Abstract:We investigated the roles of the crude antigen(CA) of Clonorchis sinensis and excretory-secretory products (ESPs) in the polarization of Th1 and Th2 cells. Bone marrow-derived cells were generated from BALB/c mice and isolated into immature DCs; immature DCs were then treated with either CA (CA stimulated group), ESPs (ESPs stimulated group), LPS (positive control group) or PBS (negative control group) for 24 hours. Then the CD4+T cells were isolated from mouse spleen by using anti-mouse-CD4 Microbeads, and further cocultured with stimulated DCs for another 72 hours. The purities of DCs and CD4+T cells were evaluated by flow cytometry and the expressing levels of T-bet mRNA and GATA-3 mRNA were detected by real-time PCR. ELISA was used to detect the levels of IFN-γ and IL-4 cytokines in the supernatant. mRNA levels of T-bet and GATA-3 in the ESPs group were higher than those in PBS-stimulated group (P<0.05). The concentrations of IFN-γ and IL-4 cytokines in the culture were increased in the ESPs group, compared with PBS stimulated group(P<0.05). IFN-γ but not IL-4 was increased in CA group (P<0.05). The results implied that CA might play a role in Th1 type immune response, and ESPs likely play roles in both Th1 and Th2 immune responses.
[1] Brindley PJ, da Costa JM,Sripa B.Why does infection with some helminths cause cancer[J].Trends Cancer, 2015, 1(3): 174-182. [2] Qian MB, Chen YD, Liang S, et al.The global epidemiology of clonorchiasis and its relation with cholangiocarcinoma[J].Infect Dis Poverty, 2012, 1(1): 4.doi:10.1186/2049-9957-1-4. [3] Cheng YJ, Yao LJ.Pathogenesis of Clonorchis sinensis and liver/gallbladder cancer[J].Chin J Zoonoses, 2010, 26 (3):275-278.(in Chinese) 程艳洁,姚丽君.华支睾吸虫与肝/胆癌发病机制[J].中国人兽共患病学报, 2010, 26 (3):275-278. [4] Qian MB, Utzinger J, Keiser J, et al.Clonorchiasis[J].Lancet, 2016, 387(10020): 800-810.doi:10.1016/S0140-6736(15)60313-0. [5] Yang GJ, Liu L, Zhu HR, et al.China’s sustained drive to eliminate neglected tropical diseases[J].Lancet Infect Dis, 2014, 14(9):881-92.doi:10.1016/S1473-3099(14)70727-3. [6] O’Shea JJ,Paul WE.Mechanisms underlying lineage commitment and plasticity of helper CD4 + T cells[J].Science, 2010, 327(5969): 1098-1102.doi:10.1126/science.1178334 [7] Hirahara K,Nakayama T.CD4 + T-cell subsets in inflammatory diseases: beyond the Th1/Th2 paradigm[J].Int Immunol, 2016, 28(4): 163-171.doi:10.1093/intimm/dxw006 [8] Pulendran B,Artis D.New paradigms in type 2 immunity[J].Science, 2012, 337(6093): 431-435.doi:10.1126/science.1221064 [9] Kapsenberg ML.Dendritic-cell control of pathogen-driven T-cell polarization[J].Nat Rev Immunol, 2003, 3(12): 984-993. [10] Liu XX, Zhu M, Xu Q, et al.Parasitic infection to dendritic cell subsets[J].Chin J Zoonoses, 2012, 28(10): 1020-1024.(in Chinese) 刘晓霞, 朱明, 徐琦, 等.寄生虫感染对树突状细胞亚群的影响[J].中国人兽共患病学报, 2012, 28(10): 1020-1024. [11] Dai QF, Fu LL, Liu YS, et al.Dendritic cells pulsed by anantigen of Clonorchis sinensis to induce immuneresponse[J].J Pathog Biol, 2009, 4(8): 582-585.(in Chinese) 戴其锋, 付琳琳, 刘宜升, 等.华支睾吸虫成虫抗原致敏树突状细胞诱导免疫应答的研究[J].中国病原生物学杂志, 2009, 4(8): 582-585. [12] Guo QQ, Fu LL, Tang RX, et al.Th1/Th2 cytokine profile in spleen mononuclear cells of mice infected with Clonorchis sinensis stimulated by excretory-secretory antigen[J].Chin Trop Med, 2009, 9(7): 1187-1189.(in Chinese) 郭倩倩, 付琳琳, 汤仁仙, 等.ESA刺激华支睾吸虫感染小鼠脾细胞分泌Th1/Th2的动态观察[J].中国热带医学, 2009, 9(7): 1187-1189. [13] Fang L, Sun L, Yang J, et al.Heat shock protein 70 from Trichinella spiralis induces protective immunity in BALB/c mice by activating dendritic cells[J].Vaccine, 2014, 32(35): 4412-4419.doi:10.1016/j.vaccine.2014.06.055 [14] Pearce EJ, C MK, Sun J, et al.Th2 response polarization during infection with the helminth parasite Schistosoma mansoni [J].Immunol Rev, 2004, 201: 117-126. [15] Mendes EA, Mendes TA, dos Santos SL, et al.Expression of IL-4, IL-10 and IFN-gamma in the liver tissue of cattle that are naturally infected with Fasciola hepatica [J].Vet Parasitol, 2013, 195(1/2): 177-182. [16] Choi YK, Yoon BI, Won YS, et al.Cytokine responses in mice infected with Clonorchis sinensis [J].Parasitol Res, 2003, 91(2): 87-93. [17] Chakir H, Wang H, Lefebvre DE, et al.T-bet/GATA- 3 ratio as a measure of the Th1/Th2 cytokine profile in mixed cell populations: predominant role of GATA- 3[J].J Immunol Methods, 2003, 278(1/2): 157-169. [18] Babu S, Kumaraswami V, Nutman TB.Transcriptional control of impaired Th1 responses in patent lymphatic filariasis by T-box expressed in T cells and suppressor of cytokine signaling genes[J].Infect Immun, 2005, 73(6): 3394-3401.