Study of the Uterine Local Immune Response in a Murine Model of Embryonic Death Due to Tritrichomonas foetus
Mariana A. Woudwyk
Laboratory of Histology and Embryology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Buenos Aires, Argentina
Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany
Search for more papers by this authorCristina E. Monteavaro
Laboratory of Clinical and Experimental Microbiology, Faculty of Veterinary Sciences, National University of the Center of Buenos Aires Province, Tandil, Buenos Aires, Argentina
Search for more papers by this authorCorresponding Author
Federico Jensen
Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany
Correspondence
Federico Jensen, Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany.
E-mail: [email protected]
Search for more papers by this authorPedro Soto
Laboratory of Clinical and Experimental Microbiology, Faculty of Veterinary Sciences, National University of the Center of Buenos Aires Province, Tandil, Buenos Aires, Argentina
Search for more papers by this authorClaudio G. Barbeito
Laboratory of Histology and Embryology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Buenos Aires, Argentina
ACZ and CGB share senior authorship.
Search for more papers by this authorAna C. Zenclussen
Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany
ACZ and CGB share senior authorship.
Search for more papers by this authorMariana A. Woudwyk
Laboratory of Histology and Embryology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Buenos Aires, Argentina
Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany
Search for more papers by this authorCristina E. Monteavaro
Laboratory of Clinical and Experimental Microbiology, Faculty of Veterinary Sciences, National University of the Center of Buenos Aires Province, Tandil, Buenos Aires, Argentina
Search for more papers by this authorCorresponding Author
Federico Jensen
Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany
Correspondence
Federico Jensen, Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany.
E-mail: [email protected]
Search for more papers by this authorPedro Soto
Laboratory of Clinical and Experimental Microbiology, Faculty of Veterinary Sciences, National University of the Center of Buenos Aires Province, Tandil, Buenos Aires, Argentina
Search for more papers by this authorClaudio G. Barbeito
Laboratory of Histology and Embryology, Faculty of Veterinary Sciences, National University of La Plata, La Plata, Buenos Aires, Argentina
ACZ and CGB share senior authorship.
Search for more papers by this authorAna C. Zenclussen
Experimental Obstetrics & Gynecology, Medical Faculty, Otto-von-Guericke University, Magdeburg, Germany
ACZ and CGB share senior authorship.
Search for more papers by this authorAbstract
Problem
Bovine tritrichomonosis is a sexually transmitted disease caused by Tritrichomonas foetus, characterized by conceptus loss. We developed a mouse model of tritrichomonosis to study the mechanisms involved in the embryonic death. We hypothesized that embryonic death may be due to an exacerbated maternal response to the pathogen that then affects embryo development.
Method of study
We infected BALB/c mice with Tritrichomonas foetus and paired them after confirming active infection. We studied the expression of pro- and anti-inflammatory cytokines, markers for T regulatory and T helper 17 cells as well as haem-oxygenase-1 expression in uterine tissue by real-time RT-PCR.
Results
As expected, TNF-α was augmented in infected animals. IL-10 and IL-4 were also up-regulated. Treg-associated genes were higher expressed in uteri of infected group. In mice that have lost their conceptus after the infection, haem-oxygenase-1 (HO-1) mRNA levels were strongly decreased, while RORγt mRNA, a reliable marker for Th17, was augmented in uterus.
Conclusion
A T effector response of type 1 and 17 may be involved in tritrichomonosis-related embryonic death. This alters protective mechanisms as HO-1. Increased regulatory T cells may facilitate embryonic death by promoting the persistence of infection.
References
- 1Skirrow SZ, Bon Durant RG: Bovine trichomoniasis. Veterinary Bulletin 1988; 58: 591–603.
- 2Bon Durant RH: Pathogenesis, diagnosis and management of trichomoniasis in cattle. Vet Clin North Am Food Anim Pract 1997; 13: 345–361.
- 3Monteavaro C, Aguirre J, Soto P, Echevarría H, Catena M, Portiansky E, Gimeno E: Interaction of Tritrichomonas foetus with the reproductive tract of experimentally infected female BALB/c mice: ultrastructural evaluation. Vet J 2007; 173: 206–210.
- 4Barbeito C, Woudwyk M, Cacciato C, Soto P, Portiansky E, Catena M, Echeverría H, Gimeno E, Monteavaro C: Tritrichomonas foetus: Experimental Infection in Pregnant BALB/c Mice. Exp Parasitol 2008; 120: 156–160.
- 5Wegmann T, Lin H, Gilbert L, Mosmann T: Bidirectional cytokine interactions in the maternal-fetal relationship: is successful pregnancy a Th2 phenomenon? Immunol Today 1993; 14: 353–356.
- 6Saito S: Cytokine network at the feto maternal interface. J Reprod Immunol 2000; 47: 87–103.
- 7Svensson L, Arvola M, Sällström MA, Holmdahl R, Mattsson R: The Th2 cytokines IL-4 and IL-10 are not crucial for the completion of allogeneic pregnancy in mice. J Reprod Immunol 2001; 51: 3–7.
- 8Aluvihare VR, Kallikourdis M, Betz AG: Regulatory T cells mediate maternal tolerance to the fetus. Nat Immunol 2004; 5: 266–271.
- 9Zenclussen AC: CD4 + CD25 + T regulatory cells in murine pregnancy. J Reprod Immunol 2005; 65: 101–110.
- 10Thuere C, Zenclussen ML, Schumacher A, Langwisch S, Schulte-Wrede U, Teles A, Paeschke S, Volk HD, Zenclussen AC: Kinetics of regulatory T cells during murine pregnancy. Am J Reprod Immunol 2007; 58: 514–523.
- 11Zenclussen AC, Gerlof K, Zenclussen ML, Sollwedel A, Zambon-Bertoja A, Ritter T, Kotsch K, Leber J, Volk HD: Abnormal T-cell reactivity against paternal antigens in spontaneous abortion: adoptive transfer of pregnancy-induced CD4 + CD25 + T regulatory cells prevents fetal rejection in a murine abortion model. Am J Pathol 2005; 166: 811–822.
- 12Schumacher A, Wafula PO, Bertoja AZ, Sollwedel A, Thuere C, Wollenberg I, Yagita H, Volk HD, Zenclussen AC: Mechanisms of action of regulatory T cells specific for paternal antigens during pregnancy. Obstet Gynecol 2007; 110: 1137–1145.
- 13Jasper MJ, Tremellen KP, Robertson SA: Primary unexplained infertility is associated with reduced expression of the T-regulatory cell transcription factor foxp3 in endometrial tissue. Mol Hum Reprod 2006; 12: 301–308.
- 14Mold JE, Michaëlsson J, Burt TD, Muench MO, Beckerman KP, Busch MP, Lee TH, Nixon DF, McCune JM: Maternal alloantigens promote the development of tolerogenic fetal regulatory T cells in utero. Science 2008; 322: 1562–1565.
- 15Zenclussen AC, Gerlof K, Zenclussen ML, Ritschel S, Zambon BertojaA, Fest S, Hontsu S, Ueha S, Matsushima K, Leber J, Volk HD: Regulatory T cells induce a privileged tolerant microenvironment at the fetal-maternal interface. Eur J Immunol 2006; 36: 82–94.
- 16Zenclussen ML, Casalis PA, El-Mousleh T, Rebelo S, Langwisch S, Linzke N, Volk HD, Fest S, Soares MP, Zenclussen AC: Haem oxygenase-1 dictates intrauterine fetal survival in mice via carbon monoxide. J Pathol 2011; 225: 293–304.
- 17Zenclussen ML, Anegon I, Zambon BertojaA, Chauveau C, Vogt K, Gerlof K, Sollwedel A, Volk HD, Ritter T, Zenclussen AC: Over-expression of heme-oxygenase-1 by adenoviral gene transfer improves pregnancy outcome in a murine model of abortion. J Reprod Immunol 2006; 69: 35–52.
- 18Fujigaki S, Saito K, Takemura M, Maekawa N, Yamada Y, Wada H, Seishima M: L-Tryptophan–L-Kynurenine Pathway Metabolism Accelerated by Toxoplasma gondii Infection Is Abolished in Gamma Interferon-Gene-Deficient Mice: Cross-Regulation between Inducible Nitric Oxide Synthase and Indoleamine-2,3-Dioxygenase. Infect Immun 2002; 70: 779–786.
- 19Mellor AL, Keskin DB, Johnson T, Chandler P, Munn DH: Cells expressing indoleamine 2,3-dioxygenase inhibit T cell responses. J Immunol 2002; 168: 3771–3776.
- 20Baban B, Chandler P, McCoola D, Marshall B, Munn DH, Mellor AL: Indoleamine 2,3-dioxygenase expression is restricted to fetal trophoblast giant cells during murine gestation and is maternal genome specific. J Reprod Immunol 2004; 61: 67–77.
- 21Aujla SJ, Dubin PJ, Kolls JK: Th17 cells and mucosal host defense. Semin Immunol 2007; 19: 377.
- 22Korn T, Bettelli E, Oukka M, Kuchroo VK: IL-17 and Th17 Cells. Annu Rev Immunol 2009; 27: 485–517.
- 23Wang WJ, Hao CF, Yi-Lin, Yin GJ, Bao SH, Qiu LH, Lin QD: Increased prevalence of T helper 17 (Th17) cells in peripheral blood and decidua in unexplained recurrent spontaneous abortion patients. J Reprod Immunol 2010; 84: 164–170.
- 24ANMAT: Reglamentación para bioterios de Laboratorios Elaboradores de Especialidades Medicinales y/o Análisis de Terceros, Cap. I, II, III, IV, V, Disposición 6344. Cary, NC, USA, Administración Nacional de Medicamentos Alimentos y Tecnología Médica, 1996.
- 25Soto P, Echevarría H, Monteavaro C, Catena M: Experimentally induced intravaginal Tritrichomonas foetus infection in a mouse model. Braz J Vet Res An Sci 2005; 25: 225–230.
- 26Allen E: The estrous cycle in the mouse. Am J Anat 1922; 30: 297.
- 27Monteavaro CE, Soto P, Gimeno EJ, Echevarría HM, Catena M, Portiansky EL, Barbeito CG: Histological and Lectin Binding Changes in the Genital Tract of Mice Infected with Tritrichomonas foetus. J Comp Path 2008; 138: 40–45.
- 28Diamond LS: Lumen-dwelling protozoa: Entamoeba, Trichomonads and Giarda. In In vitro cultivation of protozoan parasites, JB Jensen (Ed.). Boca Raton, Florida, CRC Press, 1987, pp 89–98.
- 29Parsonson IM, Clark BL, Dufty JH: Early pathogenesis and pathology of Tritrichomonas foetus infection in virgin heifers. J Comp Pathol 1976; 86: 59–66.
- 30Peck A, Mellins ED: Plasticity of T-cell phenotype and function: the T helper type 17 example. Immunology 2010; 129: 147–153.
- 31Santner-Nanan B, Peek MJ, Khanam R, Richarts L, Zhu E, Fazekas deStGrothB, Nanan R: Systemic increase in the ratio between foxp3 + and IL-17-producing CD4 + T cells in healthy pregnancy but not in preeclampsia. J Immunol 2009; 183: 7023–7030.
- 32Mellor AL, Munn DH: Extinguishing maternal immune responses during pregnancy: implications for immunosuppression. Semin Immunol 2001; 13: 213–218.
- 33Svanborg C, Godaly G, Hedlund M: Cytokine responses during mucosal infections: role in disease pathogenesis and host defence. Curr Opin Microbiol 1999; 2: 99–105.
- 34Dombrowicz D, Capron M: Eosinophils, allergy and parasites. Curr Opin Immunol 2001; 13: 716–720.
- 35Janeway CA, Travers P, Walport M, Shlomchik MJ: Immunobiology. The Immune System in Health and Disease. New York, Garland Science, 2001.
- 36Krishnan L, Guilbert LJ, Wegmann TG, Belosevic M, Mosmann TR: T helper 1 response against Leishmania major in pregnant C57BL/6 mice increases implantation failure and fetal resorptions. Correlation with increased IFN-gamma and TNF-α and reduced IL-10 production by placental cells. J Immunol 1996a; 156: 653–662.
- 37Sasaki Y, Sakai M, Miyazaki S, Higuma S, Shiozaki A, Saito S: Decidual and peripheral blood CD4 + CD25 + regulatory T cells in early pregnancy subjects and spontaneous abortion cases. Mol Hum Reprod 2004; 10: 347–353.
- 38Belkaid Y, Rouse BT: Natural regulatory T cells in infectious disease. Nat Immunol 2005; 6: 353–360.
- 39Shevach EM, DiPaolo RA, Andersson J, Zhao DM, Stephens GL, Thornton AM: The lifestyle of naturally occurring CD4 + CD25 + Foxp3 + regulatory T cells. Immunol Rev 2006; 212: 60–73.
- 40Seddon B, Mason D: Peripheral autoantigen induces regulatory T cells that prevent autoimmunity. J Exp Med 1999; 189: 877–882.
- 41Belkaid Y, Piccirillo CA, Mendez S, Shevach EM, Sacks DL: CD4 + CD25 + regulatory T cells control Leishmania major persistence and immunity. Nature 2002; 420: 502–507.
- 42Harrington LE, Hatton RD, Mangan PR, Turner H, Murphy TL, Murphy KM, Weaver CT: Interleukin 17-producing CD4 + effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nat Immunol 2005; 6: 1123–1132.
- 43Ivanov II, McKenzie BS, Zhou L, Tadokoro CE, Lepelley A, Lafaille JJ, Cua DJ, Littman DR: The orphan nuclear receptor RORγt directs the differentiation program of proinflammatory IL-17 + T helper cells. Cell 2006; 126: 1121–1133.
- 44Kelly MN, Kolls JK, Happel K, Schwartzman JD, Schwarzenberger P, Combe C, Moretto M, Khan IA: Interleukin-17/interleukin-17 receptor-mediated signaling is important for generation of an optimal polymorphonuclear response against Toxoplasma gondii infection. Infect Immun 2005; 73: 617–621.
- 45Ostojic S, Dubanchet S, Chaouat G, Abdelkarim M, Truyens C, Capron F: Demonstration of the presence of IL-16, IL-17 and IL-18 at the murine fetomaternal interface during murine pregnancy. Am J Reprod Immunol 2003; 49: 101–112.
- 46Zenclussen AC, Sollwedel A, Bertoja AZ, Gerlof K, Zenclussen ML, Woiciechowsky C, Volk HD: Heme-oxygenase as a therapeutic target in immunological pregnancy complications. Int Immunopharmacol 2005; 5: 41–51.
- 47Tachibana M, Watanabe K, Yamasaki Y, Suzuki H, Watarai M: Expression of heme-oxygenase-1 is associated with abortion caused by Brucella abortus infection in pregnant mice. Microb Pathog 2008; 45: 105–109.
- 48Taylor MW, Feng GS: Relationship between interferon-gamma, indoleamine 2,3-dioxygenase, and tryptophan catabolism. FASEB J 1991; 5: 2516–2522.
- 49Musso T, Gusella GL, Brooks A, Longo DL, Varesio L: Interleukin-4 inhibits Indoleamine 2,3-Dioxygenase expression in human monocytes. Blood 1994; 83: 1408–1411.