The role of ALS3 and SSA1 gene expression of Candida albicans in the immune mechanism of vaginal candidiasis

Abstract

Abstract:

Objective To investigate the effect of ALS3 and SSA1 genes in Candida albicans on immune response of vaginal epithelial cells. Methods Wild strains and ALS3, SSA1 gene knockout strains (SC5314, Δals3, Δssa1) of Candida albicans were cultured and their morphology was determined. Human vaginal epithelial cell line VK2/E6E7 was infected in a certain proportion. The damage of different strains to epithelial cells was evaluated by trypan blue staining and lactate dehydrogenase (LDH) activity detection. The co-culture of inflammatory cytokines and chemokines during infection was evaluated by enzyme-linked immunosorbent assay (ELISA).Results There was no significant difference in the effect of Δals3 on the germ tube length of Candida albicans, while the Δssa1 reduced the germ tube length by about 30%~40% compared with the other two strains (P<0.001). Trypan blue staining and LDH assay showed that the cell damage ability of the three strains was proportional to the fungal load when infected with epithelial cells. Compared with wild strain, the cell damage ability of Δssa1 was significantly reduced when infected with epithelial cells at the same MOI, and the difference was statistically significant (P<0.05), while the impact of Δals3 mutant was weak or even slightly higher. Detection of inflammatory cytokines and chemokines showed that the ability of mutants to induce epithelial cells to produce pro-inflammatory factors and chemokines (GM-CSF, G-CSF, IL-1a, IL-8) was significantly weakened (P<0.05).Conclusion ALS3 and SSA1 gene expression played an important role in the local immune response of vaginal epithelial cells to Candida albicans infection, and SSA1 gene expression was more significant.

Key words: vaginal epithelial cells, Candida albicans, ALS3, SSA1, immunity

CLC Number:

R519.3

GAO Ying, WANG Qiong, LIANG Guan-zhao, SHE Xiao-dong, SHI Dong-mei, SHEN Yong-nian, SU Xiao-hong, LI Dong-mei, LIU Wei-da. The role of ALS3 and SSA1 gene expression of Candida albicans in the immune mechanism of vaginal candidiasis[J]. Chinese Journal of Mycology, 2019, 14(2): 65-69.

References

[1] Sobel JD. Vulvovaginal candidosis[J]. Lancet, 2007,369(9577):1961-1971.
[2] Cenci E, Mencacci A, Spaccapelo R, et al. T helper cell type 1(Th1)-and Th2-like responses are present in mice with gastric candidiasis but protective immunity is associated with Th1 development[J]. J Infect Dis, 1995,171(5):1279-1288.
[3] Fidel PL, Jr., Lynch ME, Redondo-Lopez V, et al. Systemic cell-mediated immune reactivity in women with recurrent vulvovaginal candidiasis[J]. J Infect Dis, 1993,168(6):1458-1465.
[4] Murciano C, Moyes DL, Runglall M, et al. Evaluation of the role of Candida albicans agglutinin-like sequence (Als) proteins in human oral epithelial cell interactions[J]. PLoS One, 2012,7(3):e33362.
[5] Yang W, Yan L, Wu C, et al. Fungal invasion of epithelial cells[J]. Microbiol Res, 2014,169(11):803-810.
[6] Murciano C, Moyes D L, Runglall M, et al. Evaluation of the role of Candida albicans agglutinin-like sequence (Als) proteins in human oral epithelial cell interactions[J]. PLoS One, 2012, 7(3):e33362.
[7] Moyes D L, Runglall M, Murciano C, et al. A biphasic innate immune MAPK response discriminates between the yeast and hyphal forms of Candida albicans in epithelial cells[J]. Cell Host Microbe, 2010, 8(3):225-235.
[8] Zhao X, Oh S H, Cheng G, et al. ALS3 and ALS8 represent a single locus that encodes a Candida albicans adhesin; functional comparisons between Als3p and Als1p[J]. Microbiology, 2004, 150(Pt 7):2415-2428.
[9] Sun J N, Solis N V, Phan Q T, et al. Host cell invasion and virulence mediated by Candida albicans Ssa1[J]. PLoS Pathog, 2010, 6(11):e1001181.
[10] Tutar Y, Arslan D, Tutar L. Heat, pH induced aggregation and surface hydrophobicity of S. cerevesiae Ssa1 protein[J]. Protein J, 2010, 29(7):501-508.
[11] Murciano C, Moyes D L, Runglall M, et al. Evaluation of the role of Candida albicans agglutinin-like sequence (Als) proteins in human oral epithelial cell interactions[J]. PLoS One, 2012, 7(3):e33362.
[12] Dongari-Bagtzoglou A, Fidel PL, Jr. The host cytokine responses and protective immunity in oropharyngeal candidiasis[J]. J Dent Res, 2005,84(11):966-977.
[13] Steele C, Fidel PL, Jr. Cytokine and chemokine production by human oral and vaginal epithelial cells in response to Candida albicans[J]. Infect Immun, 2002,70(2):577-583.
[14] Lee S, Song L, Yang M, et al. Local administration of granulocyte macrophage colony-stimulating factor induces local accumulation of dendritic cells and antigen-specific CD8⁺ T cells and enhances dendritic cell cross-presentation[J]. Vaccine, 2015, 33(13):1549-1555.
[15] Carmi Y, Voronov E, Dotan S, et al. The role of macrophage-derived IL-1 in induction and maintenance of angiogenesis[J]. J Immunol, 2009,183(7):4705-4714.
[16] Homey B, Dieu-Nosjean MC, Wiesenborn A, et al. Up-regulation of macrophage inflammatory protein-3 alpha/CCL20 and CC chemokine receptor 6 in psoriasis[J]. J Immunol, 2000,164(12):6621-6632.