Source: PENNSYLVANIA STATE UNIVERSITY submitted to
PARACRINE FACTORS REGULATING IMMUNE CELL FUNCTION IN THE BOVINE CORPUS LUTEUM
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
NRI COMPETITIVE GRANT
Reporting Frequency
Annual
Accession No.
0213935
Grant No.
2004-35203-19005
Cumulative Award Amt.
(N/A)
Proposal No.
2008-03327
Multistate No.
(N/A)
Project Start Date
Mar 1, 2008
Project End Date
Jul 31, 2009
Grant Year
2008
Program Code
[41.0]- Animal Reproduction
Project Director
Pate, J. L.
Recipient Organization
PENNSYLVANIA STATE UNIVERSITY
208 MUELLER LABORATORY
UNIVERSITY PARK,PA 16802
Performing Department
(N/A)
Non Technical Summary
Reproductive efficiency in dairy cows has declined significantly, and results in substantial economic loss to farmers each year. Reproductive inefficiency can be caused by an inadequate corpus luteum, which is the ovarian structure that secretes progesterone. Proper communication between the embryo and the corpus luteum during the period known as 'maternal recognition of pregnancy' is necessary to ensure pregnancy maintenance. The purpose of this project is to evaluate the factors within the corpus luteum that may regulate function of immune cells that are present within the tissue. Understanding what factors activate or inihibit immune cell function will advance our knowledge regarding the local environment necessary to support maintenance of the corpus lutem and pregnancy.
Animal Health Component
(N/A)
Research Effort Categories
Basic
100%
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3013499102034%
3053499103033%
3053499109033%
Knowledge Area
305 - Animal Physiological Processes; 301 - Reproductive Performance of Animals;

Subject Of Investigation
3499 - Dairy cattle, general/other;

Field Of Science
1090 - Immunology; 1030 - Cellular biology; 1020 - Physiology;
Goals / Objectives
1. Determine how progesterone acts on lymphocytes to inhibit a proinflammatory response 2. Examine the potential for oxytocin to act as a signaling molecule between luteal cells and immune cells 3. Elucidate the role of ETA-1 (osteopontin) in activation of the immune response during luteolysis
Project Methods
Espression of membrane progesterone receptors in lymphocytes will be assessed by qPCR and binding of fluorecently labelled progesterone. Expression of progesterone-induced blocking factor (PIBF) by bovine lymphocytes will be determined by qPCR and western analyses, and will include the effect of stage of the estrous cycle and progesterone on PIBF expression. Also, an antibody to PIBF will be used to determine if PIBF mediates the inhibitory effect of progesterone on lymphocyte proliferation. The T cell proliferation assay will also be used to evaluate a potential shift in Th1 vs. Th2 cells, as assessed by intracellular staining of cytokines. Oxytocin receptor mRNA and binding sites in resting and activated bovine lymphocytes will be evaluated using qPCR and scatchard analysis. The effects of oxytocin and oxytocin antibody on luteal cell-induced T cell proiferation will be determined. Expression of ETA-1 mRNA and protein will be evaluated in corpora lutea at different stages of the estrous cycle and during luteal regression using qPCR and western analysis. The effect of ETA-1 on luteal cell-induced T cell proliferation will be evaluated using an antibody to ETA-1. Finally, the potential role of luteal ETA-1 to serve as a chemoattractant for immune cells will be assessed using a commercially available chemotaxis assay kit.

Progress 03/01/08 to 07/31/09

Outputs
OUTPUTS: Proposed experiments were conducted, data were analyzed, and results were presented at national meetings and published in peer-reviewed, scientific journals PARTICIPANTS: Joy Pate, PD KAlidou Ndiaye, Postdoc Koji Toyokawa, Postdoc TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The results of these experiments led to new knowledge on the regulation of lymphocyte biology as related to reproductive processes. Of primary importance was the discovery that bovine lymphocytes do not express nuclear progesterone receptors, but do express at least three distinct genes for membrane progesterone receptors. Therefore, the functional effects of progesterone on lymphocytes are likely mediated by these membrane progesterone receptors. Also, these experiments provided the first evidence that functional oxytocin receptors are present on circulating T lymphocytes in cattle. In addition to a potential role as a signaling molecule between luteal cells and T lymphocytes within the ovary, the effects of oxytocin on lymphocytes may relate to alterations in immune function during periods of peak lactation or parturition in dairy cattle. Finally, these experiments led to the discovery that SPP1 is produced by luteal cells from cows, and that there are temporal changes in processing of SSP1 protein within the CL. Overall, significant new information was obtained that helps us to better understand how ovarian cells and T lymphocytes may communicate to facilitate normal ovarian function in dairy cattle.

Publications

  • Zhang, B., P.C. Tsang, J.L. Pate, and M.A. Moses. 2006. Regulation of CYR61 by Prostaglandin F2 alpha (PGF2 ) in the Bovine Corpus Luteum. Endocrine Society Annual Meeting. P3-288
  • Bridges, G.A., M.L. Mussard, L.A. Helser, D.E. Grum, D.M. Lantz, T.L. Ott, J.L. Pate, and M.L. Day. 2006. Effect of preovulatory estradiol concentrations on conceptus and uterine development. Presented at the 7th International Ruminant Reproduction Symposium, August 13-17, 2006. Wellington, NZ. no.90. p.91 (Abstract)
  • Poole, D.H., K. Ndiaye, and J.L. Pate. 2007. Expression and Regulation of Functional Oxytocin Receptors in Bovine T Lymphocytes. Biol. Reprod. no.198 (Abstract)
  • Ndiaye, K., J. Lussier, and J.L. Pate. 2008. Characterization of DER1-Like Domain Family Member 1 (DERL1) and its Expression in Bovine Corpus Luteum. World Congress on Reproductive Biology. no.19 (Abstract)
  • Poole, D.H., K. Ndiaye, and J.L. Pate. 2008. Potential role of secreted phosphoprotein 1 in the bovine corpus luteum. World Congress on Reproductive Biology. no.34 (Abstract)
  • Ndiaye, K., D.H. Poole, T.R. Hansen, and J.L. Pate. 2008. Expression profiles of cytokine and progesterone receptor messenger ribonucleic acids (mRNA) in bovine corpus luteum during pregnancy. Biol. Reprod. no.400 (Abstract)
  • Ndiaye, K., D.H. Poole, and J.L. Pate. 2007. Expression of mRNA for membrane progesterone receptors by bovine T lymphocytes. Biol. Reprod. no. 196 (Abstract)
  • Davis, T.L. and J.L. Pate. 2007. Proliferation of major histocompatibility nonrestricted gamma delta T cells is stimulated by bovine luteal cells. Biol. Reprod. 77:914-922
  • Hou, X., E.W. Arvisais, C. Jiang, D. Chen, S.K. Roy, J.L. Pate, T.R. Hansen, B.R. Rueda, and J.S. Davis. 2008. Prostaglandin F2 (PGF2) stimulates the expression and secretion of TGFB1via induction of the early growth response 1 gene (EGR1) in the bovine corpus luteum. Mol. Endocrinol. 22:403-414
  • Ndiaye, K., D.H. Poole, and J.L. Pate. 2008. Expression and regulation of functional oxytocin receptors in bovine T lymphocytes. Biol. Reprod. 78:786-793
  • Winkelman, L.A., T.H. Elsasser, M.C. Lucy, J.L. Pate, and C.K. Reynolds. 2008. Suppressor of cytokine signaling-2 mRNA increases after parturition in the liver of dairy cows. J. Dairy Sci. 91:1080-1086
  • Ndiaye, K., M.J. Cannon, and J.L. Pate. 2009 Expression of membrane progestin receptors in bovine lymphocytes and luteal tissues and effect of progesterone on the interleukin-2/interleukin-2 receptor system. J Reprod Immunol. (Submitted)
  • Poole, D.H. and J.L. Pate. 2009 Temporal changes in SPP1 in the bovine corpus luteum and role in activation of proinflammatory T cell responses. (In Preparation)
  • Cannon, M.J. and J.L. Pate. 2004. Presence of membrane-bound progesterone receptor mRNA in bovine peripheral blood mononuclear cells. Int. Vet. Immunol. Symposium. p.206 (Abstract)
  • Winkler, J.L. and J.L. Pate. 2005. Expression of Early T Cell Activation Factor (Eta-1), also known as osteopontin, in the bovine corpus luteum (CL). Biol. Reprod. p.204 (Abstract)
  • Bridges, G.A., M.L. Mussard, J.L. Winkler, C.L. Gasser, D.E. Grum, J.L. Pate, and M.L. Day. 2005. Influence of duration of proestrus on preovulatory estradiol concentrations and uterine gene expression following induced ovulation in cattle. Biol. Reprod. p.556 (Abstract)
  • Ndiaye, K. and J.L. Pate. 2006. Evidence for Oxytocin Receptor Expression in Bovine Peripheral Blood Mononuclear Cells and T Lymphocytes. Biol. Reprod. p.74 (Abstract)