Therapy methods using ILT7-binding protein

Abstract

The present invention provides pharmaceutical compositions for use as alternative preventive measures and better therapeutic methods for treating and preventing autoimmune diseases. [Solution] A pharmaceutical composition for reducing the type I interferon gene signature (IFNGS) of a subject requiring such reduction, wherein the composition comprises a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7)-conjugated antibody, the ILT7-conjugated antibody comprising a heavy chain complementarity-determining region (HCDR) and a light chain complementarity-determining region (LCDR) comprising a specific amino acid sequence. The composition comprising the ILT7-conjugated antibody is administered to the subject when, prior to administration of the composition comprising the ILT7-conjugated antibody, the type I IFNGS is elevated by at least 1.1 times compared to the type I IFNGS of a normal subject, and the pharmaceutically effective amount is preferably 1 mg, 5 mg, 15 mg, 50 mg, 100 mg, or 150 mg.

Description

[Technical Field]

[0001] field This disclosure relates to a method for treating an autoimmune disorder in a subject, comprising administering an immunoglobulin-like transcript 7 (ILT7) binding protein to a subject exhibiting an elevated type I interferon gene signature (IFNGS). This disclosure also relates to a method for reducing pDCs in tissue, comprising administering an ILT7 binding protein to a subject in need of such reduction. [Background technology]

[0002] background The type I interferon (IFN) axis is one of the most crucial pathways in human disease, and its dysregulation is central to the pathogenesis of many chronic autoimmune diseases, including systemic lupus erythematosus (SLE). While the precise etiology of SLE and other autoimmune diseases is not fully understood, it is believed that a combination of environmental and genetic factors, coupled with the accumulation of cellular debris, leads to a breakdown of peripheral immune tolerance characterized by high levels of circulating autoreactive antibodies. Currently available methods are aimed at treating autoimmune diseases, not preventing them. Furthermore, conventional treatment options for autoimmune diseases include immunosuppressants, which have a wide range of side effects. Therefore, there is a need for alternative preventive measures and better treatments for autoimmune diseases. This disclosure addresses this need. [Overview of the project]

[0003] overview In certain embodiments, the method of the present disclosure can be used to reduce the type I interferon gene signature (IFNGS) of a subject in need. The method comprises administering a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein to a subject. The ILT7 binding protein is administered to the subject when the subject's type I IFNGS is elevated compared to that of a normal subject. In specific embodiments, the ILT7 binding protein may be administered to subjects with elevated baseline type I IFNGS compared to that of a normal subject, and such subjects are monitored for a decrease in type I IFNGS after treatment. The ILT7 binding protein binds to the same ILT7 epitope as the antibody containing the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2. In certain embodiments, the subject is monitored for a decrease in type I IFNGS after treatment.

[0004] In certain embodiments, type I IFNGS are measured in test biological samples taken from subjects. Test samples include, but are not limited to, blood, sputum, saliva, skin cells, skin biopsy samples, kidney cells, lung cells, hepatocytes, cardiac cells, brain cells, nerve tissue, thyroid cells, ocular cells, skeletal muscle cells, cartilage, bone tissue, and cultured cells.

[0005] In some embodiments, type I IFNGS are elevated at least four times in the tested biological sample compared to a normal biological sample. In certain embodiments, type I IFNGS include the collective expression levels of two or more type I interferon (IFN)-inducible genes. In some embodiments, the two or more type I interferon (IFN)-inducible genes are selected from the group consisting of SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, and USP18. In certain embodiments, type I IFNGS include all collective expression levels of SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, and USP18.

[0006] In some embodiments, type I IFNGS are determined by assaying the mRNA levels of two or more type I interferon (IFN)-inducible genes in the test specimen. In more detailed embodiments, type I IFNGS are determined by assaying the mRNA levels of 21 type I interferon (IFN)-inducible genes in the test specimen.

[0007] In some embodiments, administration of ILT7-binding protein results in a decrease in plasmacytoid dendritic cells (pDCs) in subjects. In certain embodiments, these pDCs are circulating pDCs. In detailed embodiments, the decrease in pDCs is reversible.

[0008] In some embodiments, the target autoimmune disease is treated by reducing type I IFNGS. In certain embodiments, the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, inflammatory myositis, such as dermatomyositis, inclusion body myositis, juvenile myositis and polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, pure red cell follicle, multiple sclerosis, rheumatic carditis, psoriasis, psoriatic arthritis, rheumatoid arthritis, chronic inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, acute and chronic graft-versus-host disease (GVHD), vasculitis, myocardial infarction, and type I interferonosis. In some embodiments, the autoimmune disease is SLE or CLE. In other embodiments, the autoimmune disease is Sjögren's syndrome. In yet another embodiment, the autoimmune disease is dermatomyositis. In yet another embodiment, the autoimmune disease is polymyositis. In yet another embodiment, the autoimmune disease is systemic sclerosis. In yet another embodiment, the autoimmune disease is hidradenitis suppurativa. In yet another embodiment, the autoimmune disease is vitiligo.

[0009] In some embodiments, the ILT7-binding protein is an antibody containing heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each containing the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively. In more detailed embodiments, the ILT7-binding protein is an antibody containing a variable heavy chain (VH) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1 and / or a variable light chain (VL) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. In specific embodiments, the ILT7-binding protein is an antibody containing the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2. In some embodiments, the antibody is afucosylated.

[0010] In some embodiments, the pharmaceutically effective dose of ILT7-binding protein ranges from approximately 0.1 mg to approximately 1000 mg. In specific embodiments, the pharmaceutically effective dose of ILT7-binding protein is approximately 1 mg, approximately 5 mg, approximately 15 mg, approximately 50 mg, approximately 100 mg, or approximately 150 mg. In some embodiments, the ILT7-binding protein is administered by subcutaneous injection.

[0011] In some embodiments, administration of ILT7-binding protein leads to a reduction of at least approximately 50% in type I IFNGS compared to type I IFNGS before ILT7-binding protein administration. In certain embodiments, ILT7-binding protein induces antibody-dependent cell-mediated cytotoxicity (ADCC) activity against pDCs. In some embodiments, ILT7-binding protein suppresses the release of type I interferon (IFN) from pDCs. In certain embodiments, the type I IFN is IFNα. In some embodiments, ILT7-binding protein specifically binds to ILT7. In some embodiments, ILT7 is located on the pDC.

[0012] In other embodiments, the method of the present disclosure can be used to monitor the effectiveness of treatment for conditions characterized by activated pDCs. The method comprises (a) measuring type I interferon gene signatures (IFNGS) in a biological sample taken from a subject to obtain a baseline value of type I IFNGS; and (b) measuring type I IFNGS in a biological sample taken from a subject after treatment, wherein the treatment comprises immunoglobulin-like transcript 7 (ILT7) binding protein. In some embodiments, a decrease in type I IFNGS in step (b) compared to a baseline value indicates that the treatment is effective. The ILT7 binding protein binds to the same ILT7 epitope as an antibody containing the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2.

[0013] In a further embodiment, the method of the present disclosure can be used to reduce plasmacytoid dendritic cells (pDCs) in the tissue of a subject that requires it. The method includes administering to the subject a pharmaceutically effective amount of an immunoglobulin-like transcript 7 (ILT7) binding protein. In certain embodiments, the ILT7 binding protein is an antibody comprising heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, HCDR3 and light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively.

[0014] In some embodiments, the tissue is selected from the group consisting of skin cells, skin biopsy samples, kidney cells, lung cells, liver cells, heart cells, brain cells, nerve tissue, thyroid cells, eye cells, skeletal muscle cells, cartilage, bone tissue, and airway cells. In certain embodiments, the tissue is skin cells. In some embodiments, the tissue is a skin biopsy sample.

[0015] In some embodiments, the method results in a reduction of pDCs in the tissue as compared to a baseline value. In certain embodiments, the reduction of pDCs in the tissue as compared to the baseline value ranges from about 1% to about 99%. In some embodiments, the reduction of pDCs in the tissue as compared to the baseline value is at least about 50%.

[0016] In some embodiments, the ILT7 binding protein is an antibody comprising heavy chain complementarity determining regions (HCDRs) HCDR1, HCDR2, HCDR3 and light chain complementarity determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively.

[0017] In a further embodiment, the method of the present disclosure can be used to treat an autoimmune disorder in a subject that requires it. The method includes administering to the subject a pharmaceutically effective amount of an immunoglobulin-like transcript 7 (ILT7) binding protein. In some embodiments, the pharmaceutically effective amount of the ILT7 binding protein is about 1 mg, about 5 mg, about 15 mg, about 50 mg, about 100 mg, or about 150 mg. In some embodiments, the pharmaceutically effective amount of the ILT7 binding protein is about 50 mg. In certain embodiments, the pharmaceutically effective amount of the ILT7 binding protein is about 150 mg.

[0018] In a particular embodiment, the method of the present disclosure can be used to treat an autoimmune disorder in a subject that requires it, the method includes administering to the subject a pharmaceutically effective amount of an immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the pharmaceutically effective amount of the ILT7 binding protein is about 50 mg.

[0019] In other embodiments, the method of the present disclosure can be used to treat an autoimmune disorder in a subject that requires it, the method includes administering to the subject a pharmaceutically effective amount of an immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the pharmaceutically effective amount of the ILT7 binding protein is about 150 mg.

[0020] In a further embodiment, the method of the present disclosure can be used to reduce plasmacytoid dendritic cells (pDCs) in the tissue of a subject that requires it, the method includes administering to the subject a pharmaceutically effective amount of an immunoglobulin-like transcript 7 (ILT7) binding protein. The pharmaceutically effective amount of the ILT7 binding protein is about 50 mg.

[0021] In a further embodiment, the method of the present disclosure can be used to reduce plasmacytoid dendritic cells (pDCs) in a target tissue requiring such reduction, and the method comprises administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein. The pharmaceutically effective amount of ILT7 binding protein is approximately 150 mg.

[0022] In some embodiments, the decrease in pDCs in the tissue compared to baseline ranges from approximately 1% to approximately 99%. In certain embodiments, the decrease in pDCs in the tissue compared to baseline is at least approximately 50%.

[0023] In some embodiments, subjects have elevated serum type I IFNGS levels prior to administration of ILT7-binding protein. In more detailed embodiments, subjects have elevated pDC levels in tissue biopsy material prior to administration of ILT7-binding protein.

[0024] In some embodiments, autoimmune diseases include systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, inflammatory myositis, such as dermatomyositis, inclusion body myositis, juvenile myositis, and polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, pure red cell fistula, multiple sclerosis, rheumatic carditis, psoriasis, psoriatic arthritis, rheumatoid arthritis, chronic inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, acute and chronic graft-versus-host disease (GVHD), vasculitis, myocardial infarction, and type 1 interferonosis. In some embodiments, the autoimmune disease is SLE. In other embodiments, the autoimmune disease is CLE. In some embodiments, the autoimmune disease is lupus. In certain embodiments, the subjects do not have discoid lupus erythematosus (DLE).

[0025] In some embodiments, the method of the present disclosure can be used to select patients for treatment with ILT7-binding protein, the method comprising (i) determining the patient's baseline serum type I IFNGS level, and (ii) selecting patients with high baseline serum type I IFNGS levels for treatment with ILT7-binding protein.

[0026] In certain embodiments, the method of the present disclosure relates to treating an autoimmune disorder in a subject requiring it, and the method comprises administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the subject is determined to have high serum type I IFNGS levels prior to administration of the ILT7 binding protein. In some embodiments, the ILT7 binding protein is an antibody comprising heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, respectively, comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8. In some embodiments, the ILT7-binding protein is an antibody containing a variable heavy chain (VH) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1 and / or a variable light chain (VL) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. In certain embodiments, the ILT7-binding protein is an antibody containing the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2. In some embodiments, the antibody is afucosylated. [Invention 1001] A method for reducing the type I interferon gene signature (IFNGS) of a subject in need thereof, the method comprising administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the ILT7 binding protein is administered to the subject when the type I IFNGS of the subject is elevated compared to that of a normal subject. [Invention 1002] The method of the present invention 1001, wherein the type I IFNGS is measured in a test biological sample taken from the subject, and the test sample is selected from the group consisting of blood, sputum, saliva, skin cells, skin biopsy samples, kidney cells, lung cells, liver cells, cardiac cells, brain cells, nerve tissue, thyroid cells, eye cells, skeletal muscle cells, cartilage, bone tissue, and cultured cells. [Invention 1003] The method of the present invention 1002, wherein the biological sample for examination is blood, skin cells, or a skin biopsy sample. [Invention 1004] The method of the prior art, wherein the type I IFNGS in the tested biological sample is increased to at least about four times that of a normal biological sample. [Invention 1005] Any of the prior art methods, wherein the type I IFNGS includes the collective expression levels of two or more type I interferon (IFN)-inducible genes. [Invention 1006] The method of the present invention 1005, wherein the two or more type I interferon (IFN)-inducible genes are selected from the group consisting of SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, and USP18. [Invention 1007] The method of the present invention 1005, wherein the type I IFNGS includes collective expression levels of SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, and USP18. [Invention 1008] The method of the present invention 1005, wherein the type I IFNGS is determined by assaying the mRNA levels of two or more type I interferon (IFN)-inducible genes in the test biological sample. [Invention 1009] The method of the present invention 1007, wherein the type I IFNGS is determined by assaying the mRNA levels of 21 type I interferon (IFN)-inducible genes in the biological sample being tested. [Invention 1010] A method of the prior art, wherein administering the ILT7-binding protein results in a decrease in plasmacytoid dendritic cells (pDCs) in the subject. [Invention 1011] The method of the present invention 1010, wherein the pDC is a circulating pDC. [Invention 1012] The method of the present invention 1010 or 1011, wherein the decrease of pDC is reversible. [Invention 1013] Any method of the prior art, wherein the target autoimmune disease is treated by reducing the type I IFNGS. [Invention 1014] The method of the present invention 1013, wherein the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, inflammatory myositis, such as dermatomyositis, inclusion body myositis, juvenile myositis and polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, pure red cell fistula, multiple sclerosis, rheumatic carditis, psoriasis, psoriatic arthritis, rheumatoid arthritis, chronic inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, acute and chronic graft-versus-host disease (GVHD), vasculitis, myocardial infarction, and type 1 interferonosis. [Invention 1015] The method of the present invention 1014, wherein the autoimmune disease is SLE or CLE. [Invention 1016] The method of the present invention 1014, wherein the autoimmune disease is Sjögren's syndrome. [Invention 1017] The method of the present invention 1014, wherein the autoimmune disease is dermatomyositis. [Invention 1018] The method of the present invention 1014, wherein the autoimmune disease is polymyositis. [Invention 1019] The method of the present invention 1014, wherein the autoimmune disease is systemic sclerosis. [Invention 1020] The method of the present invention 1014, wherein the autoimmune disease is hidradenitis suppurativa. [Invention 1021] The method of the present invention 1014, wherein the autoimmune disease is vitiligo. [Invention 1022] The method of the prior art, wherein the ILT7-binding protein is an antibody comprising heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each containing the amino acid sequences of SEQ ID NOs. 3, 4, 5, 6, 7, and 8, respectively. [Invention 1023] Any method of the prior art, wherein the ILT7-binding protein is an antibody comprising a variable heavy chain (VH) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1 and / or a variable light chain (VL) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. [Invention 1024] The method of the prior art, wherein the ILT7-binding protein is an antibody comprising the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2. [Invention 1025] The method according to any of the present invention 1022 to 1024, wherein the antibody is afucosylated. [Invention 1026] Any of the prior art methods, wherein the pharmaceutically effective amount of the ILT7-binding protein is in the range of about 0.1 mg to about 1000 mg. [Invention 1027] The method of the present invention 1026, wherein the pharmaceutically effective amount of the ILT7-binding protein is about 1 mg, about 5 mg, about 15 mg, about 50 mg, about 100 mg, or about 150 mg. [Invention 1028] Any of the prior art methods, wherein the ILT7-binding protein is administered by subcutaneous injection. [Invention 1029] Any method of the prior art, wherein administration of the ILT7-binding protein leads to a reduction of at least about 50% of the target type I IFNGS compared to the type I IFNGS before administration of the ILT7-binding protein. [Invention 1030] Any of the prior art methods, wherein the ILT7-binding protein induces antibody-dependent cell-mediated cytotoxicity (ADCC) activity against pDCs. [Invention 1031] The method according to any of items 1001 to 1029 of the present invention, wherein the ILT7-binding protein suppresses the release of type I interferon (IFN) from pDCs. [Invention 1032] The method of the present invention 1031, wherein the type I IFN is IFNα. [Invention 1033] Any method 1001 to 1032 of the present invention, wherein the ILT7-binding protein specifically binds to ILT7. [Invention 1034] The method of the present invention 1033, wherein the ILT7 is located on the pDC. [Invention 1035] A method for monitoring the effectiveness of treatment for a disease characterized by activated plasmacytoid dendritic cells (pDCs) in a subject, (a) A step of measuring type I interferon gene signatures (IFNGS) in biological samples taken from the subject in order to obtain baseline values ​​of type I interferon gene signatures (IFNGS); and (b) A step of measuring the type I IFNGS in a biological sample taken from the subject after treatment, wherein the treatment includes administering immunoglobulin-like transcript 7 (ILT7) binding protein to the subject. Methods that include... [Invention 1036] A method for reducing plasmacytoid dendritic cells (pDCs) in a tissue of a subject that requires such reduction, comprising administering a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein to the subject. [Invention 1037] The method of the present invention 1036, wherein the tissue is selected from the group consisting of skin cells, skin biopsy samples, kidney cells, lung cells, liver cells, cardiac cells, brain cells, nerve tissue, thyroid cells, eye cells, skeletal muscle cells, cartilage, bone tissue, and airway cells. [Invention 1038] The method of the present invention 1037, wherein the tissue is skin cells. [Invention 1039] The method of the present invention 1037, wherein the tissue is a skin biopsy sample. [Invention 1040] A method of the present invention, which causes a decrease in pDCs in the tissue compared to a baseline value, as described in any of the present invention 1036 to 1039. [Invention 1041] The method of the present invention 1040, wherein the decrease in pDCs in the tissue compared to the baseline value is in the range of about 1% to about 99%. [Invention 1042] The method of the present invention 1040 or 1041, wherein the decrease in pDCs in the tissue compared to the baseline value is at least about 50%. [Invention 1043] The method according to any one of the invention 1036 to 1042, wherein the ILT7-binding protein is an antibody containing heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each containing the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively. [Invention 1044] A method for treating an autoimmune disorder in a subject requiring such treatment, the method comprising administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the pharmaceutically effective amount of ILT7 binding protein is about 1 mg, about 5 mg, about 15 mg, about 50 mg, about 100 mg, or about 150 mg. [Invention 1045] The method according to any one of the present invention 1001 to 1034 or 1044, wherein the pharmaceutically effective amount of the ILT7-binding protein is about 50 mg. [Invention 1046] The method according to any one of the present invention 1001 to 1034 or 1044, wherein the pharmaceutically effective amount of the ILT7-binding protein is about 150 mg. [Invention 1047] A method for treating an autoimmune disorder in a subject requiring such treatment, the method comprising administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the pharmaceutically effective amount of ILT7 binding protein is about 50 mg. [Invention 1048] A method for treating an autoimmune disorder in a subject requiring such treatment, the method comprising administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the pharmaceutically effective amount of ILT7 binding protein is about 150 mg. [Invention 1049] A method for reducing plasmacytoid dendritic cells (pDCs) in a tissue of a subject that requires such reduction, the method comprising administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the pharmaceutically effective amount of ILT7 binding protein is about 50 mg. [Invention 1050] A method for reducing plasmacytoid dendritic cells (pDCs) in a tissue of a subject that requires such reduction, the method comprising administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the pharmaceutically effective amount of ILT7 binding protein is about 150 mg. [Invention 1051] The method of the present invention 1049 or 1050, wherein the decrease in pDCs in the tissue compared to the baseline value is in the range of about 1% to about 99%. [Invention 1052] The method of the present invention 1049 or 1050, wherein the decrease in pDCs in the tissue compared to the baseline value is at least about 50%. [Invention 1053] The method according to any one of items 1001 to 1052 of the present invention, wherein the subject has high blood type I IFNGS levels before administration of the ILT7-binding protein. [Invention 1054] The method according to any one of the present invention 1001 to 1053, wherein the subject has a high pDC level in the tissue biopsy material before administration of the ILT7-binding protein. [Invention 1055] The method according to any one of the 1044 to 1054 of the present invention, wherein the autoimmune disease is systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, inflammatory myositis, such as dermatomyositis, inclusion body myositis, juvenile myositis and polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, pure red cell fistula, multiple sclerosis, rheumatic carditis, psoriasis, psoriatic arthritis, rheumatoid arthritis, chronic inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, acute and chronic graft-versus-host disease (GVHD), vasculitis, myocardial infarction, and type 1 interferonosis. [Invention 1056] The method of the present invention 1055, wherein the autoimmune disease is SLE. [Invention 1057] The method of the present invention 1055, wherein the autoimmune disease is CLE. [Invention 1058] The method according to any one of the present invention 1044 to 1054, wherein the autoimmune disease is lupus. [Invention 1059] The method according to any one of the present invention 1056 to 1058, wherein the subject does not have discoid lupus erythematosus (DLE). [Invention 1060] A method for selecting patients for treatment with ILT7-binding protein, (i) Determine the baseline blood type I IFNGS level of the patient, and (ii) Select patients with high baseline serum type I IFNGS levels for treatment with the ILT7-binding protein. Methods that include... [Invention 1061] A method for treating an autoimmune disorder in a subject in need thereof, the method comprising administering to the subject a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the subject is determined to have high levels of serum type I IFNGS prior to administration of the ILT7 binding protein. [Invention 1062] The method according to any one of the invention 1044 to 1061, wherein the ILT7-binding protein is an antibody containing heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each containing the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively. [Invention 1063] The method according to any one of the present invention 1044 to 1062, wherein the ILT7-binding protein is an antibody comprising a variable heavy chain (VH) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1 and / or a variable light chain (VL) that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. [Invention 1064] The method according to any one of the present invention 1044 to 1063, wherein the ILT7-binding protein is an antibody containing the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2. [Invention 1065] The method according to any of the present invention 1060 to 1064, wherein the antibody is afucosylated. [Brief explanation of the drawing]

[0027] [Figure 1] This document outlines the overall study design of a Phase Ia randomized, blinded, placebo-controlled study evaluating the safety and tolerability of a single, escalating subcutaneous administration of ILT7-binding protein as described herein, in subjects with at least one of the following five autoimmune diseases: systemic lupus erythematosus (SLE), Sjögren's syndrome, dermatomyositis, polymyositis, or systemic sclerosis. [Figure 2] The details of the single-dose dose escalation study design are shown below. [Figure 3] The mean serum concentration profiles following a single subcutaneous administration of ILT7-binding protein using the method described herein are shown for subjects suffering from at least one of the following five autoimmune diseases: SLE, Sjögren's syndrome, dermatomyositis, polymyositis, or systemic sclerosis. [Figure 4] The following shows the pDC levels (%) over time in subjects suffering from at least one of the following five autoimmune diseases: SLE, Sjögren's syndrome, dermatomyositis, polymyositis, or systemic sclerosis, following a single subcutaneous administration (1 mg, 5 mg, 15 mg, 50 mg, or 150 mg) of ILT7-binding protein (VIB7734) as used in the method described herein, as a percentage relative to baseline levels (value using % peripheral blood mononuclear cells). [Figure 5] The following shows the pDC levels (%) over time in subjects suffering from at least one of the following five autoimmune diseases: SLE, Sjögren's syndrome, dermatomyositis, polymyositis, or systemic sclerosis, following a single subcutaneous administration (1 mg, 5 mg, 15 mg, 50 mg, or 150 mg) of ILT7-binding protein (VIB7734) as used in the method described herein, expressed as a percentage (absolute concentration) relative to baseline levels. [Figure 6] The following shows the pDC levels (absolute concentration; cells / microliter) associated with a single subcutaneous administration (1 mg, 5 mg, 15 mg, 50 mg, or 150 mg) of ILT7-binding protein (VIB7734) used in the manner described herein, in subjects suffering from at least one of the following five autoimmune diseases: SLE, Sjögren's syndrome, dermatomyositis, polymyositis, or systemic sclerosis. [Figure 7]This specification shows the percentage change in type I IFNGS (measured as a percentage of baseline) in subjects with elevated IFN levels treated with 1–150 mg of ILT7-binding protein (VIB7734) as used in the method described herein. Type I IFNGS was determined by assaying the aggregate mRNA levels of 21 type I IFN-inducible genes in biological samples taken from subjects, determining the mean or median mRNA levels of the 21 type I IFN-inducible genes, and normalizing this mean value to the mean mRNA levels of three housekeeping genes (18S rRNA, β-actin, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH)) to obtain a composite outcome. In the majority of subjects with elevated type I IFNGS, a decrease in pDC levels (Figure 7A) correlated with a decrease in type I IFNGS (reported as a percentage change of baseline) (Figure 7B). [Figure 8] In subjects with elevated baseline type I IFNGS, treatment with 15 mg of ILT7-binding protein (VIB7734) as used in the method described herein reduces type I IFNGS, but this reduction does not occur in subjects with low baseline type I IFNGS. Type I IFNGS were determined by assaying the collective mRNA levels of 21 type I IFN-inducible genes in biological samples collected from subjects, determining the mean or median mRNA levels of the 21 type I IFN-inducible genes, and then normalizing this mean value to the mean mRNA levels of three housekeeping genes (18S rRNA, β-actin, and GAPDH) to obtain a composite outcome. [Figure 9] This document outlines the overall study design of a Phase Ib randomized, blinded, placebo-controlled study evaluating the safety and tolerability of multiple doses of subcutaneously administered ILT7-binding protein (VIB7734) as used in the method described herein, in subjects with at least one of the following autoimmune diseases: systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), systemic sclerosis, polymyositis, and dermatomyositis. [Figure 10] This document presents the randomization and dose escalation scheme for multi-stage dose escalation (MAD) studies. [Figure 11] Figure 11A shows the serum concentration profiles of the ILT7-binding protein (VIB7734) used in the methods described herein, following multiple subcutaneous administrations (three doses every four weeks) of 5 mg (Cohort 1) or 50 mg (Cohort 2) of VIB7734. Figure 11B shows the serum concentration profiles of VIB7734 in Cohort 2 subjects. Figure 11B shows the mean serum concentration profiles of VIB7734 in subjects of Cohort 1 (filled circles) and Cohort 2 (filled squares). [Figure 12] The following pDC levels (%) over time are shown as a percentage relative to baseline (using % peripheral blood mononuclear cells) in whole blood subjects of Cohort 1, who had at least one of the following autoimmune diseases: SLE, CLE, systemic sclerosis, polymyositis, and dermatomyositis, following multiple subcutaneous administrations (3 doses every 4 weeks) of 5 mg of ILT7-binding protein (VIB7734) as used in the method described herein. Subjects of Cohort 1 were administered either placebo (Figure 12A) or VIB7734 (Figure 12B). [Figure 13] The following are the pDC levels (%) over time in the whole blood of subjects in Cohort 1 who had at least one of the following autoimmune diseases: SLE, CLE, systemic sclerosis, polymyositis, and dermatomyositis, following multiple subcutaneous administrations (3 doses every 4 weeks) of 5 mg of ILT7-binding protein (VIB7734) as used herein, expressed as a percentage of baseline levels (values ​​using absolute concentration). Subjects in Cohort 1 were administered either placebo (Figure 13A) or VIB7734 (Figure 13B). [Figure 14] The following shows the pDC levels (absolute concentration; cells / microliter) over time in the whole blood of subjects in Cohort 1 who had at least one of the following autoimmune diseases: SLE, CLE, systemic sclerosis, polymyositis, and dermatomyositis, following multiple subcutaneous administrations (3 doses every 4 weeks) of 5 mg of ILT7-binding protein (VIB7734) as used herein. Subjects in Cohort 1 were administered either placebo (Figure 14A) or VIB7734 (Figure 14B). [Figure 15] The pDC levels (%) over time in whole blood of subjects in cohorts 2 and 3 suffering from SLE or CLE, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg (cohort 2) or 150 mg (cohort 3) of ILT7-binding protein (VIB7734) as used in the method described herein, are shown as a percentage relative to baseline levels (values ​​using % peripheral blood mononuclear cells). Subjects in cohort 2 were administered either placebo (Figure 15A) or VIB7734 (Figure 15B). Subjects in cohort 3 were administered either placebo (Figure 15C) or VIB7734 (Figure 15D). [Figure 16] The pDC levels (%) over time in whole blood of subjects in Cohort 2 and Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg (Cohort 2) or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) as used by the method described herein, are shown as a percentage relative to baseline levels (values ​​using absolute concentration). Subjects in Cohort 2 were administered either placebo (Figure 16A) or VIB7734 (Figure 16B). Subjects in Cohort 3 were administered either placebo (Figure 16C) or VIB7734 (Figure 16D). [Figure 17] The following shows the pDC levels (absolute concentration; cells / microliter) over time in whole blood of subjects in Cohort 2 and Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg (Cohort 2) or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) as used by the method described herein. Subjects in Cohort 2 were administered either placebo (Figure 17A) or VIB7734 (Figure 17B). Subjects in Cohort 3 were administered either placebo (Figure 17C) or VIB7734 (Figure 17D). [Figure 18]The pDC levels over time, expressed as a percentage of peripheral blood mononuclear cells, are shown for whole blood subjects in Cohort 2 and Cohort 3, following multiple subcutaneous administrations (three doses every four weeks) of 50 mg (Cohort 2) or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) using the method described herein. Subjects in Cohort 2 received either placebo (Figure 18A) or VIB7734 (Figure 18B). Subjects in Cohort 3 received either placebo (Figure 18C) or VIB7734 (Figure 18D). [Figure 19] Figure 19A shows the median pDC levels over time in whole blood of subjects in Cohort 2, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) or placebo as used in the method described herein. Figure 19B shows the median absolute level of pDCs in the blood over time as a percentage (using absolute concentration) relative to baseline levels. Figure 19C shows the median pDC level in the blood over time as a percentage (using % peripheral blood mononuclear cells (PBMCs)) relative to baseline levels. Figure 19D shows the median absolute level of pDCs in the blood over time as a percentage in PBMCs. [Figure 20] Figure 20A: Median pDC levels over time in whole blood of subjects in Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) or placebo as used in the method described herein. Figure 20B: Median pDC levels (%) over time in blood, as a percentage (using absolute concentration) relative to baseline levels. Figure 20C: Median pDC levels (%) over time in blood, as a percentage (using %PBMC) relative to baseline levels. Figure 20D: Median pDC levels (cells / μL) over time in blood, as a percentage in PBMCs. [Figure 21]The following figures show the time-course type I IFNGS levels (measured as a change factor (Figures 21A and 21B) or absolute score (Figures 21C and 21D)) in the whole blood of subjects in Cohort 2 treated with 50 mg of ILT7-binding protein (VIB7734) as used in the method described herein. The type I IFNGS score was determined by assaying the aggregate mRNA levels of 21 type I IFN-inducible genes in the blood collected from the subjects, determining the mean or median mRNA levels of the 21 type I IFN-inducible genes, and normalizing this mean value to the mean mRNA levels of three housekeeping genes (18S rRNA, β-actin, and glyceraldehyde 3-phosphate dehydrogenase (GAPDH)) to obtain a composite outcome. Subjects in Cohort 2 were administered either VIB7734 (Figures 21A and 21C) or placebo (Figures 21B and 21D). [Figure 22] Figure 22A: Median blood type I IFNGS levels over time in whole blood of subjects in Cohort 2, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) or placebo as used in the method described herein. Figure 22B: Median blood type I IFNGS levels over time (measured as change rate). Figure 22C: Median blood type I IFNGS levels over time (measured as neutralization rate). [Figure 23] The CLASI activity (CLASI-A) score (measured as the change from baseline) over time is shown for subjects in Cohort 2 following multiple subcutaneous administrations (three doses every four weeks) of 50 mg of ILT7-binding protein (VIB7734) as used in the method described herein. Subjects in Cohort 2 received either placebo (Figure 23A) or VIB7734 (Figure 23B). [Figure 24-1]The CLASI-A scores over time (measured as individual plots) are shown for subjects in Cohort 2 and Cohort 3, following multiple subcutaneous administrations (three doses every four weeks) of 50 mg (Cohort 2) or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) as used by the method described herein. Subjects in Cohort 2 were administered either VIB7734 (Figure 24A) or placebo (Figure 24B). Subjects in Cohort 3 were administered either VIB7734 (Figure 24C) or placebo (Figure 24D). [Figure 24-2] See the explanation in Figure 24-1. [Figure 25] The CLASI-A scores over time (measured as the percentage of subjects with a decrease of at least 4 points from baseline) in subjects of Cohort 2, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) as used in the method described herein, compared to subjects who received placebo. [Figure 26] The CLASI-A scores over time (measured as the percentage of subjects with a decrease of at least 4 points from baseline) associated with multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) as used in the method described herein, in Cohort 3, are shown compared to subjects who received placebo. [Figure 27] The CLASI-A scores over time (measured as the percentage of subjects with a decrease of at least 4 points from baseline) in subjects from cohorts 2 and 3 combined, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg (cohort 2) or 150 mg (cohort 3) of ILT7-binding protein (VIB7734) as used by the method described herein, are shown compared to subjects from cohorts 2 and 3 who received placebo. [Figure 28]The proportion of CLASI-A score responders in Cohorts 2 and 3 following multiple subcutaneous administrations (three doses every four weeks) of 50 mg (Cohort 2) or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) as used by the method described herein is shown, compared to the proportion of subjects in Cohorts 2 and 3 who received placebo. Comparative data are shown for all subjects, subjects with discoid lupus erythematosus (DLE), and subjects without DLE. [Figure 29] The CLASI-A scores over time (measured as the percentage of subjects with at least a 50% reduction from baseline) in subjects of Cohort 2, following multiple subcutaneous administrations (three doses every four weeks) of 50 mg of ILT7-binding protein (VIB7734) as used in the method described herein, compared to subjects who received placebo. [Figure 30] This shows a comparison of CLASI-A scores (Figure 30A), pDC absolute blood levels (measured as a percentage of baseline levels) (Figure 30B), and type I IFNGS levels (measured as an absolute score) (Figure 30C) over time in subjects of Cohort 2 following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) as used in the method described herein. [Figure 31] This figure shows a comparison of CLASI-A scores (Figure 31A), pDC absolute blood levels (measured as a percentage of baseline levels) (Figure 31B), and type I IFNGS levels (measured as an absolute score) over time in subjects of Cohort 2, following multiple subcutaneous administrations of placebo (3 doses every 4 weeks). [Figure 32] This document outlines the subject-level exploratory data synthesis approach. [Figure 33-1]Figure 33A: Subjects in Cohort 2 and 3 who received the ILT7-binding protein (VIB7734) used in the method described herein (filled circles) are shown in comparison to subjects in Cohort 2 and 3 who received placebo (filled triangles). Figure 33A: Subjects in Cohort 2 received multiple subcutaneous doses of 50 mg of VIB7734 or placebo (3 doses every 4 weeks). Figure 33B: Subjects in Cohort 3 received multiple subcutaneous doses of 150 mg of VIB7734 or placebo (3 doses every 4 weeks). Unexpectedly, the median change from baseline in CLASI-A score at day 85 was higher in Cohort 3 (-9.5 in the 150 mg VIB7734 group compared to -5 in the placebo group compared to -5 in the placebo group) compared to Cohort 2 (-5 in the 50 mg VIB7734 group compared to -2.5 in the placebo group). For Cohort 2 subjects, the least squares mean difference between the VIB7734 and placebo arms at day 85 was 0.14; 95% CI (-9.86, 10.14, p=0.977). For Cohort 3 subjects, the least squares mean difference between the VIB7734 and placebo arms at day 85 was -5.12; 95% CI (-11.49, 1.24, p=0.108). Figure 33C: Percentage change from baseline (BL) in median CLASI-A score by treatment arm and number of visits for subjects in Cohort 2 and Cohort 3. [Figure 33-2] See the explanation in Figure 33-1. [Figure 34] The absolute number of pDCs (measured as the number of cells per square millimeter) in skin biopsy materials from Cohort 2, over time, is shown following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) (Figure 34B) or placebo (Figure 34A) as used by the method described herein. [Figure 35]Figure 35A: Median pDC count in skin biopsy specimens of Cohort 2 subjects, associated with multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) (filled circle) or placebo (filled triangle) as used in the method described herein. Figure 35B: Median pDC count in skin biopsy specimens of Cohort 2 subjects (measured as a percentage of baseline on day 1). Figure 35B: Median pDC count in skin biopsy specimens of Cohort 2 subjects (measured as the number of cells per square mm). The median reduction in change in pDC count in skin biopsy specimens (measured as a percentage of baseline on day 1 and the number of cells per square mm) at day 85 was 87% for Cohort 2 subjects treated with VIB7734, compared to 47% for Cohort 2 subjects treated with placebo. [Figure 36] The figures show the time course of myxovirus protein A (MxA) (positive %) in the biopsy material of skin biopsy subjects from Cohort 2, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) (Figure 36B) or placebo (Figure 36A) as used by the method described herein. [Figure 37] The median MxA (measured as a percentage of MxA-positive area; positive %) in skin biopsy materials from Cohort 2 subjects is shown, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) (filled circle) or placebo (filled triangle) as used by the method described herein. [Figure 38-1] This shows a comparison of CLASI-A scores (Figure 38A), absolute blood pDC levels (measured as cells / μL) (Figure 38B), blood type I IFNGS levels (measured as absolute scores) (Figure 38C), pDC counts in skin biopsy material (measured as the number of cells per square mm) (Figure 38D), and normalized blood type I IFNGS levels (measured as a change factor) (Figure 38E) over time in subjects of Cohort 2, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) as used in the method described herein. [Figure 38-2] See the explanation in Figure 38-1. [Figure 39-1] This figure shows a comparison of CLASI-A scores (Figure 39A), absolute blood pDC levels (measured as cells / μL) (Figure 39B), blood type I IFNGS levels (measured as absolute scores) (Figure 39C), pDC count in skin biopsy material (measured as the number of cells per square mm) (Figure 39D), and normalized blood type I IFNGS levels (measured as a change factor) (Figure 39E) in subjects of Cohort 2, following multiple subcutaneous administrations of placebo (3 doses every 4 weeks). [Figure 39-2] See the explanation in Figure 39-1. [Figure 40] This specification provides an overview of adverse effects (AEs) observed in subjects of Cohorts 1, 2, and 3 following multiple subcutaneous administrations (three doses every four weeks) of 5 mg (Cohort 1), 50 mg (Cohort 2), or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) or placebo as used by the method described herein. [Figure 41] This specification provides an overview of particularly noteworthy adverse events (AESIs) observed in subjects of Cohorts 1, 2, and 3 following multiple subcutaneous administrations (three doses every four weeks) of 5 mg (Cohort 1), 50 mg (Cohort 2), or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) or placebo as used by the method described herein. [Figure 42] This document provides an overview of the immunohistochemical (IHC) analysis methods used in skin biopsy specimens for subjects in Cohort 2 and Cohort 3. Two 4 mm punch biopsy specimens were collected from the skin of each Cohort 2 subject: a baseline biopsy specimen at the second visit (day 1) and a repeat biopsy specimen at the eleventh visit (day 85). The biopsy specimens were sectioned longitudinally. Each panel was analyzed using three sections (left, center, and right) per biopsy specimen. Three rounds of staining were performed to determine pDCs (BDCA+ / ILT7+ cells), IFN activity (MxA+ pixels), and inflammatory infiltration (CD45+ cells). [Figure 43]This document describes the analytical strategy for quantifying pDC and CD45+ cells using IHC analysis of skin biopsy materials from subjects in Cohorts 2 and 3. A 500-micron dermis area (outlined in red, Figure 43A) proximal to the dermal-epithelial junction (DEJ) was used as the analysis area for pDC and CD45+ cells. Epidermal protrusions were excluded from the analysis area (Figure 43A). The number of pDC (BDCA+ / ILT7+ cells) and CD45+ cells per square millimeter was measured as a reading. Positive cells were identified using consistent RGB values ​​(with the exception of a few cases where background staining affected detection in these settings). Figure 43B shows the analysis area without the positive cell detection algorithm. Figure 43C shows the analysis area with the positive cell detection algorithm. Red indicates positive cells; blue indicates negative cells. [Figure 44] This document describes the analytical strategy for quantifying myxoviral protein A (MxA) related to interferon (IFN) activity using IHC analysis of skin biopsy materials from subjects in Cohorts 2 and 3. The entire epidermal region (red outline, Figure 44A) was used as the analysis area for MxA. The percentage of MxA-positive area (%ROI MxA+) was measured as the reading. Positive pixels were identified using consistent RGB values. Figure 44B shows the analysis area without a positive pixel detection algorithm. Figure 44C shows the analysis area with a positive pixel detection algorithm. Red indicates positive pixels; blue indicates negative pixels. [Figure 45]The results show minimal variability within the biopsy material in terms of baseline numbers of pDCs (BDCA+ / ILT7+ cells), MxA+ pixels, and CD45+ cells for each subject in Cohort 2. A high degree of consistency was observed within the range of each skin biopsy material at baseline. Figure 45A: pDCs (BDCA+ / ILT7+ cells) from the central portion of the skin biopsy material. Figure 45B: MxA+ pixels from the central portion of the skin biopsy material. Figure 45C: CD45+ cells from the central portion of the skin biopsy material. Figure 45D: pDCs (BDCA+ / ILT7+ cells) from the right portion of the skin biopsy material. Figure 45E: MxA+ pixels from the right portion of the skin biopsy material. Figure 45F: CD45+ cells from the right portion of the skin biopsy material. Figure 45G: Baseline pDCs (measured as the number of cells per square mm) in skin biopsy materials from each subject in Cohort 2. Figure 45H: Baseline MxA+ pixels (measured as %ROI MxA+) in skin biopsy material from each subject in Cohort 2. Figure 45I: Baseline CD45+ cells (measured as the number of cells per square mm) in skin biopsy material from each subject in Cohort 2. Each point on the graph (Figures 45G-45I) represents an individual section (2-3 sections were analyzed per biopsy material). [Figure 46]Within the scope of Cohort 2, there was significant variability among biopsy specimens in terms of baseline numbers of pDCs, MxA+ pixels, and CD45+ cells. High variability was observed among skin biopsy specimens at baseline. Figure 46A: High pDC (BDCA+ / ILT7+ cells) in skin biopsy specimens. Figure 46B: High MxA+ pixel count in skin biopsy specimens. Figure 46C: High CD45+ cell count in skin biopsy specimens. Figure 46D: Median pDC (BDCA+ / ILT7+ cells) in skin biopsy specimens. Figure 46E: Median MxA+ pixel count in skin biopsy specimens. Figure 46F: Median CD45+ cell count in skin biopsy specimens. Figure 46G: Low pDC (BDCA+ / ILT7+ cells) in skin biopsy specimens. Figure 46H: Low MxA+ pixel count in skin biopsy specimens. Figure 46I: Low CD45+ cell count in skin biopsy specimens. Figure 46J: Baseline pDCs (measured as the number of cells per square mm) in skin biopsy material from each subject in Cohort 2. Subjects are categorized as having high baseline pDCs (n=5, >100 pDCs / mm2), median baseline pDCs (n=3, 10–100 pDCs / mm2), or low baseline pDCs (n=4, <10 pDCs / mm2). Figure 46K: Baseline MxA+ pixels (measured as %ROI MxA+) in skin biopsy material from each subject in Cohort 2. Subjects are categorized as having high baseline MxA+ pixels (n=5, >50% MxA+), median baseline MxA+ pixels (n=4, 5–50% MxA+), or low baseline MxA+ pixels (n=2, <5% MxA+). Figure 46L: Baseline CD45+ cells (measured as the number of cells per square mm) in skin biopsy material from each subject in Cohort 2. Subjects are classified as having high baseline CD45+ cell counts (n=3, >2000 CD45+ cells / mm2), intermediate baseline CD45+ cell counts (n=4, 500–2000 CD45+ cells / mm2), or low baseline CD45+ cell counts (n=5, <500 CD45+ cells / mm2). [Figure 47]Skin biopsy materials from cohort 2 subjects treated with placebo showed considerable variability in response to decreases in pDCs (Figure 47A), MxA+ pixels (Figure 47B), and CD45+ cells (Figure 47C) (measured by the percentage change from baseline), whereas skin biopsy materials from cohort 2 subjects treated with ILT7-binding protein (VIB7734) used in the method described herein showed a more consistent decrease in pDCs, MxA+ pixels, and CD45+ cells. [Figure 48] This demonstrates that the IHC analysis method for skin biopsy materials does not include an activity threshold. Figure 48A: Percentage change from baseline in MxA in skin biopsy materials from Cohort 2 subjects treated with placebo or ILT7-binding protein (VIB7734) used by the method described herein. Gray contours indicate skin biopsy samples with a substantial numerical multiplier increase in MxA. However, overall, very low levels of MxA maintenance were observed in skin biopsy materials from Cohort 2 subjects. Figure 48B: IHC performed on skin biopsy materials from Cohort 2 subjects following multiple subcutaneous administrations of placebo (3 doses every 4 weeks). Figure 48C: IHC performed on skin biopsy materials from Cohort 2 subjects following multiple subcutaneous administrations of 50 mg of VIB7734 (3 doses every 4 weeks). [Figure 49-1] This study shows the relationship between elevated baseline pDC count / IFN activity in skin biopsy samples from Cohort 2 subjects and their response to VIB7734. VIB7734 treatment group: In Cohort 2 subjects who received multiple subcutaneous doses of 50 mg of VIB7734 (three doses every four weeks), elevated baseline pDC count and IFN activity were observed in skin biopsy samples from 4 out of 5 responders. In non-responders, baseline pDC or IFN activity in skin biopsy samples was low. Placebo group: In Cohort 2 subjects who received multiple subcutaneous doses of placebo (three doses every four weeks), no discernible relationship was observed between pDC or IFN activity and the response. [Figure 49-2] See the explanation in Figure 49-1. [Figure 50]The CLASI-A scores over time (measured as the percentage of subjects with a decrease of at least 7 points from baseline) associated with multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) as used in the method described herein, in Cohort 2, are shown compared to subjects who received placebo. [Figure 51] The CLASI-A scores over time (measured as the percentage of subjects with a decrease of at least 7 points from baseline) associated with multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) as used in the method described herein, in Cohort 3, are shown compared to subjects who received placebo. [Figure 52] The CLASI-A scores over time (measured as the percentage of subjects with a decrease of at least 7 points from baseline) in subjects from cohorts 2 and 3 combined, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg (cohort 2) or 150 mg (cohort 3) of ILT7-binding protein (VIB7734) as used by the method described herein, are shown compared to subjects from cohorts 2 and 3 who received placebo. [Figure 53] The CLASI-A scores over time (measured as the percentage of subjects with at least a 50% reduction from baseline) associated with multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) as used in the method described herein, in subjects of Cohort 3, are shown compared to subjects who received placebo. [Figure 54] The CLASI-A scores over time (measured as the percentage of subjects with at least a 50% reduction from baseline) in subjects from cohorts 2 and 3 combined, following multiple subcutaneous administrations (three doses every four weeks) of 50 mg (cohort 2) or 150 mg (cohort 3) of ILT7-binding protein (VIB7734) as used by the method described herein, are shown compared to subjects from cohorts 2 and 3 who received placebo. [Figure 55]Figure 55A: Normalized type I IFNGS levels (measured as a change factor) over time in the whole blood of subjects in Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) or placebo as used in the method described herein. Figure 55B: Normalized type I IFNGS levels over time in the whole blood of subjects in Cohort 3 treated with VIB7734. Figure 55C: Median normalized type I IFNGS levels over time in the whole blood of subjects in Cohort 3 treated with VIB7734 (filled circle) or placebo (filled triangle). [Figure 56] The absolute number of pDCs (measured as the number of cells per square millimeter) in skin biopsy materials from Cohort 3, over time, is shown following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) (Figure 56B) or placebo (Figure 56A) as used by the method described herein. [Figure 57] The median number of pDCs (measured as the number of cells per square millimeter) in skin biopsy materials of subjects in Cohort 3 is shown over time following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg (filled circle) or placebo (filled triangle) of ILT7-binding protein (VIB7734) as used by the method described herein. The median number of pDCs (measured as a percentage of baseline at day 1) at day 85 was reduced by 99% in subjects of Cohort 3 treated with VIB7734. In contrast, the median number of pDCs (measured as a percentage of baseline at day 1) at day 85 was increased by 11% in subjects of Cohort 3 treated with placebo. [Figure 58] The figures show the time course of myxovirus protein A (MxA) (positive %) in the biopsy material (ROI area) of skin biopsy specimens from Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) (Figure 58B) or placebo (Figure 58A) as used by the method described herein. [Figure 59] The median MxA (measured as a percentage of positive area; median positive % in ROI area) of skin biopsy material over time is shown for subjects in Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) (filled circle) or placebo (filled triangle) as used in the method described herein. For subjects in Cohort 3 treated with VIB7734, the median MxA of skin biopsy material decreased from a baseline of 89.7% to 1.1% at day 85. In contrast, for subjects in Cohort 3 treated with placebo, the median MxA of skin biopsy material increased from a baseline of 1.9% to 17.7% at day 85. [Figure 60] The absolute number of CD45 cells in skin biopsy materials (measured as the number of CD45+ cells per square mm) over time is shown for multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) (Figure 60B) or placebo (Figure 60A) used in the method described herein, in skin biopsy materials from Cohort 2. [Figure 61] The median CD45 count (measured as the number of CD45+ cells per square mm) in skin biopsy materials of subjects in Cohort 2 is shown over time following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of ILT7-binding protein (VIB7734) (filled circle) or placebo (filled triangle) as used in the method described herein. The median CD45 count in skin biopsy materials for subjects in Cohort 2 treated with VIB7734 decreased from a baseline of 1119 on day 1 to 280 on day 85. In contrast, the median CD45 count in skin biopsy materials for subjects in Cohort 2 treated with placebo decreased from a baseline of 537 on day 1 to 492 on day 85. [Figure 62]The absolute number of CD45 cells in skin biopsy materials (measured as the number of CD45+ cells per square mm) over time is shown for multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) (Figure 62B) or placebo (Figure 62A) as used by the method described herein, in skin biopsy materials from Cohort 3. [Figure 63] The median CD45 count (measured as the number of CD45+ cells per square mm) in skin biopsy materials of subjects in Cohort 3 is shown over time following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) (filled circle) or placebo (filled triangle) as used in the method described herein. The median CD45 count in skin biopsy materials decreased from a baseline of 707 to 513 at day 85 in subjects of Cohort 3 treated with VIB7734. In contrast, the CD45 count in skin biopsy materials decreased from a baseline of 897 to 666 at day 85 in subjects of Cohort 3 treated with placebo. [Figure 64-1] This shows a comparison of CLASI-A scores (Figure 64A), absolute blood pDC levels (measured as cells / μL) (Figure 64B), normalized blood type I IFNGS levels (measured as a change factor) (Figure 64C), and pDC counts (measured as the number of cells per square mm) (Figure 64D) over time for subjects in Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) as used in the method described herein. [Figure 64-2] See the explanation in Figure 64-1. [Figure 65-1] This figure shows a comparison of CLASI-A scores (Figure 65A), absolute blood pDC levels (measured as cells / μL) (Figure 65B), normalized blood type I IFNGS levels (measured as a change factor) (Figure 65C), and pDC counts in skin biopsy material (measured as the number of cells per square mm) (Figure 65D) over time in subjects of Cohort 3 who received multiple subcutaneous doses of placebo (3 doses every 4 weeks). [Figure 65-2] See the explanation in Figure 65-1. [Figure 66] Both Cohort 2 and Cohort 3, treated with 50 mg and 150 mg of VIB7734 respectively, showed a decrease in cutaneous pDCs (measured as the percentage change in cell number from baseline on day 1), but pDC depletion was more consistent in Cohort 3. Figure 66A: Comparison of the decrease in pDCs in skin biopsy materials from Cohort 2 and Cohort 3 treated with VIB7734. The mean percentage decrease in pDCs from baseline in skin samples with more than 10 pDCs / mm2 at baseline was 96.31% for Cohort 3 treated with VIB7734, compared to 85.45% for Cohort 2 treated with VIB7734. Figure 66B: Comparison of the decrease in MxA+ pixels (measured as the percentage change from baseline on day 1) in skin biopsy materials from Cohort 2 and Cohort 3 treated with VIB7734. In skin samples with over 5% MxA+ at baseline, the mean reduction in MxA+ pixels from baseline on day 1 was 76.84% in cohort 3 treated with VIB7734, compared to 67.44% in cohort 2 treated with VIB7734. [Figure 67A] Figure 67A: VIB7734 treatment group: Cohort 3 subjects who received multiple subcutaneous doses of 150 mg of VIB7734 (3 doses every 4 weeks). Figure 67B: Placebo group: Cohort 3 subjects who received multiple subcutaneous doses of placebo (3 doses every 4 weeks). VIB7734 reduced pDC levels in the skin of Cohort 3 subjects. [Figure 67B] See the explanation in Figure 67A. [Figure 68] The CLASI activity (CLASI-A) score (measured as the change from baseline on day 1) over time is shown for subjects in Cohort 3 following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of ILT7-binding protein (VIB7734) as used in the method described herein. Subjects in Cohort 3 received either placebo (Figure 68A) or VIB7734 (Figure 68B). [Figure 69] The results show minimal variability within the biopsy material in terms of baseline numbers of pDCs (BDCA+ / ILT7+ cells), MxA+ pixels, and CD45+ cells for each subject in Cohort 3. A high degree of consistency was observed within the range of each skin biopsy material at baseline. Figure 69A: pDCs (BDCA+ / ILT7+ cells) from the central portion of the skin biopsy material. Figure 69B: MxA+ pixels from the central portion of the skin biopsy material. Figure 69C: CD45+ cells from the central portion of the skin biopsy material. Figure 69D: pDCs (BDCA+ / ILT7+ cells) from the right portion of the skin biopsy material. Figure 69E: MxA+ pixels from the right portion of the skin biopsy material. Figure 69F: CD45+ cells from the right portion of the skin biopsy material. Figure 69G: Baseline pDCs (measured as the number of cells per square mm) in skin biopsy materials from each subject in Cohort 3. Figure 69H: Baseline MxA+ pixels (measured as %ROI MxA+) in skin biopsy materials from each subject in Cohort 3. Figure 69I: Baseline CD45+ cells (measured as the number of cells per square mm) in skin biopsy materials from each subject in Cohort 3. Each point on the graph (Figures 69G to 69I) represents an individual section (2 to 3 sections were analyzed per biopsy material). [Figure 70]Within the scope of Cohort 3, there was considerable variability among biopsy materials in terms of baseline numbers of pDCs, MxA+ pixels, and CD45+ cells. Figure 70A: High levels of pDCs (BDCA+ / ILT7+ cells) in skin biopsy materials. Figure 70B: High levels of MxA+ pixels in skin biopsy materials. Figure 70C: High levels of CD45+ cells in skin biopsy materials. Figure 70D: Median levels of pDCs (BDCA+ / ILT7+ cells) in skin biopsy materials. Figure 70E: Median levels of MxA+ pixels in skin biopsy materials. Figure 70F: Median levels of CD45+ cells in skin biopsy materials. Figure 70G: Low levels of pDCs (BDCA+ / ILT7+ cells) in skin biopsy materials. Figure 70H: Low levels of MxA+ pixels in skin biopsy materials. Figure 70I: Low levels of CD45+ cells in skin biopsy materials. In Cohort 3 subjects, slightly increased baseline pDC and MxA signals were observed compared to Cohort 2 subjects. Figure 70J: Baseline pDCs (measured as the number of cells per square millimeter) in skin biopsy materials from subjects in Cohort 2 and Cohort 3. Subjects in Cohort 2 show high baseline pDCs (n=5, >100 pDCs / mm2), median baseline pDCs (n=3, 10–100 pDCs / mm2), or low baseline pDCs (n=4, <10 pDCs / mm2). Subjects in Cohort 3 show high baseline pDCs (n=5, >100 pDCs / mm2), median baseline pDCs (n=3, 10–100 pDCs / mm2), or low baseline pDCs (n=2, <10 pDCs / mm2). Figure 70K: Baseline MxA+ pixels (measured as %ROI MxA+) in skin biopsy materials from subjects in Cohort 2 and Cohort 3. Cohort 2 subjects exhibit either high baseline MxA+ pixel values ​​(n=5, >50% MxA+), intermediate baseline MxA+ pixel values ​​(n=4, 5-50% MxA+), or low baseline MxA+ pixel values ​​(n=2, <5% MxA+). Cohort 3 subjects exhibit either high baseline MxA+ pixel values ​​(n=6, >50% MxA+) or low baseline MxA+ pixel values ​​(n=4, <5% MxA+). Figure 70L: Baseline CD45+ cells (measured as the number of cells per square mm) in skin biopsy material from subjects in Cohort 2 and Cohort 3.Cohort 2 subjects are those with high baseline CD45+ cell counts (n=3, >2000 CD45+ cells / mm2), median baseline CD45+ cell counts (n=5, 500-2000 CD45+ cells / mm2), or low baseline CD45+ cell counts (n=4, <500 CD45+ cells / mm2). Cohort 3 subjects are those with high baseline CD45+ cell counts (n=2, >2000 CD45+ cells / mm2), median baseline CD45+ cell counts (n=4, 500-2000 CD45+ cells / mm2), or low baseline CD45+ cell counts (n=4, <500 CD45+ cells / mm2). [Figure 71] For subjects in Cohort 3, no significant effect of placebo on biopsy material markers was observed at day 85. Figure 71A: pDC cells (measured as a percentage change from baseline). Figure 71B: MxA+ pixels (measured as a percentage change from baseline). Figure 71C: CD45+ cells (measured as a percentage change from baseline). [Figure 72] This document shows that at day 85, most subjects in Cohort 3 treated with 150 mg of ILT7-binding protein (VIB7734) as used in the method described herein showed a marked decrease in pDC cells (measured as a percentage change from baseline; Figure 72A) and IFN activity (MxA+ pixels; measured as a percentage change from baseline; Figure 72B), as well as a slight decrease in inflammatory infiltration (CD45+ cells; measured as a percentage change from baseline; Figure 72C), compared to Cohort 3 subjects treated with placebo. Figure 72A: Compared to a mean increase in pDCs of 12.24 ± 37.69 (mean ± SEM) in Cohort 3 subjects treated with placebo, a mean decrease in pDCs of 80.98 ± 12.12 (mean ± SEM) was observed in Cohort 3 subjects treated with VIB7734. Figure 72B: Compared to the mean increase in MxA+ pixels of 773.6 ± 866.33 (mean ± SEM) in the placebo-treated cohort 3, the VIB7734-treated cohort 3 observed a mean decrease in MxA+ pixels of 58.29 ± 17.88 (mean ± SEM). [Figure 73]This document shows that nearly all subjects in Cohort 3 (with intermediate or high baseline signal levels) treated with 150 mg of ILT7-binding protein (VIB7734) as used in the method described herein showed a marked decrease in pDC cells (measured as a percentage of baseline on day 1; Figure 73A) and IFN activity (MxA+ pixels; measured as a percentage of baseline on day 1; Figure 73B), as well as a decrease in inflammatory infiltration (CD45+ cells; measured as a percentage of baseline on day 1; Figure 73C) at day 85 compared to the placebo-treated subjects in Cohort 3. Circles indicate samples from Cohort 3 subjects with low baseline activity. [Figure 74] Figure 74A: Changes in pDCs (measured as BDCA2+ / ILT7+ cells) from baseline (BL) to day 85 (d85) for each subject in Cohort 3 treated with 150 mg of ILT7-binding protein (VIB7734) used in the method described herein or with placebo. Figure 74B: Changes in pDCs (measured as BDCA2+ / ILT7+ cells) using IHC analysis of skin biopsy materials for cohort 3 subjects (n=8; 30010047, 10120061, 20070048, 200020040, 10010056, 10120055, 20060038, and 10140059) treated with 150 mg of VIB7734. Figure 74C: Changes in pDCs (measured as the number of cells per square mm) in skin biopsy materials for each of the cohort 3 subjects treated with VIB7734 and each of the cohort 3 subjects treated with placebo at day 85 compared to baseline. [Figure 75]This document presents combined data for subjects in Cohorts 2 and 3 treated with 50 mg (Cohort 2) or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) as used in the method described herein. VIB7734 significantly reduced cutaneous pDCs in subjects in Cohorts 2 and 3 treated with VIB7734 compared to subjects in Cohorts 2 and 3 treated with placebo. Figure 75A: Change in pDCs (measured as a percentage of baseline on day 1) in placebo-treated subjects (n=6) of Cohorts 2 and 3 compared to VIB7734-treated subjects (n=16) of Cohorts 2 and 3. The mean and median pDC reductions were 11.38% and 12.73% for the placebo-treated group in Cohorts 2 and 3, respectively, compared to the mean and median pDC reductions of 71.82% and 95.3% for the VIB7734-treated group in Cohorts 2 and 3, respectively. Figure 75B: Change in MxA+ pixels (measured as a percentage of baseline on day 1) for the placebo-treated group (n=6) in Cohorts 2 and 3 compared to the VIB7734-treated group (n=16) in Cohorts 2 and 3. The mean and median MxA+ pixel increase was 269.3% and 38.8% for the placebo-treated group in Cohorts 2 and 3, respectively, compared to the mean and median pDC reductions of 52.9% and 84.48% for the VIB7734-treated group in Cohorts 2 and 3, respectively. Figures 75C-75D: In all VIB7734-treated subjects with ≥2 pDCs / mm2 in baseline skin, a correlation was shown between the rate of change in pDCs from baseline to day 85 in skin biopsy material and MxA (Figure 75C) or CD45+ cells (Figure 75D). Spearman's correlation is shown. [Figure 76]In both cohorts 2 and 3, treated with 50 mg and 150 mg of VIB7734 respectively, cutaneous pDCs (measured as the number of cells per square mm) decreased, but pDC depletion was more consistent in cohort 3 subjects. Figure 76A: Decrease in pDCs in skin biopsy material at day 85 compared to baseline at day 1 for all VIB7734-treated cohort 2 subjects (n=8). Figure 76B: Decrease in pDCs in skin biopsy material at day 85 compared to baseline at day 1 for all VIB7734-treated cohort 3 subjects (n=8). Figure 76C: Decrease in pDCs in skin biopsy material at day 85 for VIB7734-treated cohort 2 subjects (n=6) with baseline pDC or no low IFN activity compared to baseline at day 1. Figure 76D: Decrease in pDCs in skin biopsy materials from cohort 3 subjects treated with VIB7734, where baseline pDC or IFN activity was not low, compared to baseline at day 1 (n=6). [Figure 77] The median circulating pDC levels (measured as %PBMC cells) in whole blood of subjects in Cohorts 1, 2, and 3 treated with 5 mg (Cohort 1), 50 mg (Cohort 2), or 150 mg (Cohort 3) of ILT7-binding protein (VIB7734) as used in the methods described herein are shown. A reduction in the median circulating pDC levels was evident at week 1, and this reduction persisted at least until day 85 in the VIB7734-treated subjects in Cohorts 1, 2, and 3 compared to the median circulating pDC levels in the placebo-treated subjects. [Figure 78] In the skin of subjects with cutaneous lupus, depletion of tissue-resident pDCs promotes a decrease in type I IFN activity. Figures 78A-78C: MxA staining, defined as the MxA positivity rate in the region of interest area, was quantified in skin punch biopsy materials at baseline and day 85 of the study. Each line represents an individual subject. Each point on the graph represents the mean MxA+ rate from serial sections within the biopsy material. [Figure 79]This shows that elevated baseline serum type I IFN activity is associated with a higher response rate to the ILT7-binding protein (VIB7734) used in the method described herein. Figure 79A: Percentage change in whole blood type I IFNGS at the indicated time points for placebo-treated and VIB7734-treated subjects (cohorts 1, 2, and 3) who had elevated baseline type I IFNGS scores (defined as more than four times the mean of healthy donors). The number of subjects with elevated baseline IFN activity is shown above the graph for each group (cohort 1, n=3; cohort 2, n=6; cohort 3, n=8). Median and interquartile range are shown. Figure 79B: Percentage change in serum IFNα levels for placebo-treated and VIB7734-treated subjects who had elevated baseline IFNα levels (defined as two standard deviations above the mean of healthy donors). Median and interquartile range are shown. Figure 79C: Correlation between baseline whole blood type I IFNGS and serum IFNα protein levels in subjects treated with VIB7734. Black dots indicate CLASI responders (defined as a decrease of 4 or more points on the CLASI scale), and red dots indicate CLASI non-responders. The dotted lines represent a change of four times the healthy donor mean for type I IFNGS (FC from HD mean) and a value two standard deviations above the healthy donor mean for IFNα protein. [Modes for carrying out the invention]

[0028] Detailed description of the invention Unless otherwise defined, all scientific and technical terms used herein have the same meaning as those generally understood by those skilled in the art in the field relating to this subject matter. All publications, patent applications, patents, and other references referred to herein are expressly incorporated by reference in their entirety. In case of any inconsistency, including definitions, this specification shall prevail. In addition, the materials, methods, and examples described herein are illustrative and not intended to limit the scope of the subject matter.

[0029] When used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless specifically indicated by the context.

[0030] In this specification, a range may be expressed as "approximately" from one particular value and / or "approximately" to another particular value. When such a range is expressed, an alternative embodiment includes the range from one particular value and / or the other particular value. Similarly, when a value is expressed as an approximation by using the preceding "approximately," it will be understood that the particular value constitutes an alternative embodiment. Furthermore, it will be understood that each endpoint of a range is significant both with respect to the other endpoint and independently of the other endpoint. When used herein, the term "approximately" in the context of a particular usage refers to a range of ±15% of the given number. For example, approximately 10 would include the range of 8.5 to 11.5. The term "approximately" also takes into account typical errors or inaccuracies in the measurement of the value.

[0031] This disclosure provides a method for treating an autoimmune disorder in a subject with an ILT7-binding protein. In certain embodiments, the method provides for treating an autoimmune disorder in a subject in need, where the subject is determined to have elevated serum type I interferon gene signature (IFNGS) levels. The disclosure also provides a method for reducing IFNGS in a subject in need. In some embodiments, the method involves administering a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7)-binding protein to the subject. In some embodiments, the ILT7-binding protein is administered to the subject when the subject's type I IFNGS is elevated compared to that of a normal subject. In specific embodiments, for example, the ILT7-binding protein is administered to a subject with elevated baseline type I IFNGS compared to that of a normal subject. In a particular embodiment, the method provides for selecting patients for treatment with ILT7-binding protein, the method comprising (i) determining the patient's baseline serum type I IFNGS level, and (ii) selecting patients with high baseline serum type I IFNGS levels for treatment with ILT7-binding protein. In a detailed embodiment, the ILT7-binding protein is an antibody. In a particular embodiment, the antibody is VIB7734.

[0032] In certain embodiments, type I IFNGS is a 21-gene signature. In some embodiments, the subject's type I IFNGS is elevated at least 1.5 times compared to a normal score before treatment. In some embodiments, the subject's type I IFNGS is elevated at least 2 times compared to a normal score before treatment. In certain embodiments, subjects with elevated type I IFNGS before treatment are more responsive to the treatment. In some embodiments, type I IFNGS is at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 11 times, at least about 12 times or more compared to a normal score before treatment with the ILT7 binding protein used in the method described herein. In detailed embodiments, type I IFNGS in tissue is determined from skin biopsy material. In other embodiments, type I IFNGS in tissue is determined using IFN-inducible myxovirus protein A (MxA) immunohistochemistry (IHC) testing. In a further embodiment, IFN-inducible gene expression in the epidermis is determined using skin tape stripping, RNA isolation, and gene expression profiling (https: / / dermtech.com / wp-content / uploads / Lupus-Reference.pdf).

[0033] As used herein, the terms “high” or “rising” mean, when used in conjunction with IFGNS, that type I IFNGS are at least about 1.1 to about 1000 times higher than normal type I IFNGS. “Normal type I IFNGS” refers to type I IFNGS obtained from a normal subject. The terms “high” or “rising” are used synonymously when used in conjunction with type I IFNGS. In some embodiments, type I IFNGS are “high” or “rising” when type I IFNGS used in the manner described herein are at least about 1.1, 1.2, 1.3, 1.4, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, or 100 times higher than type I IFNGS from a normal subject. In a specific embodiment, the treatment method described herein is applied when the type I IFNGS is elevated to at least about four times that of normal type I IFNGS.

[0034] In certain diseases (e.g., autoimmune diseases), activated pDCs secrete significant amounts of type I and type III interferons (IFNs). Type I IFNs are a large group of IFN proteins that help regulate the immune system. Mammalian IFNs are called IFNα, IFNβ, IFNω, IFNε, IFNκ, IFNτ, IFNδ, IFNζ, and IFNυ. In specific embodiments, IFNα is the type I IFN that produces type I IFNGS. There is no reliable method to directly measure type I IFN protein levels; however, measuring IFN-inducible genes provides a robust substitute for type I IFN protein levels. The expression levels of these type I IFN-inducible genes can be measured in biological samples (e.g., blood, skin, skeletal muscle, etc.) and analyzed as a composite outcome referred to as "type I interferon gene signature," "type I IFNGS," or "IFNGS."

[0035] In certain embodiments, type I IFNGS include the expression levels of all type I IFN-inducible genes in the biological sample. In other embodiments, type I IFNGS include the expression levels of some type I IFN-inducible genes in the biological sample.

[0036] In certain embodiments, type I IFNGS are determined by assaying the expression levels of at least 2, at least 5, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, at least 200, at least 300, at least 400, or at least 500 type I IFN-inducible genes in a biological sample. In some embodiments, type I IFNGS include the collective expression levels of two or more type I IFN-inducible genes. In certain embodiments, two or more type I interferon (IFN)-inducible genes include, but are not limited to, two or more genes selected from SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, or USP18. In certain cases, type I IFNGS are determined by assaying the collective expression levels of SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, and USP18. These gene symbols are well known in the relevant field and refer to human and non-human orthologues of the genes listed.

[0037] (Table 1) Genetic symbols and their related names TIFF2026053543000002.tif165166

[0038] In certain embodiments, the expression level of type I interferon (IFN)-inducible genes is determined by measuring the DNA level (e.g., complementary DNA, i.e., cDNA level) of type I interferon (IFN)-inducible genes in a biological sample. In certain embodiments, the expression level of type I interferon (IFN)-inducible genes is determined by measuring the messenger RNA (mRNA) level of type I interferon (IFN)-inducible genes in a biological sample. In certain embodiments, type I IFNGS includes the mRNA levels of all type I IFN-inducible genes in a biological sample. In other embodiments, type I IFNGS includes the mRNA levels of some type I IFN-inducible genes in a biological sample taken from a subject suffering from, susceptible to, or suspected to suffer from a disease, such as an autoimmune disease. In certain embodiments, type I IFNGS is determined by assaying the mRNA levels of two or more type I interferon (IFN)-inducible genes in a biological sample. In a specific embodiment, type I IFNGS are determined by assaying the mRNA levels of 21 type I interferon (IFN)-inducible genes in a biological sample. In certain embodiments, the biological sample is a test biological sample. In other embodiments, the biological sample is a normal biological sample.

[0039] In certain embodiments, type I IFNGS are measured in a test biological sample taken from the subject. In other embodiments, pDCs are measured in a test biological sample taken from the subject. Biological samples include, but are not limited to, blood, sputum, saliva, skin cells, skin biopsy samples, kidney cells, lung cells, hepatocytes, cardiac cells, brain cells, nerve tissue, thyroid cells, ocular cells, skeletal muscle cells, cartilage, bone tissue, airway cells, and cultured cells. In certain embodiments, the biological sample is blood. In other embodiments, the biological sample is tissue. In more specific embodiments, the sample is tissue containing skin cells. In other embodiments, the sample is a skin biopsy sample.

[0040] "Test specimens" refer to any specimens obtained from individuals who are suffering from, susceptible to, or suspected of suffering from a disease or condition such as autoimmune disorders, and / or, but not limited to, individuals exhibiting one or more of the symptoms, such as elevated type I IFNGS.

[0041] "Normal biological sample" refers to any biological sample obtained from a healthy subject.

[0042] As used herein, the term “subject” refers to any individual, e.g., human or non-human mammal, for which diagnosis, prognosis, or treatment is desired. The term “subject” may also mean a human or non-human mammal suffering from, susceptible to, or suspected to suffering from a disease, e.g., an autoimmune disease or condition. The terms “subject” and “patient” are used synonymously herein. While the ILT7-binding protein compositions provided herein are primarily suited for administration to humans, those skilled in the art will understand that such compositions are generally suited for administration to all kinds of subjects. In certain embodiments, the subject is a mammal. Mammals include primates such as humans, monkeys, chimpanzees, and apes, as well as non-primates such as domesticated animals including laboratory animals (rabbits and rodents, such as guinea pigs, rats, or mice) and domestic pets and farm animals (such as cats, dogs, pigs, cows, sheep, goats, horses, and rabbits), and non-domesticated animals such as wild animals, birds, reptiles, and fish.

[0043] When used herein, the term “subjects in need of it” includes subjects who may or would benefit from the methods described herein. Subjects in need of treatment include, without limitation, subjects who already have a condition or disorder, subjects prone to developing a condition or disorder, subjects suspected of having a condition or disorder, and subjects whose condition or disorder should be prevented, improved, or cured.

[0044] As used herein, the term “normal subject” refers to any healthy individual that is not suffering from any disease or is not suspected of suffering from any disease or condition, e.g., a human or non-human mammal. The term “normal subject” also refers to an individual that has not yet exhibited any symptoms associated with autoimmune disorders, e.g., a human or non-human mammal, such as elevated type I IFNGS. The normal subject is the same as the subject in need of treatment, and that subject may be the subject that has not yet exhibited any symptoms of autoimmune disorders, such as elevated type I IFNGS, but is not limited to. In other embodiments, the normal subject and the subject in need of treatment are two different individuals.

[0045] This disclosure provides a method for treating a subject with elevated type I IFNGS, comprising administering an ILT7-binding protein as described herein. A patient may exhibit elevated type I IFNGS when suffering from an autoimmune disorder. Accordingly, this disclosure provides a method for treating an autoimmune disorder when a subject exhibits elevated type I IFNGS. In some embodiments, the autoimmune disorder is otherwise asymptomatic. In certain embodiments, the method provides selecting a patient for treatment with an ILT7-binding protein, the method comprising (i) determining the patient's baseline serum type I IFNGS level, and (ii) selecting a patient with high baseline serum type I IFNGS levels for treatment with an ILT7-binding protein.

[0046] As used herein, “to treat” or “to cure” refers to the management and care of an object aimed at combating a disease, condition, or disorder, and includes the reduction of symptoms or complications of a disease, condition, or disorder, or the elimination of a disease, condition, or disorder, by administration of ILT7-binding protein used in the manner described herein. Accordingly, the term “to treat” or “to cure” refers to both therapeutic and prophylactic or defensive measures, the purpose of which is to prevent, slow (reduce), or improve the progression of a disease (e.g., an autoimmune disease). Beneficial or desired clinical outcomes include, but are not limited to, symptom reduction, reduction of the severity of the disease, stabilization of the disease state (i.e., not worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, and improvement of the disease (whether partial or complete). The term “to treat” may also include the treatment of cells in vitro or in animal models.

[0047] In some embodiments, the treatment includes the application or administration of ILT7-binding protein as used herein to a subject in need of such treatment or a subject suspected of needing such treatment, or the application or administration of ILT7-binding protein as used herein to tissues or cell lines isolated from the subject, wherein the subject has a disease (e.g., an autoimmune disease), symptoms of a disease, or a predisposition to the disease. When a subject exhibits a pathological condition or symptoms of a disease due to an excessive number or activity of pDCs, the subject may be suspected of needing the treatment described herein, even if a formal diagnosis, such as a diagnosis that the subject has SLE or CLE, has not been established. In certain embodiments, the subject suspected of needing treatment has elevated baseline serum type I IFNGS levels. In other embodiments, the treatment is also intended to include the application or administration of a pharmaceutical composition comprising an ILT7-binding protein used in the manner described herein to a subject in need of such treatment or suspected to be in need of such treatment, or the application or administration of a pharmaceutical composition comprising an ILT7-binding protein used in the manner described herein to tissues or cell lines isolated from a subject having a disease (e.g., an autoimmune disease), symptoms of a disease, or a predisposition to the disease.

[0048] Examples of autoimmune disorders that can be treated when the subject presents with elevated type I IFNGS include, but are not limited to, systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, inflammatory myositis, such as dermatomyositis, inclusion body myositis, juvenile myositis and polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, pure red cell fistula, multiple sclerosis, rheumatic carditis, psoriasis, psoriatic arthritis, rheumatoid arthritis, chronic inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, acute and chronic graft-versus-host disease (GVHD), vasculitis, myocardial infarction, and type I interferonism. In a particular embodiment, the autoimmune disease is SLE. In a further embodiment, the autoimmune disease is CLE. In certain embodiments, the autoimmune disease is lupus, but not discoid lupus erythematosus (DLE). In other embodiments, the autoimmune disease is Sjögren's syndrome. In yet another embodiment, the autoimmune disease is dermatomyositis. In yet another embodiment, the autoimmune disease is polymyositis. In yet another embodiment, the autoimmune disease is systemic sclerosis. In yet another embodiment, the autoimmune disease is hidradenitis suppurativa. In yet yet another embodiment, the autoimmune disease is vitiligo.

[0049] In further embodiments, the method of the present disclosure can be used to monitor the effectiveness of treatment for a disease or disorder by monitoring the levels of type I IFNGS and / or activated pDCs. As described above, autoimmune conditions are often characterized by elevated type I IFNGS and / or pDCs, so monitoring the effectiveness of treatment may include monitoring type I IFNGS and / or pDC levels.

[0050] Accordingly, in certain embodiments, the present disclosure provides a method for monitoring the effectiveness of treatment for an autoimmune disorder or condition, comprising the steps of (a) measuring type I interferon gene signatures (IFNGS) in a biological sample taken from a subject to obtain a baseline value of type I IFNGS; and (b) measuring type I IFNGS in a biological sample taken from a subject after treatment, wherein the treatment comprises administering ILT7-binding protein, wherein a decrease in type I IFNGS in step (b) compared to a baseline value indicates that the treatment is effective in the subject.

[0051] In certain embodiments, the treatment results in a decrease in type I IFNGS compared to baseline values. In certain embodiments, the decrease in type I IFNGS compared to baseline values ​​ranges from about 1% to about 99%. In certain embodiments, the decrease in type I IFNGS compared to baseline values ​​is at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%. In some embodiments, the decrease in type I IFNGS compared to baseline values ​​is at least about 30%. In specific embodiments, the decrease in type I IFNGS compared to baseline values ​​is at least about 50%.

[0052] In certain embodiments, the increase in type I IFNGS in the test sample compared to a normal biological sample, or in the subject requiring treatment with ILT7-binding protein compared to a normal subject, is at least about 1.1 to about 1000 times. Therefore, in some embodiments, type I IFNGS is increased by at least about 1.1, 1.2, 1.3, 1.4, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, or 100 times in the test sample compared to a normal biological sample, or in the subject requiring treatment with ILT7-binding protein compared to a normal subject. In specific embodiments, type I IFNGS is increased by at least about 4 times in the test sample compared to a normal biological sample, or in the subject requiring treatment with ILT7-binding protein compared to a normal subject.

[0053] In general, the terms “ILT7-binding protein,” “ILT7-binding molecule,” and “ILT7-binding protein as used in the methods described herein” are used synonymously to refer to a protein or molecule that specifically binds to immunoglobulin-like transcript 7 (ILT7). The terms protein and peptide are used synonymously herein. In some embodiments, the ILT7-binding protein as used in the methods described herein binds to full-length ILT7. In other embodiments, the ILT7-binding protein as used in the methods described herein binds to a fragment of ILT7. In certain embodiments, the fragment of ILT7 to which the ILT7-binding protein binds includes the extracellular domain of ILT7.

[0054] In certain embodiments, the ILT7-binding protein used in the methods disclosed herein binds to any mammalian ILT7. In specific embodiments, the ILT7-binding protein used in the methods disclosed herein binds to human ILT7 or a fragment thereof, for example, the extracellular portion of human ILT7. In other embodiments, the ILT7-binding protein used in the methods disclosed herein binds to cynomolgus monkey ILT7 or a fragment thereof, for example, the extracellular portion of cynomolgus monkey ILT7.

[0055] Examples of ILT7-binding proteins are disclosed and described in International Publication No. 2017 / 156298 (which is incorporated herein by reference in its entirety). In certain embodiments, the ILT7 to which the ILT7T-binding protein binds is located on a pDC. In specific embodiments, the ILT7-binding protein is the VIB7734 antibody or a fragment thereof. VIB7734 is described in International Publication No. 2017 / 156298 (which is incorporated herein by reference in its entirety). Specifically, VIB7734 is identified as clone ILT70137 in International Publication No. 2017 / 156298. In another embodiment, VIB7734 is also an antibody containing the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2.

[0056] In certain embodiments, the ILT7-binding protein used in the method described herein includes the heavy chain variable region (VH) of SEQ ID NO: 1. In other embodiments, the ILT7-binding protein used in the method described herein includes the light chain variable region (VL) of SEQ ID NO: 2. In certain embodiments, the ILT7-binding protein used in the method described herein includes the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO: 2. In other embodiments, the ILT7-binding protein used in the method described herein includes a VH that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1 and / or a VL that is at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2.

[0057] In a more specific embodiment, the ILT7-binding protein used in the method described herein comprises heavy chain complementarity-determining regions (HCDRs), HCDR1, HDR2, HCDR3, and light chain complementarity-determining regions (LCDRs), LCDR1, LCDR2, and LCDR3, each having the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively. In another embodiment, the ILT7-binding protein used in the method described herein comprises heavy chain complementarity-determining regions (HCDRs), HCDR1, HDR2, HCDR3, and light chain complementarity-determining regions (LCDRs), LCDR1, LCDR2, and LCDR3, each being at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NOs: 3, 4, 5, 6, 7, and 8.

[0058] In certain embodiments, the ILT7-binding protein used in the method described herein may contain a fucose moiety, or it may be afucosylated.

[0059] While not constrained by theory, the ILT7-binding protein used in the methods described herein induces antibody-dependent cell-mediated cytotoxicity (ADCC) activity against plasmacytoid dendritic cells (pDCs), thereby depleting them. In certain embodiments, ILT7-binding protein-mediated ADCC causes a decrease in circulating pDCs. In certain embodiments, ILT7-binding protein-mediated ADCC causes a decrease in local or tissue-mediated pDCs. In certain embodiments, tissues in which pDCs are reduced include, but are not limited to, skin cells, skin biopsy samples, kidney cells, lung cells, hepatocytes, cardiac cells, brain cells, nerve tissue, thyroid cells, ophthalmic cells, skeletal muscle cells, cartilage, bone tissue, and airway cells. In some embodiments, the tissue is a skin biopsy sample. In more specific embodiments, administration of ILT7-binding protein will result in a decrease in cutaneous pDCs.

[0060] Normally, pDCs do not exist in skin tissue, and immature pDCs are typically found only in blood, thymic lymphoid tissue, tonsils, and lung tissue. Therefore, the presence of pDCs in a skin biopsy sample indicates an abnormal state in which pDCs are recruited to the skin. Accordingly, the methods of the present disclosure include administering an ILT7-binding protein to a subject that requires treatment of a disease state characterized by the presence of pDCs in the skin of the subject. The methods of the present disclosure include reducing the level of pDCs in the skin of a subject by administering an ILT7-binding protein to the subject that requires such treatment.

[0061] In some embodiments, the subject exhibits an elevated or high level of pDCs in skin tissue prior to treatment. In certain embodiments, subjects with a high level of pDCs in skin tissue prior to treatment are more responsive to this treatment. In certain aspects, a subject with a high level of pDCs in skin tissue has at least about 50 pDCs / mm 2 skin tissue, at least about 60 pDCs / mm 2 skin tissue, at least about 70 pDCs / mm 2 skin tissue, at least about 80 pDCs / mm 2 skin tissue, at least about 90 pDCs / mm 2 skin tissue, at least about 100 pDCs / mm 2 skin tissue, at least about 110 pDCs / mm 2 skin tissue, at least about 120 pDCs / mm 2 skin tissue, at least about 125 pDCs / mm 2 skin tissue, at least about 150 pDCs / mm 2 skin tissue, at least about 175 pDCs / mm 2 skin tissue, at least about 200 pDCs / mm 2 skin tissue, or have a pDC level of more. In certain embodiments, a low level of pDCs in skin tissue is considered to be less than about 10 pDCs / mm 2 skin tissue. In a specific embodiment, a high level of pDCs in skin tissue is considered to be at least about 100 pDCs / mm 2 skin tissue.

[0062] In other embodiments, the methods of the present disclosure include inhibiting the release of type I IFN from pDCs, regardless of the site of the pDC, by administering an ILT7-binding protein used in the methods described herein. In other embodiments, the methods of the present disclosure include inhibiting the release of type I IFN from pDCs in the blood or circulation by administering an ILT7-binding protein. In other embodiments, the methods of the present disclosure include inhibiting the release of type I IFN from local pDCs by administering an ILT7-binding protein. In other embodiments, the methods of the present disclosure include inhibiting the release of type I IFN from pDCs in the skin of a subject by administering an ILT7-binding protein. In certain embodiments, the type I IFN whose release is inhibited is IFNα. In certain embodiments, inhibition of type I IFN release from pDCs mediated by an ILT7-binding protein results in a reduction of type I IFNGS.

[0063] The terms “reduce,” “to decrease,” or “decrease” mean making a degree, level, amount, activity, or magnitude smaller than an initial value. The decrease does not need to be statistically significant from one value to the next.

[0064] The terms “administer,” “dosage,” and “administer” refer to the contact of a compound or reagent with a subject, cell, tissue, organ, or biological sample, for example, when applied to a subject, cell, tissue, organ, or biological sample. In the context of cells, administration includes contact of the reagent with the cell (e.g., in vitro or ex vivo) and contact of the reagent with body fluids (where the body fluid is in contact with the cell). The ILT7-binding protein used in the methods described herein may be administered to a subject by various routes known in the art. Exemplary routes for administering the ILT7-binding protein used in the methods described herein include, but are not limited to, parenteral, oral, mucosal, topical, transdermal, inhalation, sublingual, buccal, rectal, vaginal, and intranasal. The term parenteral, as used herein, includes subcutaneous, intravenous, intramuscular, intrasternal injection, or infusion techniques. In certain embodiments, the ILT7-binding protein used in the methods described herein is administered intravenously. In specific embodiments, the ILT7-binding protein used in the methods described herein is administered by subcutaneous injection. The terms “administer,” “give,” or “give administration” may refer to a single or multiple administration of the ILT7-binding protein used in the methods described herein. For example, multiple administrations refer to at least two (i.e., two, three, four, five, six, seven, eight, nine, ten, or more) administrations of the ILT7-binding protein used in the methods described herein to a subject.

[0065] The “therapeutic dose,” “pharmaceutically effective dose,” or “effective dose” of a compound (e.g., ILT7-binding protein used in the methods described herein) refers to an amount sufficient to produce in a subject a response leading to the desired prevention, treatment, or improvement, or an amount sufficient to produce in a statistically significant manner the prevention or improvement of one or more symptoms of a disease or condition. When referring to an individual active ingredient administered alone, the therapeutic dose refers to that ingredient alone. When referring to a combination, the therapeutic dose refers to the amount of the combination of active ingredients that produce a therapeutic effect, whether administered sequentially or simultaneously. As used herein, the term “therapeutic dose” means that the ILT7-binding protein used in the methods described herein, when used as prescribed or directed, is capable of producing a medically beneficial effect compared to placebo (e.g., causing a reduction in elevated type I IFNGS and / or pDCs in a subject in need). The therapeutic dose will vary depending on the species and body weight of the subject being administered, but can be determined using standard techniques. In certain embodiments, the therapeutically effective dose of ILT7-binding protein used in the method described herein is in the range of about 0.1 mg to about 1000 mg. In other embodiments, the therapeutically effective dose of ILT7-binding protein used in the method described herein is in the range of about 50 mg to about 150 mg. In certain embodiments, the therapeutically effective dose of ILT7-binding protein used in the method described herein is, but is not limited to, about 1 mg, about 5 mg, about 15 mg, about 50 mg, about 100 mg, about 150 mg, about 300 mg, about 500 mg, or about 1000 mg. In certain embodiments, the therapeutically effective dose of ILT7-binding protein used in the method described herein is about 5 mg in single doses. In other embodiments, the therapeutically effective dose of ILT7-binding protein used in the method described herein is about 50 mg in single doses. In yet another embodiment, the therapeutically effective dose of ILT7-binding protein used in the method described herein is about 150 mg in single doses.The therapeutically effective dose of ILT7-binding protein used in the methods described herein may be administered to a subject in need as a single dose or multiple doses.

[0066] In certain embodiments, administration of a therapeutically effective dose of the ILT7-binding protein used in the method described herein to a subject in need leads to a reduction of approximately 1% to approximately 100% of the subject's type I IFNGS compared to the subject's type I IFNGS before administration of the ILT7-binding protein used in the method described herein. In certain embodiments, administration of a therapeutically effective dose of the ILT7-binding protein used in the method described herein to a subject in need leads to a reduction of at least approximately 1%, at least approximately 2%, at least approximately 5%, at least approximately 10%, at least approximately 20%, at least approximately 30%, at least approximately 30%, at least approximately 40%, at least approximately 50%, at least approximately 60%, at least approximately 70%, at least approximately 80%, at least approximately 90%, at least approximately 95%, at least approximately 96%, at least approximately 97%, at least approximately 98%, at least approximately 99%, or approximately 100% of the subject's type I IFNGS compared to the subject's type I IFNGS before administration of the ILT7-binding protein used in the method described herein. In a specific embodiment, administering a therapeutically effective dose of ILT7-binding protein leads to a reduction of at least approximately 50% in the target type I IFNGS.

[0067] In certain embodiments, administration of the ILT7-binding protein used in the method described herein to a subject in need leads to a reduction of at least about 50% in the subject's type I IFNGS compared to type I IFNGS before administration of the ILT7-binding protein used in the method described herein. In certain embodiments, administration of a therapeutically effective dose of the ILT7-binding protein used in the method described herein to a subject in need leads to a reduction of at least about 50% in the subject's type I IFNGS at about 8 hours, about 12 hours, about 24 hours, or about 48 hours after administration of the ILT7-binding protein.

[0068] In specific embodiments, subjects administered a therapeutically effective dose of ILT7-binding protein used in the method described herein show at least a 50% reduction in type I IFNGS at approximately 24 hours after administration of ILT7-binding protein compared to type I IFNGS in subjects before administration of ILT7-binding protein.

[0069] In certain embodiments, the decrease in type I IFNGS lasts for at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 14 days, at least about 21 days, at least about 28 days, at least about 30 days, at least about 45 days, at least about 60 days, at least about 90 days, or at least about 180 days or longer, after administering a therapeutically effective amount of ILT7-binding protein used in the manner described herein to a subject in need. In further embodiments, the decrease in type I IFNGS lasts for up to about 30 days after administering a therapeutically effective amount of ILT7-binding protein used in the manner described herein to a subject in need. Thus, in some embodiments, a therapeutically effective amount of ILT7-binding protein used in the manner described herein is administered to a subject in need at least once a month. In other embodiments, the therapeutically effective dose of the ILT7-binding protein used in the method described herein is administered to a subject at least once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, or 52 weeks. In some embodiments, the therapeutically effective dose of the ILT7-binding protein used in the method described herein is administered to a subject at least once every 4 weeks. In further embodiments, the therapeutically effective dose of the ILT7-binding protein used in the method described herein is administered to a subject at least once every 8 weeks or at least once every 12 weeks. In a further embodiment, the therapeutically effective dose of the ILT7-binding protein used in the method described herein is administered to the subject at least once every two or three months.In further embodiments, the therapeutically effective dose of the ILT7-binding protein used in the method described herein is administered to the subject at least once a year or at least once every two years.

[0070] As used herein, the terms “reduction in pDCs” or “reducing pDCs” refer to a decrease in the level of activated pDCs in or in a biological sample taken from a subject (e.g., blood and / or other tissues such as skin cells or skin biopsy samples), or a decrease in the total number of pDCs in or in a biological sample taken from a subject, or both. In some embodiments, the reduction in pDCs in a subject is about 1% to about 100% compared to the pDCs in the subject before administration of the ILT7-binding protein used in the method described herein. In certain embodiments, the reduction in pDCs in a subject is at least about 1%, at least about 2%, at least about 5%, at least about 10%, at least about 20%, at least about 30%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or about 100% compared to the pDCs in the subject before administration of the ILT7-binding protein used in the method described herein. In specific embodiments, the reduction in pDCs in a subject is at least about 50% compared to the pDCs in the subject before administration of the ILT7-binding protein used in the method described herein. Thus, in certain embodiments, administration of a therapeutically effective dose of ILT7-binding protein leads to a reduction of at least about 10% in the total number of pDCs in a subject. In further embodiments, administration of a therapeutically effective dose of ILT7-binding protein leads to a reduction of at least about 10% in the activated pDCs in a subject. In certain embodiments, pDCs are measured in a test biopsy taken from a subject. Therefore, in certain embodiments, administering a therapeutically effective amount of the ILT7-binding protein used in the method described herein to a subject in need leads to a decrease in pDCs in a test biopsy taken from the subject. In specific embodiments, the decrease in pDCs in the test biopsy taken from the subject is at least about 10% compared to the pDCs in the test biopsy before administration of the ILT7-binding protein used in the method described herein. In certain embodiments, the test biopsy is blood.In specific embodiments, the biological sample being tested is tissue, including, but not limited to, skin cells and skin biopsy specimens. In certain embodiments, pDCs are circulating pDCs. In other embodiments, pDCs are skin pDCs. In further embodiments, the reduction of pDCs is reversible.

[0071] In certain embodiments, administration of a therapeutically effective dose of ILT7-binding protein used in the method described herein to a subject in need results in a reduction of at least about 10% of pDCs in the subject at approximately 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 24 hours, or 48 hours after administration of the ILT7-binding protein. In other embodiments, administration of a therapeutically effective dose of ILT7-binding protein used in the method described herein to a subject in need results in a reduction of at least about 10% of pDCs in a test biological sample taken from the subject at approximately 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 6 hours, 12 hours, 24 hours, or 48 hours after administration of the ILT7-binding protein.

[0072] In certain embodiments, the decrease in pDCs continues for at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 14 days, at least about 21 days, at least about 28 days, at least about 30 days, at least about 45 days, at least about 60 days, at least about 90 days, or at least about 180 days or longer, after administering a therapeutically effective amount of ILT7-binding protein used in the method described herein to a subject in need. In further embodiments, the decrease in pDCs continues for at least about 30 days after administering a therapeutically effective amount of ILT7-binding protein used in the method described herein to a subject in need.

[0073] In other embodiments, the method of the present disclosure can be used to reduce the Cutaneous Lupus Erythematosus Disease Activity and Severity Index (CLASI) in a target tissue in need. The method comprises administering a pharmaceutically effective amount of ILT7-binding protein to the subject.

[0074] As used herein, the term “CLASI” refers to the Cutaneous Lupus Erythematosus Disease Activity and Severity Index. CLASI is a valid instrument for measuring the cutaneous findings of CLE. CLASI consists of two scores: the first summarizes the inflammatory activity of the disease; and the second measures the chronic damage caused by the disease. The activity score includes erythema (0-3), scaling / hypertrophy (0-2), mucosal lesions (0-1), recent alopecia (0-1), and non-scarring alopecia (0-3). The chronic damage score represents hyperpigmentation (0-1), scarring / atrophy / panniculitis (0-2), and scalp scarring (0-6). Patients are asked whether their hyperpigmentation has persisted for more than 12 months, and if so, the hyperpigmentation score is doubled. Each of the above parameters is measured at 13 different anatomical sites, specifically chosen because they are most frequently associated with CLE. Within each region, the most severe lesions are measured.

[0075] As used herein, the term “decreased CLASI” refers to a decrease in the level of CLASI activity (CLASI-A) score in or from a biological sample (e.g., skin cells, skin biopsy specimens, or other tissue issues) taken from the subject, or a decrease in the level of CLASI chronic lesion (CLASI-D) score in or from the subject, or from both.

[0076] Accordingly, in certain embodiments, the method of the present disclosure results in a reduced CLASI-A score in the subject. In certain embodiments, the reduction in the subject's CLASI-A score involves a decrease of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 points from the baseline value of the CLASI-A score. In some embodiments, the reduction in the subject's CLASI-A score involves a decrease of at least 4 points from the baseline value of the CLASI-A score. In specific embodiments, the reduction in the subject's CLASI-A score involves a decrease of at least 7 points from the baseline value of the CLASI-A score. In other embodiments, the reduction in the subject's CLASI-A score involves a decrease of at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% from the baseline value of the CLASI-A score. In specific embodiments, the reduction in the subject's CLASI-A score is accompanied by a reduction of at least 50% from the baseline value of the CLASI-A score. In certain embodiments, the baseline value is the CLASI-A score in the subject before treatment with the ILT7-binding protein used in the method described herein. In other embodiments, the method of the disclosure results in a reduced CLASI-D score in the subject. In further embodiments, the method of the disclosure results in both a reduced CLASI-A score and a reduced CLASI-D score in the subject.

[0077] Pharmaceutical composition This disclosure also relates to pharmaceutical compositions comprising ILT7-binding proteins used in the methods described herein. In certain embodiments, this disclosure provides the use of ILT7-binding proteins used in the methods described herein in the manufacture of a therapeutic pharmaceutical product of interest.

[0078] In some embodiments, the pharmaceutical compositions of this disclosure comprise an ILT7-binding protein disclosed herein and one or more pharmaceutically acceptable carriers, diluents, or excipients. In this regard, “pharmaceutically acceptable carriers, diluents, or excipients” include, but are not limited to, any adjuvants, carriers, excipients, fluidizers, sweeteners, diluents, preservatives, pigments / colorants, flavor enhancers, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers, whether or not they are approved by the U.S. Food and Drug Administration for use in humans or domestic animals. For example, suitable carriers are known to those skilled in the art and include stabilizers, diluents, and buffers. Suitable stabilizers include carbohydrates such as sorbitol, lactose, mannitol, starch, sucrose, dextran, and glucose, and proteins such as albumin or casein. Suitable diluents include physiological saline, Hanks equilibrium salts, and Ringer's solution. Suitable buffering agents include alkali metal phosphates, alkali metal carbonates, or alkaline earth metal carbonates.

[0079] In certain embodiments, the pharmaceutical compositions of the Disclosure may further contain one or more auxiliary substances, such as one or more lipids, phospholipids, carbohydrates, and lipopolysaccharides. In some embodiments, the pharmaceutical compositions of the Disclosure may optionally contain one or more additional active substances.

[0080] In certain cases, the pharmaceutical compositions of this disclosure may be prepared by techniques known to those skilled in the art. For a general overview of the formulation and / or manufacture of the pharmaceutical compositions, see, for example, Remington: The Science and Practice of Pharmacy 21st ed., Lippincott Williams & Wilkins, 2005 (in whole, incorporated herein by reference). Generally, the ILT7 binding protein or fragment thereof used in the methods described herein is mixed with a carrier to form a solution, suspension, or emulsion. One or more of the additives considered herein may be added to the carrier, or added later. The pharmaceutical compositions of this disclosure may be aqueous solutions, emulsions, or suspensions, or dry formulations. In certain embodiments, the pharmaceutical compositions of this disclosure may be dehydrated or lyophilized for storage or formulation, for example, by freeze-drying or spray-drying. This can then be reconstituted into a liquid composition by adding a suitable liquid carrier, or administered as a dry formulation using methods known to those skilled in the art. In certain embodiments, the ILT7-binding protein used in the methods described herein is stored as a lyophilized powder and subsequently reconstituted into a liquid composition before administration to a subject requiring it.

[0081] The choice of administration of a pharmaceutical composition will depend on the chosen formulation. The pharmaceutical compositions of this disclosure are administered in a manner compatible with the drug formulation and in a therapeutically effective amount. In certain embodiments, the pharmaceutical compositions of this disclosure are formulated in solid, semi-solid, liquid, or gaseous forms, including, but not limited to, tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants, gels, microspheres, and aerosols.

[0082] In certain examples, the pharmaceutical composition containing the ILT7-binding protein used in the methods described herein may be in solid or liquid form. In some embodiments, one or more carriers are granular, and therefore the composition is, for example, in the form of a tablet or powder. In other embodiments, one or more carriers are liquid, and here the composition is, for example, an oral syrup, an injection, or an aerosol useful for, for example, inhalation administration. When oral administration is intended, the pharmaceutical composition containing the ILT7-binding protein used in the methods described herein may be in either solid or liquid form, where the range of forms considered as either solid or liquid herein includes semi-solid, semi-liquid, suspension and gel forms.

[0083] In certain embodiments, as a solid composition for oral administration, the pharmaceutical composition containing the ILT7 binding protein used in the method described herein may be formulated in the form of powders, granules, compressed tablets, pills, capsules, chewing gum, or wafers. In some examples, such solid compositions will typically contain one or more inert diluents or food carriers. In certain embodiments, one or more of the following may be additionally present: binders such as carboxymethylcellulose, ethylcellulose, microcrystalline cellulose, tragacanth gum, or gelatin; excipients such as starch, lactose, or dextrin; disintegrants such as alginic acid, sodium alginate, Primogel, or corn starch; lubricants such as magnesium stearate or Sterotex; fluidizers such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; flavorings such as peppermint, methyl salicylate, or orange flavoring; and colorants.

[0084] In some embodiments, when the pharmaceutical composition of the Disclosure is in the form of a capsule, for example, a gelatin capsule, it may contain a liquid carrier such as polyethylene glycol or oil in addition to the materials disclosed herein. Oral formulations may also contain commonly used excipients, such as pharmaceutical-grade saccharin, cellulose, and magnesium carbonate.

[0085] In other embodiments, the pharmaceutical compositions of the Disclosure are in the form of liquids, such as elixirs, syrups, solutions, emulsions, or suspensions. In certain embodiments, the liquids may be for oral administration or delivery by injection. In certain embodiments, when intended for oral administration, the pharmaceutical compositions of the Disclosure contain one or more of the following: sweeteners, preservatives, colorants, and flavor enhancers, in addition to the ILT7-binding protein used in the manner described herein. In certain embodiments, pharmaceutical compositions intended for administration by injection may contain one or more of the following: surfactants, preservatives, humectants, dispersants, suspending agents, buffers, stabilizers, and isotonic agents. In certain embodiments, the pharmaceutical compositions of the Disclosure are administered intravenously to subjects requiring them. In specific embodiments, the pharmaceutical compositions of the Disclosure are administered by subcutaneous injection to subjects requiring them.

[0086] In certain cases, a liquid pharmaceutical composition containing ILT7-binding protein used in the methods described herein, whether it is a solution, suspension, or other similar form, may contain one or more of the following components: water for injection, saline solution, sterile diluents such as physiological saline, Ringer's solution, or isotonic sodium chloride; fixing oils such as synthetic monoglycerides or diglycerides that can act as a solvent or suspension medium; polyethylene glycols, glycerin, propylene glycol, or other solvents; antibacterial agents such as benzyl alcohol or methylparaben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffers such as acetates, citrates, or phosphates; and osmotic regulators such as sodium chloride or dextrose. In some cases, the formulation may be sealed in ampoules, disposable syringes, or multi-dose vials made of glass or plastic. In some embodiments, the pharmaceutical composition for injection is preferably sterile.

[0087] In other embodiments, the pharmaceutical composition containing the ILT7-binding protein used in the method described herein may be intended for topical administration, in which case the carrier may preferably comprise a solution, emulsion, ointment, or gel base. In certain embodiments, the base may comprise, for example, one or more of the following: diluents such as petrolatum, lanolin, polyethylene glycols, beeswax, mineral oil, water, and alcohol, as well as emulsifiers and stabilizers. In other embodiments, thickeners may be present in the pharmaceutical composition for topical administration. In certain embodiments, when transdermal administration is intended, the pharmaceutical composition of the ILT7-binding protein used in the method described herein may be included in a transdermal patch or iontophoresis device.

[0088] In further embodiments, the pharmaceutical composition containing the ILT7-binding protein used in the methods described herein is intended for rectal administration, for example, in the form of a suppository. The suppository may contain, for example, polyalkalene glycols or triglycerides as binders and carriers. In certain examples, the composition for rectal administration contains an oily base as a suitable non-irritating excipient. Such bases include, but are not limited to, lanolin, cocoa butter, or polyethylene glycol.

[0089] In other embodiments, a pharmaceutical composition comprising ILT7-binding protein used in the methods described herein comprises a dosage unit that can be administered as an aerosol. The term aerosol is used to refer to a variety of systems ranging from colloidal to systems consisting of pressurized packages. In certain embodiments, delivery is achieved by liquefaction or compressed gas, or by a suitable pump system for dispensing the active ingredient. In some embodiments, the aerosol of ILT7-binding protein used in the methods described herein may be delivered in a single-phase, two-phase, or three-phase system to deliver one or more active ingredients. In other embodiments, the delivery of the aerosol may include necessary containers, starters, valves, sub-containers, etc., which together may form a kit. Those skilled in the art can easily determine specific aerosol formulations and delivery methods.

[0090] The pharmaceutical compositions of this disclosure may be administered by a suitable non-toxic pharmaceutical carrier, may be contained in microcapsules, microbeads, and / or in sustained-release implants.

[0091] In some embodiments, the pharmaceutical compositions of the present disclosure include a material that forms a coating shell around an active ingredient. In some examples, the material forming the coating shell is typically inert and may be selected from, for example, sugars, shellac, and other enteric coating agents.

[0092] In yet another embodiment, the pharmaceutical compositions of this disclosure in solid or liquid form include agents that bind to ILT7-binding proteins used in the manner described herein, thereby assisting in the delivery of ILT7-binding proteins used in the manner described herein. In particular cases, preferred agents that work in this capacity include proteins or liposomes.

[0093] In certain embodiments, the pharmaceutical composition to be administered to a subject may take the form of one or more dosage units, where, for example, a tablet may be a single dosage unit, or a container of ILT7-binding protein used in the aerosol form described herein may hold multiple dosage units. Actual methods of preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The administered composition will in any case contain a therapeutically effective amount of ILT7-binding protein, or a pharmaceutically acceptable salt thereof, used in the method described herein, to assist in the treatment of the disease or condition of interest in accordance with the teachings herein.

[0094] In certain embodiments, the pharmaceutical compositions of the Disclosure comprise one or more additional therapeutic agents. In other embodiments, a therapeutically effective dose of the pharmaceutical composition of the Disclosure is administered to a subject in need of it in combination with one or more additional therapeutic agents. As used herein, “combination” means a combination comprising the ILT7 binding protein used in the manner described herein and one or more additional therapeutic agents, each of which may be administered sequentially, simultaneously, or concurrently.

[0095] The pharmaceutical compositions of this disclosure may preferably be administered at intervals of several times to maintain a sustained therapeutic effect. The pharmaceutical compositions of this disclosure may be used in conjunction with other bactericidal or bacteriostatic methods.

[0096] The descriptions of pharmaceutical compositions provided herein relate primarily to pharmaceutical compositions suitable for administration to humans, but those skilled in the art will understand that such compositions are generally suitable for administration to all kinds of subjects. In certain embodiments, the subjects are mammals. In certain embodiments, mammals include primates, such as humans, monkeys and apes, as well as non-primates, such as domesticated animals, including laboratory animals and domestic pets and agricultural animals (e.g., cats, dogs, pigs, cows, sheep, goats, horses, rabbits), and non-domesticated animals such as wild animals, birds, etc. [Examples]

[0097] In autoimmune diseases, plasmacytoid dendritic cells (pDCs), when activated by immune complexes against autologous nucleic acids, secrete significant amounts of type I and type III interferons (IFNs). pDCs make up about 0.4% of circulating leukocytes and can recognize viral nucleic acids, often bound to other proteins or immunoglobulins. While this response is thought to contribute to antiviral defense, there is growing evidence that pDCs and type I IFNs also contribute to the pathogenesis of numerous autoimmune diseases. There is no reliable method to directly measure type I IFN levels; however, binding of type I IFNs to their receptors leads to local and systemic upregulation of type I IFN-inducible genes. Messenger ribonucleic acid (mRNA) levels of these type I IFN-inducible genes can be measured in the blood and analyzed as a composite outcome called the "type I interferon gene signature" (IFNGS). We developed a test for type I IFNGS and established cutoff values ​​for scoring these signatures as "IFN test - high" and "IFN test - low." Studies using this specific gene signature have shown that elevated type I IFN signatures can be identified in some patients with systemic lupus erythromatosus, dermatomyositis, polymyositis, systemic sclerosis, and Sjögren's syndrome.

[0098] Single dose escalation (SAD) study A phase 1a randomized, center-blinded, open-label, placebo-controlled trial of a single dose of subcutaneously escalating ILT7-binding protein as used herein was conducted in five consecutive cohorts of patients with systemic lupus erythematosus (SLE), Sjögren's syndrome (SS), dermatomyositis (DM), polymyositis (PM), or systemic sclerosis (SSc). This trial evaluated the safety, drug concentration, pDC levels, anti-drug antibodies (ADA), and effects of the ILT7-binding protein (VIB7734) as used herein (as used herein) on 21 type I IFNGS.

[0099] All data were presented in the form of tables organized by cohort, treatment, and number of subjects. Summary tables of the collected data were presented by treatment group. Categorical data were summarized by frequency and proportion of subjects within each category. Proportions were calculated based on non-missing observations where applicable. Continuous variables were summarized using descriptive statistics, including the number of observations, mean, standard deviation, median, minimum, and maximum. In general, unless otherwise specified, "baseline" was defined as the last value before the first dose of VIB7734.

[0100] Example 1: SAD Study Design A total of 36 subjects were enrolled in this study. The enrolled subjects had the following diagnoses: SLE 19 cases (53%), SS 16 cases (44%), SSc 3 cases (8%), PM 2 cases (6%), and DM 2 cases (6%). Baseline demographic characteristics were well-balanced and generally similar across all VIB7734 and placebo groups. The majority of subjects were Caucasian and female (32 subjects [88.9%]). Enrolled subjects were randomized in a 3:1 ratio within 5 cohorts to receive a single subcutaneous dose of either VIB7734 or the corresponding placebo, as follows: Cohort 1: 1 mg VIB7734 (n=3) or placebo (n=1); Cohort 2: 5 mg VIB7734 (n=6) or placebo (n=2); Cohort 3: 15 mg VIB7734 (n=6) or placebo (n=2); Cohort 4: 50 mg VIB7734 (n=6) or placebo (n=2); Cohort 5: 150 mg VIB7734 (n=6) or placebo (n=2).

[0101] Figures 1 and 2 provide a schematic representation of this study. Screening visits were conducted within 42 days prior to administration. Participants received VIB7734 on day 1 and were observed overnight (at the facility) for any safety concerns. On day 2, participants were discharged after all study procedures were completed and safety concerns were confirmed. This was followed by an 85-day treatment follow-up period. During this period, participants revisited the research facility on days 4, 8, 15, 29, 57, and 85. Participants were evaluated for pDC excess. A pDC excess criterion was met if the pDC level was 50% or higher than the baseline value of the participant, or if the pDC level was higher than the lower limit of the standard range as defined in the experimental manual. If the pDC excess criterion was not met on day 85, a further 252-day follow-up (up to day 337 of the study) was conducted. If the criteria for excessive pDCs were not met by the 337th day visit, the medical monitor, in consultation with the principal investigator, determined the need for further follow-up.

[0102] Participants received only one dose of VIB7734 during this study. As shown in Figure 2, cohorts were enrolled using a dose escalation method to allow time to review safety and tolerability data before moving to the next dose level (cohort). The decision to escalate to the next dose setting cohort was made by the Dose Escalation Committee (DEC).

[0103] In each cohort, there was an interval of at least 48 hours between administrations between the first and second subjects, and between the second and third subjects. There was an interval of at least 24 hours between administrations between subsequent subjects, starting with the third subject in each cohort. Administration in cohorts 2, 3, 4, and 5 commenced after randomization and administration of VIB7734 had been completed for all subjects in the previous cohort, all evaluable subjects had completed at least 15 day visits (6th visit), cumulative safety data for all exposed subjects had been reviewed by DEC, and DEC agreed that the safety profile was acceptable.

[0104] Example 2: SAD study: Evaluation of adverse reactions One patient in the VIB7734 15 mg group (colitis) and one patient in the placebo group (death due to cerebral hemorrhage) experienced a serious adverse event. At least one adverse event (AE) was reported in 69% of patients treated with VIB7734 and 80% of patients treated with placebo. The most common AEs reported in patients treated with VIB7734 were diarrhea (12%) and upper respiratory tract infection (12%). No injection site reactions or hypersensitivity reactions occurred.

[0105] Example 3: SAD study: Immunogenicity evaluation Blood samples were collected on days 1, 2, 4, 8, 15, 29, 57, and 85 to evaluate the anti-drug antibody (ADA) response to VIB7734 in human serum. These evaluations were performed using effective electrochemiluminescence immunoassays for detection, as well as confirmation and titration of anti-drug antibodies against VIB7734 in human serum. Samples found to be negative in screening were reported to have titers of less than 30.

[0106] Baseline and post-baseline ADA results were recorded for all 26 subjects in the ILT7-binding protein group and 9 subjects in the placebo group. No positive results were observed in either treatment group. No cases of persistent or transient ADA positivity were observed in either treatment group. Therefore, overall, no safety, tolerability, or immunogenicity issues were identified with subcutaneous injection of VIB7734 in doses ranging from 1 to 150 mg.

[0107] Example 4: SAD study: Pharmacokinetic evaluation Blood samples were collected on days 1, 2, 4, 8, 15, 29, 57, and 85, and the PK of VIB7734 in serum was evaluated. The concentration of VIB7734 in human serum samples was measured using an effective enzyme-linked immunosorbent assay (ELISA) method. The effective measurement range of this assay was 0.025 μg / mL to 25.60 μg / mL. Results below the limit of quantification (LLOQ) were reported as <0.10 μg / mL.

[0108] PK analysis was performed on time-series data of VIB7734 concentrations from all 26 subjects who received any dose of VIB7734. The mean serum concentration-time profiles of VIB7734 following single subcutaneous administrations of 1, 5, 15, 50, or 150 mg are shown in Figure 3. After 1 mg administration, all concentrations were below the limit of quantification (BLQ); therefore, all summarized PK parameters are referenced to the 5–150 mg dose levels. VIB7734 PK exposure increased almost proportionally to the dose. After a single subcutaneous injection on day 1, peak concentrations were observed 5–8 days post-administration. Exposure increased almost proportionally to the dose level. The estimated half-life ranged from 13–20 days at all dose levels. The mean extravascular clearance ranged from 468–1030 mL / day. The mean extravascular volume of distribution ranged from 9.9–19.0 ​​L.

[0109] Example 5: SAD study: Pharmacodynamic evaluation (blood pDC levels) Whole blood samples for pDC levels were collected on days 1, 2, 4, 8, 15, 29, 57, and 85. Baseline pDC levels were defined as the mean of the levels measured at the first and second visits. If a screening (first visit) sample was not collected or failed for technical reasons, this was repeated, and the results had to be available before subjects could be randomized, as the results were needed to determine if subjects met all inclusion / exclusion criteria. If a day 1 (second visit) sample was not collected or failed for technical reasons, the value from the first visit sample was considered the baseline. Research facilities were blinded for post-baseline pDC levels.

[0110] In this study, pDC levels were quantified in two ways: 1) as a percentage of CD45+ peripheral blood mononuclear cells (PBMC, primary method), and 2) as pDC concentration per μL (secondary method). The primary pDC measurement is the percentage of CD45+ PBMCs, as it is directly measured by the flow cytometry assay used in this study.

[0111] At baseline, the mean pDC level in the blood was 0.13% (SD: 0.056%) of PBMCs in the VIB7734-treated population. The mean pDC concentration at baseline in the VIB7734-treated population was 2.53 cells / μL (SD: 1.24%). Figure 4 shows the level of pDCs (CD45+ cells) as a percentage and the change from baseline over time. Figure 5 shows the level of absolute pDC concentration as a percentage and the change from baseline over time. Figure 6 shows the change in the absolute level of pDCs as a percentage over time.

[0112] Following SC administration of all tested doses of VIB7734, serum pDC levels decreased. In all VIB7734 dose groups, a median decrease of at least 50% in pDC levels was evident 24 hours post-administration (first blood sample taken after administration), with a maximum decrease of 90%. Dose increases were accompanied by a non-linear increase in pDC reduction. On day 15, the median pDC levels in the VIB7734 treatment cohort changed as follows: 1 mg: -57%, 5 mg: -66%, 15 mg: -70%, 50 mg: -82%, and 150 mg: -90%, in contrast to +7.5% in the placebo treatment group.

[0113] Dose increases were generally accompanied by a prolongation of the pDC decline period. In all cases, the effect was reversible. As shown in Figures 4–6, the median pDC level returned to above 50% of baseline at the following time points for each cohort: 1 mg: day 29, 5 mg: day 57, 15 mg: day 57, 50 mg: day 85, 150 mg: day 113. Therefore, subcutaneous injection of VIB7734 in doses ranging from 1 to 150 mg caused a reversible, dose-dependent decrease in circulating pDC levels.

[0114] The maximum decrease from baseline in the median pDC level was -90%. The increase in maximum depletion appears to plateau at doses of 15–150 mg, suggesting that doses higher than 150 mg are unlikely to cause more severe maximum depletion.

[0115] Example 6: SAD study: Pharmacodynamic evaluation (Type I IFNGS) Whole blood samples were collected at screening and on days 1, 2, 4, 8, 15, 29, 57, and 85. mRNA expression of specific types of type I IFN-inducible genes was measured using a 21-gene assay. Type I IFNGS was determined by assaying the mRNA levels of 21 type I IFN-inducible genes in the biological samples collected from subjects, determining the mean or median mRNA level of the 21 type I IFN-inducible genes, and normalizing this mean value to the mean mRNA levels of three housekeeping genes (18S rRNA, β-actin, and GAPDH) to obtain a composite outcome. Type I IFNGS was reported using two methods: "median change factor" and "median target neutralization." The first method, called "median change factor," is the multiplier difference in gene product levels compared to a healthy control normalized to 1. Therefore, for a subject, a median change factor of 4 indicates that the type I IFN-inducible gene product is four times higher than that of a healthy control. Table 2 shows the median change ratio per visit for each cohort.

[0116] (Table 2) Biomarker results - as a treatment population (Type I IFNGS change ratio) TIFF2026053543000003.tif172167

[0117] The second metric, "median target neutralization," is a measure of the percentage of gene product levels compared to baseline results normalized to 100%. This is useful for comparing changes over time. For example, a median target neutralization rate of 30% at the time of visit means that the type I IFN-induced gene product is at 30% of its level at baseline.

[0118] Table 3 shows the median target neutralization rates by cohort and number of visits for subgroups of patients showing elevated baseline type I IFN signature. The median neutralization rate in the IFN-high subgroup was less than 100% on day 4 (first measurement after administration) for all VIB7734 treatment groups, compared to 100% in the placebo group. The median neutralization rate in the IFN-high subgroup was lowest at the day 8 visit (37.9% for all VIB7734 treatments vs. 118% for placebo). The median target neutralization in the IFN-high subgroup remained below 100% at all time points for all VIB7734 treatment cohorts, with the exception of the lowest dose cohort (1 mg), which exceeded 100% of baseline at the day 8, 29, and 57 visits.

[0119] (Table 3) Biomarker results - as a treatment population (neutralization rate of type I IFN signature) TIFF2026053543000004.tif108167

[0120] Figure 7 shows the percentage change in baseline IFN signature over time for each subject in the cohort with elevated baseline type 1 IFNGS. In the majority of subjects with elevated type 1 IFNGS, a decrease in pDC levels (Figure 7A) correlated with a decrease in type 1 IFNGS (reported as a percentage change relative to baseline) (Figure 7B).

[0121] Multiple dose escalation (MAD) study A phase 1b randomized, double-blind (sponsor and institutional pharmacist blinded) placebo-controlled study was conducted to evaluate the safety and tolerability of multiple doses of subcutaneous (SC) VIB7734 in addition to standard treatment in subjects with at least one of the following autoimmune diseases: systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), systemic sclerosis, polymyositis, and dermatomyositis (Figure 9). This study also evaluated the efficacy of VIB7734 against cutaneous lupus activity. The rationale for this study was to (1) evaluate the safety, PK, PD, and immunogenicity of multiple doses of VIB7734 in a relevant target population, (2) evaluate whether VIB7734 can improve cutaneous lupus findings, and (3) evaluate the effects of VIB7734 on cutaneous pDC and type I IFNGS levels. For this study, the SC administration route was selected to provide the most relevant information for future studies that will use SC formulations. The dose regimens tested were VIB7734 5 mg, 50 mg, or 150 mg SC for 4 weeks or placebo SC for 4 weeks. PK / PD models from single-dose studies indicate that doses in the range of 50–100 mg SC every 4 weeks are the minimum dose required to produce a sustained near-maximal pDC reduction. The 5 mg SC for 4 weeks (Cohort 1) dose was expected to produce a near-maximal PD effect, which should have helped define the minimum dose required for this drug to achieve the optimal PD effect. The 50 mg SC for 4 weeks (Cohort 2) dose was selected to evaluate doses within the expected target dose range. The tenfold dose increase between cohorts 1 and 2 is justified by (a) the relatively small predicted difference in maximum pDC reduction between these two dose cohorts (-78% vs. -87%), (b) the fact that no safety concerns were demonstrated for any dose tested (upper dose 150 mg SC) in the single-dose study with VIB7734, and (c) the high safety margin for all doses tested in this study. The 150 mg SC q4-week dose tested the upper limit of the candidate dose range for VIB7734, which may be particularly necessary when a higher dose than circulating is required to deplete pDCs in target tissue.This study made it possible to refine the PK / PD model and to select one or more doses to be tested in subsequent clinical trials.

[0122] Example 7: MAD Research Design A total of 31 adult subjects were enrolled in three consecutive cohorts: 8 in Cohort 1, 12 in Cohort 2, and 11 in Cohort 3. Subjects continued their standard care treatment and, in addition, received either ILT7-binding protein (VIB7734) or placebo. Randomization was not stratified. Cohorts were enrolled sequentially (Figure 10) to allow time to review safety and tolerability data before moving to the next cohort. In Cohort 1, subjects were randomized in a 3:1 ratio and received a total of three doses of VIB7734 or the corresponding placebo via SC injection every four weeks. The dose of VIB7734 administered was 5 mg SC per four weeks for three doses. In cohorts 2 and 3, participants were randomized in a 2:1 ratio and received VIB7734 50 mg (cohort 2) or 150 mg (cohort 3) or the corresponding placebo via SC injection over 4 weeks in 3 doses. Cohort 1: VIB7734, 5 mg (n=6) or placebo (n=2) SC injection every 28 days x 3 doses; Cohort 2: VIB7734, 50 mg (n=8) or placebo (n=4) SC injection every 28 days x 3 doses; Cohort 3: VIB7734, 150 mg (n=8) or placebo (n=3) SC injection every 28 days x 3 doses.

[0123] Cohort 1 enrolled a mixed disease population, including subjects with systemic lupus erythematosus (SLE) or Sjögren's syndrome, without a minimum disease activity requirement, to evaluate the safety profile of multiple doses of VIB7734 in subjects across various pDC-driven indications. Cohorts 2 and 3 recruited patients with active SLE or cutaneous lupus erythematosus (CLE) with a CLASI activity score (CLASI-A) of 8 or higher, allowing for the exploration of efficacy at the doses tested in these cohorts and the examination of effects on skin biopsy specimens. Experienced physicians-principal investigators or designated investigators performed CLASI-A assessments. Wherever possible, the same evaluator assessed subjects throughout the study. The institution also designated a preliminary evaluator for each subject.

[0124] The screening procedures for Cohorts 2 and 3 included taking digital photographs of existing skin lesions on days 29, 57, and 85. The purpose of taking skin photographs was to 1) confirm the presence of lupus-like skin lesions in subjects as part of the screening procedure, and 2) provide visual evidence of the drug's effect on the skin lesions. The principal investigator or co-investigator who examined the patient at screening determined the anatomical areas to be photographed based on the location of the lesions. These anatomical areas were documented in the screening eCRF with sufficient detail so that photographs of the same areas could be taken at different times (on days 113 and 145). Consistent lighting was maintained as much as possible during all photography visits. Photographs were uploaded and reviewed by a central reviewer to confirm the presence of lupus-like skin lesions.

[0125] The multi-dose study consisted of three study periods: screening, treatment, and extended follow-up for pDC excess (Figure 9). Screening visits were conducted within 28 days prior to administration. Randomized subjects received VIB7734 on days 1, 29, and 57. For all administrations, VIB7734 was administered in the clinic, and subjects were observed for at least 90 minutes after administration. Subjects were followed up for at least 141 days. After the visit on day 141, subjects were informed whether they met the protocol definition for sufficient pDC levels. The protocol definition for sufficient pDC levels is either higher than 50% of the subject's baseline value or higher than 0.036% of peripheral blood mononuclear cells (PBMCs). If a subject's pDC levels met the criteria for sufficient levels, the subject terminated the study. If a subject did not meet the criteria for sufficient pDC levels at the 141-day visit, they were informed of this and asked to continue returning for pDC follow-up until they met the protocol definition for sufficient pDC levels or until they reached the 337-day visit. These pDC follow-up visits were conducted regardless of whether the subject received VIB7734 or placebo, and blinding of subjects and facilities was not lifted until the study was completed.

[0126] An open-label Dose Escalation Committee (DEC) reviewed the safety of each dose cohort according to pre-specified criteria to determine whether it was safe to escalate to the next dose-setting cohort. For escalations from Cohort 1 to 2, the DEC reviewed cumulative safety data after the eighth subject in Cohort 1 completed their 15-day visit (or, if the eighth subject dropped out before 15 days, when the last evaluable Cohort 1 subject reached 15 days). For escalations from Cohort 2 to 3, the DEC reviewed cumulative safety data as soon as the nine Cohort 2 subjects completed their 15-day visit. If the data from any cohort was insufficient to determine the safety of escalating to the next cohort, the sponsor could choose to postpone the escalation until the results of a later re-evaluation of that cohort were available.

[0127] Example 8: Evaluation of adverse reactions The safety and tolerability of VIB7734 were measured by the incidence of therapeutic adverse events (TEAEs), adverse events of particular interest (AESIs), and serious adverse events (TESAEs) that occurred during treatment. As part of safety assessment, laboratory measurements, vital signs, and ECG parameters were also evaluated. Collection of adverse events (AEs) and serious adverse events (SAEs) began after the subject signed the informed consent document and continued until the final visit. TEAEs were defined as any AE that occurred after administration or after the first dose of VIB7734 until the end of follow-up. All AEs had to be coded according to the Medical Dictionary for Regulatory Activities (MedDRA). All SAEs had to be reported, regardless of whether they were considered causally related to VIB7734 or the study procedure. TEAEs, TESAEs, and TEAESIs needed to be summarized comprehensively and classified by MedDRA organ-specific major classification, basic term, severity, and relationship to VIB7734. AESIs also needed to be recorded in the eCRF within 24 hours of the event being observed, even if the event was not serious. No AEs (Figure 40) or AESIs (Figure 41) were observed in VIB7734-treated subjects in Cohorts 1, 2, and 3. The proportion of subjects experiencing adverse events was similar between the VIB7734 group and the placebo group (approximately 73% vs. 67%, respectively; Figure 41).

[0128] Example 9: Immunogenicity Evaluation Blood samples were collected on days 1, 29, 57, 85, 113, and 141, and, where applicable, on days 197, 253, and 309, to evaluate the anti-drug antibody (ADA) response to VIB7734 in human serum. ADA status and titers were summarized by treatment group. These evaluations were performed using valid immunoassay methods. No ADA response to VIB7734 in human serum was observed in subjects of Cohorts 1, 2, or 3 (data not shown).

[0129] Example 10: Pharmacokinetic evaluation Blood samples were collected on days 1, 8, 15, 29, 36, 43, 57, 64, 71, 85, 113, and 141, and, where applicable, on days 169, 197, 225, and 253, to evaluate the PK of VIB7734 in human serum. The PK of VIB7734 in serum was measured using a valid immunoassay method. Specific procedures for sample collection, processing, storage, and shipping can be found in the individual laboratory manuals provided to the facility. Non-compartmental analysis was performed on VIB7734 treatment subjects. VIB7734 concentration-time profiles were created.

[0130] Figure 11 shows the serum concentration profiles of VIB7734 following multiple subcutaneous administrations (three doses every four weeks) of 5 mg (Cohort 1) or 50 mg (Cohort 2) of VIB7734. Figure 11A shows the serum concentration profile of VIB7734 in Cohort 2 subjects. Figure 11B shows the mean serum concentration profiles of VIB7734 in subjects from Cohort 1 (filled circles) and Cohort 2 (filled squares). As shown in Figure 11B, the mean serum concentration of VIB7734 increased in Cohort 2 subjects up to day 112 compared to Cohort 1 subjects.

[0131] Example 11: Pharmacodynamic evaluation: Circulating pDC levels The effect of VIB7734 on circulating pDC levels was evaluated in blood using flow cytometry. Changes from baseline were described, and levels were summarized as a percentage of baseline. Whole blood samples were collected on days 1, 8, 15, 29, 36, 43, 57, 64, 71, 85, 113, and 141 for pDC levels (all subjects) and for CD19+ B cells (at screening, for subjects previously administered rituximab, ocrelizumab, ofatumumab, or experimental B-cell depletion mAbs). Baseline pDC levels were defined as the mean of the levels measured at the screening visit and the day 1 (pre-administration) level. If a screening (first visit) sample could not be collected or failed for technical reasons, this had to be repeated, and the results had to be available before subjects could be randomized, as the results were needed to determine if subjects met all inclusion / exclusion criteria. If only one value was available, that value was used as the baseline.

[0132] Figure 12 shows the time-course levels of pDCs (percentage of PBMC cells) and the change from baseline in the whole blood of subjects in Cohort 1. Figure 13 shows the time-course levels of absolute pDC concentrations and the change from baseline in the whole blood of subjects in Cohort 1. Figure 14 shows the time-course changes in the absolute levels of pDCs in the whole blood of subjects in Cohort 1. Consistent with the results of a single dose escalation study, subjects in Cohort 1 treated with multiple SC doses (3 doses every 4 weeks) of 5 mg of VIB7734 (Figures 12B, 13B, and 14B) showed a decrease in blood levels of pDCs compared to subjects treated with placebo (Figures 12A, 13A, and 14A).

[0133] Figure 15 shows the change in pDC levels (%) over time as a percentage (using % peripheral blood mononuclear cells) relative to baseline levels in the whole blood of subjects in Cohort 2 and Cohort 3. Figure 16 shows the absolute concentration levels and changes from baseline over time in the whole blood of subjects in Cohort 2 and Cohort 3. Figure 17 shows the change in absolute pDC levels over time in the whole blood of subjects in Cohort 2 and Cohort 3. Figure 18 shows the pDC levels and changes from baseline over time (as a percentage in PBMC cells) in the whole blood of subjects in Cohort 2 and Cohort 3. Similar to the subjects in Cohort 1, subjects in Cohort 2 treated with multiple SC doses of 50 mg VIB7734 (three doses every four weeks) (Figures 15B, 16B, 17B, and 18B) showed a decrease in blood pDC levels compared to subjects treated with placebo (Figures 15A, 16A, 17A, and 18A). Similarly, most subjects in Cohort 3 treated with multiple SC doses of 150 mg VIB7734 (three doses every four weeks) (Figures 15D, 16D, 17D, and 18D) showed a decrease in blood pDC levels compared to subjects treated with placebo (Figures 15C, 16C, 17C, and 18C). In addition, as shown in Figures 19A to 19D, the median pDC levels over time in whole blood of subjects in Cohort 2, following multiple SC administrations of 50 mg of VIB7734 (3 doses every 4 weeks), showed a significant decrease compared to the median pDC levels over time in whole blood of subjects in Cohort 2 treated with placebo. Similarly, as shown in Figures 20A to 20D, the median pDC levels over time in whole blood of subjects in Cohort 3, treated with multiple SC administrations of 150 mg of VIB7734, showed a significant decrease compared to the median pDC levels over time in whole blood of subjects in Cohort 3 treated with placebo.Furthermore, as shown in Figure 77, in the VIB7734-treated subjects of Cohort 1 (treated with 5 mg of VIB7734), Cohort 2 (treated with 50 mg of VIB7734), and Cohort 3 (treated with 150 mg of VIB7734), a clear decrease in median circulating pDC levels (measured as %PBMC cells) was observed at week 1 compared to the median circulating pDC levels in the placebo-treated subject, and this continued until at least day 85.

[0134] Example 12: Pharmacodynamic evaluation: Type I IFN signature in blood Type I IFNGS levels were measured in skin and blood using 21 gene tests. The effect of VIB7734 on blood type I IFNGS levels was evaluated in cohorts 1, 2, and 3. Whole blood was collected in PAXgene blood collection tubes on days 1, 8, 15, 29, 43, 57, 71, 85, 113, and 141, and mRNA overexpression was measured for specific types of type I IFN-inducible genes. Any residual RNA isolated from the samples was used for additional analytical studies on changes in gene expression. At baseline, 18 out of 23 subjects (78%) in cohorts 2 and 3 had elevated type I IFNGS levels in their whole blood.

[0135] Figure 21 shows the time-course type I IFNGS levels in whole blood of subjects in Cohort 2 treated with 50 mg of VIB7734 (measured as a change factor (Figures 21A and 21B) or absolute score (Figures 21C and 21D)). Subjects in Cohort 2 treated with multiple SC doses of 50 mg of VIB7734 (3 doses every 4 weeks) (Figures 21A and 21C) showed a decrease in serum type I IFNGS levels compared to subjects treated with placebo (Figures 21B and 21D). Furthermore, as shown in Figures 22A and 22C, the time-course type I IFNGS levels in whole blood of subjects in Cohort 2 treated with multiple SC doses of 50 mg of VIB7734 (3 doses every 4 weeks) are shown (change factor and absolute score, respectively). The median level (measured as) showed a significant decrease compared to the median level of type I IFNGS over time in whole blood of cohort 2 subjects treated with placebo. As shown in Figure 22C, cohort 2 subjects treated with VIB7734 experienced a reduction of over 50% in all type I IFNGS (measured as absolute scores) from the initial time point to day 85. Furthermore, as expected, the median level of type I IFNGS over time in whole blood of cohort 2 subjects treated with multiple SC doses of 50 mg VIB7734 (3 doses every 4 weeks) increased compared to the median level of pDC over time in whole blood of cohort 2 subjects treated with placebo (Figure 22B).

[0136] Figure 55 shows the normalized type I IFNGS levels (measured as a change factor) over time in the whole blood of subjects in Cohort 3, following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of VIB7734 or placebo. In the whole blood of subjects in Cohort 3 treated with multiple subcutaneous administrations of 150 mg of VIB7734 (3 doses every 4 weeks) (Figure 55A), there was a decrease in normalized type I IFNGS levels over time (days 1 to 113) compared to subjects in Cohort 3 treated with placebo (Figure 55B). In addition, as shown in Figure 55C, the median normalized type I IFNGS level (measured as a percentage change) over time (days 1 to 85) in whole blood of cohort 3 subjects treated with multiple SC doses of 150 mg VIB7734 (three doses every four weeks) was lower compared to the median pDC level over the same period in whole blood of cohort 3 subjects treated with placebo (Figure 55C). The median change in type I IFNGS level in whole blood at day 85 was -54% in the VIB7734 50 mg group (cohort 2; Figure 22C), -83% in the VIB7734 150 mg group (cohort 3; Figure 55C), and +8% in the placebo group.

[0137] To further evaluate the effects of VIB7734 on circulating type I IFN activity, gene signatures were created from whole blood RNA, and serum IFNα protein levels were measured at various time points from study participants and healthy donors. The majority of subjects demonstrated elevated circulating baseline type I IFN activity, with 73% of subjects and 44% of placebo subjects in the combined VIB7734 treatment cohort (i.e., subjects combined from cohorts 1 (n=3), 2 (n=6), and 3 (n=8)) having higher IFN-inducible gene expression than healthy subjects at baseline, and a strong correlation was observed between baseline type I IFNGS scores and IFNα protein levels (r=0.573; p=0.0203).

[0138] As shown in Figure 79, among subjects with elevated baseline serum IFN activity, treatment with VIB7734 led to both a decrease in circulating type I IFNGS score (Figure 79A) and a decrease in IFNα protein levels (Figure 79B), with the most significant and sustained decreases in these measurements observed in the highest-dose cohort, i.e., Cohort 3 (Figures 79A-79B). One month after treatment (day 29), the type I IFNGS score decreased by 75.07% in Cohort 3 (150 mg VIB7734) compared to an 11.12% increase in the placebo group at that time. Circulating IFNα protein levels also decreased dramatically with VIB7734, with the highest-dose group (Cohort 3) achieving a 95.0% decrease in serum IFNα at day 29 compared to a 33% increase in the placebo group.

[0139] In the VIB7734 treatment cohort combination, the relationship between baseline circulating type I IFN activity and clinical response to VIB7734 was also determined. As shown in Figure 79, high baseline serum type I IFN activity is associated with a higher clinical response rate to VIB7734. Of the 12 subjects demonstrating a clinical response to VIB7734 treatment, 11 had high circulating type I IFN activity at baseline, as evidenced by both type I IFNGS scores and type I IFN protein (Figure 79C, black dots). On the other hand, subjects with lower baseline type I IFN activity tended to be CLASI non-responders, with 3 out of 4 CLASI non-responders clustered in the lower baseline IFN activity group (Figure 79C, red dots). These data suggest that high baseline serum type I IFN activity is associated with a higher response rate to pDC depletion therapy.

[0140] Clinical development in SLE is challenging as numerous clinical compounds demonstrate inconsistent responses, which appear to reflect the significant heterogeneity of this disease. This study suggests that baseline serum type I IFN activity acts as a predictor of clinical response to pDC depletion, identifying patients with higher baseline levels of IFNα or type I IFNGS score who are likely to receive greater clinical benefit.

[0141] Example 13: Pharmacodynamic evaluation: Efficacy The efficacy analysis population consisted of subjects with active skin lesions and a baseline CLASI score of 8, all of whom had SLE or CLE. The CLASI activity (CLASI-A) score ranged from 0 to 70 and could be used to classify disease activity into mild (0-9), moderate (10-20), or severe (21-70). This population allowed for the testing of the hypothesis that VIB7734 reduces skin findings in SLE or CLE. The clinical efficacy endpoint was the change in the CLASI activity score. Both the CLASI-A score and the CLASI chronic lesion score were calculated on days 1, 15, 29, 43, 57, 85, 113, and 141. Subjects who newly initiated or increased the dose of oral or topical corticosteroids or immunosuppressants contrary to the protocol were considered non-responders in this responder analysis. Changes in CLASI-A from baseline were analyzed using a repeated measures mixed-effects model with covariates including treatment, baseline type I IFN signature status (low and high), number of visits, and the interaction between the number of visits and treatment. The proportion of subjects with a 4-point decrease in CLASI-A at day 85 and the proportion of subjects with a 50% decrease in CLASI-A at day 85 were analyzed using logistic regression with covariates including treatment and baseline type I IFN signature status. For exploratory purposes, subgroup analysis was performed based on baseline type I IFN signature status (low and high).

[0142] For Cohort 2 subjects (Figures 23, 24A, 24B, and 33A) and Cohort 3 subjects (Figures 68, 24C, 24D, and 33B), observed CLASI-A scores are summarized along with their change from baseline. For Cohort 2 and Cohort 3, the proportion of subjects whose CLASI-A score decreased by at least 4 points from baseline is also summarized (Figures 25-27). In addition, for Cohort 2 and Cohort 3, the proportion of subjects whose CLASI-A score decreased by at least 7 points from baseline is summarized (Figures 50-52). Furthermore, for Cohort 2 and Cohort 3, the proportion of Cohort 2 subjects whose CLASI-A score decreased by at least 50% from baseline is also summarized (Figures 29, 53, and 54).

[0143] Many subjects in Cohort 2 (Figure 24A) and all subjects in Cohort 3 (Figure 24C) showed a decrease in CLASI-A score after multiple SC administrations of VIB7734. As shown in Figure 33, for Cohort 2, the median change from baseline in CLASI-A score at day 85 was -5.0 for those treated with VIB7734, compared to -2.5 for those treated with placebo (Figure 33A). Unexpectedly, the median change from baseline in CLASI-A score at day 85 was higher for Cohort 3 subjects compared to Cohort 2 subjects. As shown in Figure 33, for Cohort 3, the median change from baseline in CLASI-A score at day 85 was -9.5 for those treated with VIB7734, compared to -5.0 for those treated with placebo (Figure 33B). For Cohort 2 subjects, the least squares mean difference between the VIB7734 and placebo arms at day 85 was 0.14; 95%Cl (-9.86, 10.14, p=0.977). In addition, for Cohort 3, the least squares mean difference between the VIB7734 arm and placebo arm at day 85 was -5.24 (95%, Cl -11.8, 1.3, p=0.11) (Figure 33B). Figure 33C shows the percentage change from baseline (BL) in the median CLASI-A score by treatment arm and number of visits for subjects in Cohorts 2 and 3.

[0144] Furthermore, at days 15, 29, 85, and 113, a higher proportion of VIB7734-treated patients in Cohort 2 showed a reduction of at least 4 points from baseline in their CLASI-A score compared to patients in Cohort 2 receiving placebo (Figure 25). At days 15, 29, 43, 57, 85, and 113, a higher proportion of VIB7734-treated patients in Cohort 3 showed a reduction of at least 4 points from baseline in their CLASI-A score compared to patients in Cohort 3 receiving placebo (Figure 26). The same trend was observed when data from Cohorts 2 and 3 were combined (Figure 27). In addition, on days 15, 29, 43, 57, 85, 113, and 141, a higher proportion of VIB7734-treated patients in Cohort 2 (Figure 50) and Cohort 3 (Figure 51) showed a reduction of at least 7 points from baseline in their CLASI-A score compared to placebo-treated patients in Cohort 2 (Figure 50) and Cohort 3 (Figure 51), respectively. The same trend was observed when data from Cohorts 2 and 3 were combined (Figure 52). Furthermore, when data from Cohorts 2 and 3 were combined, a higher proportion of CLASI-A score responders were observed in VIB7734-treated patients (75%) compared to placebo-treated patients (57.1%) (Figure 28). In a subset of subjects from cohorts 2 and 3 without discoid lupus erythematosus (DLE), the proportion of CLASI-A score responders was further increased in the VIB7734-treated subjects (91.7%) compared to the placebo-treated subjects (66.7%) (Figure 28). In addition, the proportion of subjects observing at least a 50% reduction in CLASI-A score from baseline at days 15, 29, 43, 57, 85, 113, and 141 was higher in the VIB7734-treated cohort 2 subjects compared to the placebo-treated cohort 2 subjects (Figure 29). Similarly, the proportion of subjects observing at least a 50% reduction in CLASI-A score from baseline at days 29, 43, 57, 85, 113, and 141 was higher in the VIB7734-treated cohort 3 subjects compared to the placebo-treated cohort 3 subjects (Figure 53). The same trend was observed when data from cohorts 2 and 3 were combined (Figure 54).As shown in Figure 54, at day 85, a 50% or greater improvement in CLASI-A scores was observed in 9 out of 16 patients (approximately 56%) treated with VIB7734 and in 2 out of 7 patients (approximately 29%) treated with placebo. Figures 30 and 31 summarize the observed changes in CLASI-A scores (Figure 30A), absolute blood pDC levels (Figure 30B), and blood type I IFNGS levels (measured as absolute scores) (Figure 30C) over time in subjects of Cohort 2 treated with VIB7734, compared with the changes in CLASI-A scores (Figure 31A), absolute blood pDC levels (Figure 31B), and blood type I IFNGS levels (measured as absolute scores) (Figure 31C) over time in subjects of Cohort 2 treated with placebo. Compared to the placebo group, the VIB7734 treatment group showed a decrease in CLASI-A score and absolute blood levels of pDCs, and an increase in the neutralization rate of blood type I IFNGS levels.

[0145] Example 14: Pharmacodynamic evaluation: Skin pDC and IFN-1 levels Skin biopsies were performed on subjects from cohorts 2 and 3, and the effects of VIB7734 on pDC levels and type 1 interferon (IFN-1) activity (assayed by measuring myxovirus protein A (MxA) levels) from the skin biopsy materials were evaluated. The results are presented in Figures 34-37 (cohort 2) and 56-59 (cohort 3).

[0146] One 4 mm punch biopsy sample was required for each skin biopsy sample. The anatomical sites selected for biopsy samples were areas of active inflammation as indicated by erythema or scaling. The punch biopsy site was closed with a single suture. Baseline skin biopsies were performed on day 1 before administration or during the screening period. However, baseline skin biopsies were not performed until other screening procedures had confirmed that the subjects were eligible for this study. Repeat biopsies were performed on day 85 (±14 days) or at the early discontinuation visit if the subject discontinued the study before day 85. The biopsy sample on day 85 was taken from the same anatomical site adjacent to the baseline biopsy site to avoid scarring of the previous punch biopsy sample. The effect of VIB7734 on pDCs in skin lesions was measured by evaluating the number of pDCs / mm2 in skin biopsy samples before and after drug administration. pDCs were identified using anti-ILT7 clones. The rationale for measuring changes in pDC density in the affected skin was to confirm that VIB7734 depletes pDCs in the target tissue in addition to those in the blood, and to determine whether there was a difference between the doses required to achieve the target level of pDC depletion in the blood and those required in the skin. This could aid in dose selection for subsequent clinical trials. The density of all inflammatory cells was also measured. This demonstrated whether the reduction in pDC levels led to downstream effects on the density of other inflammatory cells in the skin. The observed pDC levels and levels as a percentage of baseline were summarized.

[0147] Figure 34 shows the absolute number of pDCs (measured as the number of cells per square millimeter) in skin biopsy specimens from Cohort 2 subjects over time, following multiple subcutaneous administrations (3 doses every 4 weeks) of 50 mg of VIB7734 (Figure 34B) or placebo (Figure 34A). As shown in Figure 35, the median reduction in the change in pDCs (measured as a percentage of baseline on day 1 (Figure 35A) and the number of cells per square millimeter (Figure 35B)) at day 85 for Cohort 2 subjects was 87% for Cohort 2 subjects treated with VIB7734, compared to 47% for Cohort 2 subjects treated with placebo.

[0148] Figure 56 shows the absolute number of pDCs (measured as the number of cells per square millimeter) in skin biopsy materials from Cohort 3 subjects over time, following multiple subcutaneous administrations (3 doses every 4 weeks) of 150 mg of VIB7734 (Figure 56B) or placebo (Figure 56A). As shown in Figure 57, the median number of pDCs in skin biopsy materials at day 85 (measured as a percentage of baseline at day 1) was 99% lower in Cohort 3 subjects treated with VIB7734 compared to an 11% increase from baseline in Cohort 3 subjects treated with placebo. Combining placebo data from Cohorts 2 and 3, the median change in skin biopsy material pDC density (measured as a percentage of baseline at day 1) at day 85 was -87% for the 50 mg group (Cohort 2), -99% for the 150 mg group (Cohort 3), and -14% for the placebo group.

[0149] In skin lesions of CLE patients, type 1 interferon (IFN-1) activity is upregulated. The effect of VIB7734 on IFN-1 activity in skin lesions was also determined by assaying levels of myxoviral protein A (MxA), an interferon-regulated protein, in skin biopsy materials from subjects in Cohort 2 (Figure 37) and Cohort 3 (Figure 58). As shown in Figure 37, in VIB7734-treated subjects in Cohort 2, the median MxA in biopsy materials (measured as a percentage of MxA-positive area; positive %) decreased from a baseline of 50.5% at day 1 to 1.7% at day 85. In contrast, in Cohort 2 subjects treated with placebo, the median MxA in biopsy materials decreased from a baseline of 48.3 at day 1 to 38.0 at day 85. As further shown in Figure 59, for the VIB7734-treated subjects in Cohort 3, the median MxA (measured as the percentage of MxA-positive area; positive %) in skin biopsy material decreased from 89.7% baseline to 1.1% at day 85. In contrast, for the placebo-treated subjects in Cohort 3, the median MxA in skin biopsy material increased from 1.9% baseline to 17.7% at day 85. Combining the placebo data from Cohorts 2 and 3, the median MxA-stained area decreased with VIB7734 treatment, from 50% to 1.7% of the affected area in the 50 mg group (Cohort 2) from day 1 (baseline) to day 85; from 90% to 1.1% of the affected area in the 150 mg group (Cohort 3); and from 5.4% to 18% of the affected area in the placebo group.

[0150] Figures 38 and 39 summarize the observed changes over time in CLASI-A scores (Figure 38A), absolute blood pDC levels (Figure 38B), blood type I IFNGS levels (measured as absolute scores) (Figure 38C), number of pDCs in skin biopsy specimens (Figure 38D), and normalized blood type I IFNGS levels (measured as a change factor) (Figure 38E) for cohort 2 subjects treated with VIB7734, compared with the observed changes over time in CLASI-A scores (Figure 39A), absolute blood pDC levels (Figure 39B), blood type I IFNGS levels (measured as absolute scores) (Figure 39C), number of pDCs in skin biopsy specimens (Figure 39D), and normalized blood type I IFNGS levels (measured as a change factor) (Figure 39E) for cohort 2 subjects treated with placebo. CLASI-A score, absolute blood pDC levels, blood type I IFNGS levels, and the number of pDCs in skin biopsy samples were all lower in the VIB7734-treated cohort 2 compared to the placebo-treated cohort 2.

[0151] Figures 64 and 65 summarize the observed changes over time in CLASI-A scores (Figure 64A), absolute blood pDC levels (measured as cells / μL) (Figure 64B), normalized blood type I IFNGS levels (measured as a change factor) (Figure 64C), and pDC counts in skin biopsy material (measured as cells per square mm) (Figure 64D) for cohort 3 subjects treated with VIB7734, compared with the observed changes over time in CLASI-A scores (Figure 65A), absolute blood pDC levels (measured as cells / μL) (Figure 65B), normalized blood type I IFNGS levels (measured as a change factor) (Figure 65C), and pDC counts in skin biopsy material (measured as cells per square mm) (Figure 65D) for cohort 3 subjects treated with placebo. CLASI-A score, absolute blood pDC levels, blood type I IFNGS levels, and the number of pDCs in skin biopsy samples were all reduced in the three VIB7734-treated cohorts compared to the three placebo-treated cohorts.

[0152] In skin lesions of CLE patients, inflammatory infiltration (CD45+ cells) is upregulated. The effect of VIB7734 on inflammatory infiltration of skin lesions was also determined by assaying the level of CD45+ cells per square mm over time in skin biopsy materials from subjects in Cohort 2 (Figure 60) and Cohort 3 (Figure 62). As shown in Figure 61, in the VIB7734-treated subjects in Cohort 2, the median number of CD45 cells in skin biopsy materials decreased from a baseline of 1119 at day 1 to 280 at day 85. In contrast, in the placebo-treated Cohort 2 subjects, the number of CD45 cells in skin biopsy materials decreased from a baseline of 537 at day 1 to 492 at day 85. Also, as shown in Figure 63, in the VIB7734-treated subjects in Cohort 3, the median number of CD45 cells in skin biopsy materials decreased from a baseline of 707 to 513 at day 85. In contrast, in the placebo-treated cohort 3, the baseline CD45 count in skin biopsy material decreased from 897 to 666 on day 85. In addition, Figure 75D shows the baseline pDC / mm² of the skin. 2 This shows the correlation of the rate of change from baseline to day 85 between pDCs and CD45+ cells in VIB7734-treated subjects in a combination of cohorts 2 and 3 where the threshold was 2 or higher. In some subjects, the reduction in pDCs associated with VIB7734 treatment was achieved by a dramatic decrease in total cutaneous CD45+ cells. Overall, the median reduction in CD45+ cells in all VIB7734-treated subjects was 59.69%, compared to -23.97% in the placebo group.

[0153] VIB7734 reversibly depletes circulating pDCs with monthly administration. In addition, VIB7734 reduces type I IFNGS throughout the duration of administration. VIB7734 also reduces cutaneous MxA and cutaneous CD45 levels. CLASI scores improved with treatment with VIB7734 (particularly at a 150 mg dose). No safety issues were identified in subjects with 3 months of VIB7734 administration. No abnormal or clinically significant ECGs were observed in subjects treated with VIB7734. Furthermore, no cases of QT interval increase beyond 30 msec were observed in subjects treated with VIB7734.

[0154] Skin biopsy samples from Cohort 2 and Cohort 3 subjects were also formalin-fixed and paraffin-embedded to allow immunohistochemistry (IHC) and other analyses. Figure 42 provides an overview of the IHC analysis method for skin biopsy materials used herein. Three rounds of staining were performed to determine pDC (BDCA+ / ILT7+ cells), IFN activity (MxA+ pixels), and inflammatory infiltration (CD45+ cells). The analytical quantification strategies for pDC and CD45+ cells using the IHC analysis method for skin biopsy materials are outlined in Figures 43A-43C. The analytical quantification strategies for MxA for determining IFN activity using the IHC analysis method for skin biopsy materials are outlined in Figures 44A-44C.

[0155] As shown in Figures 45A to 45I, within the biopsy material, there was minimal variability in baseline pDC, MxA+ pixel, and CD45+ cell counts for each subject in Cohort 2. In contrast, within the Cohort 2 subjects, there was significant variability among biopsy materials in baseline pDC, MxA+ pixel, and CD45+ cell counts (Figures 46A to 46L). Figure 46J shows the baseline pDCs in skin biopsy materials from each subject in Cohort 2. The subjects had high baseline pDC levels (n=5, >100 pDCs / mm³). 2 ), baseline pDC median (n=3, 10-100 pDC / mm 2 ), or low baseline pDC (n=4, <10 pDC / mm² values). 2Figure 46K shows baseline MxA+ pixels in skin biopsy material from each subject in Cohort 2. Subjects are defined as having high baseline MxA+ pixel counts (n=5, >50% MxA+), intermediate baseline MxA+ pixel counts (n=4, 5-50% MxA+), or low baseline MxA+ pixel counts (n=2, <5% MxA+). Figure 46L shows baseline CD45+ cells in skin biopsy material from each subject in Cohort 2. Subjects are defined as having high baseline CD45+ cell counts (n=3, >2000 CD45+ cells / mm³). 2 ), baseline CD45+ cell median (n=4, 500-2000 CD45+ cells / mm³) 2 ), or low baseline CD45+ cell count (n=5, <500 CD45+ cells / mm³). 2 Figure 47 shows that there was considerable variability in the response in terms of the decrease (measured by the percentage change from baseline) of pDCs (Figure 47A), MxA+ pixels (Figure 47B), and CD45+ cells (Figure 47C) in skin biopsy material from cohort 2 subjects treated with placebo. A more consistent decrease in pDCs, MxA+ pixels, and CD45+ cells was observed in skin biopsy material from cohort 2 subjects treated with VIB7734.

[0156] Thus, the placebo group in Cohort 2 showed significant variability in cutaneous pDC count and IFN activity, both at baseline and over time. In contrast, the VIB7734 treatment group in Cohort 2 observed a more consistent decrease in cutaneous pDC count and IFN activity.

[0157] As shown in Figures 69A to 69I, there was minimal variability in the biopsy material for baseline pDC, MxA+ pixel, and CD45+ cell counts in each subject of Cohort 3. A consistent decrease in the number of pDCs in the skin was observed in the VIB7734-treated Cohort 3 group.

[0158] As shown in Figure 70, within the range of subjects in Cohort 3, there was considerable variability among biopsy materials in terms of baseline pDC, MxA+ pixels, and CD45+ cell counts (Figures 70A-70L). Compared with subjects in Cohort 2, subjects in Cohort 3 showed slightly increased baseline pDC and MxA signaling. Figure 70J shows baseline pDC (measured as the number of cells per square mm) in skin biopsy materials from subjects in Cohort 2 and Cohort 3. Subjects in Cohort 2 had high baseline pDC values ​​(n=5, >100 pDCs / mm²). 2 ), baseline pDC median (n=3, 10-100 pDC / mm 2 ), or low baseline pDC (n=4, <10 pDC / mm² values). 2 This shows the results. Cohort 3 subjects were those with high baseline pDC values ​​(n=5, >100 pDCs / mm²). 2 ), baseline pDC median (n=3, 10-100 pDC / mm 2 ), or low baseline pDC (n=2, <10 pDC / mm²) 2 Figure 70K shows baseline MxA+ pixels (measured as %ROI MxA+) in skin biopsy material from subjects in Cohort 2 and Cohort 3. Subjects in Cohort 2 show high baseline MxA+ pixel values ​​(n=5, >50% MxA+), intermediate baseline MxA+ pixel values ​​(n=4, 5-50% MxA+), or low baseline MxA+ pixel values ​​(n=2, <5% MxA+). Subjects in Cohort 3 show high baseline MxA+ pixel values ​​(n=6, >50% MxA+) or low baseline MxA+ pixel values ​​(n=4, <5% MxA+). Figure 70L shows baseline CD45+ cells (measured as the number of cells per square mm) in skin biopsy material from subjects in Cohort 2 and Cohort 3. Cohort 2 included individuals with elevated baseline CD45+ cell counts (n=3, >2000 CD45+ cells / mm³). 2 ), baseline CD45+ cell median (n=5, 500-2000 CD45+ cells / mm³) 2 ), or low baseline CD45+ cell count (n=4, <500 CD45+ cells / mm³).2 This shows the results. Cohort 3 subjects were those with elevated baseline CD45+ cell counts (n=2, >2000 CD45+ cells / mm³). 2 ), baseline CD45+ cell median (n=4, 500-2000 CD45+ cells / mm³) 2 ), or low baseline CD45+ cell count (n=4, <500 CD45+ cells / mm³). 2 This shows that, compared to the subjects in Cohort 2, the skin biopsy materials from the subjects in Cohort 3 had slightly higher baseline pDC / IFN activity.

[0159] Figure 74 shows the changes in pDCs from baseline (BL) to day 85 (d85) for each subject in Cohort 3 treated with 150 mg of VIB7734 or placebo. Figure 74A shows the changes in pDCs (measured as BDCA2+ / ILT7+ cells) using IHC analysis of skin biopsy materials for subjects in Cohort 3 treated with placebo (n=2; 10030044 and 10010052). Figure 74B shows the changes in pDCs (measured as BDCA2+ / ILT7+ cells) using IHC analysis of skin biopsy materials for cohort 3 subjects (n=8; 30010047, 10120061, 20070048, 200020040, 10010056, 10120055, 20060038, and 10140059) treated with 150 mg of VIB7734. Figure 74C shows the changes in pDCs (measured as the number of cells per square mm) in skin biopsy materials for each of the cohort 3 subjects treated with VIB7734 and each of the cohort 3 subjects treated with placebo at day 85 compared to baseline.

[0160] To investigate the contribution of pDCs to local type I IFN activity in tissues, skin samples from subjects treated with placebo or VIB7734 were also stained for well-characterized type I IFN-induced MxA protein (Figure 78). Type I IFN activity in the skin of lupus subjects varied greatly at baseline, with the majority demonstrating high tissue MxA levels, while some had little to no MxA signaling. In placebo-treated subjects, the change in MxA expression in the skin over time also varied greatly, with the median MxA stained area increasing from 5.4% at baseline to 18% at day 85 (Figure 78A). Treatment with VIB7734 reduced skin MxA levels, with a median percentage change from baseline of -84.5% at day 85 for VIB7734-treated participants (Figures 78B-78C).

[0161] Figure 71 shows that no apparent effect of placebo was observed on skin biopsy material markers pDC (measured as a percentage change from baseline on day 1; Figure 71A), MxA+ pixels (measured as a percentage change from baseline on day 1; Figure 71B), or CD45+ cells (measured as a percentage change from baseline on day 1; Figure 71C) in subjects of Cohort 3 at day 85. In contrast, as shown in Figures 72 and 73, most subjects of Cohort 3 treated with 150 mg of VIB7734 showed a significant decrease in pDC (measured as a percentage change from baseline on day 1; Figures 72A and 73A) and MxA+ pixels (measured as a percentage change from baseline on day 1; Figures 72B and 73B), as well as a decrease in CD45+ cells (measured as a percentage change from baseline on day 1; Figures 72C and 73C) at day 85 compared to the placebo-treated subjects of Cohort 3. As shown in Figure 72A, compared to the mean increase in pDC of 12.24 ± 37.69 (mean ± SEM) in the placebo-treated cohort 3, the VIB7734-treated cohort 3 observed a mean decrease in pDC of 80.98 ± 12.12 (mean ± SEM). As shown in Figure 72B, compared to the mean increase in MxA+ pixels of 773.6 ± 866.33 (mean ± SEM) in the placebo-treated cohort 3, the VIB7734-treated cohort 3 observed a mean decrease in MxA+ pixels of 58.29 ± 17.88 (mean ± SEM).

[0162] Figure 75 shows combined data for subjects from Cohort 2 and Cohort 3. VIB7734 significantly reduced pDCs in the skin of subjects in Cohorts 2 and 3 treated with VIB7734 compared to subjects in Cohorts 2 and 3 treated with placebo. As shown in Figure 75A, at day 85, the mean and median reduction in pDCs were 11.38% and 12.73% for the placebo-treated subjects (n=6) in Cohorts 2 and 3, respectively, compared to the mean and median reductions of 71.82% and 95.3%, respectively, for the VIB7734-treated subjects (n=16) in Cohorts 2 and 3. As shown in Figure 75B, on day 85, the mean and median increases in MxA+ pixels were 269.3% and 38.8% for placebo-treated subjects (n=6) in Cohorts 2 and 3, respectively, compared to the mean and median decreases in pDCs (52.9% and 84.48%, respectively) for VIB7734-treated subjects (n=16) in Cohorts 2 and 3, respectively. To better understand the relationship between changes in pDCs in the skin and changes in type I IFN activity, correlation analysis was performed. There was a highly significant correlation between the decrease in pDCs in skin biopsy samples and the decrease in MxA activity (r=0.7793, Figure 75C). Dramatic decreases in MxA activity were observed in subjects with the greatest degree of tissue pDC depletion, while incomplete / partial pDC depletion often resulted in only minimal changes in MxA. Thus, when all placebo and treatment subjects (cohorts 2 and 3) are combined, a significant effect of VIB7734 on tissue biomarkers is demonstrated. These data suggest that pDCs are a critically important source of type I IFN in lupus skin, and that when these cells in the tissue are nearly completely depleted, local IFN activity is diminished.

[0163] Figure 76 shows that while cutaneous pDCs (measured as the number of cells per square millimeter) decreased in subjects from both Cohort 2 and Cohort 3, pDC depletion was more consistent in subjects from Cohort 3. This is true for all subjects treated with VIB7734 in Cohort 2 (Figure 76A) and Cohort 3 (Figure 76B), as well as for subjects in Cohort 2 (Figure 76C) and Cohort 3 (Figure 76D) treated with VIB7734 who did not have baseline pDC or low IFN activity levels in skin biopsy samples.

[0164] Figure 66 shows that while cutaneous pDCs (measured as the percentage change in cell count from baseline on day 1) decreased in both cohort 2 and cohort 3 subjects treated with VIB7734, pDC depletion was more consistent in cohort 3 subjects. As shown in Figure 66A, at baseline, the pDC count was 10 pDCs / mm². 2 The mean percentage reduction from baseline on day 1 for pDCs in skin samples with more than 5% MxA+ at baseline was 96.31% in cohort 3 of VIB7734 treatment, compared to 85.45% in cohort 2 of VIB7734 treatment. In addition, as shown in Figure 66B, the mean percentage reduction from baseline on day 1 for MxA+ pixels in skin samples with more than 5% MxA+ at baseline was 76.84% in cohort 3 of VIB7734 treatment, compared to 67.44% in cohort 2 of VIB7734 treatment.

[0165] Figure 48 shows that the IHC analysis method for skin biopsy materials does not include an activity threshold. Figure 48A: Percentage change from baseline in MxA in skin biopsy materials from Cohort 2 subjects treated with placebo or ILT7-binding protein (VIB7734) used by the method described herein. Gray contours indicate skin biopsy samples with a substantial numerical multiplier increase in MxA. However, overall, very low levels of MxA maintenance were observed in skin biopsy materials from Cohort 2 subjects. Figure 48B: IHC performed on skin biopsy materials from Cohort 2 subjects following multiple subcutaneous administrations of placebo (3 doses every 4 weeks). Figure 48C: IHC performed on skin biopsy materials from Cohort 2 subjects following multiple subcutaneous administrations of 50 mg of VIB7734 (3 doses every 4 weeks).

[0166] Figure 49 shows the relationship between high baseline pDC count / IFN activity in skin biopsy samples from Cohort 2 subjects and their response to VIB7734. In the VIB7734 treatment group, high baseline pDC count and high IFN activity were observed in skin biopsy samples from 4 out of 5 responders. In non-responders, baseline pDC or IFN activity in skin biopsy samples was low. In the placebo group of Cohort 2 subjects, no discernible relationship was observed between pDC or IFN activity and response. Thus, high baseline pDC / IFN activity in the skin and high serum IFN levels were strongly associated with CLASI responders in the VIB7734 treatment group. On the other hand, low / intermediate baseline pDC / IFN activity in the skin was strongly associated with CLASI non-responders in the VIB7734 treatment group.

[0167] Figure 67 shows the relationship between elevated baseline pDC counts in skin biopsy material from cohort 3 subjects and the response to VIB7734. VIB7734 reduced pDC levels in the skin of cohort 3 subjects (Figure 67A).

[0168] Furthermore, in Cohort 3 of VIB7734-treated CLASI responders, a strong association was observed between intermediate / high baseline skin pDC / MxA levels and high IFN levels (note: all VIB7734-treated subjects in Cohort 3 had high baseline serum IFNγ). Depletion of pDCs with VIB7734 led to a marked decrease in type I IFN activity in CLE skin, demonstrating the essential role of these cells in IFNα production in autoimmune tissues.

[0169] Sequence Information SEQUENCE LISTING <110> VIELA BIO, INC. <120> METHODS OF TREATMENT USING ILT7 BINDING PROTEINS <150> US 63 / 109,923 <151> 2020-11-05 <150> US 63 / 083,649 <151> 2020-09-25 <150> US 63 / 024,182 <151> 2020-05-13 <150> US 63 / 023,820 <151> 2020-05-12 <150> US 62 / 944,845 <151> 2019-12-06 <160> 8 <170> PatentIn version 3.5 <210> 1 <211> 122 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 1 Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 20 25 30 Gly Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu 50 55 60 Gln Gly Arg Val Thr Met Thr Thr Asp Thr Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asn Gly Leu Trp Gly Trp Asp Ser Asp Ala Phe Asp Ile Trp 100 105 110 Gly Arg Gly Thr Leu Val Thr Val Ser Ser 115 120 <210> 2 <211> 110 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic polypeptide <400> 2 Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln 1 5 10 15 Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30 Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45 Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60 Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu 65 70 75 80 Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser 85 90 95 Ser Thr Val Val Phe Gly Gly Gly Thr Lys Val Thr Val Leu 100 105 110 <210> 3 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 3 Ser Tyr Gly Ile Ser 1 5 <210> 4 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 4 Trp Ile Ser Ala Tyr Asn Gly Asn Thr Asn Tyr Ala Gln Lys Leu Gln 1 5 10 15 Gly <210> 5 <211> 13 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 5 Asn Gly Leu Trp Gly Trp Asp Ser Asp Ala Phe Asp Ile 1 5 10 <210> 6 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 6 Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asn Tyr Val Ser 1 5 10 <210> 7 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 7 Asp Val Ser Asn Arg Pro Ser 1 5 <210> 8 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> Description of Artificial Sequence: Synthetic peptide <400> 8 Ser Ser Tyr Thr Ser Ser Ser Thr Val Val 1 5 10

Claims

1. A pharmaceutical composition for reducing the type I interferon gene signature (IFNGS) of a subject in need thereof, wherein the composition comprises a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, wherein the ILT7 binding protein is an antibody comprising heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively, wherein the composition comprising the ILT7 binding protein is administered to the subject when the type I IFNGS is at least 1.1 times higher than that of a normal subject prior to administration of the composition comprising the ILT7 binding protein, wherein the pharmaceutically effective amount is 1 mg, 5 mg, 15 mg, 50 mg, 100 mg, or 150 mg.

2. The pharmaceutical composition according to claim 1, wherein the type I IFNGS is measured in a test biological sample taken from the subject, and the test biological sample is selected from the group consisting of blood, sputum, saliva, skin biopsy, kidney cells, lung cells, liver cells, cardiac cells, brain cells, nerve tissue, thyroid cells, eye cells, skeletal muscle cells, cartilage, bone tissue, and cultured cells.

3. The pharmaceutical composition according to claim 2, wherein the biological sample for examination is blood or a skin biopsy.

4. The pharmaceutical composition according to claim 2, wherein the type I IFNGS in the test biological sample is elevated to at least about four times the baseline level of the test biological sample of a subject requiring it, which is determined by evaluating the type I IFNGS level of a normal subject or the mRNA level of the type I interferon gene.

5. The pharmaceutical composition according to claim 1, wherein the type I IFNGS is determined by a method comprising determining the mRNA levels of at least two genes selected from the group consisting of SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, and USP18.

6. The pharmaceutical composition according to claim 5, wherein the type I IFNGS is determined by a method that includes determining the mRNA levels of all of the following: SPATS2L, EPSTI1, HERC5, IFI27, IFI44, IFI44L, IFI6, IFIT1, IFIT3, ISG15, LAMP3, LY6E, MX1, OAS1, OAS2, OAS3, PLSCR1, RSAD2, RTP4, SIGLEC1, and USP18.

7. The pharmaceutical composition according to claim 1, wherein the administration is effective in reducing the following levels: i. Plasma cell-like dendritic cells (pDCs); ii. Type I IFNGS; and / or iii. The pDC and the Type I IFNGS.

8. The pharmaceutical composition according to claim 1, wherein the target population has an autoimmune disease.

9. The pharmaceutical composition according to claim 8, wherein the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, dermatomyositis, inclusion body myositis, juvenile myositis, polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, multiple sclerosis, rheumatic carditis, psoriasis, arthritis, rheumatoid arthritis, inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, and type 1 interferonosis.

10. The pharmaceutical composition according to claim 9, wherein the autoimmune disease is Sjögren's syndrome, dermatomyositis, vitiligo, polymyositis, systemic sclerosis, hidradenitis suppurativa, SLE, or CLE.

11. The pharmaceutical composition according to claim 1, wherein the ILT7-binding protein is an antibody comprising a variable heavy chain (VH) that is at least 85% identical to that of SEQ ID NO: 1 and / or a variable light chain (VL) that is at least 85% identical to that of SEQ ID NO:

2.

12. The pharmaceutical composition according to claim 1, wherein the ILT7-binding protein is an antibody comprising the heavy chain variable region (VH) of SEQ ID NO: 1 and the light chain variable region (VL) of SEQ ID NO:

2.

13. The pharmaceutical composition according to claim 1, wherein the ILT7-binding protein is an afucosylated antibody.

14. The pharmaceutical composition according to claim 7, comprising a reduced level of type I IFNGS, wherein the reduction is at least about 50% compared to the level of the ILT7-binding protein before administration.

15. A pharmaceutical composition for treating an autoimmune disorder in a subject requiring it, wherein the composition comprises a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, the pharmaceutically effective amount of the ILT7 binding protein being about 50 mg, wherein the ILT7 binding protein is an antibody comprising heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively, and further wherein the composition comprising the ILT7 binding protein is administered to the subject when, prior to administration of the composition comprising the ILT7 binding protein, the type I IFNGS is elevated by at least 1.1 times compared to the type I IFNGS of a normal subject, and the pharmaceutically effective amount is 1 mg, 5 mg, 15 mg, 50 mg, 100 mg, or 150 mg. A pharmaceutical composition.

16. A pharmaceutical composition for treating an autoimmune disorder in a subject requiring it, wherein the composition comprises a pharmaceutically effective amount of immunoglobulin-like transcript 7 (ILT7) binding protein, the pharmaceutically effective amount of the ILT7 binding protein being about 150 mg, wherein the ILT7 binding protein is an antibody comprising heavy chain complementarity-determining regions (HCDRs) HCDR1, HDR2, HCDR3 and light chain complementarity-determining regions (LCDRs) LCDR1, LCDR2, and LCDR3, each comprising the amino acid sequences of SEQ ID NOs: 3, 4, 5, 6, 7, and 8, respectively, and further wherein the composition comprising the ILT7 binding protein is administered to the subject when, prior to administration of the composition comprising the ILT7 binding protein, the level of type I IFNGS is at least 1.1 times higher than that of a normal subject.

17. The pharmaceutical composition according to claim 15, wherein the autoimmune disorder is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, dermatomyositis, inclusion body myositis, juvenile myositis, polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, multiple sclerosis, rheumatic carditis, psoriasis, arthritis, rheumatoid arthritis, inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, and type 1 interferonosis.

18. The pharmaceutical composition according to claim 16, wherein the autoimmune disorder is selected from the group consisting of systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus erythematosus (CLE), Sjögren's syndrome, dermatomyositis, inclusion body myositis, juvenile myositis, polymyositis, systemic sclerosis, diabetes mellitus, Hashimoto's disease, autoimmune adrenal insufficiency, anemia, multiple sclerosis, rheumatic carditis, psoriasis, arthritis, rheumatoid arthritis, inflammation, chronic rheumatism, vitiligo, alopecia areata, hidradenitis suppurativa, celiac disease, graft-versus-host disease (GVHD), myocardial infarction, and type 1 interferonosis.

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