Monoclonal antibody to human helper T cells, method of preparing it, hybrid cell line producing it, its diagnostic and therapeutic uses and diagnostic and therapeutic compositions comprising it

Claim 1

A monoclonal antibody of class IgG produced by a hybridoma formed by fusion of spleen cells from a mouse previously immunized with human T cells and cells from a mouse myeloma line, 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-l, 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 (OKT4 + ) which is unreactive with anti-TH 2 serum and is only minimally cytotoxic.

Claim 13

A method for preparing monoclonal antibody which: 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 (OKT4 + ) which is unreactive with anti-TH serum and is only minimally cytotoxic which comprises the steps of: i) immunizing mice with E rosette positive purified human T cells; iil removing the spleens from said mice and making a suspension of 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 the desired antibody; vi) selecting and cloning hybridomas producing the desired antibody; and vii) recovering the antibody from the supernatant above said clones.

Claim 14

A method for preparing monoclonal antibody which: 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 (OKT4 + ) which is unreactive with anti-TH serum and is only minimally cytotoxic which comprises the steps of: i) immunizing mice with E rosette positive purified human T cells; iil removing the spleens from said mice and making a suspension of 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 the desired antibody; vi) selecting and cloning hybridomas producing the desired antibody; and vii) recovering the antibody from the supernatant above said clones.

Claim 16

A method for preparing monoclonal antibody which: 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 lymphoblastlc 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 C EM , but not with HJD-1, Laz 191, or HMl; 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 (OKT4 + ) which is unreactive with anti-TH 2 serum and is only minimally cytotoxic, which comprises the steps of: il 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 the desired antibody; vi) selecting and cloning hybridomas producing the desired antibody; vii) recovering the antibody from the supernatant above said clones; viii) transferring said clones intraperitoneally into mice; and ix) harvesting the malignant ascites or serum from said mice.

Claim 17

A method for preparing monoclonal antibody which: 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 lymphoblastlc 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 C EM , but not with HJD-1, Laz 191, or HMl; 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 (OKT4 + ) which is unreactive with anti-TH 2 serum and is only minimally cytotoxic, which comprises the steps of: il 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 the desired antibody; vi) selecting and cloning hybridomas producing the desired antibody; vii) recovering the antibody from the supernatant above said clones; viii) transferring said clones intraperitoneally into mice; and ix) harvesting the malignant ascites or serum from said mice.