Monoclonal antibody to human cytotoxic and suppressor T cells and methods of preparing same

Claim 2

A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with human thymocytes, which antibody: (a) reacts with greater than 90% of cytotoxic and suppressor TH 2 + T cells (being about 20-30% of all normal human peripheral T cells), but not with normal human peripheral B cells, Null cells or macrophages; (b) reacts with about 80% of normal human thymocytes; (c) does not react with TH 2 - T cells or T cells which also react with antibody produced by hybrid cell line ATCC CRL 8002, but reacts with about 68% of T cells which do not react with antibody produced by hybrid cell line ATCC CRL 8002; (d) defines a T cell population which is strongly cytotoxic and which is lower than normal levels in primary biliary cirrhosis, multiple sclerosis, and hyper IgE, normal levels in all stages of Hodgkins disease, and higher than normal levels in Type I acquired agammaglobulinemia and acute infectious mononucleosis.

Claim 3

The monoclonal antibody of claim 1 which is of subclass IgG 1 .

Claim 4

The monoclonal antibody of claim 1 which is produced from a hybridoma formed by fusion of P3X63Ag8U1 myeloma cells and spleen cells from a CAF 1 mouse previously immunized with human thymocytes.

Claim 5

A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with human thymocytes which reacts with greater than 90% of cytotoxic and suppressor TH 2 + T cells (being about 20-30% of all normal human peripheral T cells), but not with normal human peripheral B cells, null cells, or macrophages.

Claim 6

A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of cells from a mouse myeloma line and spleen cells from a mouse previously immunized with human thymocytes which reacts with greater than 90% of cytotoxic and suppressor TH 2 + T cells (being about 20-30% of all normal human peripheral T cells), but not with normal human peripheral B cells, null cells, or macrophages.

Claim 7

Mouse monoclonal antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages.

Claim 8

Mouse monoclonal antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages.

Claim 9

A monoclonal antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages prepared by the method, which comprises the steps of: (i) immunizing mice with normal human thymocytes; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells; (vi) selecting and cloning a hybridoma producing antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages; and (vii) recovering the antibody from the supernatant above said clones.

Claim 10

A monoclonal antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages prepared by the method, which comprises the steps of: (i) immunizing mice with normal human thymocytes; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells; (vi) selecting and cloning a hybridoma producing antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages; and (vii) recovering the antibody from the supernatant above said clones.

Claim 11

A monoclonal antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages prepared by the method which comprises the steps of: (i) immunizing mice with normal human thymocytes; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells; (vi) selecting and cloning a hybridoma producing antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages; and (vii) transferring said clones intraperitoneally into mice; and (viii) harvesting the malignant ascites or serum from said mice, which ascites or serum contains the desired antibody.

Claim 12

A monoclonal antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages prepared by the method which comprises the steps of: (i) immunizing mice with normal human thymocytes; (ii) removing the spleens from said mice and making a suspension of the spleen cells; (iii) fusing said spleen cells with mouse myeloma cells in the presence of a fusion promoter; (iv) diluting and culturing the fused cells in separate wells in a medium which will not support the unfused myeloma cells; (v) evaluating the supernatant in each well containing a hybridoma for the presence of antibody to E rosette positive purified T cells; (vi) selecting and cloning a hybridoma producing antibody which reacts with greater than 90% of cytotoxic and suppressor TH 2 + human T cells but not with normal human peripheral B cells, null cells, or macrophages; and (vii) transferring said clones intraperitoneally into mice; and (viii) harvesting the malignant ascites or serum from said mice, which ascites or serum contains the desired antibody.

Claim 15

The monoclonal antibody prepared by the method of claim 8.

Claim 18

The monoclonal antibody prepared by the method of claim 10.