Monoclonal antibody to human helper 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 T cells, which antibody: (a) reacts with essentially all normal human peripheral helper T cells (being about 55% 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 leukemic cells from humans with T cell chronic lymphoblastic leukemia, B cell chronic lymphoblastic leukemia, T cell acute lymphoblastic leukemia, or null cell acute lymphoblastic leukemia; (d) reacts with the human T cell line CEM, but not with HJD-1, Laz 191, or HM1; (e) does not react with Epstein-Barr virus-transformed human B cell lines Laz 007, Laz 156, Laz 256, or SB; (f) reacts with about 55% of Rhesus monkey peripheral T cells; (g) fixes complement; and (h) defines a T cell population which is unreactive with anti-TH 2 serum and is only minimally cytotoxic.

Claim 3

The monoclonal antibody of claim 1 which is of subclass IgG 2 .

Claim 4

The monoclonal antibody of claim 1 which is produced from a hybridoma formed by fusion of P3X63Ag8Ul myeloma cells and spleen cells from a CAF 1 mouse previously immunized with E rosette purified human T cells.

Claim 5

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

Claim 6

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

Claim 9

The monoclonal antibody prepared by the method of claim 5.

Claim 12

The monoclonal antibody prepared by the method of claim 7.

Claim 13

A monoclonal antibody which reacts with essentially all normal human peripheral helper 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 helper 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 14

A monoclonal antibody which reacts with essentially all normal human peripheral helper 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 helper 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 15

A monoclonal antibody which reacts with essentially all normal human peripheral helper 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 helper 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 16

A monoclonal antibody which reacts with essentially all normal human peripheral helper 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 helper 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 17

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 helper T cells but not with normal human peripheral B cells, null cells, or macrophages.

Claim 18

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 helper T cells but not with normal human peripheral B cells, null cells, or macrophages.