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重组CagA在BALB/c小鼠模型中调节Th1/Th2平衡。

Recombinant CagA Regulates Th1/Th2 Balance in a BALB/c Murine Model.

作者信息

Paydarnia Nafiseh, Mansoori Behzad, Esmaeili Davoud, Kazemi Tohid, Aghapour Mahyar, Hajiasgharzadeh Khalil, Alizadeh Nazila, Baradaran Behzad

机构信息

Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Tabriz University of Medical Sciences, International Branch (Aras), Tabriz, Iran.

出版信息

Adv Pharm Bull. 2020 Jun;10(2):264-270. doi: 10.34172/apb.2020.031. Epub 2020 Feb 18.

Abstract

is recognized as one of the prevalent causes of human gastricinfection. In the present study, the role of mixed immunization with lipopolysaccharide(LPS) and recombinant cytotoxin-associated gene A (rCagA) as a stimulator of host immuneresponses was determined. BALB/c mice were immunized with different formulations by the systemic administrationat 14-day intervals. The effects of the formulations plus CpG adjuvants were assessed before andpost-immunization in separated studies. Moreover, the expression of Th1/Th2 cytokines wasquantified in sera of immunized mice using reverse transcription polymerase chain reaction (RTPCR)test and the protein levels confirmed with enzyme linked immunosorbent assay (ELISA).Finally, the specific antibody levels in sera were studied by ELISA and the tendency of cellularresponse was examined by IgG1/IgG2a ratio. Data of Western blotting verified the presence of constructed protein. Analysisof lymphocyte proliferation showed that CpG-conjugated rCagA increases lymphocytesproliferation compared to the control group. Also, it was shown that formulations containing LPSand rCagA promote a Th1 response indicated by interferon-gamma expression and induced Th1/Th2 balance. Additionally, the specific IgG1, total IgG and IgG2a levels elevated in response toall treatments. Ultimately, the IgG2a/IgG1 ratio in the mice immunized with rCagA-containingformulations increased. These results indicated that rCagA protein carried with CpG adjuvant not onlymaintained its antigenicity throughout the experiment but also induced robust Th1-biasedimmune responses. Therefore, it holds promise for the production of an efficient vaccine against infection.

摘要

被认为是人类胃部感染的常见原因之一。在本研究中,确定了脂多糖(LPS)和重组细胞毒素相关基因A(rCagA)混合免疫作为宿主免疫反应刺激剂的作用。以14天的间隔通过全身给药对BALB/c小鼠进行不同配方的免疫。在单独的研究中,在免疫前后评估了配方加CpG佐剂的效果。此外,使用逆转录聚合酶链反应(RT-PCR)试验对免疫小鼠血清中Th1/Th2细胞因子的表达进行定量,并通过酶联免疫吸附测定(ELISA)确认蛋白质水平。最后,通过ELISA研究血清中的特异性抗体水平,并通过IgG1/IgG2a比率检查细胞反应趋势。蛋白质印迹数据证实了构建蛋白的存在。淋巴细胞增殖分析表明,与对照组相比,CpG偶联的rCagA增加了淋巴细胞增殖。此外,含有LPS和rCagA的配方促进了以干扰素-γ表达为指标的Th1反应,并诱导了Th1/Th2平衡。另外,所有处理均使特异性IgG1、总IgG和IgG2a水平升高。最终,用含rCagA配方免疫的小鼠中IgG2a/IgG1比率增加。这些结果表明,携带CpG佐剂的rCagA蛋白不仅在整个实验过程中保持其抗原性,而且还诱导了强烈的Th1偏向性免疫反应。因此,它有望用于生产针对感染的有效疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/67e8/7191242/02e2cf3040c6/apb-10-264-g001.jpg

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