Monoclonal antibody to human T cells, and methods for 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 T cells, which antibody: (a) reacts with essentially all normal human peripheral T cells but not with normal human peripheral B cells, null cells or macrophages; (b) reacts with from about 5% to about 10% of normal human thymocytes; (c) reacts with leukemic cells from humans with T cell chronic lymphoblastic leukemia but does not react with leukemic cells from humans with T cell acute lymphoblastic leukemia; (d) exhibits a pattern of reactivity with the human T cell lines HJD-1, CEM, and HSB-2 shown in FIG. 4; and (e) does not react with Epstein-Barr virus-transformed human B cell lines.

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 fushion of P3X63AgUl myeloma cells and spleen cells from a Balb/cJ mouse previously immunized with E rosette purified human T 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 T cells which reacts with essentially 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 T cells which reacts with essentially 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 essentially all normal human peripheral T cells but not with normal human peripheral B cells, null cells, or macrophages.

Claim 8

Mouse monoclonal antibody which reacts with essentially all normal human peripheral T cells but not with normal human peripheral B cells, null cells, or macrophages.

Claim 9

A monoclonal antibody which reacts with essentially all normal human peripheral 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 E rosette positive purified human T 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; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral 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 essentially all normal human peripheral 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 E rosette positive purified human T 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; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral 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 essentially all normal human peripheral 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 E rosette positive purified human T 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; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral T cells but not with normal human peripheral B cells, null cells, or macrophages; (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 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 E rosette positive purified human T 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; (vi) selecting and cloning a hybridoma producing antibody which reacts with essentially all normal human peripheral T cells but not with normal human peripheral B cells, null cells, or macrophages; (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.