High IL-4 production related to granuloma development in high-tolerogenic mice infected with Schistosoma mansoni.
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1
Universidade do Estado do Rio de Janeiro (UERJ), Departamento de Genética (DGen), Brazil
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2
Universidade Federal Fluminense (UFF), Departamento de Imunobiologia (GIM), Brazil
Introduction:
The Schistosoma mansoni infection was studied in two strains of mice genetically selected for extreme phenotypes of susceptibility (TS) and resistance (TR) to oral tolerance (Silva, AC et al. 1998). TR strain presents good inflammatory responses and non-tolerogenic profile, while TS strain presents non-inflammatory but high-tolerogenic profile, with high percentages of CD4+CD25+Foxp3+ T regulatory cells. Furthermore, TS strain is able to produce high levels of inhibitory citokynes such as IL-10 (Silva, MF et al. 2010). The aim of this study was to correlate the cytokines production to the damage caused by the granuloma development during the infection.
Material and Methods:
TR and TS strains of mice from the F25 generation, obtained by two-way genetic selection according to susceptibility (TS) or resistance (TR) to ovalbumin oral tolerance (Silva, AC et al. 1998). TR and TS strains were exposed to 50 transcutaneous cercariae obtained from Reference Laboratory in Malacology of Instituto Oswaldo Cruz, FIOCRUZ, RJ, Brazil (Xavier, AML et al. 2010). The Committee for the Care and Use of Laboratory Animals of the Universidade do Estado do Rio de Janeiro approved the protocols of the experiments described in this work (029/2010/UERJ).
Hepatomegaly and splenomegaly were evaluated using the ratio between the weight of the organ (liver and spleen) and the body weight.
The diameter of granulomas was measured according Boros and Warren (1970) from slides containing histological sections stained with Gomori reticulin. It were used brightfield microscopy followed by capturing and processing the images with the program Axio Vision LE.
Cytokines (IL-4, IL-10 and IFN-γ) were quantified from homogenized tissues by ELISA. One hundred milligrams of tissue (ileum or liver) were homogenized in PBS containing antiproteases cocktail and 0.05% Tween 20, and quantified by ELISA following manufacturer's recommendations (Tavares, D et al. 2006).
Results:
TS infected mice showed hepatosplenomegaly more intense (Figure 1A) and larger size of hepatic granulomas (Figure 1B) than TR strain. When infected, both strains produced IFN-γ (Figure 1C), but the TS strain produced IL-10 (Figure 1D) and IL-4 in a larger amount (Figure 1E) both in the liver as well as in the ileum (Unpublished data).
Discussion:
TS strain had higher weakness, apathy, prostration and mortality (Unpublished data) due to more intense hepatosplenomegaly (Figure 1A) related to the larger size of hepatic granulomas (Figure 1B) and extensive fibrosis of these granulomas (Unpublished data).
Previous studies characterized the TS strain with a regulatory status with high frequency of Treg cells (Silva, MF et al. 2010) and high levels of a cytokine suppressive IL-10. However, IL-10 was not able to regulate the exacerbated growth of hepatic granulomas in TS strain (Figure 1D). Despite this regulatory status, this strain is not genetically prevented from developing a exacerbated granulomatous inflammatory response during the infection with S. mansoni.
A high level of IL-4 in TS strain (Figure 1E) is consistent with the exacerbation of granulomas, since IL-4, as well as IL-13, induces collagen synthesis (Mentink-Kane, MM et al. 2011, Cheever AW and Yap GS 1997) and is related to the development of fibrosis in schistosomal granuloma. Additional studies are needed to confirm our proposals and to understand the mechanisms underlying the differences in immune response of these strains in the schistosoma-host relationship.
Figure 1A. Percentage of liver weight relative to body weight of the mice. Figure 1B. Diameter of hepatic granulomas of TR and TS infected mice. The granulomas were detected by Gomori reticulin stain, analyzed in Zeiss Axioskop microscope and photographed using the Axio Vision LE program (6-10 granulomas for slide). Figure 1C-E. Concentration of cytokines in the liver. TR and TS mice in the 8th week after infection with 50 cercariae of S. mansoni and their uninfected controls (mean± sd, n = 5).
Acknowledgements
We are grateful to Dra Lygia dos Reis Corrêa, Mr Paulo Cesar dos Santos and Mr. Heloisa from Laboratório de Malacologia, IOC, Fiocruz/RJ for providing the cercariae of Schistosoma mansoni, and Drs Cid Couto and Marcia Giesta for care of the animal colonies (Laboratório de Imunobiologia). This work was supported by Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) - Grant E-26/171.423/04.
References
Boros DL, Warren KS. Delayed hypersensitivity-type granuloma formation and dermal reaction induced and elicited by a soluble factor isolated from Schistosoma mansoni eggs J Exp Med (1970) 132(3):488-507.
Cheever AW, Yap GS. Immunologic basis of disease and disease regulation in schistosomiasis. Chemical Immunology (1997) 66:159.
Silva AC, Souza KW, Machado RC, Silva MFS, Sant'Anna OA. Genetics of immunological tolerance: I. Bidirectional selective breeding of mice for oral tolerance. Res. Immunol (1998)149: 151-161.
Silva MF, Kamphorst AO, Hayashi EA, Bellio M, Carvalho CR, Faria AM, Sabino KC, Coelho MG, Nobrega A, Tavares D, Silva AC. Innate profiles of cytokines implicated on oral tolerance correlate with low- or high-suppression of humoral response. Immunology (2010) 130(3):447-57.
Mentink-Kane MM, Cheever AW, Wilson MS, Madala SK, Beers LM, Ramalingam TR, Wynn TA. Accelerated and progressive and lethal liver fibrosis in mice that lack interleukin (IL)-10, IL-12p40, and IL-13Rα2. Gastroenterology (2011) 141(6):2200-9.
Tavares D, Ribeiro RC, Silva AC. Inflammatory lesion and parasite load are inversely associated in Leishmania amazonensis infected mice genetically selected according to oral tolerance susceptibility. Microbes Infect (2006) 8(4):957-64.
Xavier AML, Magalhães JA, Cunha G dos S, Silva AC, Tavares DA, Sarro-Silva MF, de Moraes Neto AH. Morphological tegument alterations of adult Schistosoma mansoni, harbored in non anti-helminthic treated, high-immune-tolerogenic and low-inflammatory mice. Acta Trop (2010) 116(1):95-9.
Keywords:
Schistosoma mansoni,
oral tolerance,
Granuloma,
IL-4,
IL-10
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Host-pathogen interactions
Citation:
Tavares
D,
Xavier
AL and
Da Silva
A
(2013). High IL-4 production related to granuloma development in high-tolerogenic mice infected with Schistosoma mansoni..
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.01086
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Received:
28 Jun 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Prof. Daniel Tavares, Universidade do Estado do Rio de Janeiro (UERJ), Departamento de Genética (DGen), Rio de Janeiro, Rio de Janeiro, 20550-013, Brazil, tavares.uerj@gmail.com