Previous work from our laboratory has demonstrated that major histocompatibility complex (MHC) antigens are expressed on cultured bovine luteal cells following exposure to the T lymphocyte-derived cytokine, interferon-gamma (IFN-gamma). In light of these actions of IFN-gamma, it was of interest to investigate the effects of this cytokine on other aspects of luteal function. Therefore, bovine luteal cells were cultured for 7 days in the presence or absence of IFN-gamma, and luteal progesterone (P4), prostaglandin F2 alpha (PGF2 alpha), and 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) production were evaluated. After a 24-h exposure to IFN-gamma (100 U), both PGF2 alpha and 6-keto-PGF1 alpha production were decreased approximately 50% (p less than 0.05). However, as time in culture progressed, IFN-gamma markedly increased the synthesis of both prostaglandins approximately 400% above controls (p less than 0.05). Stimulation of prostaglandin production by IFN-gamma was abrogated by the addition of exogenous P4. During the period of IFN-gamma-stimulated prostaglandin synthesis, LH-stimulated P4 production was inhibited by IFN-gamma treatment. However, the suppression of P4 production by IFN-gamma was not mediated by the increase in prostaglandins since concomitant treatment with indomethacin did not reverse the inhibition of steroidogenesis. These results suggest that IFN-gamma, in addition to an indirect role in promoting immune response mechanisms, may also directly affect luteal function by enhancing luteal prostaglandin synthesis and by inhibiting luteal steroidogenesis.

To investigate immunological mechanisms that may be involved in luteal function, the presence of Class I and Class II major histocompatibility complex (MHC) antigens on cultured bovine luteal cells was examined. After 72 h in serum-free culture, Class I antigens were markedly expressed on luteal cells, as determined by indirect immunofluorescence, whereas expression of Class II antigens was limited. The expression of MHC antigens on luteal cells was increased by treatment with the T-lymphocyte factor, interferon-gamma (IFN-gamma). Class I and II antigens were elevated 25% and 370% above controls, respectively, after IFN-gamma exposure. Since the corpus luteum is regulated by luteinizing hormone (LH), luteal cells were treated with either hormone alone or hormone in addition to IFN-gamma, and antigen expression was determined. LH treatment attenuated IFN-gamma-induction of Class II antigens on bovine luteal cells. These observations are the first to demonstrate the presence of MHC antigens on bovine luteal cells and the modulation of antigen expression by the lymphokine IFN-gamma and by LH.

Summary. Corpora lutea were removed from regularly cycling dairy cows, dissociated with collagenase and cultured for 8 or 10 days in Ham's F-12 medium. In Exp. 1 treatment with insulin, or an insulin\p=n-\transferrin\p=n-\seleniumcombination (ITS), increased progesterone production from basal levels on Day 4 ofculture to 234% (P < 0\m=.\01) above controls on Day 10. LH alone increased progesterone production 45% above controls on Day 10 (P > 0\m=.\05). When LH was combined with insulin or ITS, progesterone production was stimulated to an average of 1802% (P < 0\m=.\01) above controls on Day 10 of culture. Transferrin or selenium without insulin did not allow LH to stimulate progesterone synthesis. In Exp. II, LH alone or LH plus gentamicin or penicillin\p=n-\ streptomycin increased progesterone production from basal levels on Day 2 steadily to an average of 468% (P < 0\m=.\01) above controls (no antibiotics) by Day 8 of culture. The addition of amphotericin-B, alone or in combination with the other antibiotics, inhibited all LH-stimulated progesterone synthesis, but did not affect basal progesterone levels. We conclude that insulin is essential for maximal steroidogenesis in a bovine luteal cell culture system, and that LH-stimulated progesterone production is inhibited in the presence of amphotericin-B, but is not inhibited by gentamicin or penicillin\p=n-\ streptomycin. The elimination of amphotericin-B, coupled with the addition of insulin to the cell culture system increased the responsiveness of the cells to LH. These culture conditions represent the first report in which LH increased total progesterone production for 10 days, maintaining luteal function in a chemically-defined culture system.