Hybrid cell line for producing monoclonal antibody to a human monocyte antigen, antibody, and methods

Claim 1

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 mononuclear cells, which antibody: a) reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage; b) exhibits the pattern of reactivity to normal human monocytes shown in Figure 1 and Table I; c) exhibits the pattern of reactivity with normal human E + and E- peripheral blood cells shown in Figure 2 and Table II; d) exhibits the pattern of reactivity with normal human Ig-, E-, nonadherent null cells shown in Figure 3; e) exhibits the pattern of lytic capacity shown in Figure 4 and Table III; f) reacts with leukemia cells from patients with AMML; g) defines a monocyte population necessary for a proliferative response to soluble antigens; h) defines a monocyte population (OKM1 + ) which is lower than normal levels in primary biliary cirrhosis, acute infectious mononucleosis, and graft versus host reaction, higher than normal levels in multiple sclerosis, myasthemia gravis, and abnormal Hodgkins disease, and normal levels in hyper IgE, early Hodgkins disease, and psoriasis.

Claim 9

A method for preparing monoclonal antibody which: a) reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage; b) exhibits the pattern of reactivity to normal human monocytes shown in Figure 1 and Table I; c) exhibits the pattern of reactivity with normal human E + and E- peripheral blood cells shown in Figure 2 and Table II; d) exhibits the pattern of reactivity with normal human Ig-, E-, nonadherent null cells shown in Figure 3; e) exhibits the pattern of lytic capacity shown in Figure 4 and Table III; f) reacts with leukemia cells from patients with AMML; g) defines a monocyte population necessary for a proliferative response to soluble antigens; h) defines a monocyte population (OKM1 + ) which is lower than normal levels in primary biliary cirrhosis, acute infectious mononucleosis, and graft versus host reaction, higher than normal levels in multiple sclerosis, myasthemia gravis, and abnormal Hodgkins disease, and normal levels in hyper IgE, early Hodgkins disease, and psoriasis. which comprises the steps of: i) immunizing mice with human mononuclear cells; ii) 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 10

A method for preparing monoclonal antibody which: a) reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage; b) exhibits the pattern of reactivity to normal human monocytes shown in Figure 1 and Table I; c) exhibits the pattern of reactivity with normal human E + and E- peripheral blood cells shown in Figure 2 and Table II; d) exhibits the pattern of reactivity with normal human Ig-, E-, nonadherent null cells shown in Figure 3; e) exhibits the pattern of lytic capacity shown in Figure 4 and Table III; f) reacts with leukemia cells from patients with AMML; g) defines a monocyte population necessary for a proliferative response to soluble antigens; h) defines a monocyte population (OKM1 + ) which is lower than normal levels in primary biliary cirrhosis, acute infectious mononucleosis, and graft versus host reaction, higher than normal levels in multiple sclerosis, myasthemia gravis, and abnormal Hodgkins disease, and normal levels in hyper IgE, early Hodgkins disease, and psoriasis. which comprises the steps of: i) immunizing mice with human mononuclear cells; ii) 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 11

A method for preparing mononuclear antibody which: a) reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymoblastoid cell lines, or Tumor cells of T or B cell lineage; b) exhibits the pattern of reactivity to normal human monocytes shown in Figure 1 and Table I; c) exhibits the pattern of reactivity with normal human E + and E- peripheral blood cells shown in Figure 2 and Table II; d) exhibits the pattern of reactivity with normal human Ig-, E-, nonadherent null cells shown in Figure 3; e) exhibits the pattern of lytic capacity shown in Figure 4 and Table III; f) reacts with leukemia cells from patients with AMML; g) defines a monocyte population necessary for a proliferative response to soluble antigens; h) defines a monocyte population (OKM1 + ) which is lower than normal levels in primary biliary cirrhosis, acute infectious mononucleosis, and graft versus host reaction, higher than normal levels in multiple sclerosis, myasthemia gravis, and abnormal Hodgkins disease, and normal levels in hyper IgE, early Hodgkins disease, and psoriasis. which comprises the steps of: i) immunizing mice with human mononuclear 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 12

A method for preparing mononuclear antibody which: a) reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymoblastoid cell lines, or Tumor cells of T or B cell lineage; b) exhibits the pattern of reactivity to normal human monocytes shown in Figure 1 and Table I; c) exhibits the pattern of reactivity with normal human E + and E- peripheral blood cells shown in Figure 2 and Table II; d) exhibits the pattern of reactivity with normal human Ig-, E-, nonadherent null cells shown in Figure 3; e) exhibits the pattern of lytic capacity shown in Figure 4 and Table III; f) reacts with leukemia cells from patients with AMML; g) defines a monocyte population necessary for a proliferative response to soluble antigens; h) defines a monocyte population (OKM1 + ) which is lower than normal levels in primary biliary cirrhosis, acute infectious mononucleosis, and graft versus host reaction, higher than normal levels in multiple sclerosis, myasthemia gravis, and abnormal Hodgkins disease, and normal levels in hyper IgE, early Hodgkins disease, and psoriasis. which comprises the steps of: i) immunizing mice with human mononuclear 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 19

A method of preparing monoclonal antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises culturing the hybridoma ATCC CRL 8026 in a suitable medium and recovering the antibody from the supernatant above said hybridoma.

Claim 20

A method of preparing monoclonal antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises culturing the hybridoma ATCC CRL 8026 in a suitable medium and recovering the antibody from the supernatant above said hybridoma.

Claim 21

A method of preparing monoclonal antibody which reacts with an antigen found on all normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises injecting into a mouse the hybridoma ATCC CRL 8026 and recovering the antibody from the malignant ascites or serum of said mouse.

Claim 22

A method of preparing monoclonal antibody which reacts with an antigen found on all normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises injecting into a mouse the hybridoma ATCC CRL 8026 and recovering the antibody from the malignant ascites or serum of said mouse.

Claim 23

A method for preparing monoclonal antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises the steps of: i) immunizing mice with human mononuclear 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 and E - human peripheral blood lymphocytes; vi) selecting and cloning a hybridoma producing antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage; and vii) recovering the antibody from the supernatant above said clones.

Claim 24

A method for preparing monoclonal antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises the steps of: i) immunizing mice with human mononuclear 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 and E - human peripheral blood lymphocytes; vi) selecting and cloning a hybridoma producing antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage; and vii) recovering the antibody from the supernatant above said clones.

Claim 25

A method for preparing monoclonal antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises the steps of: i) immunizing mice with human mononuclear 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 + and E human lymphocytes; vi) selecting and cloning a hybridoma producing antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage; 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 26

A method for preparing monoclonal antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage, which comprises the steps of: i) immunizing mice with human mononuclear 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 + and E human lymphocytes; vi) selecting and cloning a hybridoma producing antibody which reacts with an antigen found on normal human monocytes and granulocytes, but not with normal human peripheral T cells, B cells, Null cells, thymocytes, lymphoblastoid cell lines, or tumor cells of T or B cell lineage; 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.