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Autoimmunization of ewes against pregnancy-associated glycoproteins does not interfere with the establishment and maintenance of pregnancy

Published online by Cambridge University Press:  01 June 2009

T. E. Egen ,
A. D. Ealy ,
L. A. Landon ,
R. M. Roberts  and
J. A. Green
T. E. Egen
Affiliation:
Division of Animal Science, University of Missouri-Columbia, 920 East Campus Drive, Columbia, MO 65211, USA
A. D. Ealy
Affiliation:
Department of Animal Sciences, University of Florida, PO Box 110910, Gainesville, FL 32611, USA
L. A. Landon
Affiliation:
Division of Animal Science, University of Missouri-Columbia, 920 East Campus Drive, Columbia, MO 65211, USA
R. M. Roberts
Affiliation:
Division of Animal Science, University of Missouri-Columbia, 920 East Campus Drive, Columbia, MO 65211, USA
J. A. Green*
Affiliation:
Division of Animal Science, University of Missouri-Columbia, 920 East Campus Drive, Columbia, MO 65211, USA
*
E-mail: [email protected]
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Abstract

Pregnancy-associated glycoproteins (PAGs) are a large grouping of placental proteins that belong to the aspartic peptidase gene family. Although useful to detect pregnancy in ruminant species, the function of these molecules is unclear. Several PAGs expressed by trophoblast binucleate cells can enter the maternal circulation, suggesting that they could have a systemic role in altering maternal physiology. The objective of this work was to examine whether these circulating placental antigens were important in pregnancy by actively immunizing ewes against them. PAGs were purified by pepstatin-affinity chromatography and conjugated to the immunogenic protein, keyhole limpet hemocyanin (KLH). Ewes were immunized with PAG–KLH conjugate (n = 22) or with KLH alone (n = 9), and bred to intact rams. Blood samples, collected on Day 0 (day of estrus), Day 10, Days 15 to 25 and weekly throughout pregnancy, were analyzed for PAG by an ELISA. On Day 30, pregnancy was confirmed by ultrasound. Ewes immunized against PAG–KLH produced a range of reactive anti-PAG titers, whereas all immunized ewes had high anti-KLH immunoreactivity. PAGs became detectable in the anti-KLH (control) ewes at Day 21.6 ± 2.2 of pregnancy. Those ewes immunized against PAGs (n = 7), that had very low immunoreactivity toward PAGs, had measurable PAG by Day 22.9 ± 1.3, and their PAG serum profiles throughout pregnancy did not differ from the controls. Those exhibiting moderate to high anti-PAG immunoreactivity (n = 15), had significantly lower PAG concentrations than controls, with antigen not becoming detectable until Day 48.1 ± 15.6. The decrease in circulating PAG in the immunized animals did not correlate with changes in pregnancy rates, lamb number or lamb birth weight. These results suggest that while PAGs may play a role in maintaining pregnancy, their major contribution is likely to be at the fetal–maternal interface. Their actions at extra-placental sites are presumably of more secondary importance.

Keywords

pregnancyplacentaimmunizationtrophoblast
Type
Full Paper
Information
animal , Volume 3 , Issue 6 , June 2009 , pp. 850 - 857
Copyright
Copyright © The Animal Consortium 2009

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References

Austin, KJ, King, CP, Vierk, JE, Sasser, RG, Hansen, TR 1999. Pregnancy-specific protein B induces release of an alpha chemokine in bovine endometrium. Endocrinology 140, 542–545.CrossRefGoogle ScholarPubMed
Baldwin, MA, Medzihradszky, KF, Lock, CM, Fisher, B, Settineri, TA, Burlingame, AL 2001. Matrix-assisted laser desorption/ionization coupled with quadrupole/orthogonal acceleration time-of-flight mass spectrometry for protein discovery, identification, and structural analysis. Analytical Chemistry 73, 1707–1720.CrossRefGoogle ScholarPubMed
Belghazi, M, Bathany, K, Hountondji, C, Grandier-Vazeille, X, Manon, S, Schmitter, J-M 2001. Analysis of protein sequences and protein complexes by matrix-assisted laser desorption/ionization mass spectrometry. Proteomics 1, 946–954.3.0.CO;2-P>CrossRefGoogle ScholarPubMed
Bradford, MM 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Analytical Biochemistry 72, 248–254.CrossRefGoogle ScholarPubMed
Brandt, GA, Parks, TE, Killian, G, Ealy, AD, Green, JA 2007. A cloning and expression analysis of pregnancy-associated glycoproteins expressed in trophoblasts of the white-tail deer placenta. Molecular Reproduction and Development 74, 1355–1362.CrossRefGoogle ScholarPubMed
Clauser, KR, Baker, PR, Burlingame, AL 1999. Role of accurate mass measurement (+/−10 p.p.m.) in protein identification strategies employing MS or MS/MS and database searching. Analytical Chemistry 71, 2871–2882.CrossRefGoogle ScholarPubMed
Del Vecchio, RP, Sasser, RG, Randel, RD 1990. Effect of pregnancy-specific protein B on prostaglandin F2alpha and prostaglandin E2 release by day 16-perifused bovine endometrial tissue. Prostaglandins 40, 271–282.CrossRefGoogle ScholarPubMed
Del Vecchio, RP, Sutherland, WD, Sasser, RG 1995a. Effect of pregnancy-specific protein B on luteal cell progesterone, prostaglandin, and oxytocin production during two stages of the bovine estrous cycle. Journal of Animal Science 73, 2662–2668.CrossRefGoogle ScholarPubMed
Del Vecchio, RP, Sutherland, WD, Sasser, RG 1995b. Prostaglandin F2alpha, progesterone and oxytocin production by cultured bovine luteal cells treated with prostaglandin E2 and pregnancy-specific protein B. Prostaglandins 50, 137–150.CrossRefGoogle ScholarPubMed
Del Vecchio, RP, Sutherland, WD, Sasser, RG 1996. Bovine luteal cell production in vitro of prostaglandin E2, oxytocin and progesterone in response to pregnancy-specific protein B and prostaglandin F2alpha. Journal of Reproduction and Fertility 107, 131–136.CrossRefGoogle Scholar
Dosogne, H, Burvenich, C, Freeman, AE, Kehrli, MEJ, Detilleux, JC, Sulon, J, Beckers, JF, Hoeben, D 1999. Pregnancy-associated glycoprotein and decreased polymorphonuclear leukocyte function in early post-partum dairy cows. Veterinary Immunology and Immunopathology 67, 47–54.CrossRefGoogle ScholarPubMed
Duello, TM, Byatt, JC, Bremel, RD 1986. Immunohistochemical localization of placental lactogen in binucleate cells of bovine placentomes. Endocrinology 119, 1351–1355.CrossRefGoogle ScholarPubMed
Garbayo, JM, Green, JA, Mannekin, M, Beckers, J-F, Kiesling, DO, Ealy, AD, Roberts, RM 2000. Caprine pregnancy-associated glycoproteins (PAG): their cloning, expression and evolutionary relationship to other PAG. Molecular Reproduction and Development 57, 311–322.3.0.CO;2-F>CrossRefGoogle ScholarPubMed
Gorgani, NN, Theofilopoulos, AN 2007. Contribution of histidine-rich glycoprotein in clearance of immune complexes and apoptotic cells: implications for ameliorating autoimmune diseases. Autoimmunity 40, 260–266.CrossRefGoogle ScholarPubMed
Green, J, Roberts, R 2006. Establishment of an ELISA for the detection of native bovine pregnancy-associated glycoproteins secreted by trophoblast binucleate cells. In Placental and Trophoblast Methods and Protocols (ed. JA Hunt and MJ Soares ), pp. 321–330. Humana Press, Inc., Totowa, NJ, USA.Google Scholar
Green, JA, Xie, S, Quan, X, Bao, B, Gan, X, Mathialagan, N, Beckers, J-F, Roberts, RM 2000. Pregnancy-associated bovine and ovine glycoproteins exhibit spatially and temporally distinct expression patterns during pregnancy. Biology of Reproduction 62, 1624–1631.CrossRefGoogle ScholarPubMed
Green, JA, Parks, TE, Avalle, MP, Telugu, BP, McLain, AL, Peterson, AJ, McMillan, W, Mathialagan, N, Xie, S, Hook, RR, Roberts, RM 2005. The establishment of an ELISA for the detection of pregnancy-associated glycoproteins (PAGs) in the serum of pregnant cows and heifers. Theriogenology 63, 1481–1503.CrossRefGoogle ScholarPubMed
Hammond, J 1927. Physiology of reproduction in the cow. Cambridge University Press, Cambridge, UK.Google Scholar
Harlow, E, Lane, D 1988. Antibodies: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.Google Scholar
Hoeben, D, Burvenich, C, Massart-Leen, AM, Lenjou, M, Nijs, G, Van Bockstaele, D, Beckers, JF 1999. In vitro effect of ketone bodies, glucocorticosteroids and bovine pregnancy-associated glycoprotein on cultures of bone marrow progenitor cells of cows and calves. Veterinary Immunology and Immunopathology 68, 229–240.CrossRefGoogle ScholarPubMed
Hoeben, D, Monfardini, E, Opsomer, G, Burvenich, C, Dosogne, H, De Kruif, A, Beckers, JF 2000. Chemiluminescence of bovine polymorphonuclear leucocytes during the periparturient period and relation with metabolic markers and bovine pregnancy-associated glycoprotein. Journal of Dairy Research 67, 249–259.CrossRefGoogle ScholarPubMed
Hughes, AL, Green, JA, Garbayo, JM, Roberts, RM 2000. Adaptive diversification within a large family of recently duplicated, placentally-expressed genes. Proceedings of the National Academy of the Sciences of the United States of America 97, 3319–3323.Google ScholarPubMed
Hughes, AL, Green, JA, Piontkivska, H, Roberts, RM 2003. Aspartic proteinase phylogeny and the origin of pregnancy-associated glycoproteins. Molecular Biology and Evolution 20, 1940–1945.CrossRefGoogle ScholarPubMed
Kehrli, ME, Nonnecke, BJ, Roth, JA 1989. Alterations in bovine peripheral blood neutrophil function during the periparturient period. American Journal of Veterinary Research 50, 207–214.Google Scholar
Klisch, K, Leiser, R 2003. In bovine binucleate trophoblast giant cells, pregnancy-associated glycoproteins and placental prolactin-related protein-I are conjugated to asparagine-linked N-acetylgalactosaminyl glycans. Histochemistry and Cell Biology 119, 211–217.CrossRefGoogle ScholarPubMed
Klisch, K, Jeanrond, E, Pang, P-C, Pich, A, Schuler, G, Dantzer, V, Kowalewski, MP, Dell, A 2008. A tetraantennary glycan with bisecting N-acetylglucosamine and the SDA antigen is the predominant N-glycan on bovine pregnancy-associated glycoproteins. Glycobiology 18, 42–52.CrossRefGoogle ScholarPubMed
Liang, R, Limesand, SW, Anthony, RV 1999. Structure and transcriptional regulation of the ovine placental lactogen gene. European Journal of Biochemistry 265, 883–895.CrossRefGoogle ScholarPubMed
Link, AJ, Eng, J, Schieltz, DM, Carmack, E, Mize, GJ, Morris, DR, Garvik, BM, Yates, JRR 1999. Direct analysis of protein complexes using mass spectrometry. Nature Biotechnology 17, 676–682.CrossRefGoogle ScholarPubMed
Mathialagan, N, Hansen, TR 1996. Pepsin-inhibitory activity of the uterine serpins. Proceedings of the National Academy of the Sciences of the United States of America 93, 13653–13658.Google ScholarPubMed
Medan, MS, Wang, H, Watanabe, G, Suzuki, AK, Taya, K 2004. Immunization against endogenous inhibin increases normal oocyte/embryo production in adult mice. Endocrine 24, 115–120.CrossRefGoogle ScholarPubMed
Noakes, DE, Parkinson, TJ, England, GCW 2001. Arthur’s veterinary reproduction and obstetrics. Harcourt Publishers Ltd, London, UK.Google Scholar
Peltier, MR, Liu, WJ, Hansen, PJ 2000. Regulation of lymphocyte proliferation by uterine serpin: interleukin-2 mRNA production, CD25 expression and responsiveness to interleukin-2. Proceedings of the Society for Experimental Biology and Medicine 223, 75–81.Google ScholarPubMed
Perkins, DN, Pappin, DJC, Creasy, DM, Cottrell, JS 1999. Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20, 3551–3567.3.0.CO;2-2>CrossRefGoogle ScholarPubMed
Ranilla, M, Sulon, J, Carro, M, Mantecon, A, Beckers, J 1994. Plasmatic profiles of pregnancy-associated glycoprotein and progesterone levels during gestation in Churra and Merino sheep. Theriogenology 42, 537–545.CrossRefGoogle ScholarPubMed
Ruder, CA, Stellflug, JN, Dahmen, JJ, Sasser, RG 1988. Detection of pregnancy in sheep by radioimmunoassay of sera for pregnancy-specific protein B. Theriogenology 29, 905–912.CrossRefGoogle ScholarPubMed
Saad, AM, Concha, C, Astrom, G 1989. Alterations in neutrophil phagocytosis and lymphocyte blastogenesis in dairy cows around parturition. Journal of Veterinary Medicine, Series B 36, 337–345.CrossRefGoogle ScholarPubMed
Sasser, RG, Ruder, CA, Ivani, KA, Butler, JE, Hamilton, WC 1986. Detection of pregnancy by radioimmunoassay of a novel pregnancy-specific protein in serum of cows and a profile of serum concentrations during gestation. Biology of Reproduction 35, 936–942.CrossRefGoogle Scholar
Shevchenko, A, Wilm, M, Vorm, O, Mann, M 1996. Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Analytical Chemistry 68, 850–858.CrossRefGoogle ScholarPubMed
Shevchenko, A, Wilm, M, Mann, M 1997. Peptide sequencing by mass spectrometry for homology searches and cloning of genes. Journal of Protein Chemistry 16, 481–490.CrossRefGoogle ScholarPubMed
Simister, NE, Jacobowitz Israel, E, Ahouse, JC, Story, CM 1997. New functions of the MHC class I-related Fc receptor, FcRn. Biochemical Society Transactions 25, 481–486.CrossRefGoogle ScholarPubMed
Stevens, VC 1979. Human chorionic gonadotrophin: properties and potential immunological manipulation for clinical application. Clinical Obstetrics and Gynaecology 6, 549–566.CrossRefGoogle ScholarPubMed
Stewart, AJ, Piggott, NH, May, FE, Westley, BR 1994. Mitogenic activity of procathepsin D purified from conditioned medium of breast-cancer cells by affinity chromatography on pepstatinyl agarose. International Journal of Cancer 57, 715–718.CrossRefGoogle ScholarPubMed
Talwar, GP, Singh, O, Pal, R, Chatterjee, N, Sahai, P, Dhall, K, Kaur, J, Das, SK, Suri, S, Buckshee, K et al. 1994. A vaccine that prevents pregnancy in women. Proceedings of the National Academy of Sciences of the United States of America 91, 8532–8536.CrossRefGoogle ScholarPubMed
Vaughn, DE, Bjorkman, PJ 1998. Structural basis of pH-dependent antibody binding by the neonatal Fc receptor. Structure 6, 63–73.CrossRefGoogle ScholarPubMed
Weems, YS, Lammoglia, MA, Vera-Avila, HR, Randel, RD, Sasser, RG, Weems, CW 1998a. Effects of luteinizing hormone (LH), PGE2, 8-Epi-PGE1, 8-Epi-PGF2alpha, trichosanthin and pregnancy specific protein B (PSPB) on secretion of prostaglandin (PG) E (PGE) or F2alpha (PGF2alpha) in vitro by corpora lutea (CL) from nonpregnant and pregnant cows. Prostaglandins and other Lipid Mediators 55, 359–376.CrossRefGoogle ScholarPubMed
Weems, YS, Lammoglia, MA, Vera-Avila, HR, Randel, RD, King, C, Sasser, RG, Weems, CW 1998b. Effect of luteinizing hormone (LH), PGE2, 8-Epi-PGE1, 8-EPI-PGE2, trichosanthin, and pregnancy specific protein B on secretion of progesterone in vitro by corpora lutea (CL) from nonpregnant and pregnant cows. Prostaglandins and other Lipid Mediators 55, 27–42.CrossRefGoogle ScholarPubMed
Weems, YS, Bridges, PJ, LeaMaster, BR, Sasser, RG, Vincent, DL, Weems, CW 1999. Secretion of progesterone, estradiol-17beta, PGE, PGF2alpha, and pregnancy-specific protein B by 90-day intact and ovariectomized pregnant ewes. Prostaglandins and other Lipid Mediators 58, 139–148.CrossRefGoogle ScholarPubMed
Weems, YS, Bridges, PJ, LeaMaster, BR, Sasser, RG, Ching, L, Weems, CW 2001. Effect of the aromatase inhibitor CGS-16949A on pregnancy and secretion of progesterone, estradiol-17beta, prostaglandins E and F2alpha (PGE; PGF2alpha) and pregnancy specific protein B (PSPB) in 90-day ovariectomized pregnant ewes. Prostaglandins and other Lipid Mediators 66, 77–88.CrossRefGoogle ScholarPubMed
Weems, YS, Bridges, PJ, Sasser, RG, Ching, L, LeaMaster, BR, Vincent, DL, Weems, CW 2002. Effect of mifepristone on pregnancy, pregnancy-specific protein B (PSPB), progesterone, estradiol-17beta, prostaglandin F2alpha (PGF2alpha) and prostaglandin E (PGE) in ovariectomized 90-day pregnant ewes. Prostaglandins and other Lipid Mediators 70, 195–208.CrossRefGoogle ScholarPubMed
Weems, YS, Kim, L, Humphreys, V, Tsuda, V, Weems, CW 2003. Effect of luteinizing hormone (LH), pregnancy specific protein B (PSPB), or arachidonic acid (AA) on ovine endometrium of the estrous cycle or placental secretion of prostaglandins E2 (PGE2) and F2alpha (PGF2alpha) and progesterone in vitro. Prostaglandins and other Lipid Mediators 71, 55–73.CrossRefGoogle ScholarPubMed
Wooding, FBP, Roberts, RM, Green, JA 2005. Light and electron microscope immunocytochemical studies of the distribution of pregnancy-associated glycoproteins (PAGs) throughout pregnancy in the cow: possible functional implications. Placenta 26, 807–827.CrossRefGoogle ScholarPubMed
Wright, LM, Levy, ES, Patel, NP, Alhadeff, JA 1997. Purification and characterization of cathepsin D from normal human breast tissue. Journal of Protein Chemistry 16, 171–181.CrossRefGoogle ScholarPubMed
Xie, S, Green, J, Bao, B, Beckers, JF, Valdez, K, Hakami, L, Roberts, R 1997. Multiple pregnancy-associated glycoproteins are secreted by day 100 ovine placental tissue. Biology of Reproduction 57, 1384–1393.CrossRefGoogle ScholarPubMed
Zoli, AP, Guilbault, LA, Delahaut, P, Ortiz, WB, Beckers, JF 1992. Radioimmunoassay of a bovine pregnancy-associated glycoprotein in serum: its application for pregnancy diagnosis. Biology of Reproduction 46, 83–92.CrossRefGoogle ScholarPubMed