Murine B cell mitogens such as bacterial lipopolysaccharide (LPS), butanol-extracted water soluble adjuvant (Bu-WSA), dextran sulfate (DS), synthetic muramyl dipeptide (MDP), and its analog MDP-Lys (L18) do not show any mitogenic ability in vitro on human peripheral blood lymphocytes or mixed cell populations of purified T and B cells obtained from the lymphocytes in an ordinary culture system. However, these mitogens are capable of enhancing the mitogenic effect of concanavalin A (Con A) in the cultures.In the presence of one of these mitogens, the activity of interleukin 2 (IL 2), but not interleukin 1, in the supernatants obtained from cultures containing Con Astimulated T cell and B cell populations was higher than that of control cultures. The role of the newly produced IL 2 in the synergistic effect of the mitogens in human lymphocyte cell cultures was discussed.Concanavalin A (Con A) is one of the potent mitogens for T lymphocytes (8), but the proliferative response of T lymphocytes requires the presence of Ia-positive accessory cells such as macrophages or B lymphocytes (2, 27). Human peripheral T lymphocyte proliferation by Con A is augmented further in the presence of bacterial lipopolysaccharide (LPS) (13, 26) or butanol-extracted, water soluble adjuvant (Bu-WSA) obtained from Bacterionema matruchotii, gram positive oral bacteria (21). Although LPS (10) and Bu-WSA (19) are B cell mitogens in mice, these B cell mitogens do not appear to activate human peripheral B lymphocytes under ordinary experimental conditions (25, 26), much less peripheral T lymphocytes.The triggering of human lymphocytes and the mechanism(s) of activation are subjects of intense interest and investigation. Affects of so-called B cell mitogens on Con A-induced T cell proliferation display a complexity of mitogenic responses. The object of the present study was to investigate whether synergistic effects could be seen on Con A-induced human peripheral blood lymphocyte proliferation in vitro when other kinds of B cell mitogens were used instead of LPS or Bu-WSA and 44 1
Butanol‐extracted water‐soluble adjuvant (Bu‐WSA) obtained from Bacterionema matruchotti was mitogenic to murine splenic B lymphocytes, but not T lymphocytes. When murine splenic cells were cultured in the presence of Bu‐WSA and concanavalin A (Con A) together. [3H|thymidine uptake of the culture cells synergically increased. The mechanism of the synergy of Con A and Bu‐WSA and the participation of interleukin (IL) 1 and 2 in the synergy were studied. The proliferation cells in the synergy were I yt‐1− 23− lymphocytes.’la‐positive accessory cells were required for the response. When separated cell populations and Marbrook‐type culture vessels were used, a mixed cell population of T lymphocytes and B lymphocytes or macrophages (Mφ) produced some active factor(s) after co‐stimulation by Con A and Bu‐WSA, and the factors enhanced DNA synthesis of another Con A‐activatcd T lymphocyte population. Supernatants obtained from the spleen cell cultures or the mixed cell cultures with T lymphocytes and Mφ in the presence of Con A and Bu‐WSA contained greater amounts of IL‐1 and IL‐2 than those from cultures containing Con A or Bu‐WSA alone. An addition of exogenous IL‐1 or IL‐2 to spleen cell cultures with Con A resulted in a proliferative response like that obtained through co‐stimulation by Con A and Bu‐WSA. These results suggest that the synergistic effect of Con A and Bu‐WSA on the proliferative response in murine splenic cells is sustained by the enhancement of production of these T‐lymphocyte growth factors.
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