Mikhaĭlova N A, Nagieva F G, Grin'ko O M, Zverev V V
Zh Mikrobiol Epidemiol Immunobiol. 2010 Mar-Apr(2):69-74.
To study antiviral activity of metabolites of spore-forming strain "Pashkov" of B. pumilus on the model of enterovirus infection in vitro.
B. pumilus strain "Pashkov" isolated from environment and identified by common methods. Cell cultures: Vero-6, Vero-ECC, and Vero-E6. Enteroviruses: type 1 poliovirus, Coxsackie B virus (1-6), ECHO-3, and ECHO-6 viruses. Unfectious activity of viruses was evaluated according to their cytopathogenic effect on Vero- E6 cell line by method of serial dilutions. Cultural fluid (CF) for the study was obtained by centrifugation and sterilizing filtration of B. pumilus strain "Pashkov" biomass produced by cultivation during 72 hours on optimized nutrient medium. Cytotoxicity of CF (chronic and acute) and maximal tolerated dose were measured by effect on viability of Vero-E6 cells, which was assessed by trypan blue exclusion test of cell viability. For measurement of antiviral activity (AV-activity), two treatment schedules--therapeutic and prophylactic--were used.
The most sensitive cell lines were Vero-ECC and Vero-E6. Assessment of AV-activity showed that protective effect was observed for all dilutions of CF and lasted for 7 days from time of infection by used doses of virus. CF does not have acute and chronic cytotoxicity. CF studied in vitro with Vero-E6 cells infected with 4 types of enteroviruses provided protection against viruses and had prophylactic effect. Degree of effect of CF depended from type of enterovirus, dose used and CF dilution.
For the first time effective antiviral activity of CF, which have low cytotoxicity for Vero-E6 cell culture in vitro and is produced by strain "Pashkov" of B. pumilus, was demonstrated. Obtained data open perspectives for development of medications against enterovirus infections.
在体外肠道病毒感染模型上研究短小芽孢杆菌“帕什科夫”芽孢形成菌株代谢产物的抗病毒活性。
从环境中分离出短小芽孢杆菌“帕什科夫”菌株并通过常规方法鉴定。细胞培养物：Vero-6、Vero-ECC和Vero-E6。肠道病毒：1型脊髓灰质炎病毒、柯萨奇B组病毒（1 - 6型）、埃可病毒3型和埃可病毒6型。通过连续稀释法，根据病毒对Vero-E6细胞系的细胞病变效应评估病毒的感染活性。用于研究的培养液（CF）是通过对短小芽孢杆菌“帕什科夫”菌株在优化营养培养基上培养72小时产生的生物质进行离心和无菌过滤获得的。通过对Vero-E6细胞活力的影响来测量CF的细胞毒性（慢性和急性）以及最大耐受剂量，细胞活力通过台盼蓝排斥试验进行评估。为测量抗病毒活性（AV活性），采用了两种处理方案——治疗性和预防性。
最敏感的细胞系是Vero-ECC和Vero-E6。AV活性评估表明，CF的所有稀释液均有保护作用，且从感染所用剂量病毒之时起持续7天。CF没有急性和慢性细胞毒性。在体外对感染4种肠道病毒的Vero-E6细胞进行研究时，CF提供了抗病毒保护并有预防作用。CF的作用程度取决于肠道病毒的类型、所用剂量和CF稀释度。
首次证明了CF具有有效的抗病毒活性，其对体外Vero-E6细胞培养物的细胞毒性较低，由短小芽孢杆菌“帕什科夫”菌株产生。所得数据为开发抗肠道病毒感染药物开辟了前景。
