ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.1

年代:1981

数据来源: MAL

摘要:

ABSTRACTRat monoclonal antibodies specific for mouse kappa light chains and mouse γ heavy chains have been generated. These rat monoclonal antibodies have been biosynthetically labelled with35S methionine. The free label was dialyzed from the medium and, without further purification, the medium containing the radioactive monoclonal antibody was used in a radioimmunoassay to screen the sera of the immunized animals and hybridomas for specific mouse antibodies of the IgG class

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.5

年代:1981

数据来源: MAL

摘要:

ABSTRACTThe molecule bearing the Thy-1.2 antigen, T25, has been isolated from fractionated thymocytes as well as from cloned cytolytic and helper T cells by immune precipitation with monoclonal antibodies. Using NaDodSO4/polyacrylamide gels cross-linked with N,N′-diallytartardiamide, electrophoretic analyses of the precipitates revealed a heterogeneous pattern of polypeptide bands, ranging in apparent molecular weight from 28,000–36,000 daltons. Thymocytes separated on the basis of Thy-1.2 surface antigen density were found to differ not only in amount of Thy-1.2 expressed, but also in the structure of the surface T25 molecules isolated from each subpopulation. This structural variation was evidenced in changes detected in the relative density and electrophoretic mobility of T25 band patterns obtained on SDS gels. Differences in T25 band patterns were also observed in polypeptides isolated from cloned cytolytic or helper T cells. These data suggest that although surface T25 is structurally heterogeneous, thepatternof heterogeneity can differ from cell to cell, or from population to population. Changes in T25 structure appear to correlate with differences in either T lymphocyte maturation or T cell funct

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.13

年代:1981

数据来源: MAL

摘要:

ABSTRACTAn immunogen consisting of a 4M urea extract derived from human melanoma cells (M14), that was devoid of HLA-A.BX, HLA-DR antigens and fibronectin was adsorbed tolens culinarislectin-Sepharose 4B and used to immunize mice for production of monoclonal antibody to a melanoma-specific glycoprotein. Screening for hybridomas secreting antibodies to melanoma associated antigens was facilitated by use of a solid phase target antigen of chemically defined medium of melanoma cells (CDM). Use of these procedures allowed us to select 40 hybridomas secreting antibody which recognized determinants on melanoma cells not found on lymphoid cells. Further characterization of one of these hybridomas, 9.2.27, indicated that the antibody it secreted recognized a 240K dalton glycoprotein found on all melanoma cell lines tested but not on carcinoma, lymphoid, or fibroblastoid cultures. These results demonstrate the utility of soluble antigen preparations devoid of strongly immunogenic non tumor-specific molecules in the elicitation of tumor specific antibody. Preliminary results suggest that immunogens of this kind are superior to intact melanoma cells for production of tumor specific hybridomas.

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.27

年代:1981

数据来源: MAL

摘要:

ABSTRACTWe have established a solid-phase binding inhibition radioimmunoassay for the detection of colorectal carcinoma-specific antigens in tissue culture supernatants of human colorectal carcinoma cell lines and in serum and urine of colorectal carcinoma patients. Using the [3H]glucosamine-labeled cell membrane glycolipid antigen and colorectal carcinoma-specific monoclonal antibodies in this assay, we have been able to detect several human colorectal carcinoma membrane-specific antigens that are released from the cell membrane into tissue culture supernatants, and an antigen detected by antibodies 1116-NS-19-9 and 1116-NS-52a that is found only in the serum and urine of cancer patients.

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.37

年代:1981

数据来源: MAL

摘要:

ABSTRACTHybridoma clones producing anti-hapten antibodies were established by fusing spleen cells from mice immunized with 4-azophthalate keyhole limpet hemocyanin to drug-resistant non-producing myelomas. The affinity and homogeneity of the immunochemically purified anti-phthalate antibodies were determined by equilibrium dialysis. We report here a broad distribution of the monoclonal antibody affinities ranging from a low of 4.0 × 104to a high of 4.0 × 107(L/M). The binding data for eleven anti-phthalate antibodies described a straight line in a Scatchard plot as would be expected for homogeneous antibodies. We determined that all hybridomas, except those secreting very low affinity antibody, produced hemolytic plaques. The hybridomas made it possible to test several assumptions regarding the association of plaque morphology and plaque inhibition tion with the affinity of the antibody secreted by the plaque-forming cells. Our studies indicate that the affinity of the antibody secreted by the hybridoma clones does not correlate with either the size of the plaque or with the efficiency with which the hybridoma-produced plaques are inhibited by free hapten. By comparing hybridoma-produced plaques to plaques produced by phthalate-immune spleen cells it has been demonstrated that the plaque size within each hybridoma clone was substantially less heterogeneous than that observed for the immune spleen cells. Our results do support the assumption that the range of free hapten concentration over which PFC are inhibited is a reflection of PFC heterogeneity. The analysis of the PFC inhibition of hybridomas produced an interesting and unexpected result which is reported here. It was observed that subinhibitory doses of free hapten caused an enhancement in the number of hybridoma-produced plaques. The relevance of this finding to the recent observation and interpretation of plaque enhancement (i.e., the displacement of anti-idiotype antibodies) observed for immune spleen cells(1)is discusse

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.47

年代:1981

数据来源: MAL

摘要:

ABSTRACTMonoclonal antibodies were raised against the leukemic T cells from a patient with chronic lymphocytic leukemia. Two antibodies, termed T411 and T811, were obtained which were reactive by indirect immunofluorescence only with cells of the T cell lineage. The T411 antibody recognized a polypeptide chain of 100,000 dalton apparent molecular weight which was present on the surface of 94 ± 4% of peripheral blood T lymphocytes, but only on 20 ± 8% of thymus cells. The antibody T811 reacted with a surface molecule composed of 2 poly-peptide chains of 32,000 and 34,000 dalton apparent molecular weight, which was expressed only on 25 ± 10% of blood T lymphocytes and on 90 ± 4% of thymus cells. Functional analysis of the T811+and T811−1 T cell subsets isolated by rosetting with anti-mouse Ig coated ox erythrocytes revealed that both subpopulations were able to mount a proliferative response to allo-antigens, whereas allo-antigen induced cytotoxic cells and their precursors were only found in the T8ll+subset. The pokeweed mitogen induced in vitro differentiation of B lymphocytes into immunoglobulin secreting cells was dependent on the presence of the T811−subset, whereas the T811+T cells efficiently suppressed this differen

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.59

年代:1981

数据来源: MAL

摘要:

ABSTRACTIn order to study the heterogeneity of the anti-erythrocyte auto-antibody response in NZB mice, we have developed hybridomas producing monoclonal auto-antibodies with anti-erythrocyte activity. These monoclonals were prepared by fusion of NZB splenocytes with the P3 × 63.Ag8 myeloma and were screened for activity by indirect immunofluorescence using flow cytometry. We have produced a variety of monoclonal anti-red cell auto-antibodies that have differing antigenic reactivities and immunoglobulin isotypes. Eleven monoclonal antibodies extensively studied thus far react with intact mouse erythrocytes, whereas only two of the eleven also cross react with sheep erythrocytes and bromelain treated mouse erythrocytes. These results suggest that the fusion pattern may represent the range of anti-red cell auto-antibodies found in the intact NZB mouse, with the exception of monoclonal auto-antibodies against cryptic red cell auto-antigens, which were not demonstrated in this initial fusion study

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.71

年代:1981

数据来源: MAL

摘要:

ABSTRACTHybridomas were made between NS1/1-Ag4-1 mouse myeloma cells and spleen and lymph node cells from a sheep immunized with sheep red cells (RBC). The hybrid colonies grew well in culture but there was a substantial loss of sheep chromosomes. No hemolytic or agglutinating antibodies were detected in the culture supernatants after the 17th day following fusion, but immunofluorescence tests indicated that a few of the cells may have been expressing sheep IgG. Cytogenetic comparison of cells grown with and without HAT medium provided evidence that the enzyme HGPRT is located on the X chromosome of sheep as it is in man and mouse. Hybridoma isozyme patterns of esterase, G6PD, 6PGD, NP, LDH and SOD tested between the 63rd and 71st day of culture were like those of NS1; NP and LDH also showed zones that probably came from the sheep component.

ISSN:0272-457X    

DOI:10.1089/hyb.1.1981.1.77

年代:1981

数据来源: MAL