Expression of Toll-like receptors 2 and 9 in cells of dog jejunum and colon naturally infected with Leishmania infantum

BMC Immunology

Wagner L Tafuri³  Ana MC Faria²  Beatriz Deoti¹  Lucélia de Jesus Pinheiro³  Vítor S Barbosa³  Aldair JW Pinto³  Izabela FG Amorim³  Maria M Figueiredo³ 

[1] Departamento de Cirurgia, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Professor Alfredo Balena, 190, Belo Horizonte, Minas Gerais, Brazil;Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos, Pampulha, 6627, Belo Horizonte, Minas Gerais, Brazil;Departamento de Patologia Geral, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Antônio Carlos, Pampulha, 6627, Belo Horizonte, Minas Gerais, Brazil

关键词: Leishmania infantum;    Parasite burden;    Toll-like receptors 2 and 9;    Jejunum and colon;    Canine visceral leishmaniasis;   

Others  :  1077789
DOI  :  10.1186/1471-2172-14-22

 received in 2012-10-23, accepted in 2013-04-30,  发布年份 2013

【 摘 要 】

Background

Infection with parasite protozoa is a long-term health issue in tropical and subtropical regions throughout the world. The Toll-like receptor (TLR) signaling pathway is one of the first-responding defense systems against Leishmania. The aim of this study was to investigate the expression of TLR2 and TLR9 in jejunum and colon and its correlation with CD11c, CD11b, and CD14 receptors used as markers for dendritic cells and macrophages.

Methods

Twenty four dogs infected with Leishmania infantum were used in this study. Cytometry was carried out in lamina propria cells from jejunum and colon using markers for TLR2, TLR9, CD11b, CD11c and CD14.

Results

Cellular inflammatory exudate was diffuse in the mucosa and submucosa, predominately comprising mononuclear cells: plasma cells, macrophages, and lymphocytes. Despite the parasite load, microscopy showed no erosion was evident in the epithelial mucosa layers. The colon harbored more parasites than the jejunum. Flow cytometry revealed higher frequency of TLR2⁺ and CD11c⁺ dendritic cells in the colon than in the jejunum. Conversely, TLR9-expressing cells were more frequent in jejunum. Moreover, frequency of macrophages (CD11b⁺ and CD14⁺) expressing simultaneity TLR9 were lower in the colon than in jejunum, while CD11c⁺ cells predominated in the colon. Despite of the negative ELISA serum results, IL-10 and TNF-α were higher in jejunum than colon of infected animals. However, IL-4 was higher in colon than jejunum of infected animals. A higher expression these cytokines were demonstrated in infected dogs compared to uninfected dogs.

Conclusions

There was no correlation between clinical signs and pathological changes and immunological and parasitological findings in the gastrointestinal tract in canine visceral leishmaniasis. However, jejunum showed a lower parasite load with increased frequency and expression of CD11b, TLR9, CD14/CD11b/TLR9 receptors and IL-10 and TNF-α cytokines. Conversely, the colon showed a higher parasite load along with increased frequency and expression of TLR2, CD11c receptors, and IL-4 cytokine. Thus, Leishmania infantum is able to interfere in jejunum increased expression of TLR2, TLR9, CD11b, CD14, CD14/CD11b/TLR9 receptors, IL-10, and TNF-α; and in colon increased expression of CD11c, TLR2, TLR9, CD11b, CD14 e, CD14/CD11b/TLR9 receptors, IL-10, and TNF-α.

【 授权许可】

   
2013 Figueiredo et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 3.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Pinto AJ, Figueiredo MM, Silva FL, Martins T, Michalick MS, Tafuri WL: Histopathological and parasitological study of the gastrointestinal tract of dogs naturally infected with Leishmania infantum. Acta Vet Scand 2011, 53:67. BioMed Central Full Text
• [2]Anderson DC, Buckner RG, Glenn BL, MacVean DW: Endemic canine leishmaniasis. Vet Pathol 1980, 17(1):94-96.
• [3]Lennox WJ, Smart ME, Little PB: Canine leishmaniasis in Canada. Can Vet J 1972, 13(8):188-190.
• [4]Tryphonas L, Zawidzka Z, Bernard MA, Janzen EA: Visceral leishmaniasis in a dog: clinical, hematological and pathological observations. Can J Comp Med 1977, 41(1):1-12.
• [5]Slappendel RJ: Canine leishmaniasis. A review based on 95 cases in The Netherlands. Vet Q 1988, 10(1):1-16.
• [6]Ferrer L, Juanola B, Ramos JA, Ramis A: Chronic colitis due to Leishmania infection in two dogs. Vet Pathol 1991, 28(4):342-343.
• [7]Silva FL, Tafuri WL, Oliveira MR: Histopathological and immunohistochemical study of the gastrointestinal tract from a dog naturally infected with Leishmania (Leishmania) chagasi: a case report. Arq Bras Med Vet Zoo 2002, 54(4):340-344.
• [8]Adamama-Moraitou KK, Rallis TS, Koytinas AF, Tontis D, Plevraki K, Kritsepi M: Asymptomatic colitis in naturally infected dogs with Leishmania infantum: a prospective study. Am J Trop Med Hyg 2007, 76(1):53-57.
• [9]Keenan CM, Hendricks LD, Lightner L, Webster HK, Johnson AJ: Visceral Leishmaniasis in the German Shepherd Dog.1. Infection, Clinical-Disease, and Clinical Pathology. Vet Pathol 1984, 21(1):74-79.
• [10]Gonzalez JL, Fermin ML, Garcia P, Rollan E, Castano M: Erosive colitis in experimental canine Leishmaniasis. Zentralbl Veterinarmed B 1990, 37(5):377-382.
• [11]de Veer MJ, Curtis JM, Baldwin TM, DiDonato JA, Sexton A, McConville MJ, Handman E, Schofield L: MyD88 is essential for clearance of Leishmania major: possible role for lipophosphoglycan and Toll-like receptor 2 signaling. Eur J Immunol 2003, 33(10):2822-2831.
• [12]Tuon FF, Amato VS, Bacha HA, Almusawi T, Duarte MI, Amato Neto V: Toll-like receptors and leishmaniasis. Infect Immun 2008, 76(3):866-872.
• [13]Medzhitov R, Janeway C Jr: The Toll receptor family and microbial recognition. Trends Microbiol 2000, 8(10):452-456.
• [14]Testro AG, Visvanathan K: Toll-like receptors and their role in gastrointestinal disease. J Gastroenterol Hepatol 2009, 24(6):943-954.
• [15]Akira S: Toll-like receptors and innate immunity. Adv Immunol 2001, 78:1-56.
• [16]Hausmann M, Kiessling S, Mestermann S, Webb G, Spottl T, Andus T, Scholmerich J, Herfarth H, Ray K, Falk W, et al.: Toll-like receptors 2 and 4 are up-regulated during intestinal inflammation. Gastroenterology 2002, 122(7):1987-2000.
• [17]Takeda K, Kaisho T, Akira S: Toll-like receptors. Annu Rev Immunol 2003, 21:335-376.
• [18]Trinchieri G, Sher A: Cooperation of Toll-like receptor signals in innate immune defence. Nat Rev Immunol 2007, 7(3):179-190.
• [19]Kelly D, Conway S, Aminov R: Commensal gut bacteria: mechanisms of immune modulation. Trends Immunol 2005, 26(6):326-333.
• [20]Iwasaki A, Medzhitov R: Toll-like receptor control of the adaptive immune responses. Nat Immunol 2004, 5(10):987-995.
• [21]Macpherson AJ, Harris NL: Interactions between commensal intestinal bacteria and the immune system. Nat Rev Immunol 2004, 4(6):478-485.
• [22]Platt AM, Mowat AM: Mucosal macrophages and the regulation of immune responses in the intestine. Immunol Lett 2008, 119(1–2):22-31.
• [23]Ortega-Cava CF, Ishihara S, Rumi MA, Kawashima K, Ishimura N, Kazumori H, Udagawa J, Kadowaki Y, Kinoshita Y: Strategic compartmentalization of Toll-like receptor 4 in the mouse gut. J Immunol 2003, 170(8):3977-3985.
• [24]Triantafilou M, Triantafilou K: Lipopolysaccharide recognition: CD14, TLRs and the LPS-activation cluster. Trends Immunol 2002, 23(6):301-304.
• [25]de Amorim IFG, da Silva SM, Figueiredo MM, Moura EP, de Castro RS, Lima TKD, Gontijo ND, Michalick MSM, Gollob KJ, Tafuri WL: Toll receptors type-2 and CR3 expression of canine monocytes and its correlation with immunohistochemistry and xenodiagnosis in visceral leishmaniasis. PLoS One 2011, 6(11):1.
• [26]Tafuri WL, de Oliveira MR, Melo MN, Tafuri WL: Canine visceral leishmaniosis: a remarkable histopathological picture of one case reported from Brazil. Vet Parasitol 2001, 96(3):203-212.
• [27]dos Santos WL, David J, Badaro R, de Freitas LA: Association between skin parasitism and a granulomatous inflammatory pattern in canine visceral leishmaniosis. Parasitol Res 2004, 92(2):89-94.
• [28]Longstaffe JA, Guy MW: Leishmaniasis in dogs. Vet Annu 1985, 25:358-367.
• [29]Holmgren J, Rudim A: Mucosal immunity and bacteria. In Mucosal Immunology. Edited by Ogra PL, Mestecky J, Lmm ME, Strober W. San Diego: Academy Press; 1999:685-693.
• [30]Furrie E, Macfarlane S, Thomson G, Macfarlane GT: Toll-like receptors-2, -3 and −4 expression patterns on human colon and their regulation by mucosal-associated bacteria. Immunology 2005, 115(4):565-574.
• [31]Liew FY, Patel M, Xu D: Toll-like receptor 2 signalling and inflammation. Ann Rheum Dis 2005, 64(Suppl 4):iv104-iv105.
• [32]Pedersen G, Andresen L, Matthiessen MW, Rask-Madsen J, Brynskov J: Expression of Toll-like receptor 9 and response to bacterial CpG oligodeoxynucleotides in human intestinal epithelium. Clin Exp Immunol 2005, 141(2):298-306.
• [33]Harris G, KuoLee R, Chen W: Role of Toll-like receptors in health and diseases of gastrointestinal tract. World J Gastroenterol 2006, 12(14):2149-2160.
• [34]Lee SH, Starkey PM, Gordon S: Quantitative analysis of total macrophage content in adult mouse tissues. Immunochemical studies with monoclonal antibody F4/80. J Exp Med 1985, 161(3):475-489.
• [35]Rogler G, Hausmann M, Vogl D, Aschenbrenner E, Andus T, Falk W, Andreesen R, Scholmerich J, Gross V: Isolation and phenotypic characterization of colonic macrophages. Clin Exp Immunol 1998, 112(2):205-215.
• [36]Ishii M, Hashimoto M, Oguma K, Kano R, Moritomo T, Hasegawa A: Molecular cloning and tissue expression of canine Toll-like receptor 2 (TLR2). Vet Immunol Immunopathol 2006, 110(1–2):87-95.
• [37]Hashimoto M, Asahina Y, Sano J, Kano R, Moritomo T, Hasegawa A: Cloning of canine toll-like receptor 9 and its expression in dog tissues. Vet Immunol Immunopathol 2005, 106(1–2):159-163.
• [38]Burgener IA, Konig A, Allenspach K, Sauter SN, Boisclair J, Doherr MG, Jungi TW: Upregulation of toll-like receptors in chronic enteropathies in dogs. J Vet Intern Med 2008, 22(3):553-560.
• [39]Alves CF, de Amorim IFG, Moura EP, Ribeiro RR, Alves CF, Michalick MS, Kalapothakis E, Bruna-Romero O, Tafuri WL, Teixeira MM, et al.: Expression of IFN-gamma, TNF-alpha, IL-10 and TGF-beta in lymph nodes associates with parasite load and clinical form of disease in dogs naturally infected with Leishmania (Leishmania) chagasi. Vet Immunol Immunop 2009, 128(4):349-358.
• [40]Becker I, Salaiza N, Aguirre M, Delgado J, Carrillo-Carrasco N, Kobeh LG, Ruiz A, Cervantes R, Torres AP, Cabrera N, et al.: Leishmania lipophosphoglycan (LPG) activates NK cells through toll-like receptor-2. Mol Biochem Parasitol 2003, 130(2):65-74.
• [41]Schleicher U, Liese J, Knippertz I, Kurzmann C, Hesse A, Heit A, Fischer JA, Weiss S, Kalinke U, Kunz S, et al.: NK cell activation in visceral leishmaniasis requires TLR9, myeloid DCs, and IL-12, but is independent of plasmacytoid DCs. J Exp Med 2007, 204(4):893-906.
• [42]Zhang YW, Davis EG, Blecha F, Wilkerson MJ: Molecular cloning and characterization of equine Toll-like receptor 9. Vet Immunol Immunopathol 2008, 124(3–4):209-219.
• [43]Foley JP, Lam D, Jiang H, Liao J, Cheong N, McDevitt TM, Zaman A, Wright JR, Savani C: TLR2, TGFβ, hyaluronan and RHAMM are required for surfactant protein A-stimulated macrophage chemotaxis. J Biol Chem 2012. [Epub ahead of print]
• [44]Vivarini AC, Pereira RMS, Teixeira KLD, Calegari-Silva TC, Bellio M, Laurenti MD, Corbett CEP, Gomes CMC, Soares RP, Silva AM, et al.: Human cutaneous leishmaniasis: interferon-dependent expression of double-stranded RNA-dependent protein kinase (PKR) via TLR2. FASEB J 2011, 25(12):4162-4173.
• [45]Lima WG, Oliveira PS, Caliari MV, Goncalves R, Michalick MS, Melo MN, Tafuri WL: Histopathological and immunohistochemical study of type 3 complement receptors (CD11b/CD18) in livers and spleens of asymptomatic and symptomatic dogs naturally infected with Leishmania (Leishmania) chagasi. Vet Immunol Immunopathol 2007, 117(1–2):129-136.
• [46]Strauss-Ayali D, Baneth G, Jaffe CL: Splenic immune responses during canine visceral leishmaniasis. Vet Res 2007, 38(4):547-564.
• [47]Ettinger NA, Wilson ME: Macrophage and T-cell gene expression in a model of early infection with the protozoan Leishmania chagasi. PLoS Negl Trop Dis 2008, 2(6):e252.
• [48]Tafuri WL, Santos RL, Arantes RM, Goncalves R, de Melo MN, Michalick MS: An alternative immunohistochemical method for detecting Leishmania amastigotes in paraffin-embedded canine tissues. J Immunol Methods 2004, 292(1–2):17-23.
• [49]Melo F, Amaral M, Oliveira P, Lima W, Andrade M, Michalick M, Raso P, Tafuri W: Diffuse intralobular liver fibrosis in dogs naturally infected with Leishmania (Leishmania) chagasi. Am J Trop Med Hyg 2008, 79(2):198-204.
• [50]Figueiredo MM, Moura EP, Costa MM, Ribeiro VM, Michalick MS, Tafuri WL: Histopathological and parasitological investigations of ear healthy skin of dogs naturally and experimentally infected with Leishmania (Leishmania) chagasi. Histol Histopathol 2010, 25(7):877-887.
• [51]Figueiredo MM, Barbosa VS, de Amorim IFG, Deotti B, Michalick MSM, Faria AC, Tafuri WL: Obtaining cells from colon of dogs with leishmaniasis for flow cytometric analysis. Protocols Exchange 2011, 15: . http://www.nature.com/protocolexchange/exchange/protocols/22758 webcite