Effect of Cell Wall, Cytoplasmic Fraction and Killed-Candida albicans on Nitric Oxide Production by Peritoneal Macrophages from BALB/c Mice

Document Type: Original Article

Authors

1 Department of Immunology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Department of Mycology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

Abstract

Objective(s)
The fractions of Candida albicans have been used as an immunomodulator. The present work assessed the effect of different fractions of C. albicans on nitric oxide (NO) production by mice peritoneal macrophages. Materials and Methods
Cell wall and cytoplasmic fractions of C. albicans ATCC 10321 strain were extracted. Mice peritoneal macrophages were purified and cultured. Different concentrations of both fractions and also killed C. albicans cells were used for macrophages stimulation and evaluation of NO production. NO amount was detected in culture supernatants of macrophages by Griess reagent. Also, MTT assay was performed to assess the viability of macrophages.
Results
The results elucidated that suppressive effect of cell wall proteins on NO release was significant at the dose of 100 pg/ml (P=0.01), while cytoplasmic fraction increased NO amount at the dose of 1 pg/ml compared to the control group (P=0.003). Augmentation of NO production was statistically significant at 200 killed C. albicans per well (P=0.006).
Conclusion
According to our findings, cytoplasmic fractions and killed C. albicans have a positive effect on NO production by peritoneal macrophages, while cell wall fractions did not. Therefore, it is proposed that C. albicans fractions can be studied more as inflammation modulators.

Keywords


1.Romani L. Innate and adaptive immunity in C. albicans infections and saprophytism. J Leukoc Biol 2000; 68:175-179.

2.Crameri R, Blaser K. Allergy and immunity to fungal infections and colonization. Eur Respir J 2002;19:151-157. 

3.Fukazawa Y, Cassone A, Bistoni F, Howard DH, Kagaya K, Murphy J W, et al. Mechanisms of cell mediated immunity in fungal infections: a review. J Med Vet Mycol 1994; 32:123.

4.Shoham S, Levitz SM. Immune response to fungal infections. Br J Haematol 2005; 129:569-582.

5.Badauy CM, Barbachan JJ, Rados PV, Sant Ana Filho M, Chies JAB. Relationship between Candida infection and immune cellular response in inflammatory hyperplasia. Oral Microbiol Immunol 2005; 20:89-92.

6.Bistoni F, Cenci E, Mencacci A, Schaffolla E, Mpsci P, Puccetti P, et al. Mucosal and systemic T helper cells function after intragastric colonization of adult mice with Candida albicans. J Infect Dis 1993; 168:1449-1457.

7.Santoni G, Boccanera M, Adriani D, Lucciarini R. Immune cell-mediated protection against vaginal Candidiasis: evidence for a major role of vaginal CD4+ T cells and possible participation of other local lymphocyte effectors. Infect Immun 2002; 70:4791-4797.

8.Fidel Jr PL. Candida-host interactions in HIV disease:relationships in oropharyngeal candidiasis Adv Dent Rec. 2006; 19:80-84.

9.Fang FC, Vazquez-Torres A. Nitric oxide production by human macrophages: there's NO doubt about it. Am J Physiol Lung Cell Mol Physiol 2002; 282:L941- L943.

10.Gonzalez A, Gregori W, Velez D, Restrepo A, Cano LE. Nitric oxide participation in the fungicidal mechanism of gamma interferon-activated murine macrophages against paracoccidioides brasiliensis conidia. Infect Immun 2000; 68:2546-2552.

11.Kawakami K, Zhang T, Qureshi MH, Saito A. Cryptococcus neoformans inhibit nitric oxide production by murine peritoneal macrophages stimulated with interferon-g and lipopolysaccharide. Cell Immunol 1997; 180:47-54.

12.Elorza M, Murgui A. Dimorphism in Candida albicans; contribution of mannoprotein to the architecture of yeast and mycelial cell wall .J Gen Microbiol 1985;131:2207-2216.

13.Casanova M, Chaffin WL. Cell wall glycoproteins of Candida albicans as released by different methods. J Gen Microbiol 1991; 137:1045-1051.

14.Ribeiro-Dias F, Russo M, Nascimento FRF, Barbuto JAM, Timenetsky J, Jancar S. Thioglycollate-elicited murinemacrophages are cytotoxic to mycoplasma arginini-infected YAC-1 tumor cells. Braz J Med Biol Res 1998; 31:1425-1428.

15.Scuro LS, Simioni PU, Grabriel DL, Saviani EE, Modolo LV, Tamashiro WM, et al. Suppression of nitric oxide production in mouse macrophages by soybean flavonoids accumulated in response to nitroprusside and fungal elicitation. BMC Biochem 2004; 21; 5:5.

16.Stuehr DJ, Nathan CF. Nitric oxide: a macrophage product responsible for cytostasis and respiratory inhibition in tumor target cells. J Exp Med 1989; 169:1543-1555.

17.Sladowski D, Steer SJ, Clothier RH, Balls M. An improved MTT assay. J Immunol Methods.1993; 157: 203-7.

18.Ribeiro-Dias F, Russo M, Nascimento FRF, Barbuto JAM, Timenetsky J, Jancar S. Thioglyollate-elicited moraine macrophages are cytotocxic to mycoplasma arginini - infected YAC-1 tumor cells. Braz J Med Biol Rec. 1998; 31:1425-1428.

19.Schroppel K, Kryk M, Herrman M, Leberer E, Rollinghoff M ,Bogdan Ch. Suppression of type 2 NO-synthase activity in macrophages by Candida albicans. Int J Med Microbiol 2001; 290: 659-668.

20.Pacher P, Joseph S, Beckman JS, Liaudet L. Nitric oxide and peroxynitrite in health and disease. Physiol Rev 2007; 87:315- 424.

21.Smail EH, Kolotila MP, Ruggeri R, Diamond RD. Natural inhibitor from Candida albicans blocks release of azurophil and specific granule contents by chemotactic peptide-stimulated human neutrophils. Infect Immun 1989; 57:689-92.

22.Chinen T, Qureshi MH, Koguchi Y, Kawakami K. Candida albicans suppresses nitric oxide (NO) production by interferon-gamma (IFN-g) and lipopolysaccharide (LPS)-stimulated murine peritoneal macrophages. Clin Exp Immunol 1999; 115:491-497.

23.Lajean Chaffin W, Lopez-Ribot JL, Casanova M, Gozalbo D, Martinez JP. Cell wall and secreted proteins of Candida albicans: identification, function, and expression. Microbiol Mol Biol Rev 1998; 62:130-180.

24.Nelson RD, Shibata N, Podzorski RP, Herron MJ. Candida mannan: chemistry, suppression of cell-mediated immunity, and possible mechanisms of action. Clin Microbiol Rev 1991; 4:1-19.

25.Jouault T, Fradin C, Trinel P, Poulain D. Candida albicans-derived P-1,2-linked mannooligosaccharides induce desensitization of macrophages. Infect Immun 2000; 68:965-968.

26.Cartney-Francis NM, Allen JB, Mizel DE, Albina JE, Xie QW, Nathan CF, et al. Suppression of arthritis by an inhibitor of nitric oxide synthase. J Exp Med 1993; 178:749-754.

27.Montagnoli C, Sandini S, Bacci A, Romani L. Immunogenicity and protective effect of recombinant enolase of Candida albicans in a murine model of systemic candidiasis. Med Mycol 2004:42; 319-324.