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In vitro sensitization for human monoclonal antibody production
1Institute of Human Genetics, Polish Academy of Sciences, Poznan.
We have found that cell adhesion prior to in vitro antigenic stimulation enriched the B-cell population and diminished the CD8 lymphocyte subset. These changes in lymphocyte proportions were favourable for increasing the percentage of antibody-producing cells after culturing in vitro followed by Epstein-Barr virus (EBV) infection. The immunodepletion of leukocytes by methyl esters did not yield satisfying results under analogous culture conditions. In vitro primary antigenic stimulations with the addition of IL-2 (25 U/ml medium) and low amounts of interferon-gamma provided better recruitment of antibody-producing cells and higher binding activity of antibodies to sperm than secondary antigenic stimulation. IL-6 did not positively influence the EBV-transformed cell lines.
We have found that cell adhesion prior to in vitro antigenic stimulation enriched the B-cell population and diminished the CD8 lymphocyte subset. These changes in lymphocyte proportions were favourable for increasing the percentage of antibody-producing cells after culturing in vitro followed by Epstein-Barr virus (EBV) infection. The immunodepletion of leukocytes by methyl esters did not yield satisfying results under analogous culture conditions. In vitro primary antigenic stimulations with the addition of IL-2 (25 U/ml medium) and low amounts of interferon-gamma provided better recruitment of antibody-producing cells and higher binding activity of antibodies to sperm than secondary antigenic stimulation. IL-6 did not positively influence the EBV-transformed cell lines.