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In vitro immunization of human B lymphocytes with cultured melanoma cells (SK-MEL 28)
1Department of Biotechnology, University of Lund, Sweden.
Human peripheral blood lymphocytes (PBL) from 10 healthy donors were in vitro immunized using the human melanoma cell line SK-MEL 28 as an immunogen. The parameters necessary to obtain an optimal immune response against the tumor cells in vitro were determined. Different ratios of PBL and SK-MEL 28 were tested during the in vitro immunization period, and their effect on cell viability, specific antibody secretion, and yield of antigen-specific EBV clones was determined. A statistically significant positive dose-response relation was shown to exist between the immunogenic dose present during in vitro immunization and the number of antigen-specific EBV clones that was obtained. A ratio of PBL:SK-MEL 28 cells of 1:1 was found to be optimal and gave 2.6% positive clones from which human monoclonal antibodies specific for the melanoma cells were detected.
Human peripheral blood lymphocytes (PBL) from 10 healthy donors were in vitro immunized using the human melanoma cell line SK-MEL 28 as an immunogen. The parameters necessary to obtain an optimal immune response against the tumor cells in vitro were determined. Different ratios of PBL and SK-MEL 28 were tested during the in vitro immunization period, and their effect on cell viability, specific antibody secretion, and yield of antigen-specific EBV clones was determined. A statistically significant positive dose-response relation was shown to exist between the immunogenic dose present during in vitro immunization and the number of antigen-specific EBV clones that was obtained. A ratio of PBL:SK-MEL 28 cells of 1:1 was found to be optimal and gave 2.6% positive clones from which human monoclonal antibodies specific for the melanoma cells were detected.