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Porcine lymphocytes secrete a bioactive and immunoreactive LH-like factor in response to LHRH and concanavalin A
F E Standaert1, B P Chew, T S Wong
1Department of Animal Sciences, Washington State University, Pullman, Washington 99164-6320.
Insights
Lymphocyte conditioned medium, influenced by luteinizing hormone-releasing hormone (LHRH) and concanavalin A (con A), stimulated progesterone production in granulosa cells. LHRH antagonist blocked this LHRH-induced stimulation.
Area of Science:
- Endocrinology
- Reproductive Biology
- Cell Signaling
Background:
- Lymphocyte-conditioned media (CM) can modulate reproductive hormone functions.
- Luteinizing hormone-releasing hormone (LHRH) and its antagonist (LHRHA) play roles in hormonal regulation.
- Concanavalin A (con A) is a known immune stimulant.
Purpose of the Study:
- To investigate the effect of lymphocyte-conditioned medium (CM) with varying LHRH and LHRHA concentrations on luteinizing hormone (LH) production.
- To determine the impact of lymphocyte CM on progesterone production by porcine granulosa cells.
- To assess the role of concanavalin A (con A) in modulating these effects.
Main Methods:
- Lymphocytes were incubated with different concentrations of LHRH, LHRHA, or both, with and without con A.
- Conditioned media were analyzed for LH content.
- Porcine granulosa cells were cultured with the prepared CM, and progesterone levels were measured over time.
Main Results:
- Lymphocyte CM, but not cell-free CM, showed increased LH with higher LHRH concentrations.
- LHRHA significantly reduced LH levels in lymphocyte CM.
- Con A led to maximal LH concentrations, irrespective of LHRH or LHRHA presence.
- Lymphocyte CM with LHRH dose-dependently increased granulosa cell progesterone production, an effect blocked by LHRHA.
- Con A in lymphocyte CM also stimulated progesterone production, independent of LHRHA.
Conclusions:
- Lymphocyte CM can produce LH and modulate granulosa cell function.
- LHRH in lymphocyte CM stimulates progesterone production, mediated by LHRH receptors.
- Con A enhances LH production and granulosa cell stimulation by lymphocyte CM, suggesting complex immune-endocrine interactions.
Abstract:
Lymphocyte conditioned medium (CM) was prepared by incubating 0 (cell-free control) or 4 x 10(6) lymphocytes/ml in serum-supplemented RPMI containing 0, 10(-9), 10(-7), and 10(-5) M luteinizing hormone releasing hormone (LHRH), 10(-5) M LHRH antagonist (LHRHA), or 10(-7) M LHRH + 10(-5) M LHRHA. Treatments were applied with and without 10 micrograms/ml concanavalin A (con A), and media were analyzed for LH. Aliquots of the CM from cultures incubated for 48 h were later applied to porcine granulosa cell cultures (suspended to 1.25 x 10(5) cells in 450 mul). Thereafter, 50 mul of CM were added to granulosa cell cultures. Media were collected after 12, 24, and 48 h and progesterone determined. Immunoreactive LH increased with time of incubation in lymphocyte CM but not cell-free CM. LH content of lymphocyte CM increased as LHRH concentration increased. LHRHA significantly reduced the amount of LH measured. The presence of con A in the medium resulted in maximal concentrations of LH, irrespective of dose of LHRH or LHRHA. Cell-free CM containing LHRH, LHRHA, and/or con A did not affect progesterone production by granulosa cells at any of the time periods. Lymphocyte CM containing LHRH caused a dose-dependent increase in progesterone production at 48 h. This stimulation was blocked by lymphocyte CM containing LHRHA. Lymphocyte CM containing con A also stimulated progesterone production at all of the LHRH concentrations studied. This response was not inhibited by lymphocyte CM containing the LHRHA.(ABSTRACT TRUNCATED AT 250 WORDS)
