CpG ODN激活慢性HBV感染者外周血单个核细胞和CD4^＋T细胞功能的研究

The immunostimulatory study on peripheral blood mononuclearcell and CD4^＋ T Lymphocyte of HBV infections that were activated by CpG oligonucleotide

目的探讨人工合成硫代修饰的含CpG基序的寡脱氧核苷酸（ODN）作为佐剂对慢性HBV感染者PBMC和CD4^＋T细胞产生免疫应答的影响。方法将入选慢性HBV感染者20例分成两组，免疫耐受组和免疫清除组各10例，用CpG ODN、HBsAg、M[CpGODN＋HBsAg]分别与同一感染者的PBMC和CD4^＋T细胞孵育，采用酶联免疫斑点法（ELISPOT）检测两种细胞IFN-γ的分泌情况，以斑点数的多少来评价细胞分泌IFN-γ的能力。结果CpGODN、HBsAg、M[CpGODN＋HBsAg]与PBMC孵育后IFN-γ斑点数是3±8、339±429、375±496；与CD4^＋Tcell孵育后IFN-γ斑点数是1±4、5±16、5±12。在免疫耐受组，CpGODN、HBsAg、M[CpGODN＋HBsAg]与PBMC孵育后IFN-γ斑点数是3±8、361±153、375±276；与CD4^＋Tcell孵育后IFN-γ斑点数是0±2、2±2、4±4。在免疫清除组，CpGODN、HBsAg、M[CpGODN＋HBsAg]与PBMC孵育后IFN-γ斑点数是3±21、289±345、405±656；与CD4^＋Tcell孵育后IFN-γ斑点数是2±14、8±40、7±30。PBMC与HBsAg、M孵育后IFN-γ斑点数比较统计分析P：0．720；CD4^＋Tcell与HBsAg、M孵育后IFN-γ斑点数统计分析P=0．890，两种不同细胞中与M孵育后IFN-γ斑点数比较统计分析得P=0．000。结论在免疫耐受组和免疫清除组，CpGODN不显著增加HBsAg刺激PBMC和CD4^＋T细胞分泌IFN-γ，但CpGODN对PBMC刺激效果优于CD4^＋T细胞。

Objective To study the effects of CpG oligodeoxyribonucIeotide （ODN）as adjuvant on the immune responses in PBMC and CD4^± T cell with chronic hepatitis B virus. Methods The selected 20 infections were averagely divided two groups. The frequency of IFN-γ secreting PBMC and CD4^± T cell in immune tolerant phase and in the immune clearance phase that had stimulated by CpG ODN, HBsAg and Mixture [ CpG ODN ± HBsAg] were analyzed by enzyme linked immune spot（ELISOT）. Results The PBMC and CD4^± T cell were differently incubated by CpG ODN, HBsAg and M [ CpG ODN ± HBsAg ]. The number of IFN-γ spot differently are 3 ± 8, 339±429, 375 ±496, 1 ± 4, 5 ± 16 and 5 ± 12;the results of immume tolerance are 3 ± 8,361 ± 153, 375 ± 276, 0 ± 2, 2 ± 2 and 4 ± 4; but the results of immune clearance are 3 ± 21, 289 ± 345, 405 ± 656, 2 ± 14, 8 ± 40 and 7 ± 30. The IFN-γ spots statistical analysis of PBMC were differently incubated by HBsAg and M, the total is P = 0.720, The IFN-γ spots statistical analysis of CD4^± T cell were differently incubated by HBsAg and M, the total is P = 0. 890, The IFN-γ spots statistical analysis of PBMC and CD4^± T cell were differently incubated by M, the total is P = 0.000. Conclusions The ability that CpG ODN can not significantly increase the IFN-γ secreting of PBMC and CD4^± T cell that were incubated by HBsAg to the infection in immune tolerant phase and in the immune clearance phase,but the PBMC outweighed The CD4^± T cell.