Influence of Yersinia pestis with different plasmid composition on the erythrocyte membrane in the blood of guinea pigs

S. Klyueva, S. Bugorkova, Pavel S. Erokhin, Anastasiya Yu. Goncharova, A. L. Kravtsov
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Abstract

Introduction. Based on data on the role of blood erythrocytes in the development and implementation of the vaccine and infectious processes in plague, it was of interest to evaluate changes in erythrocyte surface architecture from the position of searching for informative criteria for the preclinical evaluation of anti-plague vaccines. Aim — using atomic force microscopy to characterize the state of the blood erythrocyte membrane of guinea pigs in response to subcutaneous administration of the vaccine strain Yersinia pestis EV NIIEG and its isogenic derivatives. Materials and methods. For immunization of animals strain Y. pestis EV NIIEG (pYT+, pYV+, pYP+) and its isogenic derivatives Y. pestis KM216 (pYT–, pYV–, pYP+), Y. pestis KM217 (pYT–, pYV+, pYP–), Y. pestis KM218 (pYT–, pYV–, pYP–) were used. Analysis of the erythrocyte membrane was carried out using a Solver P47-PRO scanning probe microscope (NT-MDT, Russia). Results. The most pronounced changes in the surface architectonics of the membrane of guinea pig erythrocytes were established during the first three days of the formation of the immune response to the Y. pestis EV strain NIIEG and its isogenic variant Y. pestis KM217, in the genome of which the pYV plasmid is preserved, administered at a dose of 5 × 108 CFU. A significant increase (p 0.05) in the proportion of transformed cell forms (43.67 ± 3.63% and 37.83 ± 7.03% versus 4.08 ± 0.86% in the control group), root mean square roughness (319 ± 8 nm and 312 ± 7 nm versus 70 ± 6 nm in the control group), Young's modulus (125.73 ± 4.48 kPa and 113.8 ± 5.41 kPa versus 53.03 ± 1.47 kPa in the control group). By the 21st day, the value of these indicators decreased by an average of 2.7, 2.0 and 1.5 times, respectively, indicating restoration of the erythrocyte membrane. Conclusion. The dependence of the changes in the erythrocyte membrane and the rate of their restoration on the plasmid composition of Y. pestis strains has been established. The data obtained contribute to the understanding of the processes of interaction of Y. pestis with the erythrocyte membrane and can be used as additional characteristics in the development of new criteria for preclinical evaluation of plague candidate vaccines.
不同质粒组成的鼠疫耶尔森菌对豚鼠血液中红血球膜的影响
导言。根据有关血液红细胞在疫苗的开发和实施以及鼠疫感染过程中的作用的数据,从寻找抗鼠疫疫苗临床前评估的信息标准的角度出发,对红细胞表面结构的变化进行评估是很有意义的。目的--使用原子力显微镜描述豚鼠血红细胞膜对皮下注射鼠疫耶尔森菌 EV NIIEG 疫苗株及其同源衍生物的反应状态。材料和方法使用鼠疫耶尔森菌 EV NIIEG 株(pYT+, pYV+, pYP+)及其同源衍生物鼠疫耶尔森菌 KM216 株(pYT-, pYV-, pYP+)、鼠疫耶尔森菌 KM217 株(pYT-, pYV+, pYP-)、鼠疫耶尔森菌 KM218 株(pYT-, pYV-, pYP-)对动物进行免疫。使用 Solver P47-PRO 扫描探针显微镜(NT-MDT,俄罗斯)对红细胞膜进行了分析。结果显示豚鼠红细胞膜表面结构的最明显变化发生在对鼠疫伊蚊 EV 株 NIIEG 及其同源变种鼠疫伊蚊 KM217(其基因组中保留了 pYV 质粒)形成免疫反应的头三天,给药剂量为 5 × 108 CFU。转化细胞的比例(43.67 ± 3.63% 和 37.83 ± 7.03% 对对照组的 4.08 ± 0.86%)、均方根粗糙度(319 ± 8 nm 和 312 ± 7 nm 对对照组的 70 ± 6 nm)、杨氏模量(125.73 ± 4.48 kPa 和 113.8 ± 5.41 kPa 对对照组的 53.03 ± 1.47 kPa)均有明显增加(P 0.05)。到第 21 天,这些指标值分别平均下降了 2.7 倍、2.0 倍和 1.5 倍,表明红细胞膜已经恢复。结论红细胞膜的变化及其恢复速度与鼠疫伊蚊菌株的质粒组成有关。获得的数据有助于了解鼠疫酵母菌与红细胞膜的相互作用过程,并可作为开发鼠疫候选疫苗临床前评估新标准的补充特征。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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