Humanized CD180 antibodies and uses thereof

Abstract

Provided are humanized anti-CD180 antibodies and methods of using such antibodies for the treatment of autoimmune and inflammatory disease or disorders.

Description

Attorney Docket No: 01434-0001-00PCTHUMANIZED CD180 ANTIBODIES AND USES THEREOFDESCRIPTION CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Application No. 63 / 776,110, filed March, 22, 2025, the entire contents of which are incorporated by reference herein for all purposes.REFERENCE TO ELECTRONIC SEQUENCE LISTING

[0002] This application contains a Sequence Listing which has been submitted electronically in .XML format and is hereby incorporated by reference in its entirety7. Said .XML copy, created on March 18, 2026, is namedL‘01434-0001-00PCT.xml” and is 88,636 bytes in size. The sequence listing contained in this .XML file is part of the specification and is hereby incorporated by reference herein in its entirety.FIELD

[0003] Humanized CD 180 Antibodies and Uses Thereof.BACKGROUND

[0004] Cluster of Differentiation 180 (CD180), also known as LY64, Ly78. or RP105, is structurally related to the toll-like receptors (TLR), a family of pattern recognition receptors. CD 180 is a cell surface molecule having extracellular leucine-rich repeats (LRR) and a short cytoplasmic tail. Expression of CD180 appears to be restricted to the antigen-presenting cells including B cells, monocytes, dendritic cells, and macrophages. The extracellular LRR is associated with a molecule called MD-1 and forms the cell surface receptor complex, CD180 / MD-1. Signaling by CD 180 / MD- 1 regulates multiple immune functions. For example, CD180 / MD-1, by working in concert with TLR4, control B cell recognition and signaling of lipopolysaccharide (LPS), a membrane constituent of Gram-negative bacteria. In addition, CD180 / MD-1 ligation inhibits TLR7- and TLR9-mediated activation of macrophages, dendritic cells, and B cells.

[0005] Current treatments for inflammatory diseases and autoimmune diseases and disorders by neutralizing pro-inflammatory cytokines such as the type 1 interferons can have a global effect, affecting the offending immune cells as well as non-immune bystander normal tissues, which may be a cause of undesired side effects. There is a need, therefore, for treatments that can specifically suppress the actions of proinflammatory immune cells to deliver targeted anti-inflammatory actions without globally suppressing normal immune functions in a patient’s body. Ligation of CD180 inhibits TLR7- and TLR9-mediated activation of the type 1 interferonAttorney Docket No: 01434-0001-00PCT pathway in CD180-expressing B cells, macrophages, and dendritic cells and specifically modulates the type 1 interferon pathway. Anti-mouse CD 180 has demonstrated therapeutic efficacy in the imiquimod-induced and MRL / lpr lupus models in mice. Accordingly, there is a need for compositions and methods to selectively modulate CD 180 activity for treating autoimmunity and inflammatory disease in humans.SUMMARY

[0006] The following numbered embodiments provide embodiments as described herein, though the embodiments recited here are not limiting:

[0007] Embodiment 1. A humanized antibody or antigen-binding fragment thereof that specifically binds to human CD 180, the antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3, wherein each CDR comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% , or 100% identity7to the amino acid sequence or 0, 1, or 2 amino acid mutations relative to the amino acid sequence set forth in:a) CDRH1: SEQ ID NO:17, 72, 50 or 77; CDRH2: SEQ ID NO:18. 84.51, or 89; and CDRH3: SEQ ID NO: 19 or 52; according to Kabat numbering; orb) CDRH1: SEQ ID NO:25, 55, 74 or 78; CDRH2: SEQ ID NO:26 or 85; and CDRH3: SEQ ID NO: 27 or 56; according to IM GT numbering;the light chain variable region comprises CDRL1, CDRL2. and CDRL3. wherein each CDR comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or 100% identity to the amino acid sequence or 0, 1, or 2 amino acid mutations relative to the amino acid sequence set forth in:a) CDRL1: SEQ ID NO:20 or 53; CDRL2: SEQ ID NO:21; and CDRL3: SEQ ID NO:22 or 54; according to Kabat numbering; orb) CDRL1: SEQ ID NO:28 or 57; CDRL2: SEQ ID NO:29; and CDRL3: SEQ ID NO:22 or 54; according to IMGT numbering;the heavy chain variable region comprises human IGVH1-03 / JH6 framework regions having an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%. 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences of SEQ ID NOs:38 and 39; the light chain variable region comprises human IGVH1-03 / JH6 framework regions having an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequences of SEQ ID NOs:40 and 41;Attorney Docket No: 01434-0001-00PCT position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering; and1. at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering.

[0008] Embodiment 2. The humanized antibody or antigen-binding fragment thereof of embodiment 1, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences set forth in one of the sets of amino acid sequences selected from:i. SEQ ID NOs: 17, 18, 19, 20, 21, and 22, according to Kabat numbering;ii. SEQ ID NOs: 17, 84, 19, 20, 21, and 22, according to Kabat numbering;iii SEQ ID NOs:72, 18, 19, 20, 21, and 22, according to Kabat numbering;iv. SEQ ID NOs: 72, 84, 19, 20, 21, and 22 according to Kabat numbering;v. SEQ ID NOs:25, 26, 27, 28, 29, and 22, according to IMGT numbering;vi. SEQ ID NOs: 25, 85, 27, 28, 29, and 22, according to IMGT numbering;vii. SEQ ID NOs:74, 26, 27, 28, 29, and 22, according to IMGT numbering;viii. SEQ ID NOs: 74, 85, 27, 28, 29, and 22, according to IMGT numbering;ix. SEQ ID NOs:50, 51, 52, 53, 21 and 54, according to Kabat numbering;x. SEQ ID NOs:50, 89, 52, 53, 21 and 54, according to Kabat numbering;xi. SEQ ID NOs:77, 51, 52, 53, 21 and 54, according to Kabat numbering;xii. SEQ ID NOs:77, 89, 52, 53, 21 and 54, according to Kabat numbering;xiii. SEQ ID NOs:55, 26, 56, 57, 29, and 54, according to IMGT numbering;xiv. SEQ ID NOs:55, 85, 56, 57, 29, and 54, according to IMGT numbering;xv. SEQ ID NOs:78, 26, 56, 57, 29, and 54, according to IMGT numbering; andxvi. SEQ ID NOs:78, 85, 56, 57, 29, and 54, according to IMGT numbering.

[0009] Embodiment 3. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 or 2, wherein:the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:3, 5, 6, 7, 9, 10, 14, 15, 16, 66, 67, 82, 64 or 65;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:4, 8, 45, 46, 47, 81, 68, 69, 70, or 71; andat least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.Attorney Docket No: 01434-0001-00PCT

[0010] Embodiment 4. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 3, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences set forth in one of the sets of amino acid sequences selected from:i. SEQ ID NOs: 17, 18, 19, 20, 21, and 22, according to Kabat numbering;ii. SEQ ID NOs:72, 18, 19, 20, 21, and 22, according to Kabat numbering;iii. SEQ ID NOs:25, 26, 27, 28, 29, and 22, according to IMGT numbering; and iv. SEQ ID NOs:74, 26, 27, 28, 29, and 22, according to IMGT numbering:the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:3, 5, 6, 7, 9, 10, 14, 15, or 16;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%. or 100% identity to an amino acid sequence set forth in SEQ ID NOs:4, 8, 45, 46, 47 and 81 ;at least one of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0011] Embodiment 5. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 4,wherein at least one of position 28 is serine, position 66 is lysine, position 67 is alanine, position 69 is leucine, and position 71 is valine in the heavy chain variable region, according to Kabat numbering; or2. position 64 is valine, position 48 is optionally valine and position 70 is optionally glutamine in the light chain variable region, according to Kabat numbering; position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0012] Embodiment 6. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 3, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences set forth one of the sets of amino acid sequences selected from:i. SEQ ID NOs:50, 51, 52, 53, 21 and 54, according to Kabat numbering;ii. SEQ ID NOs:77, 51, 52, 53, 21 and 54, according to Kabat numbering;iii. SEQ ID NOs:55, 26, 56, 57, 29, and 54, according to IMGT numbering; and iv. SEQ ID NOs:78, 26, 56, 57, 29, and 54, according to IMGT numbering;Attorney Docket No: 01434-0001-00PCT wherein the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:66, 67, 82, 64 and 65;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%,99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:68, 69, 70 and 71;3. at least one of, at least two of or all of position 48 is isoleucine, position 49 is phenylalanine or asparagine, position 64 is glycine or valine and position 70 is aspartate in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0013] Embodiment 7. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1. 2, 3, 5 and 6, wherein position 48 is isoleucine, position 49 is phenylalanine or asparagine, position 64 is glycine or valine and position 70 is aspartate in the light chain variable region, according to Kabat numbering.

[0014] Embodiment 8. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1, 2, 3, 5 and 6, wherein position 64 is valine, according to Kabat numbering.

[0015] Embodiment 9. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 5, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:17, 18, and 19, respectively, according to Kabat numbering, and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:20, 21 and 22, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0016] Embodiment 10. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 6, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:72, 18, and 19, respectively, according to Kabat numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:20, 21 and 22, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0017] Embodiment 11. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 6, wherein the HCDR1 , HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:25, 26, and 27, respectively, according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:28, 29, and 22, respectively according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.Attorney Docket No: 01434-0001-00PCT

[0018] Embodiment 12. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 6, wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:74, 26, and 27, respectively according to IMGT numbering; and LCDR1. LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:28, 29, and 22, respectively according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0019] Embodiment 13. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1. 2, 3, and 5 to 8, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:50, 51, and 52, respectively, according to Kabat numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:53, 21 and 54, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0020] Embodiment 14. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 , 2, 3, and 5 to 8, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:77, 51, and 52, respectively, according to Kabat numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:53, 21 and 54, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0021] Embodiment 15. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1, 2, 3, and 5 to 8, wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:55, 26, and 56, respectively, according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:57, 29, and 54, respectively, according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0022] Embodiment 16. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1. 2, 3, and 7 to 9, wherein the 14CDR1. HCDR2. and EICDR3 comprise the amino acid sequences set forth in SEQ ID NOs:78, 26, and 56, respectively, according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:57, 29, and 54, respectively, according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0023] Embodiment 17. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 16, wherein:a) the heavy chain variable region comprises glutamine at position 54;b) the heavy chain variable region comprises serine at position 54; orc) the heavy chain variable region comprises alanine at position 55;Attorney Docket No: 01434-0001-00PCT according to Kabat numbering.

[0024] Embodiment 18. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 17, wherein the heavy chain variable region comprises alanine at position 55; according to Kabat numbering.

[0025] Embodiment 19. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 5, and 9 to 12, wherein:a) the heavy chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%. 95%. 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:3, 5, 6, 7, 9, 10, 14, 15 or 16; andb) the light chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity' to an amino acid sequence set forth in the amino acid sequences set forth in SEQ ID NO:4. 8, 45, 46, 47 or 81.

[0026] Embodiment 20. The humanized antibody or antigen-binding fragment thereof of embodiment 19, wherein the heavy chain variable region and the light chain variable region comprise amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences set forth in:a) SEQ ID NO:6 and SEQ ID NO:8, respectively;b) SEQ ID NO: 16 and SEQ ID NO:8, respectively;c) SEQ ID NO:6 and SEQ ID NO:81, respectively;d) SEQ ID NO: 16 and SEQ ID NO:81, respectively ;e) SEQ ID NO: 14 and 8, respectively;f) SEQ ID NO: 15 and 8, respectively;g) SEQ ID NO:6 and 45, respectively;h) SEQ ID NO:6 and 46, respectively; ori) SEQ ID NO:6 and 47, respectively.

[0027] Embodiment 21. The humanized antibody or antigen-binding fragment thereof of embodiment 19 or 20, wherein the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in:a) SEQ ID NO:6 and SEQ ID NO:8, respectively;b) SEQ ID NO: 16 and SEQ ID NO:8, respectively;c) SEQ ID NO:6 and SEQ ID NO:81, respectively;d) SEQ ID NO: 16 and SEQ ID NO:81, respectively;e) SEQ ID NO: 14 and 8, respectiv ely;1) SEQ ID NO: 15 and 8, respectively;Attorney Docket No: 01434-0001-00PCT g) SEQ ID N0:6 and 45, respectively;h) SEQ ID N0:6 and 46, respectively; ori) SEQ ID NO:6 and 47, respectively.

[0028] Embodiment 22. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 3, 5 to 8, and 13 to 18, wherein:a) the heavy chain variable region comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:64, 65. 66. 67. or 82; andb) the light chain variable region comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in the amino acid sequences set forth in SEQ ID NO:68, 69, 70 or 71.

[0029] Embodiment 23. The humanized antibody or antigen-binding fragment thereof of embodiment 22, wherein the heavy’ chain variable region and the light chain variable region comprise amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences set forth in:a) SEQ ID NO:66 and SEQ ID NO:68, respectively;b) SEQ ID NO:67 and SEQ ID NO:71, respectively;c) SEQ ID NO:64 and SEQ ID NO:68, respectively;d) SEQ ID NO:82 and SEQ ID NO:68, respectively; ore) SEQ ID NO:65 and SEQ ID NO:68, respectively.

[0030] Embodiment 24. The humanized antibody or antigen-binding fragment thereof of embodiment 22 or 23, wherein the heavy’ chain vanable region and the light chain variable region comprise amino acid sequences set forth in:a) SEQ ID NO:66 and SEQ ID NO:68, respectivelyb) SEQ ID NO:67 and SEQ ID NO:71, respectively;c) SEQ ID NO:64 and SEQ ID NO:68, respectively;d) SEQ ID NO:82 and SEQ ID NO:68, respectively: ore) SEQ ID NO:65 and SEQ ID NO:68, respectively.

[0031] Embodiment 25. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 24. further comprising a heavy chain constant region attached to the heavy chain variable region and a light chain constant region attached to the light chain variable region.

[0032] Embodiment 26. The humanized antibody or antigen-binding fragment thereof of embodiment 25, wherein the heavy' chain constant region is an IgG, IgA, or IgM region and the light chain constant region is a kappa region.Attorney Docket No: 01434-0001-00PCT

[0033] Embodiment 27. The humanized antibody or antigen binding fragment thereof of embodiment 26, wherein the heavy chain constant region is an IgG region.

[0034] Embodiment 28. The humanized antibody or antigen-binding fragment thereof of embodiment 27, wherein the heavy’ chain constant region is an IgGl region.

[0035] Embodiment 29. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 26 to 28, wherein the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:42.

[0036] Embodiment 30. The humanized antibody or antigen-binding fragment thereof of embodiment 27, wherein the heavy' chain constant region is an IgG4 region, optionally wherein the IgG4 region comprises an S228P substitution.

[0037] Embodiment 31. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 29, wherein the antibody or antigen-binding fragment thereof has reduced antibody dependent cellular cytotoxicity’ (ADCC).

[0038] Embodiment 32. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 31, wherein the antibody or antigen-binding fragment thereof has reduced antibody dependent cellular phagocytosis activity (ADCP).

[0039] Embodiment 33. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 32, wherein the antibody or antigen-binding fragment thereof has reduced complement-dependent cytotoxicity7activity' (CDC).

[0040] Embodiment 34. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 33. comprising a IgGl heavy chain constant region having amino acid substitutions selected from (a) L234A L235A (LALA); (b) L234A, L235A, and P329G (LALA-PG); (c) L234A, L235E, G237A, A330S and P331S; or (d) L234A, L23 A and G237A; or comprising an IgG4 heavy chain constant region having amino acid substitutions (e) S288P.

[0041] Embodiment 35. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 25 and 31 to 34, wherein the heavy chain constant region is an Fc null.

[0042] Embodiment 36. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 31 to 35, wherein the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:43.

[0043] Embodiment 37. The antibody or antigen-binding fragment thereof of any one of embodiments 1 to 36, wherein the antibody or antigen-binding fragment thereof is a Fab, a Fab1, a F(ab')2, a Fv fragment, a diabody, a single-chain antibody, an scFv fragment or an scFv-Fc.Attorney Docket No: 01434-0001-00PCT

[0044] Embodiment 38. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 37, wherein the antibody or antigen-binding fragment thereof is monospecific, bispecific, monovalent or bivalent.

[0045] Embodiment 39. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 38, wherein the antibody or antigen-binding fragment thereof induces activation of antigen presenting cells.

[0046] Embodiment 40. The humanized antibody or antigen-binding fragment thereof of embodiment 39, wherein activation of antigen presenting cells is CD86 upregulation.

[0047] Embodiment 41. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 40, wherein the antibody or antigen-binding fragment thereof, induces increased phosphory lation of protein AKT in antigen presenting cells.

[0048] Embodiment 42. The humanized antibody or antigen-binding fragment thereof of any one of embodiments 1 to 41. wherein the antibody or antigen-binding fragment thereof, induces increased phosphorylation of protein S6 in antigen presenting cells.

[0049] Embodiment 43. A pharmaceutical composition comprising the humanized antibody or antigen-binding fragment thereof of any of embodiments 1 to 42 and a pharmaceutically acceptable carrier.

[0050] Embodiment 44. A method of treating an autoimmune or inflammatory disease or disorder comprising administering to a subject in need thereof an effective amount of the antibody or antigen binding fragment thereof of any of embodiments 1 to 42 or the pharmaceutical composition of embodiment 43.

[0051] Embodiment 45. The method of embodiment 44, wherein the subject has an inflammatory condition.

[0052] Embodiment 46. The method of embodiment 44 or 45, wherein the subject has Type 1 diabetes.

[0053] Embodiment 47. The method of embodiment 44 or 45, wherein the subject has systemic lupus erythematosus (SLE).

[0054] Embodiment 48. The method of embodiment 44 or 45, wherein the subject has autoimmune encephalomyelitis.

[0055] Embodiment 49. The method of embodiment 44 or 45, wherein the subject has scleroderma.

[0056] Embodiment 50. The method of embodiment 44 or 45, wherein the subject has psoriasis.

[0057] Embodiment 51. The method of any one of embodiments 44 to 50, wherein the administering is parenteral.Attorney Docket No: 01434-0001-00PCT

[0058] Embodiment 52. The method of embodiment 51, wherein the administering is intravenous.

[0059] Embodiment 53. The method of embodiment 51, wherein the administering is subcutaneous.

[0060] Embodiment 54. The humanized antibody of any one of embodiments 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises glutamine at position 54 according to Kabat numbering.

[0061] Embodiment 55. The humanized antibody of any one of embodiments 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises serine at position 54 according to Kabat numbering.

[0062] Embodiment 56. The humanized antibody of any one of embodiments 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises alanine at position 55 according to Kabat numbering.

[0063] Embodiment 57. The humanized antibody of any one of embodiments 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises glutamine at position 54 and alanine at position 55 according to Kabat numbering.

[0064] Embodiment 58. The humanized antibody of any one of embodiments 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises serine at position 54 and alanine at position 55 according to Kabat numbering.BRIEF DESCRIPTION OF THE DRAWINGS

[0001] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate several embodiments and together with the description, serve to explain the principles described herein.

[0002] Figure 1 shows the binding of humanized hzG28-8.1 through hzG28-8.4 antibodies to hCD180 / hMDl expressing 293T cells. The detection antibody is anti-hlgGFc-PE.

[0003] Figure 2 shows B cell activation in response to antibodies hzG28-8.1 through hzG28-8.4. Total human PBMCs were stimulated for 24 hours with indicated antibody. CD86 upregulation was determined using flow cytometry' by gating on live CD3-CD19+ cells.

[0004] Figure 3 shows B cell activation in response to antibodies hzG28-8.60 through hzG28-8.62. Total human PBMCs were stimulated for 30 minutes with indicated antibody. Protein AKT and S6 phosphorylation were determined using flow cytometry by gating on live CD3-CD19+ cells.

[0005] Figure 4 shows B cell activation in response to antibodies hzG28-8.69 and hzG28-8.71. Total human PBMCs were stimulated for 30 minutes with indicated antibody.Attomey Docket No: 01434-0001-00PCT Protein AKT and S6 phosphorylation was determined using flow cytometry by gating on live CD3-CD19+ cells.

[0006] Figure 5 shows B cell activation in response to antibodies hzG28-8.71, -8.98, -8.113 and -8.114compared to the chimeric G28-8 IgGl.1 control, as measured by protein AKT and S6 phosphorylation.

[0007] Figure 6 shows hzMHR73-l 1-1 and hzMHR73-l 1-11 binding curves as compared to the chimeric MHR73-11-1 control.

[0008] Figure 7 shows B cell activation in response to hzMHR73-l 1-1 and hzMHR73-11-11 compared to the chimeric hzMHR73-l 1 control.DETAILED DESCRIPTION

[0009] Applicants have surprisingly discovered novel humanized anti-CD180 antibodies and antigen-binding fragments thereof that specifically bind to human CD 180 and stimulate signaling by CD 180. In some embodiments, the humanized anti-CD180 antibodies described herein advantageously exhibit one or more of the following in antigen presenting cells (such as B cells): 1) equivalent or enhanced human CD180 antigen binding relative to a reference antibody (e.g., the murine parental antibody); 2) equivalent or enhanced activation of antigen presenting cells upon binding of the humanized CD 180 antibody relative to a reference antibody (e.g., a murine parental antibody); 3) equivalent or enhanced upregulation of human CD86 upon binding of the humanized CD180 antibody relative to a reference antibody (e.g., a murine parental antibody); 4) equivalent or enhanced phosphorylation of human protein S6 upon binding of the humanized CD180 antibody relative to a reference antibody (e.g., a murine parental antibody); and / or 5) equivalent or enhanced phosphorylation of human protein AKT upon binding of the humanized CD180 antibody relative to a reference antibody (e.g., a murine parental antibody).

[0010] In some embodiments, the humanized anti-CD180 antibodies and antigen binding fragments thereof described herein are useful for treating CD 180-related disorders, such as autoimmune or inflammatory diseases or disorders, as more fully described herein.

[0011] So that the disclosure may be more readily understood, certain technical and scientific terms are specifically defined below. Unless specifically defined elsewhere in this document, all other technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art.

[0012] In accordance with the description, the present disclosure is based on the discovery of novel humanized anti-CD180 antibodies and antigen-binding fragments thereof (also referred to herein as humanized CD 180 antibodies or humanized anti-CD180 antibodies or antigen binding fragments thereof). In certain aspects of the disclosure, the humanized anti-Attorney Docket No: 01434-0001-00PCT CD 180 antibodies or antigen-binding fragments thereof are derived from murine antibodies G28-8 and / or MHR73-11.

[0013] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof that specifically binds to human CD 180. the antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy7chain variable region comprises CDRH1, CDRH2, and CDRH3, wherein each CDR comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence or 0, 1, or 2 amino acid mutations relative to the amino acid sequence set forth in:a. CDRH1: SEQ ID NO: 17, 72, 50 or 77; CDRH2: SEQ ID NO:18 or 51; and CDRH3: SEQ ID NO: 19 or 52; according to Kabat numbering; or b. CDRH1: SEQ ID NO:25, 55, 74 or 78; CDRH2: SEQ ID NO:26; and CDRH3:SEQ ID NO:27 or 56; according to IMGT numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3, wherein each CDR comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence or 0, 1, or 2 amino acid mutations relative to the amino acid sequence set forth in:c. CDRL1: SEQ ID NO:20 or 53; CDRL2: SEQ ID NO:21; and CDRL3: SEQ ID NO:22 or 54; according to Kabat numbering; ord. CDRL1: SEQ ID NO:28 or 57; CDRL2: SEQ ID NO:29; and CDRL3: SEQ ID NO:22 or 54; according to IMGT numbering;the heavy chain variable region comprises human IGVH1-03 / JH6 framework regions having an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences of SEQ ID NOs:38 and 39;the light chain variable region comprises human IGVH1-03 / JH6 framework regions having an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequences of SEQ ID NOs:40 and 41; position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering; andat least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering.

[0014] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof, wherein:Attorney Docket No: 01434-0001-00PCT the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:3, 5, 6, 7, 9, 10, 14, 15, 16, 66, 67, 82, 64 or 65;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:4, 8, 45, 46, 47, 81, 68, 69, 70, or 71;at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0015] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences set forth in one of the sets of amino acid sequences selected from:a. SEQ ID NOs:17, 18, 19, 20, 21, and 22, according to Kabat numbering; b. SEQ ID NOs:72, 18, 19, 20, 21 , and 22, according to Kabat numbering; c. SEQ ID NOs:25, 26, 27, 28, 29, and 22, according to IMGT numbering; and d. SEQ ID NOs:74, 26, 27, 28, 29, and 22, according to IMGT numbering; the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:3, 5, 6, 7, 9, 10, 14, 15, or 16;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:4, 8, 45, 46, 47 and 81;at least one of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0016] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein:wherein at least one of position 28 is serine, position 66 is lysine, position 67 is alanine, position 69 is leucine, and position 71 is valine in the heavy chain variable region, according to Kabat numbering; and / orposition 64 is valine, position 48 is optionally valine and position 70 is optionally glutamine in the light chain variable region, according to Kabat numbering; position 55 in CDRH2 is optionally alanine according to Kabat numbering.Attorney Docket No: 01434-0001-00PCT

[0017] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof, wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences set forth one of the sets of amino acid sequences selected from:a. SEQ ID NOs:50, 51, 52, 53, 21 and 54, according to Kabat numbering; b. SEQ ID NOs:77, 51, 52, 53, 21 and 54, according to Kabat numbering; c. SEQ ID NOs:55, 26, 56, 57, 29, and 54, according to IMGT numbering; and d. SEQ ID NOs:78, 26, 56, 57, 29, and 54, according to IMGT numbering; wherein the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:66, 67, 82, 64 and 65;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:68, 69, 70 and 71; andat least one of, at least two of or all of position 48 is isoleucine, position 49 is phenylalanine or asparagine, position 64 is glycine or valine and position 70 is aspartate in the light chain variable region, according to Kabat numbering: and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0018] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein position 48 is isoleucine, position 49 is phenylalanine or asparagine, position 64 is glycine or valine and position 70 is aspartate in the light chain variable region, according to Kabat numbering.

[0019] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein position 64 is valine, according to Kabat numbering.

[0020] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:17, 18, and 19, respectively, according to Kabat numbering, and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:20, 21 and 22, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0021] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:72, 18, and 19, respectively, according to Kabat numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth inAttorney Docket No: 01434-0001-00PCT SEQ ID NOs:20, 21 and 22, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0022] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:25, 26, and 27, respectively, according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:28, 29, and 22, respectively according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0023] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:74, 26, and 27, respectively according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:28. 29, and 22. respectively according to IMGT numbering: wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0024] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein HCDR1 , HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:50, 51, and 52, respectively, according to Kabat numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:53, 21 and 54, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0025] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:77, 51, and 52, respectively, according to Kabat numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:53, 21 and 54, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0026] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:55, 26, and 56, respectively, according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:57, 29, and 54, respectively, according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0027] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the HCDR1 , HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:78, 26, and 56, respectively, according to IMGTAttorney Docket No: 01434-0001-00PCT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:57, 29, and 54, respectively, according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0028] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein:a) the heavy chain variable region comprises glutamine at position 54;b) the heavy chain variable region comprises serine at position 54; orc) the heavy chain variable region comprises alanine at position 55;according to Kabat numbering.

[0029] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain variable region comprises alanine at position 55; according to Kabat numbering.

[0030] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein:a) the heavy chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:3. 5, 6, 7, 9, 10, 14. 15 or 16; andb) the light chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in the amino acid sequences set forth in SEQ ID NO:4, 8, 45, 46, 47 or 81.

[0031] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain variable region and the light chain variable region comprise amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences set forth in:a. SEQ ID NO:6 and SEQ ID NO:8, respectively;b. SEQ ID NO: 16 and SEQ ID NO:8, respectively;c. SEQ ID NO:6 and SEQ ID NO:81, respectively; ord. SEQ ID NO: 16 and SEQ ID NO:81, respectively.

[0032] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in:a. SEQ ID NO:6 and SEQ ID NO:8, respectively;b. SEQ ID NO: 16 and SEQ ID NO:8, respectively;c. SEQ ID NO:6 and SEQ ID NO:81. respectively; orAttorney Docket No: 01434-0001-00PCT d. SEQ ID NO: 16 and SEQ ID NO:81, respectively.

[0033] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein:a) the heavy chain variable region comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:64. 65. 66, 67, or 82; andb) the light chain variable region comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in the ammo acid sequences set forth in SEQ ID NO:68, 69, 70 or 71.

[0034] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain variable region and the light chain variable region comprise amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences set forth in:a) SEQ ID NO:66 and SEQ ID NO:68, respectively;b) SEQ ID NO:67 and SEQ ID NO:71, respectively; orb) SEQ ID NO:64 and SEQ ID NO:68, respectively.

[0035] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain variable region and the light chain variable region comprise amino acid sequences set forth in:a) SEQ ID NO:66 and SEQ ID NO:68, respectivelyb) SEQ ID NO:67 and SEQ ID NO:71, respectively; orc) SEQ ID NO:64 and SEQ ID NO:68, respectively.

[0036] In any of the embodiments, provided is a humanized antibody or antigen-binding fragment thereof, further comprising a heavy chain constant region attached to the heavy chain variable region and a light chain constant region attached to the light chain variable region.

[0037] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain constant region is an IgG, IgA, or IgM region and the light chain constant region is a kappa region.

[0038] In some embodiments, provided is a humanized antibody or antigen binding fragment thereof as described herein, wherein the heavy chain constant region is an IgG region.

[0039] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain constant region is an IgGl region.

[0040] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:42.Attorney Docket No: 01434-0001-00PCT

[0041] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain constant region is an IgG4 region, optionally wherein the IgG4 region comprises an S228P substitution.

[0042] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof has reduced antibody dependent cellular cytotoxicity (ADCC).

[0043] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof has reduced antibody dependent cellular phagocytosis activity (ADCP).

[0044] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof has reduced complement-dependent cytotoxicity activity' (CDC).

[0045] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, comprising a IgGl heavy chain constant region having amino acid substitutions selected from (a) L234A L235A (LALA); (b) L234A, L235A, and P329G (LALA-PG); (c) L234A, L235E, G237A, A330S and P331S; or (d) L234A, L235A and G237A; or comprising an IgG4 heavy chain constant region having amino acid substitutions (e) S288P.

[0046] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain constant region is an Fc null.

[0047] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:43.

[0048] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof is a Fab, a Fab', a F(ab')2, a Fv fragment, a diabody. a single-chain antibody, an scFv fragment or an scFv-Fc.

[0049] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof is monospecific, bispecific, monovalent or bivalent.

[0050] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof induces activation of antigen presenting cells.Attorney Docket No: 01434-0001-00PCT

[0051] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein activation of antigen presenting cells is CD86 upregulation.

[0052] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof, induces increased phosphorylation of protein AKT in antigen presenting cells.

[0053] In some embodiments, provided is a humanized antibody or antigen-binding fragment thereof as described herein, wherein the antibody or antigen-binding fragment thereof, induces increased phosphorylation of protein S6 in antigen presenting cells.

[0054] In some embodiments, provided is pharmaceutical composition comprising the humanized antibody or antigen-binding fragment thereof as described herein and a pharmaceutically acceptable carrier.

[0055] In some embodiments, provided is a method of treating an autoimmune or inflammatory disease or disorder comprising administering to a subject in need thereof an effective amount of the antibody or antigen binding fragment thereof as described herein or the pharmaceutical composition thereof. In some embodiments, subject has an inflammatory condition. In some embodiments, the subject has Type 1 diabetes. In some embodiments, the subject has systemic lupus erythematosus (SLE). In some embodiments, the subject has autoimmune encephalomyelitis. In some embodiments, the subject has scleroderma. In some embodiments, the subject has psoriasis. In some embodiments, the administering is parenteral. In some embodiments, the administering is intravenous. In some embodiments, the administering is subcutaneous.

[0056] Additional objects and advantages will be set forth in part in the description which follows, and in part will be understood from the description, or may be learned by practice. The objects and advantages will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims.

[0057] It is to be understood that both the foregoing general description and the following detailed description are exemplary' and explanatory' only and are not restrictive of the claims.I. Definitions

[0058] As used herein, including the appended claims, the singular forms of words such as “a,” “an,” and “the,” include their corresponding plural references unless the context clearly dictates otherwise.Attorney Docket No: 01434-0001-00PCT

[0059] Compositions or methods “comprising” one or more recited elements may include other elements not specifically recited. For example, a composition that comprises antibody may contain the antibody alone or in combination with other ingredients.

[0060] A “polypeptide” or “polypeptide chain” refers to a polymer of amino acid residues joined by peptide bonds, whether produced naturally or synthetically. Polypeptides of less than about 10 amino acid residues are commonly referred to as “peptides.”

[0061] A “protein” is a macromolecule comprising one or more polypeptide chains. A protein may also comprise non-peptidic components, such as carbohydrate groups or other modifications. Carbohydrates and other non-peptidic substituents may be added to a protein by the cell in which the protein is produced, and will vary with the type of cell. Proteins are defined herein in terms of their amino acid backbone structures. Substituents such as carbohydrate groups are generally not specified, but may be present nonetheless.

[0062] The terms “amino-terminal” and “carboxy -terminal” denote positions within polypeptides. Where the context allows, these terms are used with reference to a particular sequence or portion of a polypeptide to denote proximity or relative position. For example, a certain sequence positioned carboxy -terminal to a reference sequence within a polypeptide is located proximal to the carboxy terminus of the reference sequence, but is not necessarily at the carboxy terminus of the complete polypeptide.

[0063] As used herein, amino acids are generally the naturally occurring L amino acid. Classes of amino acids are as follows: hydrophobic side chains: methionine (met), alanine (ala), valine (val), leucine (leu), and isoleucine (ile); neutral hydrophilic side chains: cysteine (cys), serine (ser), and threonine (thr); acidic side chains: aspartate (asp), and glutamate (glu); basic side chains: asparagine (asn), glutamine (gin), histidine (his), lysine (lys), and arginine (arg); residues influencing chain orientation: glycine (gly), and proline (pro); and aromatic side chains: tryptophan (trp), tyrosine (tyr), and phenylalanine (phe).

[0064] For purposes of classifying amino acids substitutions as conservative or nonconservative, the following amino acid substitutions are considered conservative substitutions: serine substituted by threonine, alanine, or asparagine; threonine substituted by proline or serine; asparagine substituted by aspartic acid, histidine, or serine; aspartic acid substituted by glutamic acid or asparagine; glutamic acid substituted by glutamine, lysine, or aspartic acid; glutamine substituted by arginine, lysine, or glutamic acid; histidine substituted by tyrosine or asparagine; arginine substituted by lysine or glutamine; methionine substituted by isoleucine, leucine or valine; isoleucine substituted by leucine, valine, or methionine; leucine substituted by valine, isoleucine, or methionine; phenylalanine substituted by tyrosine or tryptophan; tyrosine substituted by tryptophan, histidine, or phenylalanine; proline substituted byAttorney Docket No: 01434-0001-00PCT threonine; alanine substituted by serine; lysine substituted by glutamic acid, glutamine, or arginine; valine substituted by methionine, isoleucine, or leucine; and tryptophan substituted by phenylalanine or tyrosine. Conservative substitutions can also mean substitutions between amino acids in the same class.

[0065] Two amino acid sequences have “100% amino acid sequence identity” (or are identical) if the amino acid residues of the two amino acid sequences are the same when aligned for maximal correspondence. Sequence comparisons are typically performed using standard software programs such as the NCBI BLAST Global Align program. Other methods for comparing two nucleotide or amino acid sequences by determining optimal alignment are well-known to those of skill in the art. (See, e.g., Peruski and Peruski, The Internet and the New Biology': Tools for Genomic and Molecular Research (ASM Press, Inc. 1997); Wu et al.(eds.), “Information Superhighway and Computer Databases of Nucleic Acids and Proteins,” in Methods in Gene Biotechnology 123-151 (CRC Press, Inc. 1997); Bishop (ed.), Guide to Human Genome Computing (2nd ed.. Academic Press, Inc. 1998).) Two amino acid sequences are considered to have “substantial sequence identity” if the two sequences have at least about 80%, 85%, 90%, or 95% identity relative to each other.

[0066] Percentage sequence identities can be determined for antibody sequences maximally aligned by the Kabat numbering convention. After alignment, if a subject protein or polypeptide region (e.g., an entire variable domain of a heavy or light chain of an antibody) is being compared with the same region of a reference protein or polypeptide (e.g., a second antibody), the percentage sequence identity between the subj ect and reference regions is the number of positions occupied by the same amino acid in both the subject and reference regions divided by the total number of aligned positions of the two regions, with gaps not counted, multiplied by 100 to convert to percentage.

[0067] In the antibodies or other proteins described herein, reference to amino acid residues corresponding to those specified by SEQ ID NO: includes post-translational modifications of such residues.

[0068] The term “antibody” denotes immunoglobulin proteins produced by the body in response to the presence of an antigen and that specifically bind to the antigen, as well as antigen-binding fragments and engineered variants thereof. Hence, the term “antibody” includes, for example, intact monoclonal antibodies (e.g., antibodies produced using hybndoma technology) and antigen-binding antibody fragments, such as a F(ab')2, a Fv fragment, a diabody, a single-chain antibody, an scFv fragment, or an scFv-Fc. Genetically, engineered intact antibodies and fragments such as humanized antibodies, single-chain Fv fragments, single-chain antibodies, diabodies, minibodies, linear antibodies, multivalent or multi-specific (e.g..Attorney Docket No: 01434-0001-00PCT bispecific) hybrid antibodies, and the like, are also included. Thus, the term “antibody” is used expansively to include any protein that comprises an antigen-binding site of an antibody and is capable of specifically binding to its antigen.

[0069] An antibody can be monovalent, bivalent or multivalent. An antibody can be monospecific, bispecific or multi-specific.

[0070] The term antibody or anti gen -binding fragment thereof includes a “naked” or “unconjugated” antibody or antigen-binding fragment thereof that is not bound (i.e., covalently or non-covalently bound) to another compound (other than the target antigen of the antibody).

[0071] The term “genetically engineered antibodies” refers to an antibody in which the amino acid sequence has been varied from that of the native or parental antibody. The possible variations are many, and range from the changing of just one or a few amino acids to the complete redesign of, for example, the variable or constant region. Changes in the constant region are, in general, made to improve or alter characteristics such as, e.g.. complement binding and other effector functions. Typically, changes in the variable region are made to improve antigen-binding characteristics, improve variable region stabi I i ty . and / or reduce the risk of immunogenicity .

[0072] An “antigen-binding fragment” or “antigen binding portion” of an antibody is that portion of an antibody that is sufficient to specifically bind to its target antigen. The minimum such region is typically a variable domain or a genetically engineered variant thereof. Single domain binding molecules can be generated from camelid antibodies (see Muyldermans and Lauwereys. Mol. Recog. 12: 131-140, 1999; Nguyen et al, EMBO J. 19:921-930, 2000) or from VH domains of other species to produce single-domain antibodies (“dAbs,” see Ward et al, Nature 341 :544-546, 1989; US Patent No. 6,248,516 to Winter et al). Commonly, an antigenbinding site of an antibody comprises both a heavy chain variable (VH) domain and a light chain variable (VL) domain that bind to a common epitope. Within the context of the present disclosure, an antibody may include one or more components in addition to an antigen-binding portion and may be a monomeric or multimeric protein. Examples of molecules comprising an antigen binding site of an antibody are know n in the art and include, for example, Fv, singlechain Fv (scFv), Fab, Fab', F(ab')2, F(ab)c, diabodies, minibodies, nanobodies, Fab-scFv fusions, bispecific (scFvjHgG, and bispecific (scFvty-Fab. (See. e.g., Hu et al, Cancer Res. 56:3055-3061, 1996; Atwell et al. Molecular Immunology 33: 1301-1312, 1996; Carter and Merchant, Curr. Op. Biotechnol. 8:449-454, 1997; Zuo et al, Protein Engineering 13:361-367, 2000; and Lu et al, J. Immunol. Methods 267:213-226, 2002.)

[0073] The term “immunoglobulin” refers to a protein consisting of one or more polypeptides substantially encoded by immunoglobulin gene(s). One form of immunoglobulinAttorney Docket No: 01434-0001-00PCT constitutes the basic structural unit of native (i.e., natural or parental) antibodies in vertebrates. This form is a tetramer and consists of two pairs of immunoglobulin chains, each pair having one light chain and one heavy chain. In each pair, the light and heavy chain variable regions (VL and VH, respectively) are together primarily responsible for specific binding to an antigen, and the constant regions are primarily responsible for the antibody effector functions. Five classes of immunoglobulin protein (IgG, IgA, IgM, IgD, and IgE) have been identified in higher vertebrates. In humans, IgG immunoglobulins consist of four subclasses, designated IgGI, IgG2, IgG3. and IgG4. Each immunoglobulin heavy chain possesses a constant region that consists of constant region protein domains (CHI, hinge, CH2, and CH3; IgG3 also contains a CH4 domain) that are essentially invariant for a given subclass in a species.

[0074] DNA sequences encoding human and non-human immunoglobulin chains are known in the art. (See, e.g., Ellison et al. DNA 1: 11-18, 1981; Ellison et al, Nucleic Acids Res.10:4071-4079, 1982; Kenten et al.. Proc. Natl. Acad. Sci. USA 79:6661-6665, 1982; Seno et al., Nucl. Acids Res. 11:719-726, 1983; Riechmann et al., Nature 332:323-327, 1988; Amster et al, Nucl. Acids Res. 8:2055-2065, 1980; Rusconi and Kohler, Nature 314:330-334, 1985; Boss et al, Nucl. Acids Res. 12:3791-3806, 1984; Bothwell et al, Nature 298:380-382, 1982; van der Loo et al., Immunogenetics 42:333-341, 1995; Karlin et al, J. Mol. Evol. 22: 195-208, 1985; Kindsvogel et al., DNA 1:335-343, 1982; Breiner et al, Gene 18: 165-174, 1982; Kondo et al, Eur. J.Immunol. 23:245-249, 1993; and GenBank Accession No. J00228.) For a review of immunoglobulin structure and function, see Putnam, The Plasma Proteins, Vol V, Academic Press, Inc., 49-140. 1987; and Padlan, Mol. Immunol. 31: 169-217, 1994. The term 'Immunoglobulin” is used herein for its common meaning, denoting an intact antibody, its component chains, or fragments of chains, depending on the context.

[0075] Full-length immunoglobulin “light chains” (about 25 kDa or 214 amino acids) are encoded by a variable region gene at the amino-terminus (encoding about 110 amino acids) and by a kappa or lambda constant region gene at the carboxy 1-terminus. Full-length immunoglobulin '‘heavy chains” (about 50 kDa or 446 amino acids) are encoded by a variable region gene (encoding about 116 amino acids) and a gamma, mu, alpha, delta, or epsilon constant region gene (encoding about 330 amino acids), the latter defining the antibody’s isotype as IgG, IgM, IgA, IgD or IgE, respectively. Within light and heavy chains, the variable and constant regions are joined by a “J” region of about 12 or more amino acids, with the heavy chain also including a “D” region of about 10 more amino acids. (See generally Fundamental Immunology (Paul, ed., Raven Press, N.Y., 2nd ed. 1989), Ch. 7.)

[0076] An immunoglobulin light or heavy chain variable region (also referred to herein as a “VL region” or a "VH region”, respectively) consists of a “framework” region interruptedAttorney Docket No: 01434-0001-00PCT by three “complementarity determining regions’’ or “CDRs.” The framework regions serve to align the CDRs for specific binding to an epitope of an antigen. Thus, the term “CDR” refers to the amino acid residues of an antibody that are primarily responsible for antigen binding. From amino-terminus to carboxyl-terminus, both VL and VH domains comprise the following framework (FR) and CDR regions: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0077] The assignment of amino acids to each variable region domain is typically in accordance with the definitions of Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987 and 1991). Kabat also provides a widely used numbering convention (Kabat numbering) in which corresponding residues between different heavy chain variable regions or betw een different light chain variable regions are assigned the same number. CDRs 1 , 2 and 3 of a VL domain are also referred to herein, respectively, as CDRL1 or LI, CDRL2 or L2 and CDRL3 or L3. CDRs 1, 2 and 3 of a VH domain are also referred to herein, respectively, as CDRH1 or Hl, CDRH2 or H2 and CDRH3 or H3.

[0078] The assignment of CDRs also can be in accordance with other numbering systems, such as IMGT® numbering (Lefranc et al., Developmental & Comparative Immunology 2003, 27:55-77), Chothia (Chothia and Lesk, Canonical Structures for the Hypervariable Regions of Immunoglobulins, J. Mol. Biol. 1987, 196:901-917); AbM numbering (Abhinandan and Martin, Analysis and improvements to Kabat and structurally correct numbering of antibody variable domains, Mol. Immunol. 2008, 45(14):3832-3839) or Aho numbering (Honegger and Pluckthun, Yet Another Numbering Scheme for Immunoglobulin Variable Domains: An Automatic Modeling and Analysis Tool, J. Mol. Biol. 2001. 309:657-670) in lieu of Kabat. Unless otherwise indicated by content, the CDRs herein are designed according to Kabat.

[0079] Numbering of the heavy chain constant region is via the EU index as set forth in Kabat (Kabat, Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, MD, 1987 and 1991).

[0080] Unless the context dictates otherwise, the term “monoclonal antibody” is not limited to antibodies produced through hybridoma technology7. The term “monoclonal antibody” can include an antibody that is derived from a single clone, including any eukaryotic, prokary otic or phage clone. In particular embodiments, the antibodies described herein are monoclonal antibodies.

[0081] The terms “humanized VH domain” or “humanized VL domain” refer to an immunoglobulin VH or VL domain comprising some or all CDRs entirely or substantially from a non-human donor immunoglobulin (e.g., a mouse or rat) and variable domain framework sequences entirely or substantially from human immunoglobulin sequences. The non-humanAttorney Docket No: 01434-0001-00PCT immunoglobulin providing the CDRs is called the “donor” and the human immunoglobulin providing the framework is called the “acceptor.” In some instances, humanized antibodies will retain some non-human residues within the human variable domain framework regions to enhance proper binding characteristics (e.g.. substitutions in the frameworks may be required to preserve binding affinity when an antibody is humanized). Such residues may be referred to as “back mutations.”

[0082] A “humanized antibody” is an antibody comprising one or both of a humanized VH domain and a humanized VL domain. Immunoglobulin constant region(s) need not be present, but if they are, they are entirely or substantially from human immunoglobulin constant regions.

[0083] A humanized antibody is a genetically engineered antibody in which the CDRs from a non-human “donor” antibody are grafted into human “acceptor” antibody sequences (see, e.g., Queen, US 5,530,101 and 5,585,089; Winter. US 5,225,539; Carter, US 6.407,213; Adair. US 5,859,205; and Foote, US 6,881,557). The acceptor antibody sequences can be, for example, a mature human antibody sequence, a composite of such sequences, a consensus sequence of human antibody sequences, or a germline region sequence. Human acceptor sequences can be selected for a high degree of sequence identity in the variable region frameworks with donor sequences to match canonical forms between acceptor and donor CDRs among other criteria. Thus, a humanized antibody is an antibody having CDRs entirely or substantially from a donor antibody and variable region framework sequences and constant regions, if present, entirely or substantially from human antibody sequences. Similarly, a humanized heavy chain typically has all three CDRs entirely or substantially from a donor antibody variable region heavy chain, and a heavy chain variable region framework sequence and heavy chain constant region, if present, substantially from human heavy chain variable region framework and constant region sequences. Similarly, a humanized light chain typically has all three CDRs entirely or substantially from a donor antibody variable region light chain, and a light chain variable region framework sequence and light chain constant region, if present, substantially from human light chain variable region framework and constant region sequences. A CDR in a humanized antibody is substantially from a corresponding CDR in a non-human antibody when at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of corresponding residues (as defined by Kabat numbering or other CDR numbering system), or wherein about 100% of corresponding residues (as defined by Kabat numbering or other CDR numbering system), are identical between the respective CDRs. The variable region framework sequences of an antibody chain or the constant region of an antibody chain are substantially from a human variable region framework sequence or human constant region respectively when atAttorney Docket No: 01434-0001-00PCT least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of corresponding residues (as defined by Kabat numbering for the variable region and EU numbering for the constant region or other CDR numbering system), or about 100% of corresponding residues (as defined by Kabat numbering for the variable region and EU numbering for the constant region or other CDR numbering system) are identical.

[0084] Although humanized antibodies often incorporate all six CDRs (e.g., as defined by Kabat or IMGT® numbering) from a mouse antibody, they can also be made with fewer than all six CDRs (e.g., at least 3, 4, or 5) CDRs from a mouse antibody (see, e.g., Pascabs et al, J. Immunol. 169:3076, 2002; Vajdos et al, Journal of Molecular Biology, 320: 415-428, 2002; Iwahashi et al, Mol. Immunol. 36: 1079-1091, 1999; Tamura et al, Journal of Immunology7, 164: 1432- 1441, 2000).

[0085] Antibodies are typically provided in isolated form. This means that an antibody is typically at least about 50% w / w pure of interfering proteins and other contaminants arising from its production or purification but does not exclude the possibility that the antibody is combined with pharmaceutical acceptable carrier(s) or other vehicle intended to facilitate its use.Sometimes antibodies are at least about 60%, 70%, 80%, 90%, 95% or 99% w / w pure of interfering proteins and contaminants from production or purification.

[0086] Specific binding of an antibody to its target antigen (or specifically binds) typically refers an affinity of at least about 10’6, 10'7, 10'8, 10'9, or 10’10M. Specific binding is delectably higher in magnitude and distinguishable from non-specific binding occurring to at least one non-specific target. Specific binding can be the result of formation of bonds between particular functional groups or particular spatial fit (e.g., lock and key type), whereas nonspecific binding is typically the result of van der Waals forces.

[0087] The term ‘‘epitope” refers to a site of an antigen to which an antibody or antigen binding fragment binds. An epitope can be formed from contiguous amino acids or noncontiguous amino acids juxtaposed by tertiary folding of one or more proteins. Epitopes formed from contiguous amino acids are typically retained upon exposure of the antigen to denaturing agents, e.g., solvents, whereas epitopes formed by tertiary folding are typically lost upon treatment of the antigen with denaturing agents, e.g., solvents. An epitope typically includes at least about 3, and more usually, at least about 5, 6, 7, or 8-10 amino acids in a unique spatial conformation. Methods of determining spatial conformation of epitopes include, for example, x-ray crystallography and two-dimensional nuclear magnetic resonance. See, e.g., Epitope Mapping Protocols, in Methods in Molecular Biology, Vol. 66, Glenn E. Morris. Ed. (1996).Attorney Docket No: 01434-0001-00PCT

[0088] Antibodies that recognize the same or overlapping epitopes can be identified in a simple immunoassay showing the ability of one antibody to compete with the binding of another antibody to a target antigen. The epitope of an antibody can also be defined by X-ray crystallography of the antibody bound to its antigen to identify contact residues.

[0089] Alternatively, two antibodies have the same epitope if all amino acid mutations in the antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other (provided that such mutations do not produce a global alteration in antigen structure). Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one antibody reduce or eliminate binding of the other antibody.

[0090] Competition between antibodies can be determined by an assay in which a test antibody inhibits specific binding of a reference antibody to a common antigen (see, e.g., Junghans et al, Cancer Res. 50: 1495, 1990). A test antibody competes with a reference antibody if an excess of a test antibody inhibits binding of the reference antibody.

[0091] Antibodies identified by competition assay (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to an adjacent epitope sufficiently proximal to the epitope bound by the reference antibody for steric hindrance to occur. Antibodies identified by a competition assay also include those that indirectly compete with a reference antibody by causing a conformational change in the target protein thereby preventing binding of the reference antibody to a different epitope than that bound by the test antibody.

[0092] An antibody effector function refers to a function contributed by an Fc region or constant region. Such functions can be, for example, antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), or complement-dependent cytotoxicity (CDC). Such function(s) can be effected by, for example, binding of an Fc region to an Fc receptor on an immune cell with phagocytic or lytic activity or by binding of an Fc region to components of the complement system. Fc regions of antibodies can recruit Fc receptor (FcR)-expressing cells and juxtapose them with antibody -coated target cells. Cells expressing surface FcR for IgGs, including FcyRIII (CD16), FcyRII (CD32) and FcyRIII (CD64), can act as effector cells for the destruction of IgG-coated cells. Such effector cells include monocytes, macrophages, natural killer (NK) cells, neutrophils and eosinophils. Engagement of FcyR by IgG activates ADCC or ADCP. ADCC is mediated by CD16+ effector cells through the secretion of membrane pore-forming proteins and proteases, while phagocytosis is mediated by' CD32+ and CD64+ effector cells (see Fundamental Immunology7, 4th ed., Paul ed., Lippincott-Raven, N.Y., 1997, Chapters 3, 17 and 30; Uchida et al, J. Exp. Med. 199: 1659-69, 2004; Akewanlop et al, Cancer Res. 61:4061-65. 2001; Watanabe et al, Breast Cancer Res. Treat. 53: 199-207. 1999).Attorney Docket No: 01434-0001-00PCT

[0093] In addition to ADCC and ADCP, Fc regions of cell-bound antibodies can also activate the complement classical pathway to elicit CDC. Clq of the complement system binds to the Fc regions of antibodies when they are complexed with antigens. Binding of Clq to cellbound antibodies can initiate a cascade of events involving the proteolytic activation of C4 and C2 to generate the C3 convertase. Cleavage of C3 to C3b by C3 convertase enables the activation of terminal complement components including C5b, C6, C7, C8 and C9. Collectively, these proteins form membrane-attack complex pores on the antibody-coated cells. These pores disrupt the cell membrane integrity, killing the target cell (see Immunobiology, 6th ed.. Janeway et al, Garland Science, N. Y., 2005, Chapter 2).

[0094] The term “antibody -dependent cellular cytotoxicity” or “ADCC” refers to a mechanism for inducing cell death that depends on the interaction of antibody-coated target cells with immune cells possessing lytic activity (also referred to as effector cells). Such effector cells include natural killer cells, monocytes / macrophages and neutrophils. The effector cells attach to an Fc region of immunoglobulin bound to target cells via their antigen-combining sites. Death of the antibody-coated target cell occurs as a result of effector cell activity'. In certain exemplary embodiments, an anti-CD180 antibody mediates decreased or no ADCC relative to a parental antibody. As used herein, no ADCC means less than 1% of the activity of the parental antibody having a human IgGl Fc region.

[0095] The term “antibody -dependent cellular phagocytosis” or “ADCP” refers to the process by which antibody-coated cells are internalized, either in whole or in part, by phagocytic immune cells (e.g., by macrophages, neutrophils and / or dendritic cells) that bind to an Fc region of 1g. In certain exemplary embodiments, an anti-CD180 antibody mediates decreased or no ADCP relative to a parental antibody. As used herein, no ADCC means less than 1% of the activity' of the parental antibody having a human IgGl Fc region.

[0096] The term “complement-dependent cytotoxicity” or “CDC” refers to a mechanism for inducing cell death in which an Fc region of a target-bound antibody activates a series of enzymatic reactions culminating in the formation of holes in the target cell membrane. Typically, antigen-antibody complexes such as those on antibody-coated target cells bind and activate complement component Clq, which in turn activates the complement cascade leading to target cell death. Activation of complement may also result in deposition of complement components on the target cell surface that facilitate ADCC by binding complement receptors (e.g., CR3) on leukocytes. In certain exemplary embodiments, an anti-CD180 antibody mediates decreased or no CDC relative to a parental antibody. As used herein, no ADCC means less than 1% of the activity of the parental antibody having a human IgGl Fc region.Attorney Docket No: 01434-0001-00PCT

[0097] The term “effector function null” refers to an antibody or antigen binding fragment thereof that has no ADCP, no ADCC and no CDC. In some embodiments, an effector function null antibody or antigen-binding fragment thereof refers to an antibody or antigen binding fragment thereof that has less than 1% binding to FcR (Fc receptors) and Clq, as compared to a parental control antibody having a human IgGl Fc region.

[0098] The terms “expression unit” and “expression cassette” are used interchangeably herein and denote a nucleic acid segment encoding a polypeptide of interest and capable of providing expression of the nucleic acid segment in a host cell. An expression unit typically comprises a transcription promoter, an open reading frame encoding the polypeptide of interest, and a transcription terminator, all operably linked. In addition to a transcriptional promoter and terminator, an expression unit may further include other nucleic acid segments such as, e.g., an enhancer or a polyadenylation signal.

[0099] The term “expression vector” refers to a nucleic acid molecule, linear or circular, comprising one or more expression units. In addition to one or more expression units, an expression vector may also include additional nucleic acid segments such as, for example, one or more origins of replication or one or more selectable markers. Expression vectors are generally derived from plasmid or viral DNA, or may contain elements of both.

[0100] The term “patient” or “subject” includes human and other mammalian subjects such as non-human primates, rabbits, rats, mice, and the like, and transgenic species thereof, that receive either prophylactic or therapeutic treatment. In certain exemplary embodiments, a subject is a human patient.

[0101] The term “effective amount,” in the context of treatment of a CD180-expressing disorder by administration of an anti-CD180 antibody or antigen-binding fragment thereof as described herein, refers to an amount of such antibody or fragment that is sufficient to increase upregulation of CD86, increase phosphorylation of protein S6, and / or increase phosphorylation of protein AKT in antigen presenting cells (such as B cells). In some embodiments, an effective amount of a CD 180 antibody inhibits the occurrence of or ameliorates one or more symptoms of a CD180-related disorder, such as an autoimmune or inflammatory disease or disorder. An effective amount of an antibody is administered in an “effective regimen.” The term “effective regimen” refers to a combination of amount of the antibody being administered and dosage frequency adequate to accomplish prophylactic or therapeutic treatment of the disorder.

[0102] The term “pharmaceutically acceptable” means approved or approvable by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.Attorney Docket No: 01434-0001-00PCT The term “pharmaceutically compatible ingredient” refers to a pharmaceutically acceptable diluent, adjuvant, excipient, or vehicle with which an anti-CD180 antibody is formulated.

[0103] The phrase “pharmaceutically acceptable salt,” refers to pharmaceutically acceptable organic or inorganic salts. Exemplary’ salts include sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, acid phosphate, isonicotinate, lactate, salicylate, acid citrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate. and pamoate (i.e., 1,1'-methylene bis-(2 hydroxy -3 -naphthoate) salts. A pharmaceutically acceptable salt may further comprise an additional molecule such as, e.g., an acetate ion, a succinate ion or other counterion. A counterion may be any organic or inorganic moiety7that stabilizes the charge on the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. Instances where multiple charged atoms are part of the pharmaceutically acceptable salt can have multiple counter ions. Hence, a pharmaceutically acceptable salt can have one or more charged atoms and / or one or more counterion.

[0104] Designation of a range of values includes all integers within or defining the range.

[0105] Unless otherwise apparent from the context, when a value is expressed as “about” X or “approximately” X, the stated value of X will be understood to be accurate to ±10%.IL Anti-CD180 Antibodies and Antigen-Binding Fragments

[0106] The present disclosure provides isolated, recombinant and / or genetically engineered, humanized and / or CDR-grafted CD180 antibodies (collectively referred to herein as humanized anti-CD180 antibodies or humanized CD 180 antibodies) and antigen binding fragments thereof, as well as compositions thereof and nucleic acid molecules comprising at least one polynucleotide encoding at least a portion of a humanized CD180 antibody or antigen binding fragment thereof. The present disclosure further provides methods of making and using such antibodies, antigen binding fragments and nucleic acids for diagnostic and therapeutic compositions, methods and devices.

[0107] In some embodiments, humanized anti-CD180 antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 effector function null antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 antibodies and antigen binding fragments thereof having reduced effector function are provided. In some embodiments, humanized anti-CD180 IgGAttorney Docket No: 01434-0001-00PCT antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 effector function null IgG antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 IgG antibodies and antigen binding fragments thereof having reduced effector function are provided. In some embodiments, humanized anti-CD180 IgGl antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 effector function null IgGl antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 IgGl antibodies and antigen binding fragments thereof are provided that have reduced effector function. In some embodiments, humanized anti-CD180 IgG3 antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 effector function null IgG3 antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD180 IgG3 antibodies and antigen binding fragments thereof having reduced effector function are provided. In some embodiments, humanized anti-CD180 IgG4 antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized antiCD 180 effector function null IgG4 antibodies and antigen binding fragments thereof are provided. In some embodiments, humanized anti-CD 180 IgG4 antibodies and antigen binding fragments thereof having reduced effector function are provided.

[0108] As used herein, an effector function null humanized CD180 antibody or antigen-binding fragment thereof refers to an antibody or antigen binding fragment thereof that has less than 1% of binding to FcR (Fc receptors) and Clq, as compared to a control antibody having the same variable regions and a human IgGl Fc region.

[0109] In particular embodiments, humanized anti-CD 180 antibodies are provided that are derived from murine antibody G28-8 and / or antibody MHR73-11. In some embodiments, the humanized CD180 antibodies are derived from antibody G28-8. In some embodiments, a humanized CD 180 antibody comprises at least one CDR of antibody G28-8 and framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody is derived from murine antibody MHR73-11. In some embodiments, a humanized CD180 antibody comprises at least one CDR of antibody MHR73-11 and framework regions derived from human antibody sequences. In some embodiments, a humanized CD180 antibody is derived from murine antibody G28-8 and MHR73-11. In some embodiments, a humanized CD 180 antibody comprises at least one CDR of antibody G28-8 and one CDR of antibody MHR73-11 and framework regions derived from human antibody sequences. In any of these embodiments, human antibody sequences may be human antibody germline sequences.Attorney Docket No: 01434-0001-00PCT

[0110] In some embodiments, the humanized anti-CD180 antibodies or antigen binding fragments thereof have one or more of the following activities on antigen presenting cells (such as B cells): 1) upregulation of CD86 on antigen presenting cells (such as B cells) via CD180 binding at least 75% of, 80% of, 90% of, 95% of, or greater than the upregulation of CD86 by a reference antibody (e.g., the parental antibody) following binding of a humanized anti-CD180 antibody or antigen binding fragment to CD180 on the cell; 2) phosphorylation of human protein AKT induced by binding of a humanized anti-CD180 antibody or antigen binding fragment to CD180 on the cell of at least 75% of, 80% of, 90% of. 95% of, or greater than that of a reference antibody (e.g., the parental antibody); 3) phosphorylation of human protein S6 induced by binding of a humanized anti-CD180 antibody or antigen binding fragment to CD 180 on the cell of at least 75% of, 80% of, 90% of, 95% of, or greater than that of a reference antibody (e g., the parental antibody); 4) decreased or no antibody dependent cellular cytotoxicity (ADCC) relative to a reference antibody (e.g., the parental antibody having a human IgGl Fc region); 5) decreased or no antibody dependent cellular phagocytosis (ADCP) relative to a reference antibody (e.g., the parental antibody having a human IgGl Fc region); 6) decreased CDC activity relative to a reference antibody (e.g., the parental antibody having a human IgGl Fc region); and / or 7) specific binding to the same CD 180 epitope as a reference antibody (e.g., the parental antibody).

[0111] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to Kabat numbering, from antibody G28-8 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen- binding fragment thereof comprises a set of CDR sequences, according to IMGT numbering, from antibody G28-8 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to Kabat or IMGT numbering, from antibody G28-8 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to Chothia numbering, from antibody G28-8 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to AbM numbering, from antibody G28-8 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to AHo numbering, from antibody G28-8 and human framework regions derived from human antibody sequences. In any of theseAttorney Docket No: 01434-0001-00PCT embodiments, the human antibody sequences are derived from human antibody germline sequences.

[0112] In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to Kabat numbering, from antibody MHR73-11 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to IMGT numbering, from antibody MHR73-11 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to Kabat or IMGT numbering, from antibody MHR73-11 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to Chothia numbering, from antibody MHR73-11 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to AbM numbering, from antibody MHR73-11 and human framework regions derived from human antibody sequences. In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, according to AHo numbering, from antibody MHR73-11 and human framework regions derived from human antibody sequences. In some embodiments, the human antibody sequences are derived from human antibody germline sequences.

[0113] In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences as set forth in Table 1 or Table 2 and framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences as set forth in Table 1 or Table 2 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally serine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigenbinding fragment thereof comprises a set of CDR sequences, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a CDR set forth in Table 1 or Table 2, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1 or Table 2, and framework regions derived from human antibody sequences. In some embodiments, a humanized CD 180 antibody or antigen-Attorney Docket No: 01434-0001-00PCT binding fragment thereof comprises a set of CDR sequences, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a CDR set forth in Table 1 or Table 2. or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1 or Table 2, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, an amino acid substitution in a CDR is a conservative amino acid substitution. In some embodiments, the human antibody framework sequences are derived from human antibody germline sequences.Table 1. Heavy chain and light chain variable region CDR sequences of G28-8 antibodies Kabat IMGT Chothia AbM AHoCDRH1 GYSMN ( SEQ ID GYSFTGYT GYSFTGY GYSFTGYTM ASGYSFTGYTMN (SEQ ID (SEQ ID N ( SEQ ID (SEQ ID NO: 30 ) NO : 72 )NO : 25 ) or NO : 23 ) or NO: 36) or or ASGYTFTGYTMN GYTMN (SEQ IDGYTFTGYT GYTFGY GYTFTGYTM (SEQ ID NO: 76) NO: 17 ) (SEQ ID (SEQ ID N ( SEQ IDNO : 74 ) NO : 73 ) NO: 75)CDRH2 LINPYNGVTS YNQ INPYNGVT NPYNGV L INPYNGVT INPYNGVTSYNQKFK KFKD ( SEQ ID (SEQ ID (SEQ ID S ( SEQ ID DK (SEQ ID NO: 18 ) NO : 26 ) NO : 24 ) NO: 37 ) NO : 31) LINPYNAVTS YNQ INPYNAVT NPYNAV L INPYNAVT INPYNAVTSYNQKFK KFKD ( SEQ ID (SEQ ID (SEQ ID S ( SEQ ID DK (SEQ ID NO: 84 ) NO : 85) NO : 86) NO: 87 ) NO : 88 )CDRH3 DYNYDYFDY ARDYNYDYF DYNYDYFDY DYNYDYFDY DYNYDYFD ( SEQ (SEQ ID DY (SEQ (SEQ ID ( SEQ ID ID NO: 32 ) NO : 19 ) ID NO: 27 ) NO : 19 ) NO: 19 )CDRL1 RASEKIYSYLA EKIYSY RASEKIYSY RASEKIYSY ASEKIYSY ( SEQ (SEQ ID (SEQ ID LA SEQ ID LA (SEQ ID NO: 33 ) NO : 20 ) NO : 28 ) NO : 20 ) ID NO: 20)CDRL2 NAKTLAE (SEQ NAK (SEQ NAKTLAE NAKTLAE NAKTLAEGVPSR ID NO: 21) ID NO: 29 ) (SEQ ID ( SEQ ID (SEQ ID NO: 34 ) NO : 21 ) NO: 21)CDRL3 QHHFGSPRT QHHFGSPRT QHHFGSPRT QHHFGSPRT HFGSPR (SEQ ID (SEQ ID (SEQ ID (SEQ ID ( SEQ ID NO : 35) NO : 22 ) NO : 22 ) NO : 22 ) NO: 22 )Attorney Docket No: 01434-0001-00PCT

[0114] In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Kabat numbering as set forth in Table 1 and framework regions derived from human antibody sequences; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to IMGT numbering as set forth in Table 1 and framework regions derived from human antibody sequences; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigenbinding fragment thereof comprises a set of CDR sequences according to Kabat or IMGT numbering as set forth in Table 1 and framework regions derived from human antibody¬ sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Chothia numbering as set forth in Table 1 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to AbM numbering as set forth in Table 1 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigenbinding fragment thereof comprises a set of CDR sequences according to AHo numbering as set forth in Table 1 and framework regions derived from human antibody sequences; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, the human antibody framework sequences are derived from human antibody germline sequences.

[0115] In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR set forth in Table 1, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1, and framework regions derived from human antibodyAttorney Docket No: 01434-0001-00PCT sequences; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Kabat numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 1 or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1, and framework regions derived from human antibody sequences; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to IMGT numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 1. or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1, and framework regions derived from human antibody sequences; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Kabat or IMGT numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 1, or comprises 0, 1. or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Chothia numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%identical to the corresponding CDR as set forth in Table 1, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to AbM numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100% to the corresponding CDR as set forth in Table 1, or comprisesAttorney Docket No: 01434-0001-00PCT 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to AHo numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR, as set forth in Table 1, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 1, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, an amino acid substitution in a CDR is a conservative amino acid substitution. In some embodiments, the human antibody framework sequences are derived from human antibody germline sequences.

[0116] As used herein for G28-8 derived antibodies, position 55 in CDRH2 according to Kabat numbering corresponds to position 55 according to Chothia numbering, position 64 by IMGT numbering, position 55 according to AbM numbering, and position 66 according to AHo numbering.

[0117] As used herein for G28-8 derived antibodies, position 54 in CDRH2 according to Kabat numbering corresponds to position 54 according to Chothia numbering, position 62 by IMGT numbering, position 54 according to AbM numbering, and position 65 according to AHo numbering.Table 2. Heavy chain and light chain variable region CDR sequences of MHR73-11 antibodiesRabat IMGT Chothia AbM AHoCDRH1 DYTMN ( SEQ ID GYSFTDYT GYSFTGY GYSFTDYTM ASGYSFTDYTMN NO: 50) or (SEQ ID (SEQ ID N ( SEQ ID (SEQ ID NO: 58 ) DYSMN (SEQ ID NO : 55 ) or NO : 23 ) or NO: 63 ) or or ASGYTFTDYTMN NO : 77 ) GYTFTDYT GYTFTGY GYTFTDYTM (SEQ ID NO: 80 ) (SEQ ID (SEQ ID N ( SEQ IDNO : 78 ) NO : 73 ) NO: 79 )CDRH2 LINPYNGVTSYNP INPYNGVT NPYNGV L INPYNGVT INPYNGVTSYNPNFQ NFQG ( SEQ ID (SEQ ID (SEQ ID S ( SEQ ID GK (SEQ ID NO: 51) NO : 26 ) NO : 24 ) NO: 37 NO : 59 ) LINPYNAVTS YNP INPYNAVT NPYNAV L INPYNAVT INPYNAVTSYNPNFQAttorney Docket No: 01434-0001-00PCT NFQG ( SEQ ID (SEQ ID (SEQ ID S ( SEQ ID GK (SEQ ID NO : 89 ) NO : 85 ) NO : 86 ) NO: 87 ) NO : 90 )CDRH3 SGDYDRNYFDY ARSGDYDRN SGDYDRNYF SGDYDRNYF SGDYDRNYFD (SEQ (SEQ ID YFDY (SEQ DY (SEQ DY (SEQ ID NO: 60 ) NO: 52 ) ID NO: 56) ID NO : 52 ) ID NO: 52 )CDRL1 RASENIYSYLA ENIYSY RASENIYSY RASENIYSY ASENIYSY ( SEQ (SEQ ID (SEQ ID LA (SEQ LA (SEQ ID NO: 61) NO: 53) NO : 57 ) ID NO : 53) ID NO: 53)CDRL2 NAKTLAE (SEQ NAK (SEQ NAKTLAE NAKTLAE NAKTLAEGVPSR ID NO: 21) ID NO: 29 ) (SEQ ID ( SEQ ID (SEQ ID NO: 34 ) NO : 21 ) NO: 21)CDRL3 QHHYGS PWT QHHYGS PWT QHHYGS PWT QHHYGS PWT HYGSPW (SEQ ID (SEQ ID (SEQ ID (SEQ ID ( SEQ ID NO : 62 ) NO: 54 ) NO : 54 ) NO : 54 ) NO: 54 )

[0118] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences as set forth in Table 2 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Kabat numbering as set forth in Table 2 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally senne or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to IMGT numbering as set forth in Table 2 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Kabat or IMGT numbering as set forth in Table 2 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Chothia numbering as set forth in Table 2 and framework regions derived from human antibodyAttorney Docket No: 01434-0001-00PCT sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to AbM numbering as set forth in Table 2 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to AHo numbering as set forth in Table 2 and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering.

[0119] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Kabat numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 2, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 2, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to IMGT numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 2, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 2, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to Kabat or IMGT numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 2, or comprises 0, 1, or 2 ammo acid mutations relative to the amino acid sequence of a CDR set forth in Table 2, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-bindingAttorney Docket No: 01434-0001-00PCT fragment thereof comprises a set of CDR sequences according to Chothia numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 2, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 2, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to AbM numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 2, or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 2, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a set of CDR sequences according to AHo numbering, wherein each CDR is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the corresponding CDR as set forth in Table 2 or comprises 0, 1, or 2 amino acid mutations relative to the amino acid sequence of a CDR set forth in Table 2, and framework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering. In some embodiments, an amino acid substitution in a CDR is a conservative amino acid substitution. In some embodiments, the human antibody framework sequences are derived from human antibody germline sequences.

[0120] As used herein for MHR73-11 derived antibodies, position 55 in CDRH2 according to Kabat numbering corresponds to position 55 according to Chothia numbering, position 64 by IMGT numbering, position 55 according to AbM numbering and position 66 according to AHo numbering.

[0121] As used herein for MHR73-11 derived antibodies, position 54 in CDRH2 according to Kabat numbering corresponds to position 54 according to Chothia numbering, position 62 by IMGT numbering, position 54 according to AbM numbering and position 65 according to AHo numbering.

[0122] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:Attorney Docket No: 01434-0001-00PCT the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having the amino acid sequences set forth in one of the following sets of CDR sequences:a. CDRH1: SEQ ID NO: 17, 72, 50 or 77; CDRH2: SEQ ID NO: 18 or 51; and CDRH3: SEQ ID NO: 19 or 52; according to Kabat numbering;b. CDRH1: SEQ ID NO:25, 55, 74 or 78; CDRH2: SEQ ID NO:26; and CDRH3:SEQ ID NO:27 or 56; according to IMGT numbering;c. CDRH1: SEQ ID NO:23 or 73; CDRH2: SEQ ID NO:24; and CDRH3: SEQ ID NO: 19 or 52; according to Chothia numbering;d. CDRH1: SEQ ID NO:36, 63, 75 or 79; CDRH2: SEQ ID NO:37; and CDRH3:SEQ ID NO: 19 or 52; according to AbM numbering; ore. CDRH1: SEQ ID NO:30, 58, 76 or 80; CDRH2: SEQ ID NO:31 or 59; and CDRH3: SEQ ID NO:32 or 60 according to Aho numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having the amino acid sequences set forth in one of the following sets of CDR sequences:f CDRL1: SEQ ID NO:20 or 53; CDRL2: SEQ ID NO:21; and CDRL3: SEQ ID NO:22 or 54; according to Kabat numbering;g. CDRL1: SEQ ID NO:28 or 57; CDRL2: SEQ ID NO:29; and CDRL3: SEQ ID NO:22 or 54; according to IMGT numbering;h. CDRL1: SEQ ID NO:20 or 53; CDRL2: SEQ ID NO:21; and CDRL3: SEQ ID NO:22 or 54; according to Chothia numbering;i. CDRL1: SEQ ID NO 20 or 53; CDRL2: SEQ ID NO:21; and CDRL3: SEQ ID NO:22 or 54; according to AbM numbering; orj. CDRL1: SEQ ID NO 33 or 61; CDRL2: SEQ ID NO:34; and CDRL3: SEQ ID NO:35 or 62 according to AHo numbering; andframework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally serine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally glutamine according to Kabat numbering.

[0123] In some embodiments, CDRL3 does not have the amino acid sequence set forth in SEQ ID NO:54 when CDRL1 comprises the amino acid sequence set forth in SEQ ID NO:20, and CDRL2 comprises the amino acid sequence set forth in SEQ ID NO:21. In some embodiments CDRL3 does not have the amino acid sequence set forth in SEQ ID NO:54, when CDRL1 comprises the amino acid sequence set forth in SEQ ID NO:28, and CDRL2 comprises the amino acid sequence set forth in SEQ ID NO:29. In some embodiments, CDRL3 does notAttorney Docket No: 01434-0001-00PCT have the amino acid sequence set forth in SEQ ID NO: 54 when CDRL1 comprises the amino acid sequence set forth in SEQ ID NO:33, and CDRL2 comprises the amino acid sequence set forth in SEQ ID NO:34.

[0124] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in one of the following sets of CDR sequences:a) CDRH1: SEQ ID NO: 17, 72, 50 or 77; CDRH2: SEQ ID NO: 18 or 51; and CDRH3: SEQ ID NO: 19 or 52; according to Kabat numbering; or b) CDRH1: SEQ ID NO: 25, 55, 74 or 78; CDRH2: SEQ ID NO:26; and CDRH3:SEQ ID NO: 27 or 56; according to IMGT numbering;the light chain variable region comprises CDRL 1 , CDRL2. and CDRL3 having amino acid sequences set forth in one of the following sets of CDR sequences:c) CDRL1: SEQ ID NO:20 or 53; CDRL2: SEQ ID NO:21; and CDRL3: SEQ ID NO:22 or 54; according to Kabat numbering; ord) CDRL1: SEQ ID NO:28 or 57; CDRL2: SEQ ID NO:29; and CDRL3: SEQ ID NO:22 or 54; according to IMGT numbering; andframework regions derived from human antibody sequences, wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally serine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally glutamine according to Kabat numbering.III. Humanized G28-8 or MHR73-11 Derived Antibodies

[0125] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO: 17, 72, 50 or 77; SEQ ID NO: 18 or 51; and SEQ ID NO: 19 or 52; respectively, according to Kabat numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:20 or 53; SEQ ID NO:21; and SEQ ID NO:22 or 54; respectively; according to Kabat numbering; andAttorney Docket No: 01434-0001-00PCT heavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0126] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following amino acids: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises amino acid substitutions R66K, N61A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0127] As used herein, position 28 of the heavy chain variable region according to Kabat numbering corresponds to position 29 of IMGT numbering, position 28 of Chothia numbering, position 28 of AbM numbering, and position 29 of AHo numbering. Position 66 of the heavy chain variable region according to Kabat numbering corresponds to position 75 of IMGT numbering, position 66 of Chothia numbering, position 66 of AbM numbering and position 77 of AHo numbering. Position 67 of the heavy chain variable region according to Kabat numbering corresponds to position 76 of IMGT numbering, position 67 of Chothia numbering, position 67 of AbM numbering and position 78 of AHo numbering. Position 69 of the heavy chain variable region according to Kabat numbering corresponds to position 78 of IMGT numbering, position 69 of Chothia numbering, position 69 of AbM numbering and position 80 of AHo numbering. Position 71 of the heavy chain variable region according to Kabat numbering corresponds to position 80 of IMGT numbering, position 71 of Chothia numbering, position 71 of AbM numbering and position 82 of AHo numbering.Attorney Docket No: 01434-0001-00PCT

[0128] As used herein, position 48 of the tight chain variable region according to Kabat numbering corresponds to position 54 of IMGT numbering, position 48 of Chothia, position 48 of AbM numbering, and position 56 of AHo numbering. Position 64 of the light chain variable region according to Kabat numbering corresponds to position 78 of IMGT numbering, position 64 of Chothia numbering, position 64 of AbM numbering and position 80 of AHo numbering. Position 70 of the light chain variable region according to Kabat numbering corresponds to position 86 of IMGT numbering, position 70 of Chothia numbering, position 70 of AbM numbering and position 88 of AHo numbering.[00129J In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:25, 55, 74 or 78; SEQ ID NO:26; and SEQ ID NO:27 or 56; respectively; according to IMGT numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:28 or 57; SEQ ID NO:29; and SEQ ID NO:22 or 54; respectively; according to IMGT numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0130] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following the following amino acids: 28S, 66K, 67A, 69L, and 7 IV, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises amino acid substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chainAttorney Docket No: 01434-0001-00PCT variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0131] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:23 or 73; SEQ ID NO:24; and SEQ ID NO: 19 or 52; respectively; according to Chothia numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:20 or 53; SEQ ID NO:21; and SEQ ID NO:22 or 54; respectively; according to Chothia numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0132] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following amino acids: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises amino acid substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V.49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.Attorney Docket No: 01434-0001-00PCT

[0133] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1. CDRH2. and CDRH3 having amino acid sequences set forth in SEQ ID NO:36, 63, 75 or 79; SEQ ID NO:37; and SEQ ID NO: 19 or 52; respectively; according to AbM numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:20 or 53; SEQ ID NO:21; and SEQ ID NO:22 or 54; respectively; according to AbM numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0134] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following amino acids: 28S, 66K, 67A, I69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises amino acid substitutions R66K N61A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0135] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:Attorney Docket No: 01434-0001-00PCT the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:30, 58, 76 or 80; SEQ ID NO:31 or 59; and SEQ ID NO:32 or 60; respectively; according to AHo numbering;the light chain variable region comprises CDRL 1 , CDRL2. and CDRL3 having amino acid sequences set forth in SEQ ID NO:33 or 61; SEQ ID NO:34; and SEQ ID NO:35 or 62; respectively; according to AHo numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0136] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following amino acids: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises amino acid substitutions R66K, V67A. I69L. and R71V. according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V, 49N. and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.IV. Humanized G28-8 Derived AntibodiesIn some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:17 or 72; SEQ ID NO:18; and SEQ ID NO:19; respectively; according to Kabat numbering;Attorney Docket No: 01434-0001-00PCT the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:20; SEQ ID NO:21; and SEQ ID NO:22; respectively; according to Kabat numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0137] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0138] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NOs:25 or 74; SEQ ID NO:26; and SEQ ID NO:27; respectively; according to IMGT numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:28; SEQ ID NO:29; and SEQ ID NO:22; respectively; according to IMGT numbering; andheavy' and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0139] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67 A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region,Attorney Docket No: 01434-0001-00PCT according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0140] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:23 or 73; SEQ ID NO:24; and SEQ ID NO: 19; respectively; according to Chothia numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:20; SEQ ID NO:21; and SEQ ID NO: 22; respectively; according to Chothia numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0141] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of. at least four of, or all of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1. CDRH2. and CDRH3 having amino acid sequences set forth in SEQ ID NO:36 or 75; SEQ ID NO:37; and SEQ ID NO: 19; respectively; according to AbM numbering;wherein the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:20; SEQ ID NO:21; and SEQ ID NO:22; respectively; according to AbM numbering; andheavy and light chain variable region framew ork sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.Attorney Docket No: 01434-0001-00PCT

[0142] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0143] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1. CDRH2. and CDRH3 having amino acid sequences set forth in SEQ ID NO:30 or 76; SEQ ID NO:31; and SEQ ID NO:32; respectively; according to AHo numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:33; SEQ ID NO:34; and SEQ ID NO:35; respectively; according to AHo numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0144] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67 A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.V. Humanized MHR73-11 Derived Antibodies

[0145] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:Attorney Docket No: 01434-0001-00PCT the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:50 or 77; SEQ ID NO:51; and SEQ ID NO:52, respectively; according to Kabat numbering;the light chain variable region comprises CDRL 1 , CDRL2. and CDRL3 having amino acid sequences set forth in SEQ ID NO:53; SEQ ID NO:21; and SEQ ID NO:54; respectively; according to Kabat numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0146] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67 A, 69L, and 71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0147] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1. CDRH2. and CDRH3 having amino acid sequences set forth in SEQ ID NO:55 or 78; SEQ ID NO:26; and SEQ ID NO:56; respectively; according to IMGT numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:57; SEQ ID NO:29; and SEQ ID NO:54; respectively; according to IMGT numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.Attorney Docket No: 01434-0001-00PCT

[0148] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67 A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0149] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:23 or 73; SEQ ID NO:24; and SEQ ID NO:52; respectively; according to Chothia numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:53; SEQ ID NO:21; and SEQ ID NO:54; respectively; according to Chothia numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0150] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V, 49N. andAttorney Docket No: 01434-0001-00PCT 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0151] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1. CDRH2. and CDRH3 having amino acid sequences set forth in SEQ ID NO:63 or 79; SEQ ID NO:37; and SEQ ID NO:52; respectively; according to AbM numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:53; SEQ ID NO:21; and SEQ ID NO:54; respectively; according to AbM numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0152] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of, at least four of, or all of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region,Attorney Docket No: 01434-0001-00PCT according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.

[0153] In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein:the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 having amino acid sequences set forth in SEQ ID NO:58 or 80; SEQ ID NO:59; and SEQ ID NO:60; respectively; according to AHo numbering;wherein the light chain variable region comprises CDRL1, CDRL2, and CDRL3 having amino acid sequences set forth in SEQ ID NO:61 ; SEQ ID NO:34; and SEQ ID NO:62; respectively; according to AHo numbering; andheavy and light chain variable region framework sequences derived from the human IGVH1-03 / JH6 germline sequences and human IGVK1-02 / JK4 germline sequences, respectively.

[0154] In some embodiments, the heavy chain variable region comprises at least one of, at least two of, at least three of. at least four of, or all of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, the light chain variable region comprises substitutions at least one of 48V,49N, and 70Q, according to Kabat numbering. In some embodiments, the light chain variable region comprises at least one of, at least two of, or all of the following: 48V, 49N, and 70Q, according to Kabat numbering. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering.VI. Human Antibody Framework Regions

[0155] In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences. In some embodiments, the heavy’ chain variable region framework is derived from human 1GVH1-03 / JH6 germline sequences andAttorney Docket No: 01434-0001-00PCT has an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 80% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 85% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 90% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 91% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 92% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 93% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 94% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human 1GVH1-03 / JH6 germline sequences and has an amino acid sequence at least 95% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 96% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 97% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 98% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, the heavy chain variable region framework is derived from human IGVH1-03 / JH6 germline sequences and has an amino acid sequence at least 99% identical to the amino acid sequences set forth in SEQ ID NOs:38 and 39. In some embodiments, each of the heavy chain variable region positions T28, R66, V67, 169, and R71 are optional donor residues, according to Kabat numbering. InAttorney Docket No: 01434-0001-00PCT some embodiments, at least one of the heavy chain variable region positions T28, R66, V67, 169, and R71 (according to Kabat numbering) are substituted. In some embodiments, at least one of the heavy chain variable region positions T28, G55, R66, V67, 169, and R71 (according to Kabat numbering) are substituted with the corresponding amino acid from the parental antibody. In some embodiments, the heavy chain variable region comprises at least one of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises at least two of, at least three of, at least four of, or all of substitutions T28S. R66K, V67A, I69L, and R71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises substitutions R66K, V67A, I69L, and R71V, according to Kabat numbering.

[0156] In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 80% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 85% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human 1GVK1-02 / JK4 germline sequences and has an amino acid sequence at least 90% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 91% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 92% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 93% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 94% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 95%Attorney Docket No: 01434-0001-00PCT identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 96% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 97% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 98% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, the light chain variable region framework is derived from human IGVK1-02 / JK4 germline sequences and has an amino acid sequence at least 99% identical to the amino acid sequences set forth in SEQ ID NOs:40 and 41. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering.VII. Exemplary VH and VL Sequences

[0157] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDRH1, CDRH2. and CDRH3 as set forth in Table 1 according to the Kabat, IMGT, Chothia. AbM or AHo numbering (wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering), the heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in Table 3; and the light chain variable region comprises CDRL1, CDRL2, and CDRL3 as set forth in Table 1 according to the Kabat, IMGT, Chothia, AbM or AHo numbering, and the light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in Table 4.Attorney Docket No: 01434-0001-00PCT

[0158] In some embodiments, a humanized CD180 antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3 as set forth in Table 2 according to the Kabat, IMGT, Chothia. AbM or AHo numbering (wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or wherein position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering), the heavy chain variable framework regions comprise amino acid sequences having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences of a heavy chain variable region sequence set forth in Table 5, and the light chain variable region comprises CDRL1, CDRL2, and CDRL3 as set forth in Table 2 according to the Kabat, IMGT, Chothia, AbM or AHo numbering, and the light chain variable regions each comprise an acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences of light chain sequences set forth in Table 6.

[0159] In some embodiments, the humanized CD 180 antibody heavy chain variable region and light chain variable region have the amino acid sequences set forth in Tables 3-6 (below). In the tables, Kabat CDRs are underlined, and IMGT CDRs are bolded.Table 3. Heavy chain variable region sequences derived from the G28-8 antibody.QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQRLEWMGLINPYNGVTSYNQKFKDRVTITRDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 3 ) QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQRLEWMGLINPYNGVTSYNQKFKDRVTITRDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 5 ) QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQRLEWMGLINPYNGVTSYNQKFKDKATLTVDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 6 ) QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQRLEWMGLINPYNGVTSYNQKFKDKATLTVDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 7 ) QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQRLEWMGLINPYQGVTSYNQKFKDKATLTVDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 14 ) QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQRLEWMGLINPYNGVTSYNQKFKDRVTLTVDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 9 ) QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQRLEWMGLINPYNGVTSYNQKFKDKATITRDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 10 ) QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQRLEWMGLINPYSGVTSYNQKFKDKATLTVDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 15) QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQRLEWMGLINPYNAVTSYNQKFKDKATLTVDTS AS TAYMEL S S L RS E DTAVYYCARD YNYD YFD YWGQGT TVTVS S (SEQ ID NO : 16)Attorney Docket No: 01434-0001-00PCT Table 4. Light chain variable sequences derived from the G28-8 antibody.DIQMTQSPSSLSASVGDRVTITCRASEKIYSYLAWYQQKPGKAPKLLIYNAKTLAEGVPSRFSGSGSGTDFTLTI SSLQPEDFATYYCQHHFGSPRTFGGGTKVEIK (SEQ ID NO: 4 ) DIQMTQSPSSLSASVGDRVTITCRASEKIYSYLAWYQQKPGKAPKLLVYNAKTLAEGVPSRFSVSGSGTQFTLTI SSLQPEDFATYYCQHHFGSPRTFGGGTKVEIK (SEQ ID NO: 8 ) DIQMTQSPSSLSASVGDRVTITCRASEKIYSYLAWYQQKPGKAPKLLVYNAKTLAEGVPSRFSGSGSGTQFTLTI SSLQPEDFATYYCQHHFGSPRTFGGGTKVEIK (SEQ ID NO: 45) DIQMTQSPSSLSASVGDRVTITCRASEKIYSYLAWYQQKPGKAPKLLVYNAKTLAEGVPSRFSGSGSGTDFTLTI SSLQPEDFATYYCQHHFGSPRTFGGGTKVEIK (SEQ ID NO: 46) DIQMTQSPSSLSASVGDRVTITCRASEKIYSYLAWYQQKPGKAPKLLIYNAKTLAEGVPSRFSGSGSGTQFTLTI SSLQPEDFATYYCQHHFGSPRTFGGGTKVEIK (SEQ ID NO: 47 ) DIQMTQSPSSLSASVGDRVTITCRASEKIYSYLAWYQQKPGKAPKLLIYNAKTLAEGVPSRFSVSGSGTDFTLTI SSLQPEDFATYYCQHHFGSPRTFGGGTKVEIK (SEQ ID NO: 81)Table 5. Heavy chain variable region sequences derived from the MHR73-11 antibody.QVQL VQ S GAE VKKP GAS VKVS OKAS GYTFTD YTMNWVRQAPGQRL EWMGL INPYNGVTSYNPNFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARSG DYDRNYFDYWGQGT TVTVS S (SEQ ID NO : 66)QVQL VQ S GAE VKKP GAS VKVS OKAS GYTFTD YTMNWVRQAPGQRL EWMGL INPYNGVTSYNPNFQGKATLTVDTSASTAYMELSSLRSEDTAVYYCARSG DYDRNYFDYWGQGTTVTVSS (SEQ ID NO: 67 )QVQL VQ S GAE VKKP GAS VKVS OKAS GYTFTD YTMNWVRQAPGQRL EWMGL INPYQGVTSYNPNFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARSG DYDRNYFDYWGQGTTVTVSS (SEQ ID NO: 82 )QVQL VQ S GAE VKKP GAS VKVS C KAS GYTFTD YTMNWVRQAPGQRL EWMGL INPYSGVTSYNPNFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARSG DYDRNYFDYWGQGTTVTVSS (SEQ ID NO: 64 )QVQL VQ S GAE VKKP GAS VKVS OKAS GYTFTD YTMNWVRQAPGQRL EWMGL INPYNAVTSYNPNFQGRVTITRDTSASTAYMELSSLRSEDTAVYYCARSG DYDRNYFDYWGQGTTVTVSS (SEQ ID NO: 65 )Table 6. Light chain variable region sequences derived from the MHR73-11 antibody. DIQMTQSPSSLSASVGDRVTITCRASENIYSYLAWYQQKPGKAPKLLIYNAKTLAEGVPSRFSGSGSGTDFTLTI SSLQPEDFATYYCQHHYGSPWTFGGGTKVEIK (SEQ ID NO: 68 ) DIQMTQSPSSLSASVGDRVTITCRASENIYSYLAWYQQKPGKAPKLLVNNAKTLAEGVPSRFSGSGSGTQFTLTI SSLQPEDFATYYCQHHYGSPWTFGGGTKVEIK (SEQ ID NO: 69) DIQMTQSPSSLSASVGDRVTITCRASENIYSYLAWYQQKPGKAPKLLVNNAKTLAEGVPSRFSGSGSGTDFTLTI SSLQPEDFATYYCQHHYGSPWTFGGGTKVEIK (SEQ ID NO: 70) DIQMTQSPSSLSASVGDRVTITCRASENIYSYLAWYQQKPGKAPKLLIYNAKTLAEGVPSRFSGSGSGTQFTLTI SSLQPEDFATYYCQHHYGSPWTFGGGTKVEIK (SEQ ID NO: 71)Attorney Docket No: 01434-0001-00PCT

[0160] In some embodiments, the heavy chain variable framework regions comprise an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:3, 5, 6, 7, 9, 10, 14, 15, and 16 and 64, 65, 66, 67, and 82. In some embodiments, at least one of the heavy chain variable region positions 28, 66, 67, 69, and 71 is substituted, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises at least one of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises at least two of, at least three of, at least four of, or all of position 28 is serine, position 66 is lysine, position 67 is alanine, position 69 is leucine, and position 71 is valine, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises all of the following: position 66 is ly sine, position 67 is alanine, position 69 is leucine, and position 71 is valine, according to Kabat numbering.

[0161] In some embodiments, the light chain variable framework regions comprise amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:4, 8. 45. 46. 47 and 81, and 68, 69, 70, and 71. In some embodiments, at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate, according to Kabat numbering. In some embodiments, at least one of, at least two or all of position 48 is valine, position 64 is valine and position 70 is glutamine, according to Kabat numbering. In some embodiments, at least one of, at least two of or all of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate, according to Kabat numbering. In some embodiments, position 64 is valine, according to Kabat numbering. In some embodiments, position 64 is glycine, according to Kabat numbering.

[0162] In some embodiments, the heavy chain variable framework regions comprise amino acid sequences having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:3, 5, 6, 7, 9, 10, 14, 15, and 16; and the light chain variable framework regions comprise amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in the amino acid sequences set forth in SEQ ID NOs:4, 8, 45, 46, 47 and 81. In some embodiments, at least one of the heavy chain variable region positions 28, 66, 67, 69, and 71 is substituted, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises at least one of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises at least two of, at least three of. at least four of, or all of positionAttorney Docket No: 01434-0001-00PCT 28 is serine, position 66 is lysine, position 67 is alanine, position 69 is leucine, and position 71 is valine, according to Kabat numbering. In some embodiments, at least one of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, at least two of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering.

[0163] In some embodiments, the humanized CD 180 antibody or antigen-binding fragment thereof comprises:a) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%. 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:4;b) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 5 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%. 95%, 96%. 97%. 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NON;c) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity’ to the amino acid sequence set forth in SEQ ID NON;d) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NON and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NON;Attorney Docket No: 01434-0001-00PCT e) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 9 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:4;f) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:4;g) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:4;h) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity’ to the amino acid sequence set forth in SEQ ID NO:4;i) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:4;j) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%,Attorney Docket No: 01434-0001-00PCT 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 8;k) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 5 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 8;l) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%. 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO: 8;m) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO: 8;n) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%. 90%. 91%, 92%, 93%. 94%, 95%, 96%. 97%, 98%, 99%. 100% identity to the amino acid sequence set forth in SEQ ID NO: 9 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 8;o) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 8;p) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 14 and a light chainAttorney Docket No: 01434-0001-00PCT variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 8;q) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identity to the ammo acid sequence set forth in SEQ ID NO: 8;r) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 8;s) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:45;t) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 5 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:45;u) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity’ to the amino acid sequence set forth in SEQ ID NO:45;v) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%. 97%. 98%, 99%, or 100%Attorney Docket No: 01434-0001-00PCT identity to the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:45:w) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 9 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:45;x) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:45:y) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%. 98% or 99% identity to the amino acid sequence set forth in SEQ ID NO:45;z) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity the amino acid sequence set forth in SEQ ID NO:45;aa) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%,Attorney Docket No: 01434-0001-00PCT 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:45;bb) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:46:cc) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO: 5 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%. 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:46dd) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO:46;ee) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%. 91%. 92%, 93%, 94%, 95%, 96%. 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:46:ff) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 9 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:46;gg) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 10 and a light chainAttorney Docket No: 01434-0001-00PCT variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:46;hh) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:46;ii) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:46;jj) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:46;kk) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:47;11) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 5 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity’ to the amino acid sequence set forth in SEQ ID NO:47;mm) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%. orAttorney Docket No: 01434-0001-00PCT 100% identity to the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:47;nn) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:47;oo) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 9 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity’ to the amino acid sequence set forth in SEQ ID NO:47:pp) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:47:qq) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;rr) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:47;ss) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%,Attorney Docket No: 01434-0001-00PCT 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:47;tt) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:81:uu) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO: 5 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%. 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO: 81 ;vv) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO:81;ww) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%. 90%. 91%. 92%. 93%, 94%, 95%, 96%, 97%, 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO:81;xx) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 9 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:81;yy) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 10 and a light chainAttorney Docket No: 01434-0001-00PCT variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:81;zz) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:81;aaa) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:15 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:81; orbbb) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%. 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity' to the amino acid sequence set forth in SEQ ID NO:81.

[0164] In some embodiments, the humanized CD 180 antibody or antigen-binding fragment thereof comprises:a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NON;b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NON;c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NON;d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NON;Attorney Docket No: 01434-0001-00PCT e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:9 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:4;f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:4;g) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:4;h) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:4;i) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:4;j) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;k) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;l) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;m) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;n) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:9 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;o) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;Attorney Docket No: 01434-0001-00PCT p) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;q) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;r) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;s) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;t) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;u) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;v) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;w) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO:9 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;x) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;y) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;z) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;Attorney Docket No: 01434-0001-00PCT aa) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:45;bb) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:3 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;cc) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 5 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;dd) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;ee) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;ff) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;gg) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;hh) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;ii) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;jj) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:46;kk) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO:3 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;Attorney Docket No: 01434-0001-00PCT 11) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;mm) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;nn) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 7 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;oo) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:9 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;pp) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;qq) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;rr) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;ss) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:47;tt) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 3 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:81;uu) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:5 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:81;vv) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 81;Attorney Docket No: 01434-0001-00PCT ww) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:7 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 81;xx) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:9 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 81;yy) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:81;zz) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 14 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 81;aaa) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 15 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:81; orbbb) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:81.

[0165] In some embodiments, the humanized CD 180 antibody or antigen-binding fragment thereof comprises:a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 8;c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:6 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 81; ord) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 16 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 81.

[0166] In some embodiments, the heavy chain variable framework regions comprise amino acid sequences having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ IDAttorney Docket No: 01434-0001-00PCT NOs:64, 65, 66, 67, and 82; and the light chain variable framework regions comprise amino acid sequences having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from SEQ ID NOs:68, 69, 70 and 71. In some embodiments, at least one of the heavy’ chain variable region positions 28, 66, 67, 69, and 71 is substituted, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises at least one of the following: 28S, 66K, 67A, 69L, and 71V, according to Kabat numbering. In some embodiments, the heavy chain variable region comprises at least two of, at least three of, at least four of, or all of position 28 is serine, position 66 is lysine, position 67 is alanine, position 69 is leucine, and position 71 is valine, according to Kabat numbering. In some embodiments, the heavy7chain variable region comprises all of the following: position 66 is lysine, position 67 is alanine, position 69 is leucine, and position 71 is valine, according to Kabat numbering. In some embodiments, at least one of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, at least two of position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 48 is isoleucine, position 64 is glycine and position 70 is aspartate in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is glycine in the light chain variable region, according to Kabat numbering. In some embodiments, position 64 is valine in the light chain variable region, according to Kabat numbering. In some embodiments, position 55 in CDRH2 is optionally alanine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally serine according to Kabat numbering. In some embodiments, position 54 in CDRH2 is optionally alanine according to Kabat numbering.

[0167] In some embodiments, the humanized CD 180 antibody or antigen-binding fragment thereof comprises:a) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:68;b) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%,Attorney Docket No: 01434-0001-00PCT 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:68;c) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 82 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:68:d) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%. 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:68;e) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity7to the amino acid sequence set forth in SEQ ID NO:68;I) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%. 91%. 92%, 93%, 94%, 95%, 96%. 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:69:g) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:69;h) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 82 and a light chainAttorney Docket No: 01434-0001-00PCT variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:69;i) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:69;j) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:69;k) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:70;l) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%. 99%. or 100% identity to the amino acid sequence set forth in SEQ ID NO:70;m) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 82 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity’ to the amino acid sequence set forth in SEQ ID NO:70;n) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%. 97%. 98%, 99%, or 100%Attorney Docket No: 01434-0001-00PCT identity to the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:70:o) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:70;p) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:71:q) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:71:r) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO: 82 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:71;s) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%. 92%, 93%, 94%, 95%, 96%, 97%. 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:71; orAttorney Docket No: 01434-0001-00PCT t) a heavy chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequence set forth in SEQ ID NO:71.

[0168] In some embodiments, the humanized CD 180 antibody or antigen-binding fragment thereof comprises:a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:68;b) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:68;c) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 82 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:68;d) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:68;e) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:68;f) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:69;g) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:69;h) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:82 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:69;i) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:69;Attorney Docket No: 01434-0001-00PCT j) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:69;k) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:70;l) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:70;m) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 82 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:70;n) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:70;o) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:70;p) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:71;q) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO:67 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:71;r) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:82 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:71;s) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:64 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:71; ort) a heavy chain variable region comprising the ammo acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:71.

[0169] In some embodiments, the humanized CD 180 antibody or antigen-binding fragment thereof comprises:Attorney Docket No: 01434-0001-00PCT a) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:66 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:68; orb) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:65 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:68.

[0170] A humanized CD180 antibody can be an intact antibody or antigenbinding fragment thereof. In some embodiments, a humanized CD 180 antibody is an intact antibody. In some embodiments, a humanized CD 180 antibody is an antigen binding fragment. In some embodiments, a humanized CD 180 antibody or antigen binding fragment is monovalent. In some embodiments, a humanized CD 180 antibody or antigen binding fragment is bivalent. In some embodiments, a humanized CD 180 antibody or antigen binding fragment is multivalent. In some embodiments, a humanized CD 180 antibody or antigen binding fragment is mono-specific. In some embodiments, a humanized CD180 antibody or antigen binding fragment is bi-specific. In some embodiments, a humanized CD 180 antibody or antigen binding fragment is multispecific. In some embodiments, a humanized CD 180 antibody is unconjugated. In some embodiments, a humanized CD 180 antigen binding fragment is, for example, an Fv, single-chain Fv (scFv), a Fab, a Fab', a F(ab')2, a F(ab)c, a diabody, a minibody, a nanobody, a Fab-scFv fusion, a bispecific (scFv)4-IgG, or bispecific (scFv)2-Fab.

[0171] In some embodiments, a humanized CD180 antibody comprises an “Fc region’" (the C-terminal region of an immunoglobulin heavy chain). In some embodiments, a humanized CD 180 antibody or antigen-binding fragment thereof comprises a heavy chain constant region attached to the heavy chain variable region and a light chain constant region attached to the light chain variable region. In some embodiments, the heavy chain constant region is an IgG, IgA, or IgM region. In some embodiments, the heavy chain constant region is an IgG region. In some embodiments, the heavy chain constant region is an IgGl region. In some embodiments, the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:42. In some embodiments, the heavy chain constant region is an IgG4 region. In some embodiments, the heavy chain constant region is an IgG4 region. In some embodiments, the light chain constant region is a kappa constant region. In some embodiments, the light chain constant region is a lambda constant region.VIII. GLYCOSYLATION MODIFICATION

[0172] The humanized CD 180 antibodies and antigen- binding fragments thereof optionally may be glycosylated at conserved positions in their constant regions (Jefferis andAttorney Docket No: 01434-0001-00PCT Lund, (1997) Chem. Immunol. 65: 111-128; Wright and Momson, (1997) TibTECH 15:26-32). The oligosaccharide side chains of the immunoglobulins affect the protein’s function (Boyd et al., (1996) Mol. Immunol. 32: 1311-1318; Wittwe and Howard, (1990) Biochem. 29:4175-4180), and the intramolecular interaction between portions of the glycoprotein which can affect the conformation and presented three-dimensional surface of the glycoprotein (Jefferis and Lund, supra; Wyss and Wagner, (1996) Current Op. Biotech. 7:409-416). Oligosaccharides may also serve to target a given glycoprotein to certain molecules based upon specific recognition structures. For example, it has been reported that in agalactosylated IgG, the oligosaccharide moiety 'flips’ out of the inter-CH2 space and terminal N-acetylglucosamine residues become available to bind mannose binding protein (Malhotra et al, (1995) Nature Med. 1:237-243). Removal by glycopeptidase of the oligosaccharides from CAMPATH- 1H (a recombinant humanized murine monoclonal IgGl antibody which recognizes the CDw52 antigen of human lymphocytes) produced in Chinese Hamster Ovary (CHO) cells resulted in a complete reduction in complement mediated lysis (CMCL or CDC) (Boyd et al., (1996) Mol. Immunol. 32: 1311-1318), while selective removal of sialic acid residues using neuraminidase resulted in no loss of CMCL. Glycosylation of antibodies has also been reported to affect antibody-dependent cellular cytotoxicity’ (ADCC). In particular, CHO cells with tetracycline-regulated expression of a(l,4)-N-acetylglucosaminyltransferase III (GnTIII), a glycosyltransferase catalyzing formation of bisecting GlcNAc, was reported to have improved ADCC activity7(Umana et al. (1999) Mature Biotech. 17: 176-180).

[0173] Glycosylation of antibodies is typically either N-linked or O-linked. N-linked refers to the attachment of the carbohydrate moiety to the side chain of an asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are the recognition sequences for enzy matic attachment of the carbohydrate moiety to the asparagine side chain. Thus, the presence of either of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose, or xylose to a hydroxyamino acid, most commonly serine or threonine, although 5 -hydroxy proline or 5-hydroxylysine may also be used.

[0174] Glycosylation variants of antibodies are variants in which the glycosylation pattern of an antibody is altered as compared to a parental or reference antibody. By altering is meant deleting one or more carbohydrate moieties found in the antibody, adding one or more carbohydrate moieties to the antibody, changing the composition of glycosylation (glycosylation pattern), the extent of glycosylation, etc.Attorney Docket No: 01434-0001-00PCT

[0175] Addition of glycosylation sites to a humanized anti-CDl 80 antibody or an antigen-binding fragment thereof can be accomplished by altering the amino acid sequence such that it contains one or more of the above-described tripeptide sequences (for N-linked glycosylation sites). The alteration may also be made by the addition of, or substitution by. one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites). Similarly, removal of glycosylation sites can be accomplished by amino acid alteration within the native glycosylation sites of the antibody.

[0176] The amino acid sequence is typically altered by changing the underlying nucleic acid sequence. These methods include isolation from a natural source (in the case of naturally-occurring amino acid sequence variants) or preparation by oligonucleotide-mediated (or site-directed) mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlier prepared variant or anon-variant version of the antibody.

[0177] The glycosylation (including glycosylation pattern) of antibodies may also be altered without changing the amino acid sequence or the underlying nucleotide sequence. Glycosylation largely depends on the host cell used to express the antibody. Since the cell type used for expression of recombinant glycoproteins, e.g., antibodies, as potential therapeutics is rarely the native cell, significant variations in the glycosylation pattern of the antibodies can be expected. See, e.g., Hse et al, (1997) J. Biol. Chem. 272:9062-9070. In addition to the choice of host cells, factors which affect glycosylation during recombinant production of antibodies include grow th mode, media formulation, culture density, oxygenation, pH, purification schemes and the like. Various methods have been proposed to alter the glycosylation pattern achieved in a particular host organism including introducing or overexpressing certain enzymes involved in oligosaccharide production (see, e.g., U.S. Patent Nos. 5047335; 5510261; and 5278299).Glycosylation, or certain ty pes of glycosylation, can be enzymatically removed from the glycoprotein, for example using endoglycosidase H (Endo H). In addition, the recombinant host cell can be genetically engineered, e.g., make defective in processing certain types of polysaccharides. These and similar techniques are well known in the art.

[0178] The glycosylation structure of antibodies can be readily analyzed by conventional techniques of carbohydrate analysis, including lectin chromatography, NMR, mass spectrometry, HPLC. GPC, monosaccharide compositional analysis, sequential enzymatic digestion, and HPAEC-PAD, which uses high pH anion exchange chromatography to separate oligosaccharides based on charge. Methods for releasing oligosaccharides for analytical purposes are also known, and include, without limitation, enzymatic treatment (commonly performed using peptide-N-glycosidase F / endo-P-galactosidase), elimination using harsh alkalineAttorney Docket No: 01434-0001-00PCT environment to release mainly O-linked structures, and chemical methods using anhydrous hydrazine to release both N-and O-linked oligosaccharides.

[0179] In some embodiments, a form of modification of glycosylation of antibodies is reduced core fucosylation. "‘Core fucosylation" refers to addition of fucose (“fucosylation”) to N-acetylglucosamine (“GlcNAc”) at the reducing terminal of an N-linked glycan.

[0180] A “complex N-gly coside-linked sugar chain’' is ty pically bound to asparagine 297 (according to the number of Kabat). As used herein, the complex N-gly coside-linked sugar chain has a biantennary composite sugar chain, mainly having the following structure:'i'AFucrxl ♦AGa^t — * 4GicNAc[l1 — *2MaiwGlcNAs[) 1 - 1 - — *- 4GlcNAc31 ► 4GlcN Ac3♦AGalH — ► 4G1cNA.o[51 — *where + / - indicates the sugar molecule can be present or absent, and the numbers indicate the position of linkages between the sugar molecules. In the above structure, the sugar chain terminal which binds to asparagine is called a reducing terminal (at right), and the opposite side is called a non-reducing terminal. Fucose is usually bound to N-acetylglucosamine (“GlcNAc”) of the reducing terminal, typically by an al, 6 bond (the 6-position of GlcNAc is linked to the 1 -position of fucose). “Gal” refers to galactose, and “Man” refers to mannose.

[0181] A “complex N-glycoside-linked sugar chain” includes: 1) a complex type, in which the non-reducing terminal side of the core structure has one or more branches of galactose-N-acetylglucosamine (also referred to as “gal-GlcNAc") and the non-reducing terminal side of Gal-GlcNAc optionally has a sialic acid, bisecting N-acetylglucosamine or the like; or 2) a hybrid type, in which the non-reducing terminal side of the core structure has both branches of a high mannose N-glycoside-linked sugar chain and complex N-glycoside-linked sugar chain. In some embodiments, the “complex N-glycoside-linked sugar chain” includes a complex type in which the non-reducing terminal side of the core structure has zero, one or more branches of galactose-N-acetylglucosamine (also referred to as “gal-GlcNAc”) and the non-reducing terminal side of Gal-GlcNAc optionally further has a structure such as a sialic acid, bisecting N-acetylglucosamine or the like.Attorney Docket No: 01434-0001-00PCT Methods of making non-fucosylated antibodies by incubating antibody -producing cells with a fucose analogue are described, e.g., in W02009 / 135181. Briefly, cells that have been engineered to express a humanized CD 180 antibody or antigen binding fragment are incubated in the presence of a fucose analogue or an intracellular metabolite or product of the fucose analog. An intracellular metabolite can be, for example, a GDP-modified analog or a fully or partially de-esterified analog. A product can be, for example, a fully or partially de-esterified analog. In some embodiments, a fucose analogue can inhibit an enzyme(s) in the fucose salvage pathway. For example, a fucose analog (or an intracellular metabolite or product of the fucose analog) can inhibit the activity of fucokinase, or GDP-fucose-pyrophosphorylase. In some embodiments, a fucose analog (or an intracellular metabolite or product of the fucose analog) inhibits fucosyltransferase (e.g., a 1,6-fucosyltransferase, e.g., the FUT8 protein). In some embodiments, a fucose analog (or an intracellular metabolite or product of the fucose analog) can inhibit the activity of an enzyme in the de novo synthetic pathway for fucose. For example, a fucose analog (or an intracellular metabolite or product of the fucose analog) can inhibit the activity of GDP-mannose 4,6-dehydratase or / or GDP-fucose synthetase. In some embodiments, the fucose analog (or an intracellular metabolite or product of the fucose analog) can inhibit a fucose transporter (e.g., GDP-fucose transporter).In some embodiments, the fucose analogue is 2-flurofucose. Methods of using fucose analogues in growth medium and other fucose analogues are disclosed, e.g., in W02009 / 135181, which is herein incorporated by reference.

[0182] In certain embodiments, only a minor amount of a fucose analog (or a metabolite or product of the fucose analog) is incorporated into the complex N -glycoside-linked sugar chain(s). For example, in various embodiments, less than about 60%, less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, or less than about 3% of the humanized CD180 antibodies have core fucosylation by a fucose analog or a metabolite or product of the fucose analog. In some embodiments, about 2% of the CD 180 antibodies have core fucosylation by a fucose analog or a metabolite or product of the fucose analog.

[0183] Other methods for engineering cell lines to reduce core fucosylation included gene knock outs, gene knock-ins and RNA interference (RNAi). In gene knock-outs, the gene encoding FUT8 (alpha 1,6- fucosyltransferase enzyme) is inactivated. FUT8 catalyzes the transfer of a fucosyl residue from GDP-fucose to position 6 of Asn-linked (N-linked) GlcNac of an N-glycan. FUT8 is reported to be the only enzyme responsible for adding fucose to the N-linked biantennary carbohydrate at Asn297. Gene knock-ins add genes encoding enzy mes such as GNTIII or a Golgi alpha mannosidase II. An increase in the levels of such enzymes in cellsAttorney Docket No: 01434-0001-00PCT diverts monoclonal antibodies from the fucosylation pathway (leading to decreased core fucosylation), and having increased amount of bisecting N-acetylglucosamines. RNAi typically also targets FUT8 gene expression, leading to decreased mRNA transcript levels or knocking out gene expression entirely. Any of these methods can be used to generate a cell line that would be able to produce a non-fucosylated antibody.

[0184] Many methods are available to determine the amount of fucosylation on an antibody. Methods include, e.g., LC-MS via PLRP-S chromatography and electrospray ionization quadrupole TOF MS.IX. Fc MODIFICATION

[0185] In some embodiments, an Fc region of a humanized CD 180 antibody or an antigen binding fragment thereof is altered by replacing at least one amino acid residue in the Fc region with a different amino acid residue to alter the effector function(s) of the antibody. For example, one or more amino acids can be replaced with a different amino acid residue such that the antibody has an altered affinity for an effector ligand but retains the antigen-binding ability of the parent antibody.

[0186] As used herein, the terms “isotype constant region’7or “Fc region” are used interchangeably to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. The human IgG heavy chain Fc region is generally defined as comprising the amino acid residue from position C226 or from P230 to the carboxyl-terminus of the IgG antibody, wherein the numbering is according to the EU numbering system. The C-terminal lysine (residue K447) of the Fc region may be removed, for example, during production or purification of the antibody or its corresponding codon deleted in the recombinant constructs. Accordingly, a composition of the humanized CD 180 antibodies of the disclosure 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.

[0187] In some specific embodiments, the hinge region of CHI is modified such that the number of cysteine residues in the hinge region is altered, e.g., increased or decreased. This approach is described further in U.S. Patent No. 5,677.425 by Bodmer et al. In some embodiments, the number of cysteine residues in the hinge region of CHI is altered to, for example, facilitate assembly of the light and heavy chains or to increase or decrease the stability of the antibody.

[0188] In some embodiments, the Fc hinge region of an antibody is modified to decrease the biological half-life of the antibody. More specifically, one or more amino acidAttorney Docket No: 01434-0001-00PCT substitutions are introduced into the CH2-CH3 domain interface region of the Fc-hinge fragment such that the antibody has impaired Staphylococcyl protein A (SpA) binding relative to native Fc-hinge domain SpA binding. This approach is described in further detail in U.S. Patent No. 6.165,745 by Ward et al.

[0189] An Fc region may interact with different types of Fc receptors (FRs). The different types of FcRs may include, for example, FcgRI, FcgRIIA, FcgRIIB, FcgRIIIA, FcgRIIIB, FcaRI, FcpR, FceRI, FceRII, and FcRn. FcRs may be located on the membrane of certain immune cells including, for example, B lymphocytes, natural killer cells, macrophages, neutrophils, follicular dendritic cells, eosinophils, basophils, platelets, and mast cells. Once the FcR is engaged by the Fc region, the FcR may initiate functions including, for example, clearance of an antigen-antibody complex via receptor-mediated endocytosis, antibodydependent cell-mediated cytotoxicity (ADCC), antibody dependent cell-mediated phagocytosis (ADCP), and ligand-triggered transmission of signals across the plasma membrane that can result in alterations in secretion, exocytosis, and cellular metabolism. FcRs may deliver signals when FcRs are aggregated by antibodies and multivalent antigens at the cell surface.

[0190] In some embodiments, an Fc region of the antibody can exhibit increased binding affinity to one or more Fc receptors. In some embodiments, an Fc region can exhibit increased binding affinity to one or more Fcgamma receptors. In some embodiments, an Fc region can exhibit increased binding affinity to FcRn receptors. In some embodiments, an Fc region can exhibit increased binding affinity to Fcgamma and FcRn receptors.

[0191] In some embodiments, an Fc region of the antibody can exhibit reduced binding affinity to one or more Fc receptors. In some embodiments, an Fc region can exhibit reduced binding affinity to one or more Fcgamma receptors. In some embodiments, an Fc region exhibit reduced binding affinity to FcRn receptors. In some embodiments, an Fc region can exhibit reduced binding affinity to Fcgamma and FcRn receptors. In some embodiments, an Fc region is an Fc null. In some embodiments, an Fc region can exhibit reduced binding affinity to FcRn receptors, but have the same or increased binding affinity to one or more Fcgamma receptors as compared to a wildtype IgG. In some embodiments, an Fc region can exhibit increased binding affinity' to FcRn receptors, but have the same or decreased binding affinity to one or more Fcgamma receptors. As used herein, an “Fc null” refers to an Fc region or a domain thereof that exhibits weak to no binding to any of the Fcgamma receptors. In some embodiments, an Fc null exhibits a reduction in binding affinity (e.g., increase in Kd) to Fc gamma receptors of at least 1000-fold. In some embodiments, an Fc region is an effector function null. As used herein, an “effector function null” exhibits essentially no ADCC, ADCP and CDC activity(i.e.. less than 1% of the parental antibody having a human IgGl Fc region).Attorney Docket No: 01434-0001-00PCT

[0192] In some embodiments, an Fc region may have one or more, two or more, three or more, or four or more amino acid substitutions that decrease binding of the Fc region to an Fc receptor. In certain embodiments, an Fc region has decreased binding affinity for one or more of FcgRI (CD64), FcgRIIA (CD32), FcgRIIIA (CD 16a), FcgRIIIB (CD 16b), or any combination thereof. In order to decrease binding affinity of an Fc region to an Fc receptor, the Fc region may comprise one or more amino acid substitutions that reduces the binding affinity of the Fc region to an Fc receptor.

[0193] In some embodiments, the one or more substitutions comprise any one or more of IgGl heavy chain substitutions corresponding to E233P, L234V, L234A, L235A, L235E, AG236, G237A, E318A, K320A, K322A, A327G, A330S, orP331S, according to the EU index of Kabat numbering.

[0194] In some embodiments, the Fc region can comprise a sequence of an IgG isoform that has been modified from the wild-type IgG sequence. In some embodiments, the Fc region can comprise a sequence of the IgGl isoform that has been modified from the wild-type IgGl sequence. In some embodiments, the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc region to all Fcg receptors. In some embodiments, a modification by be L234A and L235A (“LALA”) according to the EU index of Kabat. In some embodiments, a modification can be L234A, L235A, and P329G ( "L ALA-PG "). according to the EU index of Kabat. In some embodiments, a modification can be substitution of E233, L234 and L235, such as E233P, L234V and L235A or E233P, L234V, L235A, and D236G, according to the EU index of Kabat. In some embodiments, a modification can be L234A, L235E, G237A, A330S and P331S, according to the EU index of Kabat. In some embodiments, a modification can be L234A, L235E, G237A, A330S and P331S in an IgGl Fc region, according to the EU index of Kabat. In some embodiments, a modification can be L234A, L235E and G237A, according to the EU index of Kabat. In some embodiments, a modification can be L234A, L235E and G237A in an IgGl Fc region, according to the EU index of Kabat. A modification can be a substitution of P238, such as P238A; substitution of D265, such as D265A; substitution of N297, such as N297A; substitution of A327, such as A327Q; or substitution of P329, such as P329A, according to the EU index of Kabat. In some embodiments, the heavy chain variable region has the amino acid sequence set forth in SEQ ID NO:43 (IgGl.l). In some embodiments, the heavy chain variable region has the amino acid sequence set forth in SEQ ID NO:83 (IgGl.3). In some embodiments, a modification can be S228P in an IgG4 Fc region, according to the EU index of Kabat.

[0195] In some embodiments, an IgG Fc region comprises at least one amino acid substitution that reduces its binding affinity to FcgRI. as compared to a wild-type or referenceAttorney Docket No: 01434-0001-00PCT IgG Fc region. A modification can comprise a substitution at F241, such as F241 A; substitution at F243, such as F243A; substitution at V264, such as V264A; or substitution at D265, such as D265 A according to the EU index of Kabat.

[0196] In some embodiments, an IgG Fc region comprises at least one amino acid substitution that increases its binding affinity to FcgRl, as compared to a wild-type or reference IgG Fc region. A modification can comprise a substitution at A327 and P329, such as A327Q / P329A, according to the EU index of Kabat.

[0197] In some embodiments, the modification comprises substitution of one or more amino acids that reduce binding affinity of an IgG Fc region to FcgRII and FcgRIIIA receptors. A modification can be a substitution of D270, such as D270A; substitution of Q295, such as Q295A; or substitution of A327, such as A237S, according to the EU index of Kabat.

[0198] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc region to FcgRII and FcgRIIIA receptors. A modification can be a substitution of T256, such as T256A; substitution of K290, such as K290A.

[0199] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc region to FcgRII receptor. A modification can be a substitution of R255, such as R255A; substitution of E258, such as E258A; substitution of S267, such as S267A; substitution of E272, such as E272A; substitution of N276, such as N276A; substitution of D280, such as D280A; substitution of H285, such as H285A; substitution of N286, such as N286A; substitution of T307, such as T307A; substitution of L309, such as L309A; substitution of N315, such as N315A; substitution of K326, such as K326A; substitution of P331, such as P331A; substitution of S337, such as S337A; substitution of A378, such as A378A; or substitution of E430, such as E430, according to the EU index of Kabat.

[0200] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc region to FcgRII receptor and reduces the binding affinity to FcgRIIIA receptor. A modification can be a substitution of H268, such as H268A; substitution of R301, such as R301A; or substitution of K322, such as K322A, according to the EU index of Kabat.

[0201] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc region to FcgRII receptor but does not affect the binding affinity to FcgRIIIA receptor. A modification can be a substitution of R292; or substitution of K414, such as K414A, according to the EU index of Kabat.Attorney Docket No: 01434-0001-00PCT

[0202] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc region to FcgRIIIA receptor. A modification can be substitution of F241 and F243, such as F241S / F243S or F241I / F243I, according to the EU index of Kabat.

[0203] In some embodiments, the modification comprises substitution of one or more amino acids that decreases binding affinity of an IgG Fc region to FcgRIIIA receptor and does not affect the binding affinity to FcgRII receptor. A modification can be a substitution of S239. such as S239A; substitution of E269, such as E269A; substitution of E293. such as E293A; substitution of Y296, such as Y296F; substitution of V303, such as V303A; substitution of A327, such as A327G; substitution of K338, such as K338A; or substitution of D376, such as D376A, according to the EU index of Kabat.

[0204] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc region to FcgRIIIA receptor and does not affect the binding affinity to FcgRII receptor. A modification can be a substitution of E333, such as E333A; substitution of K334, such as K334A; substitution of A339, such as A339T; or substitution of S239 and 1332, such as S239D / I332E according to the EU index of Kabat.

[0205] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity7of an IgG Fc region to FcgRIIIA receptor. A modification can be substitution of L235, F243, R292, Y300 and P396, such as L235V / F243L / R292P / Y1300L / P396L (IgGIVLPLL) according to the EU index of Kabat. A modification can be substitution of S298, E333 and K334, such as S298A / E333 A / K334A, according to the EU index of Kabat. A modification can be substitution of K246, such as K246F, according to the EU index of Kabat.

[0206] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc region or region to FcgRII receptor and increases the binding affinity' to FcgRIIIA receptor. A modification can be a substitution of S298, such as S298A; substitution of S239, 1332 and A330, such as S239D / I332E / A330L or substitution of S239 and 1332, such as S239D / I332E.

[0207] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc region or region to a FcgRII receptor and decreases binding to a FcgRIII receptor such as L234A / L235A / G237A / K322A / S267E / L328F.Attorney Docket No: 01434-0001-00PCT

[0208] In some embodiments, the modification comprises substitution of one or more amino acids that increases binding affinity of an IgG Fc region or region to a FcgRII receptor decreases binding to a FcgRIII receptor such as S267E / L328F.

[0209] Other substitutions in an IgG Fc region that affect its interaction with one or more Fcg receptors are disclosed in U.S. Patent Nos.7, 317, 091 and 8,969,526 (the disclosures of which are incorporated by reference herein).

[0210] In some embodiments, an IgG Fc region comprises at least one amino acid substitution that reduces the binding affinity to FcRn, as compared to a wild-type or reference IgG Fc region. A modification can comprise a substitution at H435, such as H435A; a substitution at 1253, such as I253A; a substitution at H310, such as H310A or substitutions at 1253, H310 and H435, such as I253A / H310A / H435A according to the EU index of Kabat.

[0211] In some embodiments, a modification can comprise a substitution of one amino acid residue that increases the binding affinity of an IgG Fc region for FcRn, relative to a wildtype or reference IgG Fc region. A modification can comprise a substitution at V308, such as V308P; at M428, such as M428L; at N434, such as N434A; at T250 and M428, such as T250Q and M428L; at M428 and N434, such as M428L and N434S, N434A or N434H; at M252, S254 and T256. such as M252Y / S254T / T256E; or substitution of one or more amino acids selected from P257L, P257N, P257I, V279E, V279Q, V279Y, A281S, E283F, V284E, L306Y, T307V, V308F, Q31 IV, D376V, and N434H according to the EU index of Kabat. Other substitutions in an IgG Fc region that affect its interaction with FcRn are disclosed in U.S. Patent No.9,803,023 (the disclosure of which is incorporated by reference herein).

[0212] In other embodiments, the Fc region is modified to decrease the ability of the antibody to mediate antibody dependent cellular cytotoxicity (ADCC) and / or to decrease the affinity of the antibody for an Fey receptor by modifying one or more amino acids. Such antibodies with decreased effector functions, and in particular decreased ADCC include silent antibodies.

[0213] In certain embodiments, the Fc region of the IgGl isotype is used. In some specific embodiments, a mutant variant of the IgGl Fc fragment is used, e.g. a silent IgGl Fc which reduces or eliminates the ability of the humanized CD 180 antibody or antigen binding fragment thereof to mediate antibody dependent cellular cytotoxicity (ADCC) and / or to bind to an Fey receptor.

[0214] In certain embodiments, the Fc region of the IgG4 isotype is used. In some specific embodiments, a mutant variant of the IgG4 Fc fragment is used, e.g. a silent IgG4 Fc which reduces or eliminates the ability of the humanized CD 180 antibody or antigen bindingAttorney Docket No: 01434-0001-00PCT fragment to mediate antibody dependent cellular cytotoxicity (ADCC) and / or to bind to an Fey receptor.

[0215] Silenced effector functions can be obtained by mutation in the Fc region of the antibodies and have been described above and in the art (see, e.g., Baudino et al., J.Immunol. 2008; Strohl, Biotechnology 202009). Examples of silent IgGl antibodies comprise the triple mutant variant IgGl L247F L248E P350S. Examples of silent IgG4 antibodies comprise the double mutant variant IgG4 S241P L248E.

[0216] In certain embodiments, the Fc region is a silent Fc mutant preventing glycosylation at position 314 of the Fc region. For example, the Fc region contains an amino acid substitution of asparagine at position 314. An example of such amino acid substitution is the replacement ofN314 by a glycine or an alanine.

[0217] The effector ligand to which affinity is altered can be, for example, an Fc receptor or the Cl component of complement. This approach is described in further detail in U.S. Patent Nos. 5,624,821 and 5,648,260, both by Winter et al.X. Antibody Molecule Expression

[0218] Nucleic acids of the present disclosure can be expressed in a host cell that contains endogenous DNA encoding a humanized CD 180 antibody or antigen binding fragment thereof of the present disclosure. Such methods are well known in the art, e.g., as described in U.S. Pat. Nos. 5,580,734, 5,641,670, 5,733,746, and 5,733,761. Also see, e.g., Sambrook et al, (2001) Molecular Cloning: A Laboratory Manual. 3rd Edition, Vol. 1, Cold Spring Harbor Laboratory Press, New York, and Ausubel et al., (2003) Current Protocols in Molecular Biology, Wiley. Those of ordinary skill in the art are knowledgeable in the numerous expression systems available for expression of a nucleic acid encoding a humanized CD 180 antibody or antigen binding fragment thereof. Illustrative of cell cultures useful for the production of the antibodies, and specified portions or variants thereof, are mammalian cells. Mammalian cell systems often will be in the form of monolayers of cells although mammalian cell suspensions or bioreactors can also be used. A number of suitable host cell lines capable of expressing intact glycosylated proteins have been developed in the art, and include the COS-1 (e.g., ATCC CRL 1650), COS-7 (e g., ATCC CRL-1651). HEK293, BHK21 (e.g., ATCC CRL-10), CHO (e g., ATCC CRL 1610) and BSC-1 (e.g., ATCC CRL-26) cell lines, hep G2 cells, P3X63Ag8.653, SP2 / 0-Agl4, HeLa cells and the like, which are readily available from, for example, American Type Culture Collection (ATCC), Manassas, VA. Yeast and bacterial host cells may also be used and are w ell known to those of skill in the art. Other cells useful for production of nucleic acids or proteins ofAttorney Docket No: 01434-0001-00PCT the present disclosure are known and / or available, for instance, from the American Type Culture Collection Catalogue of Cell Lines and hybridomas or other know n or commercial sources.

[0219] Expression vectors can include one or more of the following expression control sequences, such as. but not limited to an origin of replication; a promoter (e.g., late or early SV40 promoters, the CMV promoter (U.S. Pat. Nos. 5,168,062; and 5,385,839), an HSV tk (thymidine kinase) promoter, a pgk (phosphoglycerate kinase) promoter, an EF-1 alpha promoter (U.S. Pat. No. 5,266,491), at least one human immunoglobulin promoter; an enhancer, and / or processing information sites, such as ribosome binding sites, RNA splice sites, polyadenylation sites (e.g., an SV40 large T Ag poly A addition site), and transcriptional terminator sequences). See, e.g., Ausubel et al, supra; Sambrook, et al, supra.

[0220] Expression vectors optionally include at least one selectable marker. Such markers include, e.g., but are not limited to, methotrexate (MTX). dihydrofolate reductase (DHFR, U.S. Pat. Nos. 4,399,216; 4.634,665; 4.656,134; 4,956.288; 5,149.636; and 5,179.017), ampicillin, neomycin (G418), mycophenolic acid, or glutamine synthetase (GS; U.S. Pat. Nos.5,122,464; 5,770,359; and 5,827,739), resistance for eukaryotic cell culture, and tetracycline or ampicillin resistance genes for culturing in E. coli and other bacteria or prokaryotes. Appropriate culture media and conditions for the above-described host cells are known in the art. Suitable vectors will be readily apparent to the skilled artisan. Introduction of a vector construct into a host cell can be effected by calcium phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection or other known methods. Such methods are described in the art, such as Sambrook, supra: Ausubel. supra.

[0221] In some embodiments, the nucleic acid insert is operatively linked to an appropriate promoter. The expression constructs may further contain sites for transcription initiation, termination and, in the transcribed region, a ribosome binding site for translation. The coding portion of the mature transcripts expressed by the constructs may include a translation initiating sequence at the beginning and a termination codon (e.g., UAA, UGA or UAG) appropriately positioned at the end of the mRNA to be translated. In some embodiments, the translation initiation sequence is UAA or UAG. In some embodiments, the expression constructs are used for mammalian or eukaryotic cell expression.

[0222] When eukaryotic host cells are employed, polyadenylation or transcription terminator sequences are typically incorporated into the vector. An example of a terminator sequence is the polyadenylation sequence from the bovine growth hormone gene. Sequences for accurate splicing of the transcript can also be included. An example of a splicing sequence is the VP1 intron from SV40 (Sprague, et al. (1983) J. Virol. 45:773-781). Additionally, geneAttorney Docket No: 01434-0001-00PCT sequences to control replication in the host cell can be incorporated into the vector, as known in the art.XI. Antibody Isolation and Purification

[0223] Humanized anti-CDl 80 antibodies and antigen binding fragments thereof described herein can be recovered and purified from recombinant cell cultures by well-known methods including, but not limited to, protein A purification, ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. High performance liquid chromatography (HPLC) can also be employed for purification. See, e.g., Colligan, Current Protocols in Immunology’, or Current Protocols in Protein Science, John Wiley & Sons, New York. N.Y., (1997-2001).

[0224] Humanized CD 180 antibodies and antigen binding fragment thereof described herein can include purified products, products of chemical synthetic procedures, and products produced by recombinant techniques from a eukary otic host, including, for example, yeast, higher plant, insect and mammalian cells. Depending upon the host employed in a recombinant production procedure, the humanized CD 180 antibody or antigen binding fragment thereof of the present disclosure can be glycosylated or can be non-glycosylated. In some embodiments, the humanized CD 180 antibody or antigen binding fragment thereof is glycosylated. Such methods are described in many standard laboratory manuals, such as Sambrook, supra; Ausubel, supra.XII. Nucleic Acid Molecules

[0225] Nucleic acid molecules of the present disclosure can be in the form of RNA, such as mRNA or any other form, or in the form of DNA, including, but not limited to, cDNA and genomic or other DNA obtained by cloning or produced synthetically, or any combinations thereof. The DNA can be double-stranded or single-stranded, or any combination thereof. Any portion of at least one strand of the DNA or RNA can be the coding strand, also known as the sense strand, or it can be the non-coding strand, also referred to as the anti-sense strand.

[0226] Isolated nucleic acid molecules of the present disclosure can include nucleic acid molecules comprising an open reading frame (ORF), optionally with one or more introns, e.g., but not limited to. at least one specified portion of at least one CDR. as CDR1, CDR2 and / or CDR3 of at least one heavy chain or light chain; nucleic acid moleculesAttorney Docket No: 01434-0001-00PCT comprising the coding sequence for a humanized anti-CD180 antibody or a humanized antiCD 180 antibody variable region or antigen binding fragment thereof; and nucleic acid molecules which comprise a nucleotide sequence substantially different from those described above but which, due to the degeneracy of the genetic code, still encode at least one humanized anti-CD180 antibody or antigen binding fragment thereof as described herein and / or as known in the art. Given that the genetic code is well-known in the art, it is routine for one skilled in the art to generate such degenerate nucleic acid variants that code for specific humanized anti-CD180 antibodies or antigen binding fragment thereof of the present disclosure.[00227J As indicated herein, nucleic acid molecules of the present disclosure which comprise a nucleic acid encoding a humanized anti-CD180 antibody molecule or antigen binding fragment thereof can include, but are not limited to, those encoding the amino acid sequence of a humanized CD 180 antibody fragment, by itself; the coding sequence for the entire antibody or a portion thereof; the coding sequence for a humanized CD180 antibody, fragment or portion, as well as additional sequences, or such as the coding sequence of at least one signal leader or fusion peptide, with or without the aforementioned additional coding sequences, such as at least one intron, together with additional, non-coding sequences, including but not limited to, non-coding 5' and 3' sequences, such as the transcribed, non-translated sequences that play a role in transcription, mRNA processing, including splicing and polyadenylation signals (for example — ribosome binding and stability of mRNA); an additional coding sequence that codes for additional amino acids, such as those that provide additional functionalities. In some embodiments, the sequence encoding a humanized CD 180 antibody or antigen binding fragment thereof can be fused to a marker sequence, such as a sequence encoding a peptide that facilitates purification of the fused antibody comprising a humanized CD 180 antibody fragment or portion.XIII. Construction of Nucleic Acids

[0228] The isolated nucleic acids encoding a humanized CD 180 antibody or antigen binding fragment thereof can be made using (a) recombinant methods, (b) synthetic techniques, (c) purification techniques, or combinations thereof, as well-known in the art. The nucleic acids can conveniently comprise sequences in addition to a polynucleotide of the present disclosure. For example, a multi-cloning site comprising one or more endonuclease restriction sites can be inserted into the nucleic acid to aid in isolation of the polynucleotide. Also, translatable sequences can be inserted to aid in the isolation of the translated polynucleotide of the present disclosure. For example, a hexa-histidine marker sequence provides a convenient means to purify the proteins of the present disclosure. The nucleic acid of the present disclosure — excluding the coding sequence — is optionally a vector, adapter, or linker forAttorney Docket No: 01434-0001-00PCT cloning and / or expression of a polynucleotide of the present disclosure. Additional sequences can be added to such cloning and / or expression sequences to optimize their function in cloning and / or expression, to aid in isolation of the polynucleotide, or to improve the introduction of the polynucleotide into a cell. Use of cloning vectors, expression vectors, adapters, and linkers are well known in the art. (See, e.g., Ausubel, supra; or Sambrook, supra.)

[0229] The isolated nucleic acid compositions encoding a humanized CD 180 antibody or antigen binding fragment thereof, such as RNA, cDNA, genomic DNA, or any combination thereof, can be obtained from biological sources using any number of cloning methodologies known to those of skill in the art. In some embodiments, oligonucleotide probes that selectively hybridize, under stringent conditions, to the polynucleotides of the present disclosure are used to identify the desired sequence in a cDNA or genomic DNA 1 i brary . The isolation of RNA, and construction of cDNA and genomic libraries, is well known to those of ordinary skill in the art. (See, e.g., Ausubel, supra; or Sambrook, supra.)XIV. Therapeutic Applications

[0230] Provided herein are methods of treating disorders associated with cells that express CD180, e.g.. autoimmune and inflammatory diseases and disorders. The cells may or may not express elevated levels of CD 180 relative to cells that are not associated with a disorder of interest. As a result, the disclosure provides a method of treating a subject, for example, a subject having an autoimmune or inflammatory disease or disorder, using the humanized anti-CD180 antibodies or antigen binding fragment thereof described herein. The method comprises administering an effective amount of a humanized anti-CD180 antibody or antigen binding fragment thereof or a composition comprising a humanized anti-CD180 antibody or antigen binding fragment thereof to a subject in need thereof.

[0231] As used herein, the terms “subject’" and “patient" refer to organisms to be treated by the methods of the present disclosure with a humanized CD 180 antibody or antigen binding fragment thereof. Such organisms may include, but are not limited to, mammals (e.g., murines, simians, equines, bovines, porcines, canines, felines, and the like), and humans. In some embodiments, the organism is a human. As used herein, the terms, “treat"’, “treatment” and “treating” include any effect, e.g., lessening, reducing, modulating, ameliorating or eliminating, that results in the improvement of the condition, disease, disorder, and the like, or ameliorating a symptom thereof, such as for example, an autoimmune or inflammatory disease or disorder.

[0232] As used herein, the term “effective amount” refers to the amount of a compound (e g., a humanized anti-CD180 antibody or antigen binding fragment thereof) sufficient to effect beneficial or desired results. An effective amount can be administered in oneAttorney Docket No: 01434-0001-00PCT or more administrations, applications or dosages and is not intended to be limited to a particular formulation or administration route. Generally, a therapeutically effective amount of active component is in the range of 0.1 mg / kg to 100 mg / kg, e.g., 1 mg / kg to 100 mg / kg, 1 mg / kg to 10 mg / kg. The dosage administered can vary depending upon known factors, such as the pharmacodynamic characteristics of the particular agent, and its mode and route of administration; the age, health, and weight of the recipient; the type and extent of disease or indication to be treated, the nature and extent of symptoms, kind of concurrent treatment, frequency of treatment, and the effect desired. The initial dosage can be increased beyond the upper level in order to rapidly achieve the desired blood-level or tissue-level. Alternatively, the initial dosage can be smaller than the optimum, and the daily dosage may be progressively increased during the course of treatment. Dosing frequency can vary7, depending on factors such as route of administration, dosage amount, serum half-life of the humanized CD 180 antibody or antigen binding fragment, and the disease being treated. Exemplary dosing frequencies are once per day, once per week and once every two weeks. Formulation of monoclonal antibody-based drugs is within ordinary skill in the art. In some embodiments, a monoclonal antibody is lyophilized, and then reconstituted in buffered saline, at the time of administration.

[0233] In certain exemplary’ embodiments, the present disclosure provides a method for treating an autoimmune or inflammatory disease or disorder in a subject or patient. In particular embodiments, the present disclosure provides a method for treating an autoimmune or inflammatory' disease or disorder in a human. Exemplary’ examples include, but are not limited to, autoimmune or inflammatory diseases or disorder such as Type 1 diabetes; systemic lupus erythematosus (SLE); autoimmune encephalomyelitis, scleroderma, or psoriasis.

[0234] The methods and compositions of the disclosure can be used in combination with other therapeutic agents and / or modalities. The term administered “in combination,” as used herein, is understood to mean that two (or more) different treatments are delivered to the subject during the course of the subject’s affliction with the disorder, such that the effects of the treatments on the patient overlap at a point in time. In certain embodiments, the delivery' of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as “simultaneous” or “concurrent delivery.” In other embodiments, the delivery of one treatment ends before the delivery' of the other treatment begins. In some embodiments of either case, the treatment is more effective because of combined administration. For example, the second treatment is more effective, e.g., an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in the absence of the first treatment, or the analogous situation is seen with the firstAttorney Docket No: 01434-0001-00PCT treatment. In some embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive (i.e., a synergistic response). The delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered.

[0235] In some embodiments, the methods of the disclosure include administering to the subject a humanized anti-CD180 antibody or antigen binding fragment thereof as described herein, e.g., a composition or preparation, in combination with one or more additional therapies. In one embodiment, the additional therapy may include an antiinflammatory, anti-angiogenic, anti-fibrotic, or anti-proliferative compound, e.g., asteroid, a biologic immunomodulatory, such as an inhibitor of an immune checkpoint molecule, a monoclonal antibody, an antibody fragment, an aptamer, an siRNA, an antisense molecule, a fusion protein, a cytokine, a cytokine receptor, a bronchodilator, a statin, an anti-inflammatory agent (e.g. methotrexate), or an NSAID. In another embodiment, the additional therapy could include combining therapeutics of different classes. The humanized anti-CD180 antibody or antigen binding fragment thereof preparation and the additional therapy can be administered simultaneously or sequentially.

[0236] Throughout the description, where compositions and kits are described as having, including, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are compositions and kits of the present disclosure that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present disclosure that consist essentially of, or consist of, the recited processing and method steps.XV. Pharmaceutical Compositions and Formulations

[0237] For therapeutic use. a humanized anti-CD 180 antibody or antigen binding fragment thereof may be combined with a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” means buffers, carriers, and excipients suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. The carrier(s) should be “acceptable” in the sense of being compatible with the other ingredients of the formulations and not deleterious to the recipient. Pharmaceutically acceptable carriers include buffers, solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is known in the art.Attorney Docket No: 01434-0001-00PCT

[0238] Accordingly, compositions of humanized anti-CD180 antibody or antigen binding fragment thereof can comprise at least one of any suitable excipients, such as, but not limited to, diluent, binder, stabilizer, buffer, salt, lipophilic solvent, preservative, adjuvant or the like. In some embodiments, the excipient is a pharmaceutically acceptable excipient. Nonlimiting examples of, and methods of preparing such sterile solutions are well known in the art, such as, but not limited to, those described in Gennaro, Ed., Remington’s Pharmaceutical Sciences, 18th Edition, Mack Publishing Co. (Easton, Pa.) 1990. Pharmaceutically acceptable carriers can be routinely selected that are suitable for the mode of administration, solubility and / or stability of the antibody molecule, fragment or variant composition as well known in the art or as described herein.

[0239] Pharmaceutical excipients and additives useful in the present composition include but are not limited to proteins, peptides, amino acids, lipids, and carbohydrates (e.g., sugars, including monosaccharides, di-, tri-, tetra-, and oligosaccharides; derivatized sugars such as alditols, aldonic acids, esterified sugars and the like; and polysaccharides or sugar polymers), which can be present singly or in combination, comprising alone or in combination 1-99.99% by weight or volume. Exemplary' protein excipients include serum albumin such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative components, which can also function in a buffering capacity, include alanine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, and the like.

[0240] Carbohydrate excipients suitable for use include, for example, monosaccharides such as fructose, maltose, galactose, glucose, D-mannose, sorbose, and the like; disaccharides, such as lactose, sucrose, trehalose, cellobiose, and the like; polysaccharides, such as raffinose, melezitose, maltodextrins, dextrans, starches, and the like; and alditols, such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol), myoinositol and the like. In some embodiments, the carbohydrate excipient(s) are mannitol, trehalose, and raffinose.

[0241] Antibody compositions can also include a buffer or a pH adjusting agent; ty pically, the buffer is a salt prepared from an organic acid or base. Representative buffers include organic acid salts such as salts of citric acid, acetic acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, or phthalic acid; tris, tromethamine hydrochloride, or phosphate buffers.

[0242] Additionally, antibody molecule compositions can include polymeric excipients / additives such as polyvinylpyrrolidones, ficolls (a polymeric sugar), dextrins (e.g., cyclodextrins, such as 2-hydroxypropyl-cyclodextrin), polyethylene glycols, flavoring agents, antimicrobial agents, sweeteners, antioxidants, antistatic agents, surfactants (e.g.. polysorbatesAttorney Docket No: 01434-0001-00PCT such as TWEEN 20 and TWEEN 80), lipids (e.g., phospholipids, fatty acids), steroids (e.g., cholesterol), and chelating agents (e.g., EDTA).

[0243] These and additional known pharmaceutical excipients and / or additives suitable for use in the compositions according to the disclosure are known in the art, e.g.. as listed in “Remington: The Science & Practice of Pharmacy,’’ 19th ed., Williams & Williams, (1995), and in the “Physician’s Desk Reference,” 52nd ed., Medical Economics, Montvale, N.J. (1998). In some embodiments, the carrier or excipient materials are carbohydrates (e.g., saccharides and alditols) and buffers (e.g., citrate) or polymeric agents.[00244J The present disclosure provides for stable compositions, comprising at least one humanized anti-CD180 antibody or antigen binding fragment thereof in a pharmaceutically acceptable formulation. In some embodiments, the formulation includes a preservative. Preserved formulations contain at least one known preservative or optionally selected from at least one phenol, m-cresol, p-cresol, o-cresol. chlorocresol, benzyl alcohol, phenylmercuric nitrite, phenoxyethanol, formaldehyde, chlorobutanol, magnesium chloride (e.g., hexahydrate), alkylparaben (methyl, ethyl, propyl, butyl and the like), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate and thimerosal, or mixtures thereof in an aqueous diluent. Any suitable concentration or mixture can be used as known in the art, such as 0.001-5%, or any range or value therein. Non-limiting examples include, no preservative, 0.1-2% m-cresol (e.g., 0.2, 0.3, 0.4, 0.5, 0.9, or 1.0%), 0.1-3% benzyl alcohol (e.g., 0.5, 0.9, 1.1., 1.5, 1.9, 2.0, or 2.5%), 0.001-0.5% thimerosal (e.g., 0.005 or 0.01%), 0.001-2.0% phenol (e.g., 0.05, 0.25, 0.28, 0.5, 0.9. or 1.0%), 0.0005-1.0% alkylparaben(s) (e.g., 0.00075, 0.0009, 0.001. 0.002, 0.005, 0.0075, 0.009. 0.01. 0.02. 0.05, 0.075, 0.09, 0.1, 0.2. 0.3. 0.5, 0.75, 0.9, or 1.0%), and the like.

[0245] Pharmaceutical compositions containing a humanized anti-CD180 antibody or antigen binding fragment thereof as disclosed herein can be presented in a unit dosage form and can be prepared by any suitable method. A pharmaceutical composition should be formulated to be compatible with its intended route of administration. Examples of routes of administration are intravenous (IV), intradermal, inhalation, transdermal, topical, transmucosal, and rectal administration. In some embodiments, the route of administration for the humanized anti-CD180 antibody or antigen binding fragment thereof or a pharmaceutical composition thereof is IV infusion. Useful formulations can be prepared by methods known in the pharmaceutical art. For example, see Remington’s Pharmaceutical Sciences (1995) supra.Formulation components suitable for parenteral administration include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens;Attorney Docket No: 01434-0001-00PCT antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as EDTA; buffers such as acetates, citrates or phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose.

[0246] For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL™ (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). The carrier should be stable under the conditions of manufacture and storage, and should be preserved against microorganisms. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof.

[0247] In some embodiments, the pharmaceutical formulations are sterile.Sterilization can be accomplished by any suitable method, e.g., filtration through sterile filtration membranes. Where the composition is lyophilized, filter sterilization can be conducted prior to or following lyophilization and reconstitution.

[0248] The compositions of this disclosure may be in a variety of forms. These include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, and liposomes. The form depends on the intended mode of administration and therapeutic application. Typical compositions are in the form of injectable or infusible solutions. In some embodiments, the mode of administration is parenteral (e.g., intravenous, subcutaneous, intraocular, intraperitoneal, intramuscular). In some embodiments, the preparation is administered by intravenous infusion or injection. In other embodiments, the preparation is administered by intramuscular or subcutaneous injection.

[0249] The phrases "parenteral administration’7and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, subcutaneous, intraarterial, intrathecal, intracapsular, intraorbital, intravitreous, intracardiac, intradermal, intraperitoneal, transtracheal, inhaled, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrastemal injection and infusion.

[0250] The present disclosure also provides a kit, comprising packaging material and at least one vial comprising a powder (e.g., a lyophilizate) or solution of at least one humanized anti-CD180 antibody or antigen binding fragment thereof with the prescribed buffers and / or preservatives, optionally in an aqueous diluent. The aqueous diluent optionally further comprises a pharmaceutically acceptable preservative. Preservatives include those selected from phenol, m-cresol, p-cresol, o-cresol, chlorocresol, benzy l alcohol, alkydparaben (methy l, ethyl, propyl, butyl and the like), benzalkonium chloride, benzethonium chloride, sodium dehydroacetate and thimerosal, or mixtures thereof. The concentration of preservative used in theAttorney Docket No: 01434-0001-00PCT formulation is a concentration sufficient to yield an anti-microbial effect. Such concentrations are dependent on the preservative selected and are readily determined by the skilled artisan.

[0251] Other excipients, e.g. isotonicity' agents, buffers, antioxidants, preservative enhancers, can be optionally added to the diluent. An isotonicity agent, such as glycerin, is commonly used at known concentrations. A physiologically tolerated buffer can be added to provide improved pH control. The formulations can cover a wide range of pHs, such as from about pH 4.0 to about pH 10.0, from about pH 5.0 to about pH 9.0, or about pH 6.0 to about pH 80[00252J Other additives, such as a pharmaceutically acceptable solubilizers like TWEEN 20 (polyoxyethylene (20) sorbitan monolaurate), TWEEN 40 (polyoxyethylene (20) sorbitan monopalmitate), TWEEN 80 (polyoxyethylene (20) sorbitan monooleate), Pluronic F68 (polyoxyethylene poly oxy propylene block copolymers), and PEG (polyethylene glycol) or nonionic surfactants such as polysorbate 20 or 80 or poloxamer 184 or 188. Pluronic® polyls, other block co-polymers, and chelators such as EDTA and EGTA can optionally be added to the formulations or compositions to reduce aggregation. These additives are particularly useful if a pump or plastic container is used to administer the formulation. The presence of pharmaceutically acceptable surfactant mitigates the propensity for the protein to aggregate.

[0253] Various delivery systems can be used to administer humanized antiCD 180 antibodies or antigen binding fragment thereof to a subj ect. In certain exemplary embodiments, administration of a humanized anti-CD180 antibody or antigen binding fragment thereof is by intravenous infusion. In some embodiments, administration is by a two hour intravenous infusion.

[0254] Any of the formulations described above can be stored in a liquid or frozen form and can be optionally subjected to a preservation process. In some embodiments, the formulations described above are lyophilized, i.e., they are subjected to lyophilization. In some embodiments, the formulations described above are subjected to a preservation process, for example, lyophilization, and are subsequently reconstituted with a suitable liquid, for example, water. By lyophilized, it is meant that the composition has been freeze-dried under a vacuum. Lyophilization typically is accomplished by freezing a particular formulation such that the solutes are separated from the solvent(s). The solvent is then removed by sublimation (i.e., primary drying) and next by desorption (i.e., secondary drying).

[0255] The formulations of the present disclosure can be used with the methods described herein or with other methods for treating disease. The humanized anti-CD180 antibody or antigen binding fragment thereof formulations may be further diluted before administration to a subject. In some embodiments, the formulations will be diluted with saline and held in IV bagsAttorney Docket No: 01434-0001-00PCT or syringes before administration to a subject. Accordingly, in some embodiments, the methods for treating an autoimmune or inflammatory7disease or disorder in a subj ect will comprise administering to a subject in need thereof a weekly dose of a pharmaceutical composition comprising a humanized anti-CD180 antibody or antigen binding fragment thereof.

[0256] It will be readily apparent to those skilled in the art that other suitable modifications and adaptations of the methods described herein may be made using suitable equivalents without departing from the scope of the embodiments disclosed herein. Having now described certain embodiments in detail, the same will be more clearly understood by reference to the following examples, which are included for purposes of illustration only and are not intended to be limiting. All patents, patent applications and references described herein are incorporated by reference in their entireties for all purposes.EXAMPLESExample 1: Antibody Generation

[0257] Humanized versions of the murine G28-8 antibody (having heavy and light chain variable region sequences set forth in SEQ ID NOs: 1 and 2, respectively) and the MHR73-11 antibody (having heavy and light chain variable region sequences set forth in SEQ ID NOs:48 and 49, respectively) were prepared. These humanized antibodies are designated hzG28-8.X and hzMHR73-l 1-X, respectively. For humanization, the murine antibody framework sequences were compared to the sequences of human antibody germline genes in V-Base (MRC, Center for Protein Engineering). Human germline frameworks with the highest framework homology, VH1-03 (SEQ ID NO:38) and VKI-02 (SEQ ID NO:40), were selected for Kabat-defined CDR-grafting. Three-dimensional structural models were generated to assist in the selection of residues in the murine framework regions that may be necessary to restore any decreased binding or functional activity resulting from the CDR-grafting. Several amino acid substitutions (back mutations) were evaluated in the heavy chain variable region (T28S, R66K, V67A, I69L, R71V) and light chain variable region (I48V, D70Q). Substitutions G64V and Y49N were also evaluated in G28-8 and MHR73-11 variable light regions, respectively. The heavy chain variable regions contain a potential deamidation site in CDRH2 (by Kabat numbering). Substitutions N54S, N54Q and G55A were included in hzG28-8 and hzMHR73-l 1 humanized sequences individually to test the effects of removing this motif.

[0258] Antibodies were generated using either a contract research organization or produced using in-house facilities. Humanized variable region sequences were ordered from Integrated DNA Technologies Inc. and cloned into vector pcDNA3.4 containing either theAttorney Docket No: 01434-0001-00PCT human kappa light chain (SEQ ID NO:44) or IgGl.l (SEQ ID NO:43) sequences. IgGl.l is a null effector variant of wild-type IgGl that has substitutions resulting in the reduction of Fey receptor binding and abi lity to fix complement (Gross et al., Immunity, 15:289-302 (2001)). The variable domains were fused to antibody constant domains using NEBuilder® HiFi DNA Assembly and transformed into ToplO Chemically Competent E. coli. Colonies were selected and expanded in Luria Broth and isolated using Qiagen’s HiSpeed Plasmid Maxi Kit according to the manufacturer’s instructions. Maxi preps were sequenced and the desired clones were selected. Plasmids were transfected into ExpiCho cells (ThermoFisher) according to manufacturer’s standard protocol instructions and supernatants were harvested for purification. Antibodies were purified from the filtered conditioned media using a combination of Mab Select PrismA™ affinity chromatography resin and SUPERDEX® 200 Size exclusion chromatography. Yields were estimated by absorbance at UV-A280 nm, and quality of the antibody preparations was evaluated by analytical size exclusion high performance liquid chromatography.

[0259] The murine and humanized antibodies are described in the following Tables 7 and 8. The prefix “hz” indicates a humanized VH or VL sequence.Table 7. Parental and Humanized G28-8 AntibodiesName VH VL VH Seq VL ID No. Seq ID No. G28-8.C G28-8VH G28-8VL 1 2 hzG28-8.1 hzG28-8 VH1 -03 hzG28-8 VKI-O2 3 4 hzG28-8.2 hzG28-8 VH1 -03 T28S hzG28-8 VKI-O2 5 4 hzG28-8.3 hzG28-8 VH1 -03 R66K, V67A, I69L, hzG28-8 VKI-O2 6 4R71VhzG28-8.4 hzG28-8 VH1 -03 T28S, R66K, V67A, hzG28-8 VKI-O2 7 4I69L, R71VhzG28- hzG28-8 VH1 -03 G28-8VL 3 2 8.60hzG28- hzG28-8 VH1 -03 T28S G28-8VL 5 2 8.61hzG28- hzG28-8 VH1 -03 R66K, V67A, I69L, G28-8VL 6 2 8.62 R71VhzG28- hzG28-8 VH1 -03 T28S, R66K, V67A, G28-8VL 7 2 8.63 I69L, R71VhzG28- G28-8VH hzG28-8 VKI-O2 1 4 8.64hzG28- hzG28-8 VH1 -03 hzG28-8 VKI-O2 I48V, G64V, D70Q 3 8Attorney Docket No: 01434-0001-00PCT 8.69hzG28- hzG28-8 VH1 -03 R66K, V67A, I69L, hzG28-8 VKI-O2 I48V, G64V, D70Q 6 8 8.71 R71VhzG28- G28-8VH hzG28-8 VKI-O2 I48V, G64V, D70Q 1 8 8.81hzG28- hzG28-8 VH1 -03 I69L, R71V hzG28-8 VKI-O2 I48V, G64V, D70Q 9 8 8.87hzG28- hzG28-8 VH1 -03 R66K, V67A hzG28-8 VKI-O2 I48V, G64V, D70Q 10 8 8.88G28-8.L1 G28-8VH N54S G28-8VL 11 2 G28-8.L2 G28-8VH N54Q G28-8VL 12 2 G28-8.L3 G28-8VH G55A G28-8VL 13 2 hzG28- hzG28-8 VH1 -03 N54Q, R66K, V67A, hzG28-8 VKI-O2 I48V, G64V, D70Q 14 8 8.93 I69L, R71VhzG28- hzG28-8 VH1 -03 N54S, R66K, V67A, hzG28-8 VKI-O2 I48V, G64V, D70Q 15 8 8.97 I69L, R71VhzG28- hzG28-8 VH1 -03 G55A, R66K, V67A, hzG28-8 VKI-O2 I48V, G64V, D70Q 16 8 8.98 I69L, R71VhzG28- hzG28-8 VH1 -03 R66K, V67A, I69L, hzG28-8 VKI-O2 I48V, D70Q 6 45 8.99 R71VhzG28- hzG28-8 VH1 -03 R66K, V67A, I69L, hzG28-8 VKI-O2 I48V 6 46 8.100 R71VhzG28- hzG28-8 VH1 -03 R66K, V67A, I69L, hzG28-8 VKI-O2 D70Q 6 47 8.101 R71VhzG28- hzG28-8 VH1 -03 R66K, V67A, I69L, hzG28-8 VKI-O2 G64V 6 81 8.113 R71VhzG28- hzG28-8 VH1 -03 G55A, R66K, V67A, hzG28-8 VKI-O2 G64V 16 81 8.114 I69L, R71VTable 8. Parental and Humanized MHR73-11 Antibodies Name VH VL VH VLSeq Seq ID ID MHR73-11 MHR73-11 VH MHR73-11 VL 48 49 hzMHR73-11-1 hzMHR73-11 VH1-03 hzMHR73-11 VKI-O2 66 68 hzMHR73-11-2 hzMHR73-11 VH1-03 hzMHR73-11 VKI-O2 I48V, Y49N, 66 69 D70QhzMHR73-11-3 hzMHR73-11 VH1-03 hzMHR73-11 VKI-O2 I48V, Y49N 66 70 hzMHR73-11-4 hzMHR73-11 VH1-03 hzMHR73-11 VKI-O2 D70Q 66 71 hzMHR73-11-5 hzMHR73-11 VH1 -03 R66K, V67A, hzMHR73-11 VKI-O2 67 68 I69L, R71VhzMHR73-11-6 hzMHR73-11 VH1-03 R66K, V67A, hzMHR73-11 VKI-O2 I48V, Y49N, 67 69 I69L, R71V D70QAttorney Docket No: 01434-0001-00PCT hzMHR73-11-7 hzMHR73-11 VH1-03 R66K, V67A, hzMHR73-11 VKI-O2 I48V, Y49N 67 70 I69L, R71VhzMHR73-11-8 hzMHR73-11 VH1 -03 R66K, V67A, hzMHR73-11 VKI-O2 D70Q 67 71 I69L, R71VhzMHR73-11-9 hzMHR73-11 VH1-03 N54Q hzMHR73-11 VKI-O2 82 68 hzMHR73-11- hzMHR73-11 VH1-03 N54S hzMHR73-11 VKI-O2 64 68 10hzMHR73-11- hzMHR73-11 VH1 -03 G55A hzMHR73-11 VKI-O2 65 68 11Example 2: Binding of CD 180 antibodies to a human CD 180 / MDI expressing cell line

[0260] Select humanized anti-CD180 antibodies were tested for the ability to bind to 293T cells co-expressing human CD 180 and human MD1. Cells were stained with a titration of each anti-CD180 antibody. Bound antibodies were detected with a fluorescently conjugated anti-human IgG polyclonal secondary antibody (anti-hlgG PE or APC). The fluorescence intensity was measured using an Attune NxT flow cytometer (ThermoFisher). The geometric mean fluorescence intensity7(gMFI) of anti-CD180 antibody staining was calculated for live cells (FlowJo™ software). EC50s for antibody binding were calculated using GraphPad Prism software. Initial studies were carried out using antibodies in the form of supernatants from cultures of host cells transfected with recombinant antibody expression vectors.

[0261] As shown in Table 9, the anti-CD180 antibodies derived from parental antibody G28-8 bound to CD180 / MDI expressing 293T cells with nanomolar EC50s. Within the variability7of the assay, no significant loss in binding yvas observed across any of the antibodies tested. Referring to Table 9 and Figure 1, a series of antibodies having changes in the heavy chain variable region framework sequences and no changes in the light chain variable region framework sequences were evaluated. These antibodies yvere hzG28-8.1 (having a CDR- grafted human VH1-03 heavy chain variable region (no framework back mutations) and a CDR- grafted light chain variable region (no framework back mutations)). hzG28-8.2 (having a CDR- grafted human heavy chain variable region containing a T28S framework back mutation and a CDR-grafted light chain variable region (no framework back mutations)), hzG28-8.3 (having a CDR-grafted human heavy chain variable region containing R66K, V67A, I69L and R71V framework back mutations and a CDR-grafted light chain variable region (no framework back mutations)), and hzG28-8.4 (having a CDR-grafted human heavy chain variable region containing the T28S, R66K, V67A, I69L and R71V framework back mutations and a CDR- grafted light chain variable region (no framework back mutations). These antibodies had comparable binding.Attorney Docket No: 01434-0001-00PCT

[0262] Referring to Table 10, combining the same CDR grafted heavy chain variable regions with the murine light chain variable region produced similar results (see antibodies hzG28-8.60 to hzG28-8.63). When the murine G28-8 heavy chain variable region was combined with a CDR-grafted human light chain variable region with no back mutations (see antibody hzG28-8.64), the resulting antibody had an EC50 comparable to the parental chimeric antibody.

[0263] The binding of antibody hzG28-8.69 (containing a CDR-grafted human heavy chain variable region (no framework back mutations) and a CDR grafted light chain variable region containing three framework back mutations, I48V, G64V and D70Q) was compared to the binding of antibody hzG28-8.71 (having a CDR-grafted human heavy chain variable regions containing R66K, V67A, I69L and R71V framework back mutations and the CDR grafted light chain variable region containing three framework back mutations, I48V. G64V and D70Q), and to the binding of antibody hzG28-8.81 (having the murine G28-8 heavy chain variable region and a CDR grafted light chain variable region containing three framework back mutations, I48V, G64V and D70Q). All three antibodies, antibodies hzG28-8.69, hzG28-8.71 and hzG28-8.81, exhibited similar binding.

[0264] Antibodies hzG28-8.87 and hzG28-8.88 were then assayed for binding to determine if a reduced set of framework back mutations in the heavy chain variable region (I69L and R71V, or R66K and V67A, respectively) in combination with a light chain variable region containing three framework back mutations (I48V, G64V and D70Q) to determine their EC50 values. Both antibodies exhibited similar binding as compared to antibody hzG28-8.71 (containing all four framework back mutations in the heavy chain variable region).

[0265] A deamidation site is present in CDRH2 (Kabat). To determine whether the deamidation site could be removed without affecting binding, antibodies G28-8.L1 to G28-8.L3 were tested. These antibodies contained N54S, N54Q and G55A substitutions, respectively, in CDRH2 in the murine heavy chain variable region. Antibody G28-8.L1 had reduced binding while antibodies G28-8.L2 and G28-8.L3 exhibited binding similar to the parental antibody. These three CDRH2 substitutions were then introduced into the hzG28-8.71 heavy chain variable region to yield antibodies hzG28-8.93 (N54Q), hzG28-8.97 (N54S) and hzG28-8.98 (G55A). Antibodies hzG28-8.93 (N54Q) and hzG28-8.98 (G55A) showed good binding activity compared to antibody hzG28-8.97 (N54S), indicating that some substitutions removing the deamidation site could be removed while retaining good binding activity.Attorney Docket No: 01434-0001-00PCTTable 9. Characterization of humanized G28-8 antibodies Name Cell Cell CD86 pAKT pS6Binding binding Upreg*. Upreg* Upreg*ECso, Curve MaxnN (%chimeric)G28-8.C 0.7-1.9 100 +++ +++ +++hzG28- 1.6-1.9 96-98 ++ + +8.1hzG28- 1.8-2 97-98 ++ + +8.2hzG28- 1.8-2.3 98-102 ++ ++ ++8.3hzG28- 1.5-1.8 97-102 ++ ++ ++8.4*“+++” within 10% activity chimeric, “++” less than 10% but not less than30% of the activity of chimeric, “+” less than 30% activity chimericTable 10. Characterization of humanized G28-8 antibodies Name Cell Cell CD86 pAKT pAKT pS6 pS6Binding binding Upreg*. EC50 Curve Max EC50 Curve Max ECso Curve Max nM (% chimeric) nM (% nM (% chimeric) chimeric)G28-8.C 0.7-1.9 100 +++ 1.2-1 .8 100 3.5-3.7 100 hzG28- 1.7 100 +++ ND ND ND ND8.60hzG28- 1.6 100 +++ ND ND ND ND8.61hzG28- 1.6 97 +++ 1.2 111 3.3 1358.62hzG28- 1.5 100 +++ 1.1 107 2.6 1068.63hzG28- 1.3 96 ++ ND ND ND ND8.64hzG28- 1.5 98 ND 1.2 88 2.9 828.69hzG28- 0.7-1.5 95 +++ 1.0-1 .2 112-114 2.5-3.0 143-147 8.71hzG28- 1.0 95 ND ND ND ND NDAttorney Docket No: 01434-0001-00PCT 8.81hzG28- 0.9 97 ND 1.2 96 3.3 1178.87hzG28- 0.8 92 ND 1.2 84 3.8 978.88G28-8.L1 1.4 82 ND ND ND ND NDG28-8.L2 1.6 94 ND ND ND ND NDG28-8.L3 2.2 97 ND ND ND ND ND hzG28- 0.8 85 + None None None8.93hzG28- 0.7 72 + None 9 None 148.97hzG28- 1.0 99 +++ 1.7 64 4.0 988.98ND means not determined; none means no activity was detected; “+++” within 10% activity chimeric, “++” less than 10% but not less than 30% of the activity of chimeric, “+” less than 30% activity chimeric.

[0266] Referring to Table 11, antibodies hzG28-8.99, hzG28-8.100 and hzG28-8.101 were assayed to determine whether antibodies having a reduced set of framework back mutations in the light chain variable region would retain binding activity when combined with the heavy chain variable region containing the framework back mutations R66K, V67A, I69L and R71V. Antibody hzG28-8.99, containing the I48V and D70Q substitutions, exhibited similar binding to antibodies having the I48V or D70Q substitutions alone (hzG28-8.100 and hzG28-8.101, respectively).Table 11. Characterization of humanized G28-8 antibodies Name Cell Cell binding pAKT pAKT pS6 pS6Binding Curve Max (% ECso Curve Max ECso Curve Max ECso, nM chimeric) nM (%chimeric) nM (%chimeric) G28-8.C 0.410 100 0.94 100 2.42 100hzG28-8.71 0.477 89 0.99 118 2.21 134hzG28-8.99 0.495 90 3.28 70 7.59 79hzG28- 8.100 0.569 88 3.55 64 7.62 74hzG28- 8.101 0.590 93 2.93 64 6.53 74Attorney Docket No: 01434-0001-00PCT

[0267] Referring to Table 12, the binding activity of antibodies hzG28-8.113 and hzG28-8.114 (having a heavy chain variable region containing four framework back mutations (R66K. V67A, I69L and R71V or the R66K, V67A. I69L and R71V and the G55A substitution modifying the deamidation site, respectively) and a light chain variable region having only the G64V back mutation) were compared to that of antibodies hzG28-8.71 and hzG28-8.98.Antibody hzG28-8.113 had comparable cell binding activity to that of antibody hzG28-8.71.Table 12. Characterization of humanized G28-8 antibodies Name Cell Cell Binding CD86* pAKT pAKT pS6 pS6 Binding Curve Max Upreg ECso Curve Max ECso Curve Max ECso, nM (%chimeric) nM (%chimeric) nM (%chimeric) G28-8.C 1.28 100 +++ 1.2 100 2.63 100 hzG28- 1.01 98-100+++ 1.3 112 2.7 124 8.71hzG28- 0.98 87-98+++ 1.2 113 2.5 127 8.98hzG28- 0.75 98-100+++ 1.2 117 2.5 137 8.113hzG28-8- 1.06 98-100+++ 1.3 103 2.8 124 114■■+++” within 10% activity chimeric. “++” less than 10% but not less than 30% of the activity of chimeric, “+” less than 30% activity' chimeric

[0268] Similar binding studies were conducted on humanized versions of murine MHR73-11 antibody. These antibodies bound to CD180 / MD1 expressing 293T cells with nanomolar EC50s. Referring to Table 13, the cell binding activity of antibody hzMHR73-ll-l (having a CDR grafted heavy and light chain variable region with no framework back mutations) was compared to that of antibody hzMHR73-l 1-2 (having a CDR grafted heavy chain variable region (no framework back mutations) and a light chain variable region having I48V. Y49N and D70Q framework back mutations). The 48V, Y49N and D70Q framework back mutations were predicted to restore any decreased binding. Antibody hzMHR73-l 1-1, having no back mutations, exhibited similar cell binding to antibody hzMHR73-l 1-2.Antibodies hzMHR73-l 1-3 and hzMHR73-l 1-4 were assayed to determine the effect of a reduced set of light chain framework back mutations on binding. Reducing the number of light chain variable region framework back mutations somewhat improved binding activity; antibody hzMHR73-l 1-3 (having I48V and Y49N back mutations) had similar binding to antibodyAttorney Docket No: 01434-0001-00PCT hzMHR73-l 1-4 (having a D70Q back mutation alone). Antibodies hzMHR73-l 1-5 to hzMHR73-ll-8 were then assayed to compare the binding activities of antibodies containing the same heavy chain variable region framework back mutations as in antibody hzG28-8.71 (R66K, V67A, I69L and R71V) in combination with a light chain variable region containing no framework back mutations, the I48V, Y49N and D70Q framework back mutations or the reduced sets of back mutations, respectively. Antibodies hzMHR73-l 1-6 and hzMHR73-l 1-7, containing all three back mutations or the I48V and Y49N back mutations, exhibited the similar and slightly better binding.Referring to Table 14 and Figure 6, antibody hzMHR73-l 1-1 (having a CDR grafted heavy and light chain variable region with no framework back mutations) was compared to hzMHR73-l 1-9 (having aN54Q substitution in the heavy chain variable region and a light chain variable region with no back mutations) and hzMHR73-l 1-11 (having a G55A substitution in the heavy chain variable region and a light chain variable region with no back mutations). The cell binding of antibodies hzMHR73-l 1 -9 and hzMHR73-l 1-11 was similar to that of hzMHR73-l 1-1.Table 13. Characterization of humanized MHR73-11 antibodies Name Cell Binding Cell Binding CD86* pAKT pAKT pS6 pS6ECso nM Curve Max Upreg ECso Curve Max ECso Curve Max (° / ochimeric) nM (% chimeric) nM (° / ochimeric) MHR73-11 1.2 100 +++ 1.4-2.0 100 3.1-3.7 100 hzMHR73-11- 1.41 93 +++ 1.3-1 .4 100-101 2.93 103-1051hzMHR73-11- 1.57 86 ND 1.48 98 3.19 1012hzMHR73-11- 1.34 92 ND 1.72 98 3.59 993hzMHR73-11- 1.27 93 ND 1.24 95 2.8 974hzMHR73-11- 1.58 99 ND 1.5 101 3.15 1065hzMHR73-11- 1.36 93 ND 1.32 99 2.84 1036hzMHR73-11- 1.24 91 ND 1.94 96 3.7 1027hzMHR73-11- 1.22 92 ND 1.1...

Claims

Attorney Docket No: 01434-0001-00PCT What is claimed:

1. A humanized antibody or antigen-binding fragment thereof that specifically binds to human CD 180. the antibody or antigen-binding fragment comprising a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises CDRH1, CDRH2, and CDRH3, wherein each CDR comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% , or 100% identity to the amino acid sequence or 0, 1, or 2 amino acid mutations relative to the amino acid sequence set forth in:a) CDRH1: SEQ ID NO:17, 72, 50 or 77; CDRH2: SEQ ID NO:18, 84,51, or 89; and CDRH3: SEQ ID NO: 19 or 52; according to Kabat numbering; or b) CDRH1: SEQ ID NO:25, 55, 74 or 78; CDRH2: SEQ ID NO:26 or 85; and CDRH3: SEQ ID NO:27 or 56; according to IMGT numbering;the light chain variable region comprises CDRL1, CDRL2, and CDRL3, wherein each CDR comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%, or 100% identity to the amino acid sequence or 0, 1, or 2 amino acid mutations relative to the amino acid sequence set forth in:c) CDRL1: SEQ ID NO:20 or 53; CDRL2: SEQ ID NO:21; and CDRL3: SEQ ID NO:22 or 54; according to Kabat numbering; ord) CDRL1: SEQ ID NO:28 or 57; CDRL2: SEQ ID NO:29; and CDRL3: SEQ ID NO:22 or 54; according to IMGT numbering;the heavy chain variable region comprises human IGVH1-03 / JH6 framework regions having an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences of SEQ ID NOs:38 and 39; the light chain variable region comprises human IGVH1-03 / JH6 framework regions having an amino acid sequence having at least 80%. 85%. 90%. 91%, 92%. 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequences of SEQ ID NOs:40 and 41; position 55 in CDRH2 is optionally alanine according to Kabat numbering and / or position 54 in CDRH2 is optionally serine or glutamine according to Kabat numbering; andat least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering.

2. The humanized antibody or antigen-binding fragment thereof of claim 1, w herein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 have the amino acid sequences set forth in one of the sets of amino acid sequences selected from:Attorney Docket No: 01434-0001-00PCT i. SEQ ID NOs: 17, 18, 19, 20, 21, and 22, according to Kabat numbering; ii. SEQ ID NOs: 17, 84, 19, 20, 21, and 22, according to Kabat numbering; iii. SEQ ID NOs:72, 18, 19, 20, 21, and 22, according to Kabat numbering; iv. SEQ ID NOs: 72, 84, 19, 20, 21, and 22 according to Kabat numbering; v. SEQ ID NOs:25, 26, 27, 28, 29, and 22, according to IMGT numbering; vi. SEQ ID NOs: 25, 85, 27, 28, 29, and 22, according to IMGT numbering; vii. SEQ ID NOs: 74, 26, 27, 28, 29, and 22, according to IMGT numbering; viii. SEQ ID NOs: 74, 85, 27, 28, 29, and 22, according to IMGT numbering; ix. SEQ ID NOs:50, 51, 52, 53, 21 and 54, according to Kabat numbering; x. SEQ ID NOs:50, 89, 52, 53, 21 and 54, according to Kabat numbering; xi. SEQ ID NOs:77, 51, 52, 53, 21 and 54, according to Kabat numbering; xii. SEQ ID NOs:77, 89, 52, 53, 21 and 54, according to Kabat numbering; xiii. SEQ ID NOs:55, 26, 56, 57, 29, and 54, according to IMGT numbering; xiv. SEQ ID NOs:55, 85, 56, 57, 29, and 54, according to IMGT numbering; xv. SEQ ID NOs:78, 26, 56, 57, 29, and 54, according to IMGT numbering;andxvi. SEQ ID NOs:78, 85, 56, 57, 29, and 54, according to IMGT numbering.

3. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 or 2, wherein:the heavy chain variable region comprises an amino acid sequence having at least 80%.85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:3, 5, 6, 7, 9, 10, 14, 15, 16, 66, 67, 82, 64 or 65;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:4, 8, 45, 46, 47, 81, 68, 69, 70, or 71; and at least one of position 48 is valine or isoleucine, position 64 is valine or glycine and position 70 is glutamine or aspartate in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

4. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 3, wherein HCDR1, HCDR2, HCDR3, LCDR1. LCDR2 and LCDR3 have the amino acid sequences set forth in one of the sets of amino acid sequences selected from:Attorney Docket No: 01434-0001-00PCT i. SEQ ID NOs: 17, 18, 19, 20, 21, and 22, according to Kabat numbering; ii. SEQ ID NOs:72, 18, 19, 20, 21, and 22, according to Kabat numbering; iii. SEQ ID NOs:25, 26, 27, 28, 29, and 22, according to IMGT numbering;andiv. SEQ ID NOs:74, 26, 27, 28, 29, and 22, according to IMGT numbering; the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:3, 5, 6, 7. 9, 10, 14, 15. or 16;the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NOs:4, 8, 45, 46, 47 and 81;at least one of position 48 is valine, position 64 is valine and position 70 is glutamine in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

5. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 4. wherein at least one of position 28 is serine, position 66 is lysine, position 67 is alanine, position 69 is leucine, and position 71 is valine in the heavy chain variable region, according to Kabat numbering; orposition 64 is valine, position 48 is optionally valine and position 70 is optionally glutamine in the light chain variable region, according to Kabat numbering; position 55 in CDRH2 is optionally alanine according to Kabat numbering.

6. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 3, wherein HCDR1, HCDR2, HCDR3, LCDR1. LCDR2 and LCDR3 have the amino acid sequences set forth one of the sets of amino acid sequences selected from:i. SEQ ID NOs:50, 51, 52, 53, 21 and 54, according to Kabat numbering; ii. SEQ ID NOs:77, 51, 52, 53, 21 and 54, according to Kabat numbering; iii. SEQ ID NOs:55, 26, 56, 57, 29, and 54, according to IMGT numbering;andiv. SEQ ID NOs:78, 26, 56, 57, 29, and 54, according to IMGT numbering; wherein the heavy chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:66, 67, 82. 64 and 65;Attorney Docket No: 01434-0001-00PCT the light chain variable region comprises an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence selected from SEQ ID NOs:68, 69, 70 and 71;at least one of, at least two of or all of position 48 is isoleucine, position 49 is phenylalanine or asparagine, position 64 is glycine or valine and position 70 is aspartate in the light chain variable region, according to Kabat numbering; and position 55 in CDRH2 is optionally alanine according to Kabat numbering.

7. The humanized antibody or antigen-binding fragment thereof of any one of claims 1, 2, 3, 5 and 6, wherein position 48 is isoleucine, position 49 is phenylalanine or asparagine, position 64 is glycine or valine and position 70 is aspartate in the light chain variable region, according to Kabat numbering.

8. The humanized antibody or antigen-binding fragment thereof of any one of claims 1, 2, 3, 5 and 6, wherein position 64 is valine, according to Kabat numbering.

9. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 5, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:17, 18, and 19, respectively, according to Kabat numbering, and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:20, 21 and 22, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

10. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 6, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:

72. 18, and 19. respectively, according to Kabat numbering; and LCDR1, LCDR2. and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:20, 21 and 22, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

11. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 6, wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:25, 26, and 27, respectively, according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:28, 29, and 22,Attorney Docket No: 01434-0001-00PCT respectively according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

12. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 6. wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:74, 26, and 27, respectively according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:28, 29, and 22, respectively according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

13. The humanized antibody or antigen-binding fragment thereof of any one of claims 1, 2, 3, and 5 to 8, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:50, 51, and 52, respectively, according to Kabat numbering; and LCDR1.LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:53, 21 and 54, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

14. The humanized antibody or antigen-binding fragment thereof of any one of claims 1, 2, 3, and 5 to 8, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:77, 51, and 52, respectively, according to Kabat numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:

53. 21 and 54, respectively, according to Kabat numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

15. The humanized antibody or antigen-binding fragment thereof of any one of claims 1, 2, 3, and 5 to 8. wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:55, 26, and 56, respectively, according to IMGT numbering; and LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:57, 29, and 54, respectively, according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

16. The humanized antibody or antigen-binding fragment thereof of any one of claims 1, 2, 3, and 7 to 9, wherein the HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:78, 26, and 56, respectively, according to IMGT numbering; and LCDR1, LCDR2. and LCDR3 comprise the amino acid sequences set forth in SEQ ID NOs:

57.

29. andAttorney Docket No: 01434-0001-00PCT 54, respectively, according to IMGT numbering; wherein position 55 in CDRH2 is optionally alanine according to Kabat numbering.

17. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 16, wherein:a) the heavy chain variable region comprises glutamine at position 54;b) the heavy chain variable region comprises serine at position 54; orc) the heavy chain variable region comprises alanine at position 55;according to Kabat numbering.

18. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 17, wherein the heavy chain variable region comprises alanine at position 55; according to Kabat numbering.

19. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 5, and 9 to 12, wherein:a) the heavy chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:3, 5, 6, 7, 9, 10, 14, 15 or 16; andb) the light chain variable region comprises an amino acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in the amino acid sequences set forth in SEQ ID NO:4, 8, 45, 46, 47 or 81.

20. The humanized antibody or antigen-binding fragment thereof of claim 19, wherein the heavy chain variable region and the light chain variable region comprise amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences set forth in:a) SEQ ID NO:6 and SEQ ID NO:8, respectively;b) SEQ ID NO: 16 and SEQ ID NO:8, respectively;c) SEQ ID NO:6 and SEQ ID NO:81, respectively;d) SEQ ID NO: 16 and SEQ ID NO:81, respectively;e) SEQ ID NO: 14 and 8, respectively;f) SEQ ID NO: 15 and 8, respectively;g) SEQ ID NO:6 and 45, respectively;Attorney Docket No: 01434-0001-00PCT h) SEQ ID NO:6 and 46, respectively; ori) SEQ ID NO:6 and 47, respectively.

21. The humanized antibody or antigen-binding fragment thereof of claim 19 or 20, wherein the heavy chain variable region and the light chain variable region comprise the amino acid sequences set forth in:a) SEQ ID NO:6 and SEQ ID NO:8, respectively;b) SEQ ID NO: 16 and SEQ ID NO:8, respectively;c) SEQ ID NO:6 and SEQ ID NO:81, respectively;d) SEQ ID NO: 16 and SEQ ID NO:81, respectively;e) SEQ ID NO: 14 and 8, respectively;f) SEQ ID NO: 15 and 8, respectively;g) SEQ ID NO:6 and 45, respectively;h) SEQ ID NO:6 and 46, respectively; ori) SEQ ID NO:6 and 47, respectively.

22. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 3.5 to 8, and 13 to 18, wherein:a) the heavy chain variable region comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in SEQ ID NO:64, 65.

66. 67, or 82; andb) the light chain variable region comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to an amino acid sequence set forth in the amino acid sequences set forth in SEQ ID NO:68, 69, 70 or 71.

23. The humanized antibody or antigen-binding fragment thereof of claim 22, wherein the heavy chain variable region and the light chain variable region comprise amino acid sequences having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to the amino acid sequences set forth in:a) SEQ ID NO:66 and SEQ ID NO:68, respectively;b) SEQ ID NO:67 and SEQ ID NO:71, respectively;c) SEQ ID NO:64 and SEQ ID NO:68, respectively;d) SEQ ID NO:82 and SEQ ID NO:68, respectively; ore) SEQ ID NO:65 and SEQ ID NO:68, respectively.Attorney Docket No: 01434-0001-00PCT 24. The humanized antibody or antigen-binding fragment thereof of claim 22 or 23, wherein the heavy chain variable region and the light chain variable region comprise amino acid sequences set forth in:a) SEQ ID NO:66 and SEQ ID NO:68, respectivelyb) SEQ ID NO:67 and SEQ ID NO:71, respectively:c) SEQ ID NO:64 and SEQ ID NO:68, respectively;d) SEQ ID NO:82 and SEQ ID NO:68, respectively; ore) SEQ ID NO:65 and SEQ ID NO:68, respectively.

25. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 24, further comprising a heavy chain constant region attached to the heavy chain variable region and a light chain constant region attached to the light chain variable region.

26. The humanized antibody or antigen-binding fragment thereof of claim 25, wherein the heavy chain constant region is an IgG, IgA, or IgM region and the light chain constant region is a kappa region.

27. The humanized antibody or antigen binding fragment thereof of claim 26, wherein the heavy chain constant region is an IgG region.

28. The humanized antibody or antigen-binding fragment thereof of claim 27, wherein the heavy chain constant region is an IgGl region.

29. The humanized antibody or antigen-binding fragment thereof of any one of claims 26 to 28, wherein the heavy chain constant region comprises the amino acid sequence set forth in SEQ ID NO:42.

30. The humanized antibody or antigen-binding fragment thereof of claim 27, wherein the heavy chain constant region is an IgG4 region, optionally wherein the IgG4 region comprises an S228P substitution.

31. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 29, wherein the antibody or antigen-binding fragment thereof has reduced antibody dependent cellular cytotoxicity (ADCC).Attorney Docket No: 01434-0001-00PCT 32. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 31, wherein the antibody or antigen-binding fragment thereof has reduced antibody dependent cellular phagocytosis activity (ADCP).

33. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 32, wherein the antibody or antigen-binding fragment thereof has reduced complementdependent cytotoxicity' activity7(CDC).

34. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 33, comprising a IgGl heavy chain constant region having amino acid substitutions selected from (a) L234A L235A (LALA); (b) L234A, L235A, and P329G (LALA-PG); (c) L234A, L235E, G237A, A330S and P331S; or (d) L234A, L235A and G237A; or comprising an IgG4 heavy chain constant region having amino acid substitutions (e) S288P.

35. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 25 and 31 to 34, wherein the heavy chain constant region is an Fc null.

36. The humanized antibody or antigen-binding fragment thereof of any one of claims 31 to 35, wherein the heavy7chain constant region comprises the amino acid sequence set forth in SEQ ID NO:43.

37. The antibody or antigen-binding fragment thereof of any one of claims 1 to 36, wherein the antibody7or antigen-binding fragment thereof is a Fab, a Fab', a F(ab')2, a Fv fragment, a diabody, a single-chain antibody, an scFv fragment or an scFv-Fc.

38. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 37, wherein the antibody or antigen-binding fragment thereof is monospecific, bispecific, monovalent or bivalent.

39. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 38, wherein the antibody or antigen-binding fragment thereof induces activation of antigen presenting cells.

40. The humanized antibody or antigen-binding fragment thereof of claim 39, wherein activation of antigen presenting cells is CD86 upregulation.Attorney Docket No: 01434-0001-00PCT41. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 40, wherein the antibody or antigen-binding fragment thereof, induces increased phosphorylation of protein AKT in antigen presenting cells.

42. The humanized antibody or antigen-binding fragment thereof of any one of claims 1 to 41, wherein the antibody or antigen-binding fragment thereof, induces increased phosphorylation of protein S6 in antigen presenting cells.

43. A pharmaceutical composition comprising the humanized antibody or antigen-binding fragment thereof of any of claims 1 to 42 and a pharmaceutically acceptable carrier.

44. A method of treating an autoimmune or inflammatory disease or disorder comprising administering to a subject in need thereof an effective amount of the antibody or antigen binding fragment thereof of any of claims 1 to 42 or the pharmaceutical composition of claim 43.

45. The method of claim 44, wherein the subject has an inflammatory condition.

46. The method of claim 44 or 45, wherein the subject has Type 1 diabetes.

47. The method of claim 44 or 45, wherein the subject has systemic lupus erythematosus (SLE).

48. The method of claim 44 or 45, wherein the subject has autoimmune encephalomyelitis.

49. The method of claim 44 or 45, wherein the subject has scleroderma.

50. The method of claim 44 or 45, wherein the subject has psoriasis.

51. The method of any one of claims 44 to 50, wherein the administering is parenteral.

52. The method of claim 51, wherein the administering is intravenous.

53. The method of claim 51, wherein the administering is subcutaneous.Attorney Docket No: 01434-0001-00PCT54. The humanized antibody of any one of claims 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises glutamine at position 54 according to Kabat numbering.

55. The humanized antibody of any one of claims 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises serine at position 54 according to Kabat numbering.

56. The humanized antibody of any one of claims 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises alanine at position 55 according to Kabat numbering.

57. The humanized antibody of any one of claims 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy chain variable region comprises glutamine at position 54 and alanine at position 55 according to Kabat numbering.

58. The humanized antibody of any one of claims 1 to 5, 9 to 12, 19 to 21, or 25 to 42, wherein the heavy7chain variable region comprises serine at position 54 and alanine at position 55 according to Kabat numbering.

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