Monoclonal antibody to a human T cell antigen 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 leukemic cells from a human with T-ALL, which antibody: (a) reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells; (b) reacts with early, common, and mature human thymocytes and with inducer and cytotoxic/suppressor human T cells, but not with human prothymocytes; (c) defines a T cell population which is lower than normal levels in myasthenia gravis and multiple sclerosis; higher than normal levels in acute graft versus host reaction, hyper IgE, acute infectious mononucleosis, and primary biliary cirrhosis; and completely absent in all stages of Hodgkins disease and psoriasis.

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 leukemic T-ALL cells.

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 leukemic T-ALL cells which reacts with essentially all normal human peripheral T cells and with approximately 95 percent of normal human thymocytes, but not with normal human B cells or null cells.

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 leukemic T-ALL cells which reacts with essentially all normal human peripheral T cells and with approximately 95 percent of normal human thymocytes, but not with normal human B cells or null cells.

Claim 7

Mouse monoclonal antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells.

Claim 8

Mouse monoclonal antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells.

Claim 9

A monoclonal antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes but not with normal human B cells or Null cells prepared by the method, which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (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 or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells; and (vii) recovering the antibody from the supernatant above said clones.

Claim 10

A monoclonal antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes but not with normal human B cells or Null cells prepared by the method, which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (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 or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells; and (vii) recovering the antibody from the supernatant above said clones.

Claim 11

A monoclonal antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells prepard by the method, which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (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 or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells; (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 essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells prepard by the method, which comprises the steps of: (i) immunizing mice with human leukemic T-ALL cells; (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 or human thymocytes; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral T cells and with approximately 95% of normal human thymocytes, but not with normal human B cells or Null cells; (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.