In vitro activation of mouse lymphocytes in serum‐free medium: effect of T and B cell mitogens on proliferation and antibody synthesis

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Abstract
Mouse spleen cells were successfully activated by mitogens in serum‐free medium. The T cell mitogen, concanavalin A (ConA), was found to activate DNA synthesis in the presence or absence of serum to the same extent, although the dose response curve was shifted. Thus, concentrations ten times lower were optimally stimulating in serum‐free medium. The strain differences existing with regard to Con A activation were the same whether serum was present or not. B cell mitogens, such as lipopolysaccharide (LPS) and purified protein derivative (PPD) of tuberculin, were equally active in the presence or absence of serum with regard to dose response curves and kinetics for the induction of DNA synthesis.When added to normal spleen cells in culture in the absence of serum, Con A also activated antibody synthesis against a variety of antigens and haptens. It had no such effect on spleen cells from nude animals, indicating that it exerted its B cell‐activating effect via T cells. It was also effective when DNA synthesis in the lymphocytes had been suppressed with mitomycin.LPS and PPD activated antibody synthesis to haptens in normal and nude spleen cells, the response in nude animals being markedly stronger.Since there is virtually no background antibody synthesis in unstimulated cultures in the absence of serum, but a substantial background in serum, it follows that the factor of activation caused by T and B cell mitogens is more pronounced in the absence of serum.

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