References

mimmun

Медицинская иммунология

Medical Immunology (Russia)

1563-06252313-741X

SPb RAACI

10.15789/1563-0625-TGA-3138

mimmun-3138

Research Article

ОРИГИНАЛЬНЫЕ СТАТЬИ

ORIGINAL ARTICLES

Таргетная вакцинация против стрептококка группы A: рекомбинантная вакцина с фрагментами ScpA и SpeA

Targeting Group A Streptococcus with a Recombinant Chimeric Vaccine: Integrating ScpA and SpeA Fragments

Дуплик

Н. В.

Duplik

N. V.

Дуплик Н.В. – к.б.н., научный сотрудник отдела молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Duplik N.V., PhD (Biology), Researcher, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

nadezhdaduplik@gmail.com

Леонтьева

Г. Ф.

Leontieva

G. F.

Леонтьева Г.Ф. – к.б.н., ведущий научный сотрудник отдела молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Leontieva G.F., PhD (Biology), Leading Researcher, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

Крамская

Т. А.

Kramskaya

T. A.

Крамская Т.А. – старший научный сотрудник отдела молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Kramskaya T.A., Senior Researcher, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

Богатырева

К. П.

Bogatireva

K. P.

Богатырева К.П. – младший научный сотрудник отдела молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Bogatireva K.P., Junior Researcher, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

Гупалова

Т. В.

Gupalova

T. V.

Гупалова Т.В. – д.б.н., ведущий научный сотрудник отдела молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Gupalova T.V., PhD, MD (Biology), Leading Researcher, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

Бормотова

Е. А.

Bormotova

E. A.

Бормотова Е.А. – научный сотрудник отдела молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Bormotova E.A., Researcher, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

Королева

И. В.

Koroleva

I. V.

Королева И.В. – к.б.н., старший научный сотрудник отдела молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Koroleva I.V., PhD (Biology), Senior Researcher, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

Суворов

А. Н.

Suvorov

A. N.

Суворов А.Н. – д.м.н., профессор, член-корр. РАН, заведующий отделом молекулярной микробиологии

197376, Санкт-Петербург, ул. Акад. Павлова, 12.

Suvorov A.N., PhD, MD (Medicine), Professor, Corresponding Member, Russian Academy of Sciences, Нead, Molecular Microbiology Department

12 Acad. Pavlov St St. Petersburg 197376

ФГБНУ «Институт экспериментальной медицины»РоссияInstitute of Experimental MedicineRussian Federation

2025

15102025

2759851000

2025

Дуплик Н.В., Леонтьева Г.Ф., Крамская Т.А., Богатырева К.П., Гупалова Т.В., Бормотова Е.А., Королева И.В., Суворов А.Н.

Duplik N.V., Leontieva G.F., Kramskaya T.A., Bogatireva K.P., Gupalova T.V., Bormotova E.A., Koroleva I.V., Suvorov A.N.

Данная работа распространяется под лицензией Creative Commons Attribution 4.0.

This work is licensed under a Creative Commons Attribution 4.0 License.

https://www.mimmun.ru/mimmun/article/view/3138

Цель – разработка вакцины против стрептококков группы А (S. pyogenes, СГА) – возбудителя широкого спектра инфекций различной степени тяжести. Для клонирования генов рекомбинантных белков использовали экспрессионные вектора pET27 и pQE30. Для иммунизации использовали аффинно-очищенные полипептиды. Самок мышей иммунизировали дважды подкожно полипептидами в дозе 20 мкг/мышь с адъювантом Alum (2:1) с интервалом в 3 недели. Образцы иммунных сывороток исследовали методом ИФА. Для оценки специфической защитной эффективности иммунного ответа через 3 недели после последней инъекции мышей внутрибрюшинно заражали СГА M1 серотипа в дозе 5 × 107 КОЕ/мышь. Защитную эффективность вакцинации оценивали путем сравнения скорости бактериального клиренса у вакцинированных и контрольных животных, о чем судили по бактериальной нагрузке в селезенке через 3 и 15 часов после заражения. Конструкция вакцины представляет собой гибридную молекулу, состоящую из фрагментов, полученных из двух ключевых белков СГА: пептидазы C5a (ScpA) и экзотоксина SpeA. Используя биоинформатический анализ, мы идентифицировали и выбрали для включения в состав рекомбинантного белка Т- и В-клеточные эпитопы в консервативных регионах, общих для разных серотипов СГА. Посредством стратегии интеграции этих фрагментов предполагается усилить защиту от штаммов СГА, несущих экзотоксин, особенно тех, которые связаны с инвазивными инфекциями. Оценка иммуногенности на моделях мышей продемонстрировала сильный гуморальный иммунный ответ после парентеральной вакцинации, направленный на оба компонента гибридной молекулы. Последующая оценка защитной эффективности против внутрибрюшинного введения СГА выявила ускорение бактериального клиренса у вакцинированных животных, при этом фрагмент SpeA проявлял значительный защитный эффект. Представленные результаты демонстрируют потенциал полученной рекомбинантной химерной вакцины как многообещающего кандидата для разработки вакцины против стрептококков группы А, удовлетворяющей острую потребность в эффективных профилактических стратегиях против патологий, вызванных СГА.

Our objective was to develop a vaccine against group A streptococci (Streptococcus pyogenes, GAS), the causative agent of a broad spectrum of infections with varying severity. The expression vectors pET27 and pQE30 were used to clone genes encoding recombinant proteins, which were subsequently affinity-purified. Female mice were immunized subcutaneously twice with the purified polypeptides (20 μg/mouse) formulated with Alum adjuvant (2:1) at three-week intervals. Immune sera were analyzed using ELISA to evaluate antigen-specific responses. Three weeks after the final immunization, mice were challenged intraperitoneally with GAS M1 serotype at a dose of 5 × 107 CFU/mouse. Vaccination efficacy was determined by comparing bacterial clearance in vaccinated versus control animals, assessed by bacterial loads in the spleen at 3 and 15 hours post-infection. The vaccine candidate is a hybrid recombinant protein comprising fragments from two essential GAS virulence factors: C5a peptidase (ScpA) and SpeA exotoxin. T and B cell epitopes from conserved regions, common across multiple GAS serotypes, were identified and included into the construct using bioinformatics tools. The integration of these epitopes is designed to confer broad-spectrum protection against GAS strains carrying exotoxin, particularly those linked to invasive infections. Immunogenicity studies in mice revealed a robust humoral immune response targeting both components of the hybrid protein. Further evaluation of protective efficacy demonstrated accelerated bacterial clearance in vaccinated animals, with the SpeA fragment playing a significant protective role. These findings emphasize the potential of this recombinant chimeric vaccine as a promising candidate for the prevention of group A streptococcal infections, addressing the critical need for effective prophylactic strategies against GAS-associated diseases.

стрептококк группы АC5a-пептидазаэкзотоксин SpeAрекомбинантная химерная вакцинаиммуногенностьпротективность

group A StreptococcusC5a peptidaseexotoxin SpeArecombinant chimeric vaccineimmunogenicityprotectivity

The study was supported by the Ministry of Science and Higher Education of the Russian Federation (Project No. 122020300194-0).

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The authors declare that there are no conflicts of interest present.