants of the antibodies provided herein are contemplated. For example, it may be desirable to improve the binding affinity and / or other biological properties of the antibody. Amino acid sequence variants of an antibody may be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody, or by peptide synthesis. Such modifications include, for example, deletions from, and / or insertions into and / or substitutions of residues within the amino acid sequences of the antibody. Any combination of deletion, insertion, and substitution can be made to arrive at the final construct, provided that the final construct possesses the desired characteristics, e.g., antigen-binding.a) Substitution, Insertion, and Deletion Variants

[0190] In certain embodiments, antibody variants having one or more amino acid substitutions are provided. Sites of interest for substitutional mutagenesis include the HVRs and FRs.Conservative substitutions are shown in Table A under the heading of "preferred substitutions."92MOFO-360876942Attorney Docket No.14639-20729.40More substantial changes are provided in Table A under the heading of "exemplary substitutions," and as further described below in reference to amino acid side chain classes. Amino acid substitutions may be introduced into an antibody of interest and the products screened for a desired activity, e.g., retained / improved antigen binding, decreased immunogenicity, or improved ADCC or CDC.TABLE A

[0191] Amino acids may be grouped according to common side-chain properties:93MOFO-360876942Attorney Docket No.14639-20729.40(1) hydrophobic: Norleucine, Met, Ala, Val, Leu, Ile;(2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;(3) acidic: Asp, Glu;(4) basic: His, Lys, Arg;(5) residues that influence chain orientation: Gly, Pro;(6) aromatic: Trp, Tyr, Phe.

[0192] Non-conservative substitutions will entail exchanging a member of one of these classes for another class.

[0193] One type of substitutional variant involves substituting one or more hypervariable region residues of a parent antibody (e.g. a humanized or human antibody). Generally, the resulting variant(s) selected for further study will have modifications (e.g., improvements) in certain biological properties (e.g., increased affinity, reduced immunogenicity) relative to the parent antibody and / or will have substantially retained certain biological properties of the parent antibody. An exemplary substitutional variant is an affinity matured antibody, which may be conveniently generated, e.g., using phage display-based affinity maturation techniques such as those described herein. Briefly, one or more HVR residues are mutated and the variant antibodies displayed on phage and screened for a particular biological activity (e.g. binding affinity).

[0194] Alterations (e.g., substitutions) may be made in HVRs, e.g., to improve antibody affinity. Such alterations may be made in HVR “hotspots,” i.e., residues encoded by codons that undergo mutation at high frequency during the somatic maturation process (see, e.g., Chowdhury, Methods Mol. Biol.207:179-196 (2008)), and / or residues that contact antigen, with the resulting variant VH or VL being tested for binding affinity. Affinity maturation by constructing and reselecting from secondary libraries has been described, e.g., in Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O’Brien et al., ed., Human Press, Totowa, NJ, (2001).) In some embodiments of affinity maturation, diversity is introduced into the variable genes chosen for maturation by any of a variety of methods (e.g., error-prone PCR, chain shuffling, or oligonucleotide-directed mutagenesis). A secondary library is then created. The library is then screened to identify any antibody variants with the desired affinity. Another method to introduce diversity involves HVR-directed approaches, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding94MOFO-360876942Attorney Docket No.14639-20729.40may be specifically identified, e.g., using alanine scanning mutagenesis or modeling. CDR-H3 and CDR-L3 in particular are often targeted.

[0195] In certain embodiments, substitutions, insertions, or deletions may occur within one or more HVRs so long as such alterations do not substantially reduce the ability of the antibody to bind antigen. For example, conservative alterations (e.g., conservative substitutions as provided herein) that do not substantially reduce binding affinity may be made in HVRs. Such alterations may, for example, be outside of antigen contacting residues in the HVRs. In certain embodiments of the variant VH and VL sequences provided above, each HVR either is unaltered, or contains no more than one, two or three amino acid substitutions.

[0196] A useful method for identification of residues or regions of an antibody that may be targeted for mutagenesis is called "alanine scanning mutagenesis" as described by Cunningham and Wells (1989) Science, 244:1081-1085. In this method, a residue or group of target residues (e.g., charged residues such as arg, asp, his, lys, and glu) are identified and replaced by a neutral or negatively charged amino acid (e.g., alanine or polyalanine) to determine whether the interaction of the antibody with antigen is affected. Further substitutions may be introduced at the amino acid locations demonstrating functional sensitivity to the initial substitutions.Alternatively, or additionally, a crystal structure of an antigen-antibody complex to identify contact points between the antibody and antigen. Such contact residues and neighboring residues may be targeted or eliminated as candidates for substitution. Variants may be screened to determine whether they contain the desired properties.

[0197] Amino acid sequence insertions include amino- and / or carboxyl-terminal fusions ranging in length from one residue to polypeptides containing a hundred or more residues, as well as intrasequence insertions of single or multiple amino acid residues. Examples of terminal insertions include an antibody with an N-terminal methionyl residue. Other insertional variants of the antibody molecule include the fusion to the N- or C-terminus of the antibody to an enzyme (e.g. for ADEPT) or a polypeptide which increases the serum half-life of the antibody.b) Glycosylation variants

[0198] In certain embodiments, an antibody provided herein is altered to increase or decrease the extent to which the antibody is glycosylated. Addition or deletion of glycosylation sites to an antibody may be conveniently accomplished by altering the amino acid sequence such that one or more glycosylation sites is created or removed.95MOFO-360876942Attorney Docket No.14639-20729.40

[0199] Where the antibody comprises an Fc region, the carbohydrate attached thereto may be altered. Native antibodies produced by mammalian cells typically comprise a branched, biantennary oligosaccharide that is generally attached by an N-linkage to Asn297 of the CH2 domain of the Fc region. See, e.g., Wright et al. TIBTECH 15:26-32 (1997). The oligosaccharide may include various carbohydrates, e.g., mannose, N-acetyl glucosamine (GlcNAc), galactose, and sialic acid, as well as a fucose attached to a GlcNAc in the “stem” of the biantennary oligosaccharide structure. In some embodiments, modifications of the oligosaccharide in an antibody of the invention may be made in order to create antibody variants with certain improved properties.

[0200] In one embodiment, antibody variants are provided having a carbohydrate structure that lacks fucose attached (directly or indirectly) to an Fc region. For example, the amount of fucose in such antibody may be from 1% to 80%, from 1% to 65%, from 5% to 65% or from 20% to 40%. The amount of fucose is determined by calculating the average amount of fucose within the sugar chain at Asn297, relative to the sum of all glycostructures attached to Asn 297 (e. g. complex, hybrid and high mannose structures) as measured by MALDI-TOF mass spectrometry, as described in WO 2008 / 077546, for example. Asn297 refers to the asparagine residue located at about position 297 in the Fc region (Eu numbering of Fc region residues); however, Asn297 may also be located about ± 3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300, due to minor sequence variations in antibodies. Such fucosylation variants may have improved ADCC function. See, e.g., US Patent Publication Nos. US 2003 / 0157108 (Presta, L.); US 2004 / 0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to “defucosylated” or “fucose-deficient” antibody variants include: US 2003 / 0157108; WO 2000 / 61739; WO 2001 / 29246; US 2003 / 0115614; US 2002 / 0164328; US 2004 / 0093621; US 2004 / 0132140; US 2004 / 0110704; US 2004 / 0110282; US 2004 / 0109865; WO 2003 / 085119; WO 2003 / 084570; WO 2005 / 035586; WO 2005 / 035778; WO2005 / 053742; WO2002 / 031140; Okazaki et al. J. Mol. Biol.336:1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng.87: 614 (2004). Examples of cell lines capable of producing defucosylated antibodies include Lec13 CHO cells deficient in protein fucosylation (Ripka et al. Arch.Biochem. Biophys.249:533-545 (1986); US Pat Appl No US 2003 / 0157108 A1, Presta, L; and WO 2004 / 056312 A1, Adams et al., especially at Example 11), and knockout cell lines, such as alpha-1,6-fucosyltransferase gene, FUT8, knockout CHO cells (see, e.g., Yamane-Ohnuki et al.96MOFO-360876942Attorney Docket No.14639-20729.40Biotech. Bioeng.87: 614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94(4):680-688 (2006); and WO2003 / 085107).

[0201] Antibodies variants are further provided with bisected oligosaccharides, e.g., in which a biantennary oligosaccharide attached to the Fc region of the antibody is bisected by GlcNAc. Such antibody variants may have reduced fucosylation and / or improved ADCC function.Examples of such antibody variants are described, e.g., in WO 2003 / 011878 (Jean-Mairet et al.); US Patent No.6,602,684 (Umana et al.); and US 2005 / 0123546 (Umana et al.). Antibody variants with at least one galactose residue in the oligosaccharide attached to the Fc region are also provided. Such antibody variants may have improved CDC function. Such antibody variants are described, e.g., in WO 1997 / 30087 (Patel et al.); WO 1998 / 58964 (Raju, S.); and WO 1999 / 22764 (Raju, S.).c) Fc region variants

[0202] In certain embodiments, one or more amino acid modifications may be introduced into the Fc region of an antibody provided herein, thereby generating an Fc region variant. The Fc region variant may comprise a human Fc region sequence (e.g., a human IgG1, IgG2, IgG3 or IgG4 Fc region) comprising an amino acid modification (e.g. a substitution) at one or more amino acid positions.

[0203] In certain embodiments, the invention contemplates an antibody variant that possesses some but not all effector functions, which make it a desirable candidate for applications in which the half life of the antibody in vivo is important yet certain effector functions (such as complement and ADCC) are unnecessary or deleterious. In vitro and / or in vivo cytotoxicity assays can be conducted to confirm the reduction / depletion of CDC and / or ADCC activities. For example, Fc receptor (FcR) binding assays can be conducted to ensure that the antibody lacksFc R binding (hence likely lacking ADCC activity), but retains FcRn binding ability. Theprimary cells for mediating ADCC, NK cells, express Fc RIII only, whereas monocytes expressFc RI, Fc RII and Fc RIII. FcR expression on hematopoietic cells is summarized in Table 3 onpage 464 of Ravetch and Kinet, Annu. Rev. Immunol.9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest is described in U.S. Patent No.5,500,362 (see, e.g. Hellstrom, I. et al. Proc. Nat’l Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I et al., Proc. Nat’l Acad. Sci. USA 82:1499-1502 (1985); 5,821,337 (see97MOFO-360876942Attorney Docket No.14639-20729.40Bruggemann, M. et al., J. Exp. Med.166:1351-1361 (1987)). Alternatively, non-radioactive assays methods may be employed (see, for example, ACTI™ non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, CA; and CytoTox 96®non-radioactive cytotoxicity assay (Promega, Madison, WI). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in a animal model such as that disclosed in Clynes et al. Proc. Nat’l Acad. Sci. USA 95:652-656 (1998). C1q binding assays may also be carried out to confirm that the antibody is unable to bind C1q and hence lacks CDC activity. See, e.g., C1q and C3c binding ELISA inWO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, a CDC assay may be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, M.S. et al., Blood 101:1045-1052 (2003); and Cragg, M.S. and M.J. Glennie, Blood 103:2738-2743 (2004)). FcRn binding and in vivo clearance / half life determinations can also be performed using methods known in the art (see, e.g., Petkova, S.B. et al., Int’l. Immunol.18(12):1759-1769 (2006)).

[0204] Antibodies with reduced effector function include those with substitution of one or more of Fc region residues 238, 265, 269, 270, 297, 327 and 329 (U.S. Patent No.6,737,056). Such Fc mutants include Fc mutants with substitutions at two or more of amino acid positions 265, 269, 270, 297 and 327, including the so-called “DANA” Fc mutant with substitution of residues 265 and 297 to alanine (US Patent No.7,332,581).

[0205] In certain embodiments, the Fc variants described herein further comprise one or more amino acid modifications for attenuating effector function (such as CDC and / or ADCC). In exemplary embodiments, the modification to attenuate effector function is a modification that does not alter the glycosylation pattern of the Fc region. In certain embodiments, the modification to attenuate effector function reduces or eliminates binding to human effector cells, binding to one or more Fc receptors, and / or binding to cells expressing an Fc receptor. In an exemplary embodiment, the Fc variants described herein comprise the following modifications: L234A, L235A and P329G in the Fc region of human IgG1, that result in attenuated effector function. Substitutions L234A, L235A, and P329G (the L234A / L235A / P329G triple variant is referred to as LALAPG) have previously been shown to reduce binding to Fc receptors and complement (see e.g., US Publication No.2012 / 0251531).98MOFO-360876942Attorney Docket No.14639-20729.40

[0206] In various embodiments, Fc variants having reduced effector function refer to Fc variants that reduce effector function (e.g., CDC, ADCC, and / or binding to FcR, etc. activities) by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, 99% or more as compared to the effector function achieved by a wild-type Fc region (e.g., an Fc region not having a mutation to reduce effector function, although it may have other mutations). In certain embodiments, Fc variants having reduced effector function refer to Fc variants that eliminate all detectable effector function as compared to a wild-type Fc region. Assays for measuring effector function are known in the art and described below.

[0207] In vitro and / or in vivo cytotoxicity assays can be conducted to confirm the reduction / depletion of CDC and / or ADCC activities. For example, Fc receptor (FcR) bindingassays can be conducted to ensure that the antibody lacks Fc R binding (hence likely lackingADCC activity). The primary cells for mediating ADCC, NK cells, express Fc RIII only,whereas monocytes express Fc RI, Fc RII and Fc RIII. FcR expression on hematopoietic cellsis summarized in Ravetch and Kinet, Annu. Rev. Immunol.9:457-492 (1991). Non-limiting examples of in vitro assays to assess ADCC activity of a molecule of interest is described in U.S. Patent No.5,500,362 (see, e.g. Hellstrom, I. et al. Proc. Nat’l Acad. Sci. USA 83:7059-7063 (1986)) and Hellstrom, I et al., Proc. Nat’l Acad. Sci. USA 82:1499-1502 (1985); 5,821,337 (see Bruggemann, M. et al., J. Exp. Med.166:1351-1361 (1987)). Alternatively, non-radioactive assays methods may be employed (see, for example, ACTI™ non-radioactive cytotoxicity assay for flow cytometry (CellTechnology, Inc. Mountain View, CA; and CytoTox 96®non-radioactive cytotoxicity assay (Promega, Madison, WI). Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells.Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes et al. Proc. Nat’l Acad. Sci. USA 95:652-656 (1998). C1q binding assays may also be carried out to confirm that the antibody is unable to bind C1q and hence lacks CDC activity. See, e.g., C1q and C3c binding ELISA in WO 2006 / 029879 and WO 2005 / 100402. To assess complement activation, a CDC assay may be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, M.S. et al., Blood 101:1045-1052 (2003); and Cragg, M.S. and M.J. Glennie, Blood 103:2738-2743 (2004)).99MOFO-360876942Attorney Docket No.14639-20729.40

[0208] Certain antibody variants with improved or diminished binding to FcRs are described. (See, e.g., U.S. Patent No.6,737,056; WO 2004 / 056312, and Shields et al., J. Biol. Chem.9(2): 6591-6604 (2001).)

[0209] In certain embodiments, an antibody variant comprises an Fc region with one or more amino acid substitutions which improve ADCC, e.g., substitutions at positions 298, 333, and / or 334 of the Fc region (EU numbering of residues).

[0210] In some embodiments, alterations are made in the Fc region that result in altered (i.e., either improved or diminished) C1q binding and / or Complement Dependent Cytotoxicity (CDC), e.g., as described in US Patent No.6,194,551, WO 99 / 51642, and Idusogie et al. J. Immunol. 164: 4178-4184 (2000).

[0211] Antibodies with increased half lives and improved binding to the neonatal Fc receptor (FcRn), which is responsible for the transfer of maternal IgGs to the fetus (Guyer et al., J.Immunol.117:587 (1976) and Kim et al., J. Immunol.24:249 (1994)), are described in US2005 / 0014934A1 (Hinton et al.). Those antibodies comprise an Fc region with one or more substitutions therein which improve binding of the Fc region to FcRn. Such Fc variants include those with substitutions at one or more of Fc region residues: 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or 434, e.g., substitution of Fc region residue 434 (US Patent No.7,371,826).

[0212] See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Patent No.5,648,260; U.S. Patent No.5,624,821; and WO 94 / 29351 concerning other examples of Fc region variants.d) Cysteine engineered antibody variants

[0213] In certain embodiments, it may be desirable to create cysteine engineered antibodies, e.g., “thioMAbs,” in which one or more residues of an antibody are substituted with cysteine residues. In particular embodiments, the substituted residues occur at accessible sites of the antibody. By substituting those residues with cysteine, reactive thiol groups are thereby positioned at accessible sites of the antibody and may be used to conjugate the antibody to other moieties, such as drug moieties or linker-drug moieties, to create an immunoconjugate, as described further herein. In certain embodiments, any one or more of the following residues may be substituted with cysteine: V205 (Kabat numbering) of the light chain; A118 (EU numbering) of the heavy chain; and S400 (EU numbering) of the heavy chain Fc region. Cysteine engineered antibodies may be generated as described, e.g., in U.S. Patent No.7,521,541.100MOFO-360876942Attorney Docket No.14639-20729.40e) Antibody Derivatives

[0214] In certain embodiments, an antibody provided herein may be further modified to contain additional nonproteinaceous moieties that are known in the art and readily available. The moieties suitable for derivatization of the antibody include but are not limited to water soluble polymers. Non-limiting examples of water soluble polymers include, but are not limited to, polyethylene glycol (PEG), copolymers of ethylene glycol / propylene glycol, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinyl pyrrolidone, poly-1, 3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyaminoacids (either homopolymers or random copolymers), and dextran or poly(n-vinyl pyrrolidone)polyethylene glycol, propropylene glycol homopolymers, prolypropylene oxide / ethylene oxide co-polymers, polyoxyethylated polyols (e.g., glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde may have advantages in manufacturing due to its stability in water. The polymer may be of any molecular weight, and may be branched or unbranched. The number of polymers attached to the antibody may vary, and if more than one polymer are attached, they can be the same or different molecules. In general, the number and / or type of polymers used for derivatization can be determined based on considerations including, but not limited to, the particular properties or functions of the antibody to be improved, whether the antibody derivative will be used in a therapy under defined conditions, etc.

[0215] In another embodiment, conjugates of an antibody and nonproteinaceous moiety that may be selectively heated by exposure to radiation are provided. In one embodiment, the nonproteinaceous moiety is a carbon nanotube (Kam et al., Proc. Natl. Acad. Sci. USA 102: 11600-11605 (2005)). The radiation may be of any wavelength, and includes, but is not limited to, wavelengths that do not harm ordinary cells, but which heat the nonproteinaceous moiety to a temperature at which cells proximal to the antibody-nonproteinaceous moiety are killed.A. Recombinant Methods and Compositions

[0216] Antibodies may be produced using recombinant methods and compositions, e.g., as described in U.S. Patent No.4,816,567. In one embodiment, isolated nucleic acid encoding an anti-CD20 antibody described herein is provided. Such nucleic acid may encode an amino acid sequence comprising the VL and / or an amino acid sequence comprising the VH of the antibody (e.g., the light and / or heavy chains of the antibody). In a further embodiment, one or more101MOFO-360876942Attorney Docket No.14639-20729.40vectors (e.g., expression vectors) comprising such nucleic acid are provided. In a further embodiment, a host cell comprising such nucleic acid is provided. In one such embodiment, a host cell comprises (e.g., has been transformed with): (1) a vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antibody and an amino acid sequence comprising the VH of the antibody, or (2) a first vector comprising a nucleic acid that encodes an amino acid sequence comprising the VL of the antibody and a second vector comprising a nucleic acid that encodes an amino acid sequence comprising the VH of the antibody. In one embodiment, the host cell is eukaryotic, e.g. a Chinese Hamster Ovary (CHO) cell or lymphoid cell (e.g., Y0, NS0, Sp20 cell). In one embodiment, a method of making an anti-CD20 antibody is provided, wherein the method comprises culturing a host cell comprising a nucleic acid encoding the antibody, as provided above, under conditions suitable for expression of the antibody, and optionally recovering the antibody from the host cell (or host cell culture medium).

[0217] For recombinant production of an anti-CD20 antibody, nucleic acid encoding an antibody, e.g., as described above, is isolated and inserted into one or more vectors for further cloning and / or expression in a host cell. Such nucleic acid may be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of the antibody).

[0218] Suitable host cells for cloning or expression of antibody-encoding vectors include prokaryotic or eukaryotic cells described herein. For example, antibodies may be produced in bacteria, in particular when glycosylation and Fc effector function are not needed. For expression of antibody fragments and polypeptides in bacteria, see, e.g., U.S. Patent Nos.5,648,237, 5,789,199, and 5,840,523. (See also Charlton, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ, 2003), pp.245-254, describing expression of antibody fragments in E. coli.) After expression, the antibody may be isolated from the bacterial cell paste in a soluble fraction and can be further purified.

[0219] In addition to prokaryotes, eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for antibody-encoding vectors, including fungi and yeast strains whose glycosylation pathways have been “humanized,” resulting in the production of an antibody with a partially or fully human glycosylation pattern. See Gerngross, Nat. Biotech. 22:1409-1414 (2004), and Li et al., Nat. Biotech.24:210-215 (2006).102MOFO-360876942Attorney Docket No.14639-20729.40

[0220] Suitable host cells for the expression of glycosylated antibody are also derived from multicellular organisms (invertebrates and vertebrates). Examples of invertebrate cells include plant and insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells, particularly for transfection of Spodoptera frugiperda cells.

[0221] Plant cell cultures can also be utilized as hosts. See, e.g., US Patent Nos.5,959,177, 6,040,498, 6,420,548, 7,125,978, and 6,417,429 (describing PLANTIBODIESTMtechnology for producing antibodies in transgenic plants).

[0222] Vertebrate cells may also be used as hosts. For example, mammalian cell lines that are adapted to grow in suspension may be useful. Other examples of useful mammalian host cell lines are monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Virol.36:59 (1977)); baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod.23:243-251 (1980)); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary tumor (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N.Y. Acad. Sci.383:44-68 (1982); MRC 5 cells; and FS4 cells. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216 (1980)); and myeloma cell lines such as Y0, NS0 and Sp2 / 0. For a review of certain mammalian host cell lines suitable for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol.248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ), pp.255-268 (2003).B. Assays

[0223] Anti-CD20 antibodies provided herein may be identified, screened for, or characterized for their physical / chemical properties and / or biological activities by various assays known in the art.1. Binding assays and other assays

[0224] In one aspect, an antibody of the invention is tested for its antigen binding activity, e.g., by known methods such as ELISA, Western blot, etc. CD20 binding may be determined using methods known in the art and exemplary methods are disclosed herein. In one embodiment,103MOFO-360876942Attorney Docket No.14639-20729.40binding is measured using radioimmunoassay. An exemplary radioimmunoassay is provided below. CD20 antibody is iodinated, and competition reaction mixtures are prepared containing a fixed concentration of iodinated antibody and decreasing concentrations of serially diluted, unlabeled CD20 antibody. Cells expressing CD20 (e.g., BT474 cells stably transfected with human CD20) are added to the reaction mixture. Following an incubation, cells are washed to separate the free iodinated CD20 antibody from the CD20 antibody bound to the cells. Level of bound iodinated CD20 antibody is determined, e.g., by counting radioactivity associated with cells, and binding affinity determined using standard methods. In another embodiment, ability of CD20 antibody to bind to surface-expressed CD20 (e.g., on B cell subsets) is assessed using flow cytometry. Peripheral white blood cells are obtained (e.g., from human, cynomolgus monkey, rat or mouse) and cells are blocked with serum. Labeled CD20 antibody is added in serial dilutions, and T cells are also stained to identify T cell subsets (using methods known in the art). Following incubation of the samples and washing, the cells are sorted using flow cytometer, and data analyzed using methods well known in the art. In another embodiment, CD20 binding may be analyzed using surface plasmon resonance. An exemplary surface plasmon resonance method is exemplified in the Examples.

[0225] In another aspect, competition assays may be used to identify an antibody that competes with any of the anti-CD20 antibodies disclosed herein for binding to CD20. In certain embodiments, such a competing antibody binds to the same epitope (e.g., a linear or a conformational epitope) that is bound by any of the anti-CD20 antibodies disclosed herein. Detailed exemplary methods for mapping an epitope to which an antibody binds are provided in Morris (1996) “Epitope Mapping Protocols,” in Methods in Molecular Biology vol.66 (Humana Press, Totowa, NJ).

[0226] In an exemplary competition assay, immobilized CD20 is incubated in a solution comprising a first labeled antibody that binds to CD20 (e.g., rituximab, a GA101 antibody, etc.) and a second unlabeled antibody that is being tested for its ability to compete with the first antibody for binding to CD20. The second antibody may be present in a hybridoma supernatant. As a control, immobilized CD20 is incubated in a solution comprising the first labeled antibody but not the second unlabeled antibody. After incubation under conditions permissive for binding of the first antibody to CD20, excess unbound antibody is removed, and the amount of label associated with immobilized CD20 is measured. If the amount of label associated with104MOFO-360876942Attorney Docket No.14639-20729.40immobilized CD20 is substantially reduced in the test sample relative to the control sample, then that indicates that the second antibody is competing with the first antibody for binding to CD20. See Harlow and Lane (1988) Antibodies: A Laboratory Manual ch.14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, NY).2. Activity assays

[0227] Anti-CD20 antibodies of the present disclosure (e.g., a type II antibody) may be identified and / or characterized by one or more activity assays known in the art. For example, a complement-dependent cytotoxicity (CDC) and / or antibody-dependent cellular cytotoxicity (ADCC) may be used, as described herein.

[0228] It is understood that any of the above assays may be carried out using an immunoconjugate of the invention in place of or in addition to an anti-CD20 antibody.

[0229] It is understood that any of the above assays may be carried out using anti-CD20 antibody and an additional therapeutic agent.Methods of Administering a type II anti-CD20 antibody

[0230] Provided herein are methods for treating lupus nephritis (LN) in an individual that has lupus, wherein the methods comprise administering to the individual at least a first antibody exposure to a type II anti-CD20 antibody and a second antibody exposure to the type II anti-CD20 antibody; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure; wherein the first antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; wherein the second antibody exposure comprises one or two doses of the type II anti-CD20 antibody, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody; wherein the type II anti-CD20 antibody comprises a heavy chain that comprises a heavy chain variable (VH) region comprising HVR-H1 sequence of SEQ ID NO:1, HVR-H2 sequence of SEQ ID NO:2, and HVR-H3 sequence of SEQ ID NO:3, and a light chain that comprises a light chain variable (VL) region comprising HVR-L1 sequence of SEQ ID NO:4, HVR-L2 sequence of SEQ ID NO:5, and HVR-L3 sequence of SEQ ID NO:6. In some embodiments, the methods further comprise105MOFO-360876942Attorney Docket No.14639-20729.40administering to the individual a third antibody exposure to the type II anti-CD20 antibody, wherein the third antibody exposure comprises one or two doses of the type II anti-CD20 antibody, and wherein the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure. In some embodiments, the individual has a baseline UPCR of greater than or equal to about 2.0 g / g or greater than or equal to about 3.0 g / g. In some embodiments, the type II anti-CD20 antibody is obinutuzumab.

[0231] LN is known in the art as a manifestation of lupus (e.g., systemic lupus erythematosus, drug-induced lupus, neonatal lupus, or discoid lupus) in the kidney(s). The most common type of lupus that manifests in the kidneys is systemic lupus erythematosus (SLE). It is thought that 25-50% of SLE patients have abnormalities in the urine and / or renal function early in the course of their disease, with up to 60% of adults and 80% of children eventually developing LN (for more details, see Cameron, J.S. (1999) J. Am. Soc. Nephrol.10:413-424). LN is thought to account for at least 50% of the morbidity and mortality associated with SLE.

[0232] In addition, renal manifestations have also been noted in other types of lupus, such as discoid (Roujeau, J.C. et al. (1984) Acta Derm. Venereol.64:160-163) and drug-induced lupus (Smith, P.R. et al. (1999) Rheumatology (Oxford) 38:1017-1018). In some embodiments, the individual has SLE, discoid lupus, or drug-induced lupus.

[0233] Diagnosis of SLE may be according to current American College of Rheumatology (ACR) criteria. Active disease may be defined by one British Isles Lupus Activity Group's (BILAG) “A” criteria or two BILAG “B” criteria; SLE Disease Activity Index (SLEDAI); or systemic lupus erythematosus (SLE) responder index (SRI) as noted in the Examples below and descrived in Furie et al., Arthritis Rheum.61(9):1143-51 (2009). Some signs, symptoms, or other indicators used to diagnose SLE adapted from: Tan et al. “The Revised Criteria for the Classification of SLE” Arth Rheum 25 (1982) may be malar rash such as rash over the cheeks, discoid rash, or red raised patches, photosensitivity such as reaction to sunlight, resulting in the development of or increase in skin rash, oral ulcers such as ulcers in the nose or mouth, usually painless, arthritis, such as non-erosive arthritis involving two or more peripheral joints (arthritis in which the bones around the joints do not become destroyed), serositis, pleuritis or pericarditis, renal disorder such as excessive protein in the urine (greater than 0.5 gm / day or 3+ on test sticks) and / or cellular casts (abnormal elements derived from the urine and / or white cells and / or kidney tubule cells), neurologic signs, symptoms, or other indicators, seizures (convulsions), and / or106MOFO-360876942Attorney Docket No.14639-20729.40psychosis in the absence of drugs or metabolic disturbances that are known to cause such effects, and hematologic signs, symptoms, or other indicators such as hemolytic anemia or leukopenia (white blood count below 4,000 cells per cubic millimeter) or lymphopenia (less than 1,500 lymphocytes per cubic millimeter) or thrombocytopenia (less than 100,000 platelets per cubic millimeter). The leukopenia and lymphopenia must be detected on two or more occasions. The thrombocytopenia must be detected in the absence of drugs known to induce it. The invention is not limited to these signs, symptoms, or other indicators of lupus.

[0234] The presence of autoantibodies may be tested as an indication for lupus.Autoantibodies may include without limitation anti-dsDNA antibodies, anti-complement antibodies, and antinuclear antibodies (e.g., an ENA panel). ENA refers to Extractable Nuclear Antigens, i.e., a group of nuclear antigens including, e.g., RNP, Ro / SS-A, La / SS-B, Sm, SCL-70, Jo-1, as described in McNeilage et al., J., Clin. Lab. Immunol.15:1-17 (1984); Whittingham, Ann. Acad. Med.17(2):195-200 (1988); Wallace and Hahn, DUBOIS’ LUPUS ERYTHEMATOSUS, 7THED. LIPPINCOTT (2007); Tang et al., Medicine 89(1): 62-67 (2010). Antibodies to ENA have been correlated to lupus. McNeilage et al., 1984; Whittingham 1988; Asherson et al., Medicine 68(6): 366-374 (1989); and Tang et al., 2010. Reduced complement activity may also be associated with lupus, e.g., as measured by C3 levels, C4 levels, and / or a CH50 assay.

[0235] As described above in reference to SLE, it is known in the art that LN often manifests progressively in patients with lupus (e.g., systemic lupus erythematosus, drug-induced lupus, neonatal lupus, or discoid lupus). That is to say, a patient may be diagnosed with lupus without a clinical or pathological manifestation of one or more LN symptoms. Nonetheless, the patient may still be considered to be at risk for developing LN due to the high frequency of lupus patients that eventually develop LN. Therefore, in some embodiments, the methods of the present disclosure may find use in delaying progression of LN, or preventing LN, in a patient with lupus. In some embodiments, the methods of the present disclosure may find use in postponing or preventing the onset of LN in a patient with lupus (e.g., a form of lupus that lacks a manifestation in the kidney(s)). The terms “individual” and “patient” may be used interchangeably herein.

[0236] LN pathology may be classified according to the International Society of Nephrology / Renal Pathology Society (ISN / RPS) 2003 classification system, as shown in the107MOFO-360876942Attorney Docket No.14639-20729.40table below (see Markowitz GS, D’Agati VD (2007) Kidney Int 71:491–495 and Weening, JJ (2004) Kidney Int 65:521-530 for further descriptions and definitions of terms).Table 3. ISN / RPS 2003 Classification of Lupus Nephritis.108MOFO-360876942Attorney Docket No.14639-20729.40LN lupus nephritis; A = active; C = chronic; G = global; S = segmental.Note: Class V may occur in combination with Class III or IV, in which case both will be diagnosed. Class V LN may show advanced sclerosis.

[0237] In some embodiments, the patient has class III or class IV LN. In some embodiments, the patient has class III LN. For example, in some embodiments, the patient has class III(A) or class III(A / C) LN. In some embodiments, the patient has class IV LN. For example, in some embodiments, the patient has class IV-S(A), IV-G(A), IV-S(A / C), or IV-G(A / C) LN. As shown in Table 3 above, class V LN may also occur concomitantly with class III or class IV LN. In some embodiments, the methods of the present disclosure are used to treat a patient with class III or class IV LN and concomitant class V LN. In some embodiments, the patient does not have class V LN. In some embodiments, the patient has biopsy-proven proliferative LN. In some embodiments, the patient has been diagnosed with Class III or IV LN as evidenced by renal biopsy, e.g., performed within 6 months prior to treatment. In some embodiments, the patient has concomitant class V LN. In some embodiments, the patient has at least 10, at least 12, at least 13, or at least 15 B cells per mm2tissue in a kidney biopsy sample obtained prior to treatment. In some embodiments, the patient has an AI of greater than or equal to 4 or greater than or equal to 6 in a kidney biopsy sample obtained prior to treatment.

[0238] As discussed above, a high frequency of patients with lupus (e.g., SLE) eventually develop LN. In some embodiments, the patient is at risk for developing LN. In some embodiments, the patient is at risk for developing class III or class IV LN. In some embodiments, the patient is at risk for developing class III or class IV LN with concomitant class V LN.

[0239] In some embodiments, the patient does not have class III(C) LN (e.g., as described in Table 3 above). In some embodiments, the patient does not have class IV(C) LN, such as class IV-S(C) or IV-G(C) LN (e.g., as described in Table 3 above).109MOFO-360876942Attorney Docket No.14639-20729.40

[0240] In some embodiments, the patient has a urine to protein creatinine ratio (UPCR) ofe.g. e.g. , e.g. prior to treatment, e.g., on a24-hour urine collection.treatment, e.g., on a 24-hour urine collection.

[0241] In some embodiments, the patient has received at least one dose of pulse methylprednisolone (e.g., 500-1000mg IV) prior to treatment. In some embodiments, the patient has received an ACE inhibitor or angiotensin-days prior to treatment.

[0242] In some embodiments, the patient does not have severe renal impairment or need for dialysis or renal transplantation, e.g., prior to treatment as described herein. In some embodiments, the patient does not have sclerosis in >50% of glomeruli on renal biopsy, e.g., prior to treatment as described herein. In some embodiments, the patient does not have active central nervous system SLE, e.g., prior to treatment as described herein. In some embodiments, the patient does not have history of progressive multifocal leukoencephalopathy (PML), e.g., prior to treatment as described herein. In some embodiments, the patient does not have positive hepatitis C serology, hemoglobin < 7g / dL (unless caused by autoimmune hemolytic anemia resulting from SLE), platelet count <20,000 / uL, or positive serum human chorionic gonadotropin, e.g., prior to treatment as described herein. In some embodiments, the patient does not have known HIV infection, e.g., prior to treatment as described herein. In some embodiments, the patient has not been treated with one or more of: cyclophosphamide, calcineurin inhibitor, JAK inhibitor, BTK inhibitor, TYK2 inhibitor, or IV antibiotic prior to treatment as described herein (e.g., 3 months prior to treatment as described herein).

[0243] Several lab tests known in the art may be used to diagnose and / or monitor the presence, progression, and / or response to treatment in lupus nephritis. In some embodiments, serum creatinine may be measured. In some embodiments, the normal range for serum creatinine may be from about 0.6 to about 1.3 mg / dL, with some variation seen by age, between men and women, and from lab to lab. In some embodiments, the presence of urinary sediment and / or casts may be measured, e.g., by microscopic examination of urine. For example, the number of red blood cells in a urine sample may be assayed by microscopic examination. In some embodiments, a normal value for urinary sediment may be about 4 red blood cells (RBC) or less per high power field (HPF). Urinary casts may include without limitation red blood cell casts,110MOFO-360876942Attorney Docket No.14639-20729.40white blood cell casts, renal tubular epithelial cell casts, waxy casts, hyaline casts, granular casts, and fatty casts. In some embodiments, the patient has active urinary sediment as evidenced by 10 RBCs / HPF or the presence of red cell casts.

[0244] In some embodiments, a urinary protein to creatinine ratio (UPCR) may be measured. The presence of protein in the urine (proteinuria) may also be assayed by tests including without limitation a urine albumin to creatinine ratio (UACR) and dipstick urinalysis. Other tests and / or measures that may be useful for examining renal function include without limitation a renal panel, creatinine clearance, sodium, potassium, chloride, bicarbonate, phosphorus, calcium, albumin, blood urea nitrogen (BUN), creatinine, glucose, estimated glomerular filtration rate (eGFR), BUN / creatinine ratio, and anion gap, and may include a measurement of the above parameters in the blood and / or urine, where appropriate. For more detailed description, see, e.g., the American College of Rheumatology Guidelines for Screening, Case Definition, Treatment and Management of Lupus Nephritis (Hahn, B. et al. (2012) Arthritis Care Res.64:797-808).

[0245] In some embodiments, the patient has a UPCR of greater than or equal to about 2.0 g / g or greater than or equal to about 3.0 g / g prior to a treatment of the present disclosure. In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.7 g / g, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure). In some embodiments, the methods of the present disclosure result in a UPCR of less than or equal to about 0.5 g / g, e.g., at about 52 weeks, about 76 weeks, or about 104 weeks after commencement of treatment, e.g., after administration of the first antibody exposure (e.g., after administration of a first dose of a first antibody exposure of the present disclosure).

[0246] In some embodiments, an eGFR may be measured. In some embodiments, the patient has an eGFR that is greater than or equal to about 80% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure) after a treatment of the present disclosure. In some embodiments, the patient has an eGFR that is greater than or equal to about 60mL / min or greater than or equal to about 80% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure) after a treatment of the present111MOFO-360876942Attorney Docket No.14639-20729.40disclosure. In some embodiments, the patient has an eGFR that is greater than or equal to about 85% of a baseline eGFR (e.g., an eGFR of the individual prior to administration of the first antibody exposure, e.g., prior to administration of a first dose of a first antibody exposure of the present disclosure) after a treatment of the present disclosure.

[0247] In some embodiments, the methods of the present disclosure include administering to the individual a first antibody exposure to a type II anti-CD20 antibody of the present disclosure, a second antibody exposure to the type II anti-CD20 antibody of the present disclosure, and a third antibody exposure to the type II anti-CD20 antibody of the present disclosure. In some embodiments, the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure. In some embodiments, the second antibody exposure is not provided until about 18 weeks after the first antibody exposure, about 19 weeks after the first antibody exposure, about 20 weeks after the first antibody exposure, about 21 weeks after the first antibody exposure, about 22 weeks after the first antibody exposure, about 23 weeks after the first antibody exposure, about 24 weeks after the first antibody exposure, about 25 weeks after the first antibody exposure, or about 26 weeks after the first antibody exposure. In some embodiments, the second antibody exposure is not provided until less than about any of the following weeks after the first antibody exposure: 26, 25, 24, 23, 22, 21, 20, or 19. In some embodiments, the second antibody exposure is not provided until greater than about any of the following weeks after the first antibody exposure: 18, 19, 20, 21, 22, 23, 24, or 25. That is, the second antibody exposure is not provided until any of a range of weeks having an upper limit of 26, 25, 24, 23, 22, 21, 20, or 19 and an independently selected lower limit of 18, 19, 20, 21, 22, 23, 24, or 25, wherein the lower limit is less than the upper limit. In some embodiments, the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure. In some embodiments, the third antibody exposure is not provided until about 24 weeks after the second antibody exposure, about 25 weeks after the second antibody exposure, about 26 weeks after the second antibody exposure, about 27 weeks after the second antibody exposure, about 28 weeks after the second antibody exposure, about 29 weeks after the second antibody exposure, about 30 weeks after the second antibody exposure, about 31 weeks after the second antibody exposure, or about 32 weeks after the second antibody exposure. In some embodiments, the third antibody exposure is not provided until less than about any of the following weeks after the second antibody exposure: 32, 31, 30, 29, 28, 27, 26, or 25. In112MOFO-360876942Attorney Docket No.14639-20729.40some embodiments, the third antibody exposure is not provided until greater than about any of the following weeks after the second antibody exposure: 24, 25, 26, 27, 28, 29, 30, or 31. That is, the third antibody exposure is not provided until any of a range of weeks having an upper limit of 32, 31, 30, 29, 28, 27, 26, or 25 and an independently selected lower limit of 24, 25, 26, 27, 28, 29, 30, or 31, wherein the lower limit is less than the upper limit.

[0248] In some embodiments, the methods of the present disclosure include administering to the individual a first antibody exposure to a type II anti-CD20 antibody of the present disclosure and a second antibody exposure to the type II anti-CD20 antibody of the present disclosure. In some embodiments, the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure. In some embodiments, the second antibody exposure is not provided until about 18 weeks after the first antibody exposure, about 19 weeks after the first antibody exposure, about 20 weeks after the first antibody exposure, about 21 weeks after the first antibody exposure, about 22 weeks after the first antibody exposure, about 23 weeks after the first antibody exposure, about 24 weeks after the first antibody exposure, about 25 weeks after the first antibody exposure, or about 26 weeks after the first antibody exposure. In some embodiments, the second antibody exposure is not provided until less than about any of the following weeks after the first antibody exposure: 26, 25, 24, 23, 22, 21, 20, or 19. In some embodiments, the second antibody exposure is not provided until greater than about any of the following weeks after the first antibody exposure: 18, 19, 20, 21, 22, 23, 24, or 25. That is, the second antibody exposure is not provided until any of a range of weeks having an upper limit of 26, 25, 24, 23, 22, 21, 20, or 19 and an independently selected lower limit of 18, 19, 20, 21, 22, 23, 24, or 25, wherein the lower limit is less than the upper limit.

[0249] The dosing regimens described herein use a consistent system for tracking time between doses whereby the first dose is administered to the patient on Day 1 or week 0. As described herein, an antibody exposure of the present disclosure may include one or two doses. In cases where the antibody exposures contain one dose, references to a second antibody exposure not provided until a period of time has elapsed after a first antibody exposure (as described herein) refer to the amount of time elapsed between the dose of the first antibody exposure (e.g., Day 1 or week 0) and the dose of the second antibody exposure. If the first antibody exposure includes two doses, the first dose of the first antibody exposure is provided on Day 1 or week 0. In cases where the antibody exposures contain two doses, references to a113MOFO-360876942Attorney Docket No.14639-20729.40second antibody exposure not provided until a period of time has elapsed after a first antibody exposure (as described herein) refer to the amount of time elapsed between the first of the two doses of the first antibody exposure (e.g., Day 1 or week 0) and the first dose of the two doses of the second antibody exposure. For example, if a method of the present disclosure includes a first antibody exposure with two doses and a second antibody exposure with two doses, and the second antibody exposure is not provided until about 22 weeks after the first antibody exposure, then the interval between the first dose of the first antibody exposure and the first dose of the second antibody exposure is about 22 weeks.

[0250] In some embodiments, a first antibody exposure of the present disclosure includes one or two doses of a type II anti-CD20 antibody of the present disclosure. In some embodiments, the first antibody exposure contains a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody. In some embodiments, the first antibody exposure contains a total exposure of about 1800mg, about 1900mg, about 2000mg, about 2100mg, or about 2200mg of the type II anti-CD20 antibody. In some embodiments, the individual weighs greater than or equal to 45kg.

[0251] In some embodiments, the first antibody exposure includes two doses. In some embodiments, the first antibody exposure includes a first dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody and a second dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody. In some embodiments, the first dose of the first antibody exposure contains about 1000mg of the type II anti-CD20 antibody. In some embodiments, the second dose of the first antibody exposure contains about 1000mg of the type II anti-CD20 antibody. In some embodiments, the individual weighs greater than or equal to 45kg.

[0252] In some embodiments, the second dose of the first antibody exposure is not provided until about 1.5 weeks to about 2.5 weeks after the first dose of the first antibody exposure. In some embodiments, the second dose of the first antibody exposure is not provided until about 2 weeks after the first dose of the first antibody exposure.

[0253] In some embodiments, a second antibody exposure of the present disclosure includes one or two doses of a type II anti-CD20 antibody of the present disclosure. In some embodiments, the second antibody exposure contains a total exposure of between about 1800mg and about 2200mg of the type II anti-CD20 antibody. In some embodiments, the second114MOFO-360876942Attorney Docket No.14639-20729.40antibody exposure contains a total exposure of about 1800mg, about 1900mg, about 2000mg, about 2100mg, or about 2200mg of the type II anti-CD20 antibody. In some embodiments, the individual weighs greater than or equal to 45kg.

[0254] In some embodiments, the second antibody exposure includes two doses. In some embodiments, the second antibody exposure includes a first dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody and a second dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody. In some embodiments, the first dose of the second antibody exposure contains about 1000mg of the type II anti-CD20 antibody. In some embodiments, the second dose of the second antibody exposure contains about 1000mg of the type II anti-CD20 antibody. In some embodiments, the individual weighs greater than or equal to 45kg.

[0255] In some embodiments, the second dose of the second antibody exposure is not provided until about 1.5 weeks to about 2.5 weeks after the first dose of the second antibody exposure. In some embodiments, the second dose of the second antibody exposure is not provided until about 2 weeks after the first dose of the second antibody exposure.

[0256] In some embodiments, a third antibody exposure of the present disclosure includes one or two doses of a type II anti-CD20 antibody of the present disclosure. In some embodiments, the third antibody exposure contains a total exposure of between about 800mg and about 1200mg of the type II anti-CD20 antibody. In some embodiments, the third antibody exposure contains a total exposure of about 800mg, about 900mg, about 1000mg, about 1100mg, or about 1200mg of the type II anti-CD20 antibody. In some embodiments, the individual weighs greater than or equal to 45kg.

[0257] In some embodiments, the third antibody exposure includes a single dose. In some embodiments, the third antibody exposure includes a single dose of between about 900mg and about 1100mg of the type II anti-CD20 antibody. In some embodiments, the single dose of the third antibody exposure contains about 1000mg of the type II anti-CD20 antibody. In some embodiments, the individual weighs greater than or equal to 45kg.

[0258] In some embodiments, a type II anti-CD20 antibody of the present disclosure is administered intravenously (e.g., by IV infusion).

[0259] In some embodiments, the methods of the present disclosure further include administering a standard of care treatment (e.g., in conjunction with a type II anti-CD20 antibody115MOFO-360876942Attorney Docket No.14639-20729.40as described herein). In some embodiments, a standard of care treatment may be administered before, during, or after administration of a type II anti-CD20 antibody of the present disclosure, e.g., for treating or preventing one or more symptoms of lupus. In certain embodiments, a standard of care treatment may be administered after a second antibody exposure of the present disclosure. In certain embodiments, a standard of care treatment may be administered after a third antibody exposure of the present disclosure. For example, a type II anti-CD20 antibody of the present disclosure may be administered as described herein to a patient as an induction therapy, then the patient may be treated according to standard of care as a maintenance therapy. Standard of care treatments for lupus are well known in the art and include without limitation an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin-receptor blocker, cyclophosphamide, azathioprine, a glucocorticoid or corticosteroid, and mycophenolic acid or a derivative or salt thereof, including without limitation mycophenolate mofetil (e.g., at a dose as described herein, such as 2.0-2.5 g / day). For example, an ACE inhibitor or angiotensin-receptor blocker can be titrated to adequate blood pressure control for age and sex, e.g., as recommended by the KDIGO (Kidney Disease: Improving Global Outcomes) Blood Pressure Work Group for chronic kidney disease (Becker et al. (2012) Kidney International Supplements 2:337-414). In some embodiments, MMF is administered to the individual at a target dose of about 2.0g / day to about 2.5g / day and / or oral prednisone is administered to the individual at about 0.5mg / kg / day, with a maximum of about 60mg / day and tapered to about 5mg / day beginning at Week 2, as part of standard of care treatment.

[0260] In some embodiments, the methods of the present disclosure further include administering an effective amount of an...

Claims

1. Attorney Docket No.14639-20729.402.CLAIMS3.What is claimed is:

1. A method for depleting kidney-resident B cells in an individual that has lupus nephritis, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure;5.wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab;6.wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab;7.wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab;8.wherein the individual is a human; and9.wherein treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure.

2. The method of claim 1, wherein kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure have an average of at least 10, at least 12, at least 13, or at least 15 B cells per mm2tissue.

3. A method for depleting kidney-resident B cells in an individual that has lupus nephritis, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure;12.14013.MOFO-360876942 Attorney Docket No.14639-20729.4014.wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab;15.wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab;16.wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab;17.wherein the individual is a human; and18.wherein treatment with obinutuzumab in a plurality of individuals results in a 95% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure.

4. A method for depleting kidney-resident B cells in an individual that has lupus nephritis, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the second antibody exposure is not provided until from about 18 weeks to about 26 weeks after the first antibody exposure, and the third antibody exposure is not provided until from about 24 weeks to about 32 weeks after the second antibody exposure;20.wherein the first antibody exposure comprises one or two doses of obinutuzumab, the first antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab;21.wherein the second antibody exposure comprises one or two doses of obinutuzumab, the second antibody exposure comprising a total exposure of between about 1800mg and about 2200mg of obinutuzumab;22.wherein the third antibody exposure comprises one or two doses of obinutuzumab, each dose of the third antibody exposure comprising between about 900mg and about 1100mg of obinutuzumab;23.wherein the individual is a human; and24.14125.MOFO-360876942 Attorney Docket No.14639-20729.4026.wherein a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1.

5. The method of claim 4, wherein a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0.

6. The method of claim 4 or claim 5, wherein a kidney biopsy sample obtained from the individual prior to administration of the first antibody exposure exhibits an AI of greater than or equal to 4 or greater than or equal to 6.

7. The method of any one of claims 1-6, wherein the kidney-resident B cells are CD79a+ / CD138- B cells.

8. The method of any one of claims 1-7, wherein, prior to administration of the first antibody exposure, the individual has one or more of the following:31.(a) a urine protein-to-creatinine ratio (UPCR) of greater than or equal to about 1.0 g / g, greater than or equal to about 2.0 g / g, or greater than or equal to about 3.0 g / g;32.(b) an estimated glomerular filtration rate (eGFR) that is less than or equal to 90 mL / min / 1.73m2;33.(c) a serum sample that is positive for anti-double-stranded DNA (anti-dsDNA) antibodies; (d) a serum sample that has a low C3 complement level;34.(e) a serum sample that has a low C4 complement level;35.(f) no prior history or diagnosis of lupus and / or lupus nephritis;36.(g) concomitant class V lupus nephritis; and / or37.(h) is female.

9. The method of any one of claims 1-8, wherein, prior to administration of the first antibody exposure, the individual has active lupus nephritis.

10. The method of any one of claims 1-9, wherein the individual has class III or class IV lupus nephritis.

11. The method of any one of claims 1-9, wherein the individual has class III (C) or class IV (C) lupus nephritis.

12. The method of any one of claims 1-11, wherein the individual has concomitant class V lupus nephritis.42.14243.MOFO-360876942 Attorney Docket No.14639-20729.4013. The method of any one of claims 1-12, wherein the first antibody exposure comprises a first dose of about 1000mg of obinutuzumab and a second dose of about 1000mg of obinutuzumab.

14. The method of any one of claims 1-13, wherein the first antibody exposure comprises a first dose of obinutuzumab and a second dose of obinutuzumab, and wherein the second dose of the first antibody exposure is not provided until about 2 weeks after the first dose of the first antibody exposure.

15. The method of any one of claims 1-14, wherein the second antibody exposure comprises a first dose of about 1000mg of obinutuzumab and a second dose of about 1000mg of obinutuzumab.

16. The method of any one of claims 1-15, wherein the second antibody exposure comprises a first dose of obinutuzumab and a second dose of obinutuzumab, and wherein the second dose of the second antibody exposure is not provided until about 2 weeks after the first dose of the second antibody exposure.

17. The method of any one of claims 1-16, wherein the third antibody exposure comprises a single dose of about 1000mg of obinutuzumab.

18. The method of claim 17, wherein the single dose of the third antibody exposure is not provided until about 52 weeks after the first dose of the first antibody exposure or until about 28 weeks after the first dose of the second antibody exposure.

19. The method of any one of claims 1-16, wherein the third antibody exposure comprises two doses of about 1000mg of obinutuzumab.

20. The method of claim 19, wherein the first dose of the third antibody exposure is not provided until about 50 weeks after the first dose of the first antibody exposure or until about 26 weeks after the first dose of the second antibody exposure, and wherein the second dose of the third antibody exposure is not provided until about 52 weeks after the first dose of the first antibody exposure or until about 28 weeks after the first dose of the second antibody exposure.

21. The method of any one of claims 1-12, wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab on week 52 of treatment.

22. The method of any one of claims 1-12, wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on weeks 0 and 2 of treatment; wherein the second antibody54.14355.MOFO-360876942 Attorney Docket No.14639-20729.4056.exposure comprises two doses of 1000mg of obinutuzumab on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab on weeks 50 and 52 of treatment.

23. The method of any one of claims 1-12, wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on days 1 and 15 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on days 168 and 182 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab on day 364 of treatment.

24. The method of any one of claims 1-12, wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab on days 1 and 15 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab on days 168 and 182 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab on days 350 and 364 of treatment.

25. The method of any one of claims 1-24, further comprising administering to the individual an effective amount of a standard of care treatment.

26. The method of claim 25, wherein the standard of care treatment comprises treatment with one or more of an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin-receptor blocker, cyclophosphamide, azathioprine, a glucocorticoid or corticosteroid, and mycophenolic acid or a derivative or salt thereof.

27. The method of claim 26, wherein the standard of care treatment comprises mycophenolate mofetil (MMF).

28. The method of claim 27, wherein MMF is administered to the individual at a target dose of about 2.0g / day to about 2.5g / day.

29. The method of any one of claims 1-28, further comprising administering to the individual an effective amount of a glucocorticoid or corticosteroid.

30. The method of claim 29, wherein the glucocorticoid or corticosteroid comprises prednisone.

31. The method of claim 30, comprising administering to the individual an effective amount of oral prednisone.66.14467.MOFO-360876942 Attorney Docket No.14639-20729.4032. The method of claim 31, comprising administering to the individual about 0.5mg / kg / day oral prednisone, with a maximum of about 60mg / day and tapered to about 5mg / day beginning at Week 2.

33. The method of any one of claims 1-32, further comprising administering to the individual an effective amount of a glucocorticoid or corticosteroid prior to one or more dose(s) of obinutuzumab.

34. The method of claim 33, wherein the glucocorticoid or corticosteroid comprises methylprednisolone.

35. The method of claim 34, comprising intravenously administering an effective amount of methylprednisolone to the individual prior to one or more dose(s) of obinutuzumab.

36. The method of claim 35, comprising intravenously administering 80mg methylprednisolone to the individual prior to one or more dose(s) of obinutuzumab.

37. The method of any one of claims 1-36, further comprising administering to the individual an effective amount of acetaminophen prior to one or more dose(s) of obinutuzumab.

38. The method of claim 37, comprising administering to the individual about 650mg to about 1000mg of acetaminophen prior to one or more dose(s) of obinutuzumab.

39. The method of any one of claims 1-38, further comprising administering to the individual an effective amount of an antihistamine prior to one or more dose(s) of obinutuzumab.

40. The method of claim 39, wherein the antihistamine comprises diphenhydramine.

41. The method of claim 40, comprising administering to the individual about 50mg of diphenhydramine prior to one or more dose(s) of obinutuzumab.

42. The method of any one of claims 1-41, wherein the first, second, and / or third antibody exposure(s) is / are administered intravenously to the individual.

43. A method for depleting kidney-resident B cells in an individual that has lupus nephritis, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment;80.wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment;81.14582.MOFO-360876942 Attorney Docket No.14639-20729.4083.wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment;84.wherein the individual is a human; and85.wherein:86.(a) treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure;87.(b) treatment with obinutuzumab in a plurality of individuals results in a 95% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure; and / or88.(c) a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1, optionally a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0.

44. A method for depleting kidney-resident B cells in an individual that has lupus nephritis, comprising administering to the individual at least a first antibody exposure to obinutuzumab, a second antibody exposure to obinutuzumab, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment;90.wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment;91.wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment;92.wherein the individual is a human; and93.wherein:94.(a) treatment with obinutuzumab in a plurality of individuals results in an average of less than one B cell per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure;95.14696.MOFO-360876942 Attorney Docket No.14639-20729.4097.(b) treatment with obinutuzumab in a plurality of individuals results in a 95% or greater reduction in average number of B cells per mm2tissue in kidney biopsy samples obtained at about 76 weeks to about 80 weeks after administration of the first antibody exposure, as compared to average number of B cells per mm2tissue in kidney biopsy samples obtained from the plurality of individuals prior to administration of the first antibody exposure; and / or98.(c) a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an activity index (AI) of less than or equal to 1, optionally a kidney biopsy sample obtained from the individual at about 76 weeks to about 80 weeks after administration of the first antibody exposure exhibits an AI of 0.

45. The method of claim 43 or claim 44, further comprising administering to the individual an effective amount of a standard of care treatment.

46. The method of claim 45, wherein the standard of care treatment comprises mycophenolate mofetil (MMF).

47. The method of claim 46, wherein MMF is administered to the individual at a target dose of about 2.0g / day to about 2.5g / day.

48. The method of any one of claims 43-47, further comprising administering to the individual an effective amount of a glucocorticoid or corticosteroid.

49. The method of claim 48, wherein the glucocorticoid or corticosteroid comprises prednisone.

50. The method of claim 49, comprising administering to the individual about 0.5mg / kg / day oral prednisone, with a maximum of about 60mg / day and tapered to about 5mg / day beginning at Week 2.

51. The method of any one of claims 43-50, further comprising, prior to one or more dose(s) of obinutuzumab, administering to the individual an effective amount of a glucocorticoid or corticosteroid, acetaminophen, and / or an antihistamine.

52. The method of claim 51, comprising intravenously administering 80mg methylprednisolone to the individual prior to one or more dose(s) of obinutuzumab.

53. The method of claim 51 or claim 52, comprising administering to the individual about 650mg to about 1000mg of acetaminophen prior to one or more dose(s) of obinutuzumab.

54. The method of any one of claims 51-53, comprising administering to the individual about 50mg of diphenhydramine prior to one or more dose(s) of obinutuzumab.109.147110.MOFO-360876942 Attorney Docket No.14639-20729.4055. A kit for depleting kidney-resident B cells in an individual in need thereof, comprising:112.(a) a container comprising obinutuzumab; and113.(b) a package insert with instructions for depleting kidney-resident B cells in an individual, wherein the individual is a human, and wherein the instructions indicate that obinutuzumab is to be administered to the individual in a first antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment, a second antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment, and a third antibody exposure that comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment.

56. A kit for depleting kidney-resident B cells in an individual in need thereof, comprising:115.(a) a container comprising obinutuzumab; and116.(b) a package insert with instructions for depleting kidney-resident B cells in an individual, wherein the individual is a human, and wherein the instructions indicate that obinutuzumab is to be administered to the individual in a first antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment, a second antibody exposure that comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment, and a third antibody exposure that comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment.

57. The kit of claim 55 or claim 56, wherein the individual has or has been diagnosed with lupus nephritis.

58. The kit of any one of claims 55-57, further comprising a container that comprises:119.(c) a second medicament, wherein obinutuzumab is a first medicament; and120.wherein the package insert further comprises instructions for administering the second medicament to the individual.

59. The kit of claim 58, wherein the second medicament is a standard-of-care medicament for treating lupus nephritis.

60. The kit of claim 59, wherein the second medicament is mycophenolate mofetil (MMF).

61. Obinutuzumab for use in the method according to any one of claims 1-54.124.148125.MOFO-360876942 Attorney Docket No.14639-20729.4062. Obinutuzumab for use in a method of depleting kidney-resident B cells in an individual that has lupus nephritis, said method comprising administering intravenously to the individual a first, a second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment.

63. Obinutuzumab for use in a method of depleting kidney-resident B cells in an individual that has lupus nephritis, said method comprising administering intravenously to the individual a first, a second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment.

64. Use of obinutuzumab in the manufacture of a medicament for depleting kidney-resident B cells in an individual that has lupus nephritis, wherein the obinutuzumab is to be administered intravenously to the individual in a first, a second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises one dose of 1000 mg of obinutuzumab intravenously administered to the individual on week 52 of treatment.

65. Use of obinutuzumab in the manufacture of a medicament for depleting kidney-resident B cells in an individual that has lupus nephritis, wherein the obinutuzumab is to be administered intravenously to the individual in a first, a second, and a third antibody exposure to obinutuzumab; wherein the first antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered to the individual on weeks 0 and 2 of treatment; wherein the second antibody exposure comprises two doses of 1000mg of obinutuzumab intravenously administered130.149131.MOFO-360876942 Attorney Docket No.14639-20729.40132.to the individual on weeks 24 and 26 of treatment; and wherein the third antibody exposure comprises two doses of 1000 mg of obinutuzumab intravenously administered to the individual on weeks 50 and 52 of treatment.133.150134.MOFO-360876942

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