Monoclonal antibody to human T cells, and methods for preparing same
AssigneeOrtho Pharmaceutical Corporation
InventorsPatrick C. Kung, Gideon Goldstein
Life of this patent
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Abstract
Hybrid cell line for production of monoclonal antibody to an antigen found on all normal human T cells. The hybrid is formed by fusing splenocytes from immunized Balb/cJ mice with P3X63Ag8U1 myeloma cells. Diagnostic and therapeutic uses of the monoclonal antibody are also disclosed.
Claims11
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;
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- (e) does not react with Epstein-Barr virus-transformed human B cell lines.
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.
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.
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;
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- (vii) recovering the antibody from the supernatant above said clones.
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.
A method of preparing monoclonal antibody which reacts with essentially all normal human peripheral T cells but not with normal human peripheral B cells, null cells, or macrophages, which comprises culturing the hybridoma ATCC CRL 8000 in a suitable medium and recovering the antibody from the supernatant above said hybridoma.
A method of preparing monoclonal antibody which reacts with essentially all normal human peripheral T cells but not with normal human peripheral B cells, null cells, or macrophages, which comprises injecting into a mouse the hybridoma ATCC CRL 8000 and recovering the antibody from the malignant ascites or serum of said mouse.
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