Адгезивность Поринов Ompf И Ompс Yersinia Pseudotuberculosis К Макрофагам J774

Byvalov, A. A.; университет, Вятский государственный; Konyshev, I. V.; Novikova, O. D.; Портнягина, О. Ю.; Belozerov, Vladislav S.; Khomenko, V. A.; Давыдова, В. Н.; Ран, Институт физиологии Коми Нц УрО; Ран, Тихоокеанский институт биоорганической химии Дво

doi:10.1134/s0006302918050101

Cited by 3 publications

(4 citation statements)

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“…До использования в эксперименте препараты поринов хранили при температуре 4-6°С в фосфатном буферном растворе (ФБР), рН 7,2-7,4, содержащем 0,1% SDS. Cенсибилизацию микросфер препаратами поринов, а также валидацию факта указанной сенсибилизации проводили по методикам, описанным в работе [1].…”

Section: материалы и методыunclassified

“…Мышиные антисыворотки к поринам получали, как описано в работе [1]. Титры антител в полученных антипориновых сыворотках определяли с помощью метода непрямого твердофазного иммуноферментного анализа (ТИФА), используя микропланшеты Costar (США).…”

Section: материалы и методыunclassified

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Immunochemical activity of <i>Yersinia pseudotuberculosis</i> ompF and ompC porins evaluated by optical trapping

Konyshev¹,

Novikova²,

Portnyagina³

et al. 2022

Russian Journal of Infection and Immunity

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Introduction. Study of features for interacting antigen-antibody system is of great importance for developing new modern tools for diagnostics and therapy of infectious diseases. In this regard, it is of great interest to study the rupture force between bacterial antigens and antibodies using modern biophysical methods including optical trapping. The importance of surface antigens in the immunochemical activity of Yersinia pseudotuberculosis assessed by such method has not been evaluated yet. In this work we examined an opportunity to evaluate the interaction of hydrophobic Y. pseudotuberculosis porins OmpF and OmpC with specific antibodies using optical trapping method. Materials and methods. Polystyrene microspheres (d = 1 m) were coated by passive adhesion with purified preparations of OmpF and OmpC porins; microsphere sensitization was verified by enzyme immunoassay. Antibodies from mouse sera were adsorbed onto the glass surface by chemical linking. The rupture force in the porins-antibodies system was determined using a laser trap according to the previously developed algorithm. Results. Using a model system including polystyrene microspheres sensitized with the proteins and aminated glass substrate coated with immune or nonimmune serum, significant differences in binding strength of OmpF and OmpC porins to homologous immune versus nonimmune sera were detected. The average forces of interaction with immune sera was 60 pN for OmpF microspheres (control 40 pN) and 69 pN for OmpC microspheres (control 49 pN). The proportion of irreversible substrate binding of the microspheres coated by the antigens to the treated with immune vs. non-immune sera was significantly higher. The results of assessing the average interaction force, as well as the predominance of the proportion of irreversible binding of antigen-coated microspheres with sera-treated substrates, indicates that specific interactions contribute significantly to the force of interaction. The aforementioned method can be used to evaluate the forces of intermolecular interaction in similar model systems using other microbial antigens.

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Section: материалы и методыunclassified

Immunochemical activity of <i>Yersinia pseudotuberculosis</i> ompF and ompC porins evaluated by optical trapping

Konyshev¹,

Novikova²,

Portnyagina³

et al. 2022

Russian Journal of Infection and Immunity

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“…С помощью метода оптической ловушки получены экспериментальные данные, свидетельствующие о способности порина OmpF, продуцируемого клетками Y. pseudotuberculosis при низкой температуре культивирования, связываться с мембраной макрофагов J774. Порин OmpC, экспрессируемый этими бактериями при температуре 37°С, указанным свойством не обладает [3].…”

Section: иные белковые адгезиныunclassified

“…Вышеизложенное позволяет предположить, что в организм человека в естественных условиях per os могут попасть бактерии этого возбудителя, экспрессирующие в функционально значимом количестве только два адгезина -InvA и «холодовой» ЛПС. Роль порина OmpF, продуцируемого клетками Y. pseudotuberculosis при низких значениях температуры, в адгезии микроба к эукариотическим клеткам, предположительно, относительно мала, учитывая его меньшую удельную адгезивность в модельных экспериментах [3], а также существенно более низкое содержание в наружной мембране по сравнению с ЛПС. Ни малая продолжительность периода прохождения инфекта через верхние отделы пищеварительного тракта до своих входных ворот -эпителия тонкого кишечника, ни соответствующие неблагоприятные внешние условия на этом пути, по-видимому, не позволяют микробным клеткам не только пролиферировать, но и изменять свои поверхностные структуры до фенотипа 37-градусных, «тепловых» культур возбудителя, способных к продукции иных адгезинов.…”

Section: адгезивность Y Pseudotuberculosis в патогенезе заболеванияunclassified

Yersinia pseudotuberculosis-derived adhesins

Byvalov

Konyshev

2019

Russian Journal of Infection and Immunity

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Around fifteen surface components referred to adhesins have been identified in Yersinia pseudotuberculosis combining primarily microbiological, molecular and genetic, as well as immunochemical and biophysical methods. Y. pseudotuberculosis-derived adhesins vary in structure and chemical composition but they are mainly presented by protein molecules. Some of them were shown to participate not only in adhesive but in other pathogen-related physiological functions in the host-parasite interplay. Adhesins can mediate bacterial adhesion to eukaryotic cell either directly or via the extracellular matrix components. These adhesion molecules are encoded by chromosomal DNA excepting YadA protein which gene is located in the calcium-dependence plasmid pYV common for pathogenic yersisniae. An optimum temperature for adhesin biosynthesis is located close to the body temperature of warm-blooded animals; however, at low temperature only invasin InvA, full-length smooth lipopolysaccharide and porin OmpF are produced in Y. pseudotuberculosis. Several adhesins (Psa, InvA) can be expressed at low pH (corresponds to intracellular content), thereby defining pathogenic yersiniae as facultative intracellular parasites. Three human Yersinia genus pathogens differ by ability to produce adhesins. Y. pseudotuberculosis adherence to host cells or extracellular matrix components is determined by a cumulative adhesion-based activity, which expression depends on chemical composition and physicochemical environmental conditions. It’s proposed that at the initial stage of infectious process adherence of Y. pseudotuberculosis to intestinal epithelium is mediated by InvA protein and “smooth” LPS form. These adhesins are produced in bacterial cells at low (lower than 30°С) temperature occurring in environment from which a pathogen invades into the host. At later stages of pathogenesis, after penetrating through intestinal epithelium, bacterial cells produce other adhesins, which promote survival and dissemination primarily into the mesenteric lymph nodes and, possibly, liver and spleen. At later stages of pathogenesis, after penetrating through intestinal epithelium, bacterial cells produce other adhesins, which promote survival and dissemination primarily into the mesenteric lymph nodes and, perhaps, liver and spleen. Qualitative and quantitative spectrum of Y. pseudotuberculosis adhesins is determined by environmental parameters (intercellular space, intracellular content within the diverse eukaryotic cells).

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