【摘要】：目的探討含非甲基化胞嘧啶和鳥(niǎo)嘌呤核苷酸序列的寡脫氧核糖核苷酸(CpG-ODN)對(duì)卡介苗(BCG)的免疫增強(qiáng)作用,為提高BCG的免疫效果提供實(shí)驗(yàn)依據(jù)。方法 (1)細(xì)胞實(shí)驗(yàn):分空白組、低濃度組和高濃度組,均接種小鼠巨噬細(xì)胞株Ana-1預(yù)培養(yǎng)。分別加入培養(yǎng)液、低濃度CpG-ODN和高濃度CpG-ODN,培養(yǎng)后測(cè)定上清液中IL-12、IL-18和IFN-γ含量。(2)動(dòng)物實(shí)驗(yàn):分對(duì)照組(A組)、感染組(B組)、BCG預(yù)防組(C組)和BCG+CpG-ODN預(yù)防組(D組)。A組常規(guī)飼養(yǎng),B、C、D組分別注射無(wú)菌PBS、BCG、BCG和CpG-ODN混合液,2周后B、C、D組注射結(jié)核分枝桿菌,再2周后處死所有小鼠,取肺組織病檢,收集血清測(cè)定IL-12、IL-18和IFN-γ含量。結(jié)果 (1)細(xì)胞實(shí)驗(yàn):CpG-ODN刺激培養(yǎng)下,Ana-1分泌IL-12、IL-18和IFN-γ均呈CpG-ODN劑量依賴性,加入CpG-ODN含量越高,Ana-1分泌上述三種Th1型細(xì)胞因子含量越多。(2)動(dòng)物實(shí)驗(yàn):A組肺組織未見(jiàn)炎癥細(xì)胞浸潤(rùn);B組肺組織可見(jiàn)大量炎癥細(xì)胞浸潤(rùn);C組和D組肺組織可見(jiàn)少量炎癥細(xì)胞浸潤(rùn)。B組小鼠血清中IL-12、IL-18和IFN-γ含量均顯著低于A組小鼠(均P0.05)。C組和D組小鼠血清中IL-12、IL-18和IFN-γ含量均顯著高于A組(均P0.05)。D組小鼠血清中IL-12、IL-18和IFN-γ含量均顯著高于C組(均P0.05)。結(jié)論 CpG-ODN能夠增強(qiáng)BCG的免疫預(yù)防效果,該免疫增強(qiáng)作用可能與CpG-ODN上調(diào)Th1型細(xì)胞因子含量有關(guān)。
[Abstract]:Aim to investigate the immunoenhancement effect of oligodeoxyribonucleotide (CpG-ODN) containing unmethylated cytosine and guanine nucleotide sequence on BCG (BCG) in order to provide experimental basis for improving the immune effect of BCG. Methods (1) Cell experiment: divided into three groups: blank group, low concentration group and high concentration group, all of them were inoculated with mouse macrophage cell line Ana-1 pre-culture. The contents of IL-12,IL-18 and IFN- 緯 in the supernatant were determined after culture with low concentration of CpG-ODN and high concentration of CpG-ODN, respectively. (2) Animal experiment: divided into control group (group A), infection group (group B), control group (group A) and infection group (group B). BCG prevention group (group C) and BCG CpG-ODN prevention group (group D,). A, B, C, D) were injected with aseptic PBS,BCG,BCG and CpG-ODN mixture respectively. Two weeks later, Mycobacterium tuberculosis was injected into group B, C and D, respectively. After 2 weeks, all the mice were killed for lung pathological examination and serum levels of IL-12,IL-18 and IFN- 緯 were determined. Results (1) Cell experiment: under the stimulation of CpG-ODN, Ana-1 secreted IL-12,IL-18 and IFN- 緯 in a dose-dependent manner, and the higher the content of CpG-ODN, the higher the content of CpG-ODN. Ana-1 secreted more Th1 cytokines. (2) Animal experiment: there was no infiltration of inflammatory cells in lung tissue in group A; In group B, a large number of inflammatory cell infiltration was observed in lung tissue. The levels of IL-12,IL-18 and IFN- 緯 in serum of mice in group B were significantly lower than those in group A (P0.05,). C and D, P 0.05), and the levels of IL-12, in serum of mice in group B were significantly lower than those in group A (P 0.05). The levels of IL-18 and IFN- 緯 in serum of group A were significantly higher than those of group A (P0.05). The levels of IL-12,IL-18 and IFN- 緯 in serum of). D group were significantly higher than those of group C (P0.05). Conclusion CpG-ODN can enhance the immunoprophylactic effect of BCG, which may be related to the up-regulation of Th1 cytokine content by CpG-ODN.
【作者單位】： 深圳市羅湖區(qū)慢性病防治院;深圳市羅湖區(qū)婦幼保健院;
【基金】：深圳市羅湖區(qū)科技計(jì)劃項(xiàng)目(編號(hào):2014040)
【分類號(hào)】：R392

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