伊朗肠毒血症疫苗的非动物试验:产气荚膜梭菌毒素的细胞培养试验。

IF 1.5 4区 生物学 Q4 CELL BIOLOGY
Anahita Emadi, Lida Abdolmohammadi Khiav, Sina Soleimani, Mohsen Lotfi, Faranak Abnaroodheleh, Maryam Dadar
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引用次数: 0

摘要

产气荚膜梭菌D型产生的Epsilon毒素是第三强的梭菌毒素。它会引起绵羊和羔羊的肠毒血症。梭状芽孢杆菌疫苗已被用于治疗梭状芽孢杆菌病,其有效性以血清中和(SN)试验为金标准进行评价。研究人员担心用体外试验取代体内试验。本研究的目的是评价细胞培养法检测产气荚膜梭菌毒素中和抗体的可行性。用纯化的epsilon毒素对Madin-Darby犬肾(MDCK)和非洲绿猴肾(Vero)细胞株进行显微镜检查和3-[4,5-imethylthiazol-2-yl]-2,5二苯基溴化四氮唑(MTT)染色,观察细胞反应。在细胞培养试验中计算抗体,并使用Pearson相关分析SN结果。根据我们的结果,只有MDCK对epsilon毒素敏感。细胞病变效应在这个细胞培养中是圆形的。毒素浓度与细胞活力的关系表明,毒素浓度的增加显著降低了细胞活力。SN与体外测定结果具有良好的相关系数(r = 0.987)
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Non-animal testing of Iranian enterotoxemia vaccine: cell culture assay for Clostridium perfringens epsilon toxin.

Epsilon toxin produced by Clostridium perfringens type D is the third most potent clostridial toxin. It causes enterotoxemia in sheep and lambs. The clostridial vaccine has been used against clostridial disease, and its efficacy is evaluated using the serum neutralization (SN) assay as a gold standard. Researchers are concerned about replacing in vivo tests with in vitro tests. Our study aimed to evaluate the cell culture assay to measure the neutralizing antibodies against Clostridium perfringens epsilon toxin as an alternative SN assay. Madin-Darby canine kidney (MDCK) and African green monkey kidney (Vero) cell lines were used to monitor the cell line response after treatment with purified epsilon toxin through microscopic examination and 3-[4,5-imethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) staining. Antibodies were calculated in cell culture assays, and SN results were analyzed using Pearson's correlation. Based on our results, only MDCK was sensitive to the epsilon toxin. The cytopathic effect in this cell culture was rounded. The relationship between toxin concentration and cell viability showed that increasing toxin concentrations significantly decreased cell viability. Good correlation coefficients were obtained between SN and the in vitro assay (r = 0.987) (p < 0.01). The antibody titers obtained by SN were within the range of the cytotoxicity assay and had high reproducibility. Therefore, cell culture may be a suitable alternative for SN assays. Cell culture is one of the tools used in toxicity testing, resulting in consistent and reproducible results.

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来源期刊
CiteScore
3.70
自引率
4.80%
发文量
96
审稿时长
3 months
期刊介绍: In Vitro Cellular & Developmental Biology - Animal is a journal of the Society for In Vitro Biology (SIVB). Original manuscripts reporting results of research in cellular, molecular, and developmental biology that employ or are relevant to organs, tissue, tumors, and cells in vitro will be considered for publication. Topics covered include: Biotechnology; Cell and Tissue Models; Cell Growth/Differentiation/Apoptosis; Cellular Pathology/Virology; Cytokines/Growth Factors/Adhesion Factors; Establishment of Cell Lines; Signal Transduction; Stem Cells; Toxicology/Chemical Carcinogenesis; Product Applications.
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