Abstract:In this study, we designed a siRNA-plasmid to lower the expression of CAP10 gene. A mouse model of Cryptococcus neoformans infection was established by inhalation of Cryptococcus neoformans through one nostril. On day 7 after inoculation (acute phase), the infected lung were observed by histopathological analysis. Th1-Th2 cytokines (IFN-γ, TNF-α and IL-10) were detected by ELISA. In acute phase, the levels of these cytokines were (19.24±1.31)pg/mL, (36.94±2.04)pg/mL, (18.32±3.00) pg/mL in control, (14.34±1.26)pg/mL, (25.37±1.37)pg/mL, (72.96±8.83)pg/mL in Wild-Type(WT), (14.63±0.95)pg/mL, (26.22±1.55)pg/mL, (38.73±4.61)pg/mL in siRNA-CAP10 respectively. The level of IFN-γ and TNF-α were significantly lower in WT and siRNA-CAP10 groups than those in control mice, while the level of IL-10 was significantly higher in WT and siRNA-CAP10 groups than in control mice (P<0.01). Compared to siRNA-CAP10 groups, the level of IL-10 were significantly higher in WT (P<0.01), while the level of IFN-γ and TNF-α were not statistically significant difference. Using the ratio of Th1/Th2 to evaluate the status of immune response, the ratios of IFN-γ/IL-10 and TNF-α/IL-10 were 1.08±0.21, 2.07±0.34 in control, 0.20±0.03, 0.35±0.05 in WT, and 0.38±0.04, 0.69±0.09 in siRNA-CAP10. The level of IFN-γ/IL-10 and TNF-α/IL-10 were significantly lower in WT and siRNA-CAP10 groups than in control mice, and the level of IFN-γ/IL-10 and TNF-α/IL-10 were significantly lower in WT than in siRNA-CAP10 mice (P<0.01). Results showed CAP10 gene expression related to antifungal immune response in Cryptococcus neoformans infection in mice. The down-regulation of CAP10 gene may control the spread of Cryptococcus neoformans and turned the ratio of Th1/Th2 back to balance. CAP10 gene played a very important role in regulating inflammatory response, which may became new molecular targets for treatment.
吴赞艺, 林旎, 林梨平, 欧启水, 林东红, 陈敏, 徐建萍. 应用RNAi技术探讨CAP10基因在新型隐球菌感染小鼠中对Th1-Th2型免疫的影响[J]. 中国人兽共患病学报, 2016, 32(10): 893-897.
WU Zan-yi, LIN Ni, LIN Li-ping, OU Qi-shui, LIN Dong-hong, CHEN Min, XU Jian-ping. RNAi technology application for evaluating effect of CAP10 gene in Th1-Th2 immune of mice infection with Cryptococcus neoformans. Chinese Journal of Zoonoses, 2016, 32(10): 893-897.
[1] Zaragoza O, Rodrigues ML, De Jesus M, et al. The capsule of the fungal pathogen Cryptococcus neoformans [J]. Adv Appl Microbiol, 2009, 68: 133-216. doi:10.1016/S0065-2164(09)01204-0 [2] Teresa R, O’Meara J, Alspaugh A. The Cryptococcus neoformans capsule: a sword and a shield[J]. Clin Microbiol Rev, 2012, 25(3): 387-408. doi:10.1128/CMR.00001-12 [3] Zaragoza O, Telzak A. The polysaccharide capsule of the pathogenic fungus Cryptococcus neoformans enlarges by distal growth and is rearranged during budding[J]. Mol Microbiol, 2006, 59(1): 67-83. doi:10.1111/j.1365-2958.2005.04928.x [4] Voelz K, May RC. Cryptococcal interactions with the host immune system[J]. Eukaryot Cell, 2010, 9(6): 835-846. doi:10.1128/EC.00039-10 [5] Okabayashi K, Kano R, Watanabe S, et al. Expression of capsule-associated genes of Cryptococcus neoformans [J]. Mycopathologia, 2005, 160: 1-7. [6] Wozniak KL, Kolls JK, Wormley FL Jr. Depletion of neutrophils in a protective model of pulmonary cryptococcosis results in increased IL-17A production by gammadelta T cells[J]. BMC Immunol, 2012, 13: 65. doi:10.1186/1471-2172-13-65 [7] Jarvis JN, Casazza JP, Stone HH, et al. The phenotype of the Cryptococcus -specific CD4 + memory T-cell response is associated with disease severity and outcome in HIV-associated cryptococcal meningitis[J]. J Infect Dis, 2013, 207(12): 1817-1828. doi:10.1093/infdis/jit099 [8] Wozniak KL, Ravi S, Macias S, et al. Insights into the mechanisms of protective immunity against Cryptococcus neoformans infection using a mouse model of pulmonary cryptococcosis[J]. PLoS One, 2009, 4(9): e6854. doi:10.1371/journal.pone.0006854 [9] Wozniak KL, Hardison SE, Kolls JK, et al. Role of IL-17A on resolution of pulmonary C. neoformans infection[J]. PLoS One, 2011, 6: e17204. doi:10.1371/journal.pone.0017204 [10] Qiu Y, Davis MJ, Dayrit JK, et al. Immune modulation mediated by cryptococcal laccase promotes pulmonary growth and brain dissemination of virulent Cryptococcus neoformans in mice[J]. PLoS One, 2012, 7(10): e47853. doi:10.1371/journal.pone.0047853 [11] Alanio A, Desnos-Ollivier M, Dromer F. Dynamics of Cryptococcus neoformans -macrophage interactions reveal that fungal background influences outcome during cryptococcal meningoencephalitis in humans[J]. mBio, 2011, 2(4): e00158-00111. doi:10.1128/mBio.00158-11