In vitro lymphoproliferative response and cytokine production in mice with experimental disseminated candidiasis

Document Type : Original Article

Authors

1 Mycology Research Center, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Pathobiology, Faculty of Veterinary Medicine, Amol University of Special Modern Technologies, Amol, Iran

3 Department of Food Sciences, Qazvin, Iranian Veterinary Organization

Abstract

Objective(s): Systemic candidiasis is an infection of Candida albicans (C. albicans) causing disseminated disease and sepsis, invariably when host defenses are compromised. We investigated the histopathological changes as well as the lymphoproliferative responses and cytokine production of splenic cells after stimulation with Concanavalin A (Con A) and Pokeweed mitogen (PWM) in mice with disseminated candidiasis.
Materials and Methods:Lymphoproliferative responses were stimulated in vitro with Con A (1 µg/ml) and PWM (1 µg/ml) mitogens in Roswell Park Memorial Institute (RPMI) 1640 media, and the production of interferon (IFN)-γ and interleukin-4 (IL-4) in the supernatants was measured by enzyme-linked immunosorbent assay (ELISA).
Results: The results revealed that C. albicans organisms multiplied to a greater extent in the kidneys than in the liver and spleen of infected mice. The most predominant forms of C. albicans in different parts of the kidneys were yeast mixed with hyphal forms. Infected mice had a significantly increased proliferative response when splenocytes were stimulated with PWM (2.0±0.16) and Con A (1.9±0.19) (P<0.05). PWM and Con A-stimulated production of IFN-γ significantly tended to be higher in infected mice (PWM: 68.4±14.0 pg/ml; Con A: 53.7±17.3 pg/ml) when compared to controls (P<0.05). Stimulation with PWM and Con A showed no differences in IL-4 production between infected mice and controls.
Conclusion: These findings demonstrated a significant increase in both cell proliferation and IFN-γ secretion in supernatants of PWM and Con A- stimulated splenocyte cultures obtained from mice with disseminated candidiasis.

Keywords


1. Calderone RA. Candida and Candidiasis. New York: American Society for Microbiology; 2001.
2. Garber G. An overview of fungal infections. Drugs 2001; 61:1-12.
3. Romani L, Mencacci A, Cenci E, Del Sero G, Bistoni F, Puccetti P. An immunoregulatory role for neutrophils in CD41 T helper subset selection in mice with candidiasis. J Immunol 1997; 158:2356-2362.
4. Vasquez-Torres A, Balish E. Macrophages in resistance to candidiasis. Microbiol Mol Rev 1997; 61:170-192.
5. O’Garra A. Cytokines induce the development of functionally heterogeneous T helper cell subsets. Immun 1998; 8:275-283.
6. Cenci E, Mencacci A, Del Sero G, Fed Ostiani C, Mosci P, Kopf M. IFN-γ is required for IL-12 responsiveness in mice with Candida albicans infection. J Immunol 1998; 161:3543-3550.
7. Chin VK, Foong KJ, Maha A, Rusliza B, Norhafizah M, Chong PP. Multi-step pathogenesis and induction of local immune response by systemic Candida albicans infection in an intravenous challenge mouse model.Int J Mol Sci2014; 15:14848-14867.
8. Lin L, Ibrahim AS, Xu X, Farber JM, Avanesian V, Baquir B, et al. Th1-Th17 Cells mediate protective adaptive immunity against Staphylococcus aureus and Candida albicans infection in Mice. PLoS Pathog 2009; 5:e1000703.
9. Mencacci A, Cenci E, Del Sero G, Fe`d’Ostiani C, Mosci P, Bistoni F. Defective co-stimulation and impaired Th1 development in tumor necrosis factor/lymphotoxin-a double-deficient mice infected with Candida albicans. Int Immunol 1998; 10:37-48.
10. Ashman RB, Vijayan D, Wells CA. IL-12 and related cytokines: function and regulatory implications in Candida albicans infection. Clin Dev Immunol 2011; 2011:1-9.
11. Heizmann P, Klefisch F, Heizmann WR. Basic research-significance of detection and clinical impact of Candida albicans in non-immunosuppressed patients. Pharmacol Pharm 2011; 2:354-360.
12. Netea MG, Brown GD, Kullberg BJ, Gow NA. An integrated model of the recognition of Candida albicans by the innate immune system. Nat Rev Microbiol 2008; 6:67-78.
13. Khosravi AR, Mardjanmehr H, Shokri H, Naghshineh R, Rostamibashman M, Naseri A. Mycological and histopathological findings of experimental disseminated candidiasis in dogs. Iran J Vet Res 2009; 10:228-234.
14. Soltani M, Khosravi AR, Asadi F, Shokri H. Evaluation of protective efficacy of Spirulina platensis in BALB/C mice with candidiasis. J Mycol Méd 2012; 22:329-334.
15. Jimenez-Valera M, Moreno E, Amat MA, Ruiz-Bravo A. Modification of mitogen-driven lymphoproliferation by ceftriaxone in normal and immunocompromised mice. Int J Antimicrob Agents 2003; 22:607-612.
16. Brown MR, Thompson CA, Mohamed FM. Systemic candidiasis in an apparently immunocompetent dog. J Vet Diagn Invest 2005; 17:272-276.
17. Fatahinia M, Khosravi AR, Shokri H. Propolis efficacy on TNF-α, IFN-γ and IL2 cytokines production in old mice with and without systemic candidiasis. J Mycol Méd 2012; 22:237-242.
18. Elahi S, Pang G, Clancy R, Ashman RB. Cellular and cytokine correlates of mucosal protection in murine model of oral candidiasis. Infect Immun 2000; 68:5771-5777.
19. Katial RK, Sachanandani D, Pinney C, Lieberman MM. Cytokine production in cell culture by peripheral blood mononuclear cells from immunocompetent hosts. Clin Diagn Lab Immunol 1998; 1:78-81.
20. Mencacci A, Del Sero G, Cenci E, d’Ostiani CF, Bacci A, Montagnoli C. Endogenous interleukin 4 is required for development of protective CD41 T helper type 1 cell responses to Candida albicans. J Exp Med 1998; 187:307-317.
21. Seder RA, Le Gros G, Ben-Sasson SZ, Urban J, Finkelman FD, Paul WE. Increased frequency of interleukin 4-producing T cells as a result of polyclonal priming. Use of a single-cell assay to detect interleukin 4-producing cells. Eur J Immunol 1991; 21:1241-1247.
22. Kaposzta R, Tree P, Marodi L, Gordon S. Characteristics of invasive candidiasis in gamma interferon and interleukin-4-deficient mice: role of macrophages in host defense against Candida albicans. Infect Immun 1998; 66:1708-1717.
23. Vazquez-Torres A, Jones-Carson J, Warner WT, Balish E. Early resistance of IL-10 knockout mice to acute systemic candidiasis. Infect Immun 1999; 67:670-674.
24. Romani L, Mencacci A, Grohmann U, Mocci S, Mosci P, Pucetti P. Neutralizing antibody to interleukin 4 induces systemic protection and T helper type 1-associated immunity in murine candidiasis. J Exp Med 1992; 176:19-25.
25. Simon MM, Hochgeschwender U, Brugger U, Landolfo S. Monoclonal antibodies to interferon-γ inhibit interleukin 2-dependent induction of growth and maturation in lectin/antigen-reactive cytolytic T lymphocyte precursors. J Immunol 1996; 136:2755-2762.
26. Oykhman P, Mody CH. Direct microbicidal activity of cytotoxic T-lymphocytes. J Biomed Biotechnol 2010; 2010:1-9.
27. Norian R, Delirezh N, Azadmehr A. Evaluation of proliferation and cytokines production by mitogen- stimulated bovine peripheral blood mono nuclear cells. Vet Res Forum 2015; 6:265-271.