OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO

The formation of immunity against infectious diseases is accompanied by an immunoallergic alteration of the organism, while the intensity of the allergic reaction is associated with the presence of specific immunity. Skin allergotesting is often used to determine the intensity of sensitization of th...

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Autores principales:	S. E. Gostischeva, N. V. Abzaeva, D. A. Kovalev, D. G. Ponomarenko, Yu. V. Siritsa, E. L. Rakitina, E. N. Afanasyev, M. V. Kostuchenko
Formato:	article
Lenguaje:	RU
Publicado:	Sankt-Peterburg : NIIÈM imeni Pastera 2018
Materias:	allergen pestine pp аntiplague immunity vaccination spectrophotometry flow cytometry chromatographic analysis Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/91f72717b34f4c689940f6d7bffdeb4a
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spelling	oai:doaj.org-article:91f72717b34f4c689940f6d7bffdeb4a2021-11-22T07:09:49ZOPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO2220-76192313-739810.15789/2220-7619-2018-1-85-90https://doaj.org/article/91f72717b34f4c689940f6d7bffdeb4a2018-05-01T00:00:00Zhttps://www.iimmun.ru/iimm/article/view/677https://doaj.org/toc/2220-7619https://doaj.org/toc/2313-7398The formation of immunity against infectious diseases is accompanied by an immunoallergic alteration of the organism, while the intensity of the allergic reaction is associated with the presence of specific immunity. Skin allergotesting is often used to determine the intensity of sensitization of the body. When determining the immunity of vaccines against the plague, previously an allergen was suggested as a pestin PP — a polypeptide polysaccharide complex of a plague microbe. The authors optimized the technique for obtaining the preparation of pestin with preservation of its chemical composition, high specificity and allergenic activity. It is known that a lack of allergic test in vivo is a high risk of formation of adverse reactions. A method for estimating adaptive antiplague immunity with the allergen pestin in antigen-specific cellular tests in vitro is proposed. The preparation of the allergen by a modified procedure was carried out by hydrolysis of the biomass of the vaccine strain of the plague microbe Yersinia pestis EV of the NIIEG line, followed by filtration and lyophilization of the precipitate. In the preparation obtained, the pH and the protein concentration were determined. To check the specificity, the samples were subjected to spectrophotometric and chromatographic analysis. To assess specific activity, blood samples of 17 people immunized with the plague live vaccine from the Yersinia pestis EV strain of the NIIEG line were used for epidemic indications. As a comparative control, a sterile isotonic sodium chloride solution was used. The contingent was examined before vaccination on days 7, 21 and 3 months after immunization by evaluating the expression intensity of basophils CD63. Biochemical analysis of the obtained by the modified procedure of the pestin and derivatives allowed to judge the qualitative composition, to show the absence of impurities of the protein nature, as well as to determine the carbohydrate profile. The use of the drug as an allergen to assess the formation of antiplague immunity in the vaccinated contingent confirmed its specificity. The obtained data showed the possibility and prospect of using the Pestin PP as a test allergen for the establishment of the reaction of activation of basophils in vitro.S. E. GostischevaN. V. AbzaevaD. A. KovalevD. G. PonomarenkoYu. V. SiritsaE. L. RakitinaE. N. AfanasyevM. V. KostuchenkoSankt-Peterburg : NIIÈM imeni Pasteraarticleallergen pestine ppаntiplague immunityvaccinationspectrophotometryflow cytometrychromatographic analysisInfectious and parasitic diseasesRC109-216RUInfekciâ i Immunitet, Vol 8, Iss 1, Pp 85-90 (2018)
institution	DOAJ
collection	DOAJ
language	RU
topic	allergen pestine pp аntiplague immunity vaccination spectrophotometry flow cytometry chromatographic analysis Infectious and parasitic diseases RC109-216
spellingShingle	allergen pestine pp аntiplague immunity vaccination spectrophotometry flow cytometry chromatographic analysis Infectious and parasitic diseases RC109-216 S. E. Gostischeva N. V. Abzaeva D. A. Kovalev D. G. Ponomarenko Yu. V. Siritsa E. L. Rakitina E. N. Afanasyev M. V. Kostuchenko OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO
description	The formation of immunity against infectious diseases is accompanied by an immunoallergic alteration of the organism, while the intensity of the allergic reaction is associated with the presence of specific immunity. Skin allergotesting is often used to determine the intensity of sensitization of the body. When determining the immunity of vaccines against the plague, previously an allergen was suggested as a pestin PP — a polypeptide polysaccharide complex of a plague microbe. The authors optimized the technique for obtaining the preparation of pestin with preservation of its chemical composition, high specificity and allergenic activity. It is known that a lack of allergic test in vivo is a high risk of formation of adverse reactions. A method for estimating adaptive antiplague immunity with the allergen pestin in antigen-specific cellular tests in vitro is proposed. The preparation of the allergen by a modified procedure was carried out by hydrolysis of the biomass of the vaccine strain of the plague microbe Yersinia pestis EV of the NIIEG line, followed by filtration and lyophilization of the precipitate. In the preparation obtained, the pH and the protein concentration were determined. To check the specificity, the samples were subjected to spectrophotometric and chromatographic analysis. To assess specific activity, blood samples of 17 people immunized with the plague live vaccine from the Yersinia pestis EV strain of the NIIEG line were used for epidemic indications. As a comparative control, a sterile isotonic sodium chloride solution was used. The contingent was examined before vaccination on days 7, 21 and 3 months after immunization by evaluating the expression intensity of basophils CD63. Biochemical analysis of the obtained by the modified procedure of the pestin and derivatives allowed to judge the qualitative composition, to show the absence of impurities of the protein nature, as well as to determine the carbohydrate profile. The use of the drug as an allergen to assess the formation of antiplague immunity in the vaccinated contingent confirmed its specificity. The obtained data showed the possibility and prospect of using the Pestin PP as a test allergen for the establishment of the reaction of activation of basophils in vitro.
format	article
author	S. E. Gostischeva N. V. Abzaeva D. A. Kovalev D. G. Ponomarenko Yu. V. Siritsa E. L. Rakitina E. N. Afanasyev M. V. Kostuchenko
author_facet	S. E. Gostischeva N. V. Abzaeva D. A. Kovalev D. G. Ponomarenko Yu. V. Siritsa E. L. Rakitina E. N. Afanasyev M. V. Kostuchenko
author_sort	S. E. Gostischeva
title	OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO
title_short	OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO
title_full	OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO
title_fullStr	OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO
title_full_unstemmed	OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO
title_sort	optimization of the method of obtaining pestine pp and studying its specific activity in vitro
publisher	Sankt-Peterburg : NIIÈM imeni Pastera
publishDate	2018
url	https://doaj.org/article/91f72717b34f4c689940f6d7bffdeb4a
work_keys_str_mv	AT segostischeva optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro AT nvabzaeva optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro AT dakovalev optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro AT dgponomarenko optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro AT yuvsiritsa optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro AT elrakitina optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro AT enafanasyev optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro AT mvkostuchenko optimizationofthemethodofobtainingpestineppandstudyingitsspecificactivityinvitro
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OPTIMIZATION OF THE METHOD OF OBTAINING PESTINE PP AND STUDYING ITS SPECIFIC ACTIVITY IN VITRO

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