Graduate Thesis Or Dissertation
 

In vitro regulation and cultivation of salmonid lymphocytes

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/z316q526w

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  • The foundation of the research compilation presented here began with the derivation of an improved method for obtaining optimal in vitro mitogenic responses in salmonid lymphocyte cultures by utilizing autologous or homologous plasma as the primary serum supplement. It was observed that lymphocytes which were previously unresponsive to mitogenic challenge in vitro, when cultured in fetal bovine serum, responded well when cultured in the presence of homologous plasma. Salmonid plasma sources not only repeatedly enhanced the mitogen-specific proliferation of the lymphocyte cultures, but enhanced the antibody response as well. A prolonged kinetic response further supported the contention that former conditions of salmonid lymphocyte culture, employing only fetal bovine serum, not only fail to provide the optimal conditions for cell growth, but in many cases the essential conditions. Enhancement of the mitogenic response was observed for three distinct species, using a common plasma source, suggesting utilization of plasma as an alternate serum supplement has broad applications and may be adapted to many fish systems. With this improved culture system, examination of the regulation of lymphocytes, specifically B cells, was undertaken. Evidence for the existence of a natural regulatory cell population located in the anterior kidney is presented. Addition of anterior kidney cells to either autologous peripheral blood or spleen cell cultures resulted in significant suppression of the mitogen response. The degree of suppressor activity appears to be correlated with the anterior kidney lymphocyte's ability to respond to mitogenic stimulus. It is demonstrated that a decrease in the anterior kidney mitogen response correlates significantly with an increase in the suppressor activity observed upon coculture of these same cells with peripheral blood lymphocytes. Interestingly, while addition of anterior kidney cells to spleen plaque forming cell cultures also resulted in suppression, anterior kidney cells had either no effect or enhanced the antibody response of the peripheral blood lymphocyte cultures. It is postulated that the mediator of this anterior kidney activity is a suppressor cell population which may possess an important immunoregulatory function.
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