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

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Abstract

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.

About the authors

S. E. Gostischeva

Stavropol Plague Control Research Institute

Author for correspondence.
Email: chumnpl@yandex.ru

Svetlana E. Gostischeva - Researcher, Scientific and Industrial Laboratory of Plague Vaccine.

355035, Stavropol, Sovietskya str., 13–15, Phone/fax: +7 (8652) 26-20-50 (office)

Russian Federation

N. V. Abzaeva

Stavropol Plague Control Research Institute

Email: fake@neicon.ru

PhD (Biology), Head of the Scientific and Industrial Laboratory of Plague Vaccine Research and Production

Russian Federation

D. A. Kovalev

Stavropol Plague Control Research Institute

Email: fake@neicon.ru
PhD (Chemistry), Head of the Laboratory of Biochemistry Russian Federation

D. G. Ponomarenko

Stavropol Plague Control Research Institute

Email: fake@neicon.ru

PhD (Biology), Head of the Laboratory of Brucellosis

Russian Federation

Yu. V. Siritsa

Stavropol Plague Control Research Institute

Email: fake@neicon.ru

Junior Researcher, Laboratory of Biochemistry

Russian Federation

E. L. Rakitina

Stavropol Plague Control Research Institute

Email: fake@neicon.ru

PhD (Medicine), Leading Researcher, Department of Clinical Immunology and Pathomorphology of Especially Dangerous Infectious Diseases, Laboratory of Brucellosis

Russian Federation

E. N. Afanasyev

Stavropol Plague Control Research Institute

Email: fake@neicon.ru

PhD, MD (Medicine), Chief Researcher, Scientific and Industrial Laboratory of Drugs for Diagnosis of Especially Dangerous Infectious Diseases

Russian Federation

M. V. Kostuchenko

Stavropol Plague Control Research Institute

Email: fake@neicon.ru

Researcher, Department of Clinical Immunology and Pathomorphology of Especially Dangerous Infectious Diseases, Laboratory of Brucellosis

Russian Federation

References

  1. Бахрах Е.Э., Тараненко Т.М. Изучение химического состава аллергена пестина ПП. Очистка пестина фильтрацией через гель сефадекса // Проблемы особо опасных инфекций. 1968. № 2 С. 146–153.
  2. Белобородов Р.А., Тараненко Т.М., Бахрах Е.Э., Муравьева Н.К., Дудкова В.К. Эффективность компонентов пестина ПП при определении корреляции аллергической реактивности и приобретенной резистентности к чуме // Проблемы особо опасных инфекций. 1974. № 6 (40). С. 51–54.
  3. Коробкова Е.И. Живая противочумная вакцина. М.: Медгиз, 1956. 205 с.
  4. Кравцов А.Л., Шмелькова Т.П., Щуковская Т.Н. Влияние противочумной вакцинации на функциональную активность клеток врожденного иммунитета человека // Проблемы особо опасных инфекций. 2011. № 107. С. 77–80.
  5. Медуницин Н.В. Вакцинология. М.: Триада-Х, 2010. 507 с. [Medunitsin N.V. Vaktsinologiya [Vaccination]. Moscow: Triada-X, 2010. 507 p.]
  6. Патент 2574207 Российская Федерация, МПК G01N33/48. Способ дифференциации поствакцинного и инфекционного бруцеллезного процессов по степени повышенной чувствительности организма к бруцеллам в условиях in vitro / Пономаренко Д.Г., Ракитина Е.Л., Логвиненко О.В., Саркисян Н.С., Костюченко М.В., Куличенко А.Н., Лямкин Г.И., Голубь О.Г., Бердникова Т.В. Заявитель и патентообладатель: ФКУЗ Ставропольский научно-исследовательский противочумный институт Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека; опубл. 10.02.2016.
  7. Тараненко Т.М. Изучение химического состава аллергена пестина ПП. Исследование пестина методом электрофореза // Проблемы особо опасных инфекций. 1968. № 2. С. 154–157.
  8. Alpenfels W.F. A rapid and sensitive method for the determination of monosaccharides as their dansyl hydrazones by high-performance liquid chromatography. Anal. Biochem., 1981, vol. 114, no. 1, pp. 153–157. doi: 10.1016/0003-2697(81)90466-8
  9. Avigad G. Dansyl hydrazine as a fluorimetric reagent for thin-layer chromatographic analysis of reducing sugars. J. Chromatogr. A, 1977, vol. 139, no. 2, pp. 343–347. doi: 10.1016/S0021-9673(00)89330-9
  10. Yuan X., Zeng Y., Nie K., Luo D., Wang Z. Extraction optimization, characterization and bioactivities of a major polysaccharide from Sargassum thunbergii. PloS ONE, 2015, vol. 10, no. 12:e0144773. doi: 10.1371/journal.pone.0144773

Copyright (c) 2018 Gostischeva S.E., Abzaeva N.V., Kovalev D.A., Ponomarenko D.G., Siritsa Y.V., Rakitina E.L., Afanasyev E.N., Kostuchenko M.V.

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