Probiotic lactobacillus and estrogen effects on vaginal epithelial gene expression responses to Candida albicans

Journal of Biomedical Science

Shemedia J Johnson¹  R Doug Wagner¹ 

[1] Microbiology Division, National Center for Toxicological Research, 3900 NCTR Rd, Jefferson, AR, 72079, USA

关键词: Estrogen;    Candidiasis;    Signal transduction genes;    Gene expression;    Epithelial cells;    Probiotic;   

Others  :  824890
DOI  :  10.1186/1423-0127-19-58

 received in 2012-02-21, accepted in 2012-06-20,  发布年份 2012

【 摘 要 】

Background

Vaginal epithelial cells have receptors, signal transduction mechanisms, and cytokine secretion capabilities to recruit host defenses against Candida albicans infections. This research evaluates how probiotic lactobacilli affect the defensive epithelial response.

Methods

This study used quantitative reverse transcription-polymerase chain reaction assay (qRT-PCR), flow cytometry, and a multiplex immunoassay to observe changes in the regulation of gene expression related to cytokine responses in the VK2 (E6/E7) vaginal epithelial cell line treated with 17β-estradiol, exposed to probiotic Lactobacillus rhamnosus GR-1® and Lactobacillus reuteri RC-14® and challenged with C. albicans. Data were statistically evaluated by repeated measures analysis of variance and paired t-tests where appropriate.

Results

C. albicans induced mRNA expression of genes related to inflammatory cytokine responses associated with nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signal transduction pathways. 17β-estradiol suppressed expression of interleukin-1α (IL-1α), IL-6, IL-8, and tumor necrosis factor alpha (TNFα) mRNA. Probiotic lactobacilli suppressed C. albicans-induced nuclear factor-kappa B inhibitor kinase kinase alpha (Iκκα), Toll-like receptor-2 (TLR2), TLR6, IL-8, and TNFα, also suggesting inhibition of NF-κB signaling. The lactobacilli induced expression of IL-1α, and IL-1β mRNA, which was not inhibited by curcumin, suggesting that they induce an alternate inflammatory signal transduction pathway to NF-κB, such as the mitogen activated protein kinase and activator protein-1 (MAPK/AP-1) signal transduction pathway. Curcumin inhibited IL-13 secretion, suggesting that expression of this cytokine is mainly regulated by NF-κB signaling in VK2 cells.

Conclusions

The results suggest that C. albicans infection induces pro-inflammatory responses in vaginal epithelial cells, and estrogen and lactobacilli suppress expression of NF-κB-related inflammatory genes. Probiotic lactobacilli may induce IL-1α and IL-1β expression by an alternate signal transduction pathway, such as MAPK/AP-1. Activation of alternate signaling mechanisms by lactobacilli to modify epithelial cell cytokine production may be a mechanism for probiotic modulation of morbidity in vulvovaginal candidiasis.

【 授权许可】

   
2012 Wagner and Johnson.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Falagas ME, Betsi GI, Athanasiou S: Probiotics for prevention of recurrent vulvovaginal candidiasis: a review. J Antimicrob Chemother 2006, 58:266-272.
• [2]Sobel JD: Vulvovaginal candidosis. Lancet 2007, 369:1961-1971.
• [3]Reid G, Bruce AW: Urogenital infections in women: can probiotics help? Postgrad Med J 2003, 79:428-432.
• [4]Barousse MM, Espinoza T, Dunlap K, Fidel PL: Vaginal epithelial cell anti-Candida albicans activity is associated with protection against symptomatic vaginal candidiasis. Infect Immun 2005, 73:7765-7767.
• [5]Fidel PE: History and update on host defense against vaginal candidiasis. Am J Reproduct Immunol 2007, 57:2-12.
• [6]Cremel M, Berlier W, Hamzeh H, Cognasse F, Lawrence P, Genin C, Bernengo J-C, Lambert C, Dieu-Nosjean M-C, Delézay O: Characterization of CCL20 secretion by human epithelial vaginal cells: involvement in Langerhans cell precursor attraction. J Leukoc Biol 2005, 78:158-166.
• [7]Peterson ML, Ault K, Kremer MJ, Klingelhutz AJ, Davis CC, Squier CA, Schlievert PM: The innate immune system is activated by stimulation of vaginal epithelial cells with Sataphylococcus aureus and toxic shock syndrome toxin 1. Infect Immun 2005, 73:2164-2174.
• [8]Pivarcsi A, Nagy I, Koreck A, Kis K, Kenderessy-Szabo A, Szell M, Dobozy A, Kemeny L: Microbial compounds induce the expression of pro-inflammatory cytokines, chemokines and human β-defensin-2 in vaginal epithelial cells. Microbes Infect 2005, 7:1117-1127.
• [9]McGowin CL, Ma L, Martin DH, Pyles RB: Mycoplasma genitalium-encoded MG309 activates NF-κB via toll-like receptors 2 and 6 to elicit proinflammatory cytokine secretion from human genital epithelial cells. Infect Immun 2009, 77:1175-1181.
• [10]Wira CR, Fahey JV, Ghosh M, Patel MV, Hickey DK, Ochiel DO: Sex hormone regulation of innate immunity in the female reproductive tract: the role of epithelial cells in balancing reproductive potential with protection against sexually transmitted pathogens. Am J Reprod Immunol 2010, 63:544-565.
• [11]Fichrova RN, Rheinwald JG, Anderson DJ: Generation of papillomavirus-immortalized cell lines from normal human ectocervical, endocervical, and vaginal epithelium that maintain expression of tissue-specific differentiation proteins. Biol Reproduct 1997, 57:847-855.
• [12]Schaller M, Zakikhany K, Naglik JR, Weindl G, Hube B: Models of oral and vaginal candidiasis based on in vitro reconstituted human epithelia. Nat Protocol 2006, 6:2767-2773.
• [13]Kinsman OS, Collard AE: Hormonal factors in vaginal candidiasis in rats. Infect Immun 1986, 53:498-504.
• [14]Geshnizgani AM, Onderedonk AB: Defined medium simulation genital tract secretions for growth of vaginal microflora. J Clin Microbiol 1992, 30:1323-1326.
• [15]Motulsky H: Choosing an appropriate sample size. Oxford University Press, New York; 1995:195-204.
• [16]Abad CL, Safdar N: The role of Lactobacillus probiotics in the treatment or prevention of urogenital infections – a systematic review. J Chemother 2009, 21:243-252.
• [17]Martinez RCR, Franceschini SA, Patta MC, Quintana SM, Candido RC, Ferreira JC, De Martinis ECP, Reid G: Improved treatment of vulvovaginal candidiasis with fluconazole plus probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14. Lett Appl Microbiol 2009, 48:269-274.
• [18]Netea MG, van de Veerdonk F, Verschueren I, van der Meer JWM, Kullberg BJ: Role of TLR1 and TLR6 in the host defense against disseminated candidiasis. FEMS Immunol Med Microbiol 2008, 52:118-123.
• [19]Bahri R, Saidane-Mosbahi D, Rouabhia M: Candida famata modulates toll-like receptor, β-defensin, and proinflammatory cytokine expression by normal human epithelial cells. J Cell Physiol 2009, 222:209-218.
• [20]Martinez RCR, Seney SL, Summers KL, Nomizo A, De Martinis ECP, Reid G: Effect of Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 on the ability of Candida albicans to infect cells and induce inflammation. Microbiol Immunol 2009, 53:487-495.
• [21]Basso B, Giménez F, López C: IL-1b, IL-6 and IL-8 levels in gyneco-obstetric infections. Infect Dis Obstet Gynecol 2005, 13:207-211.
• [22]Coutreau MM, Chennathukuzhi VM, Harris HA, Han L, Dorner AJ, Apseloff G, Varadarajan U, Hatstat E, Zakaria M, Strahs AL, Crabtree JS, Winneker RC, Jelinsky SA: A study of 17β-estradiol-regulated genes in the vagina of postmenopausal women with vaginal atrophy. Maturitas 2007, 58:366-376.
• [23]Fahey JV, Wright JA, Shen L, Smith JM, Ghosh M, Rossoll RM, Wira CR: Estradiol selectively regulates innate immune function by polarized human uterine epithelial cells in culture. Mucosal Immunol 2008, 1:317-325.
• [24]Pahl R, Brunke G, Steubesand N, Schubert S, Böttner M, Wedel T, Jürgensen C, Hampe J, Schäfer H, Zeissig S, Schreiber S, Rosenstiel P, Reiss K, Arlt A: IL-1β and ADAM17 are central regulators of β-defensin expression in Candida esophagitis. Am J Physiol 2011, 300:G547-G553.