HUMORAL IMMUNE RESPONSE TO SHIGA TOXIN 2 (STX2) IN CHILDREN WITH ESCHERICHIOSIS WITH HEMOLYTIC-UREMIC SYNDROME



Cite item

Full Text

Abstract

Shiga toxin-producing Escherichia coli (STEC) causes acute intestinal infections and also causes acute renal failure, especially in children. Shiga toxins (Stx) occupy a central place in the pathogenesis of hemolytic uremic syndrome (HUS) in Escherichiosis. The presented work analyzes the effectiveness of laboratory diagnostics of enterohemorrhagic escherichiosis in patients at the stage of manifestation of HUS and/or acute renal failure using microbiological, immunological research methods and PCR analysis. The study used clinical material from 30 patients in the pediatric intensive care unit of the St. Vladimir Children's City Clinical Hospital in Moscow with symptoms of HUS aged from 8 months to 5 years. Blood sera from 20 healthy donors were used as control. As a result of PCR analysis, stx2 DNA was detected in 23.3% of cases. Bacteriological research made it possible to sow a pure culture of Escherichia coli O157:H7 in only 3.3% of cases. Since the development of HUS begins in patients with acute intestinal infection caused by Shiga toxin-producing microorganisms starting from the 5th day of the disease, when antibiotic therapy is already carried out, the bacteria can be completely destroyed, which makes it difficult to identify them by bacteriological methods, as well as to detect genes encoding Shiga toxin in PCR analysis. Typically, patients with HUS are admitted to the intensive care unit 5-7 days after the onset of the disease, when class G immunoglobulins specific to the pathogen are already beginning to circulate in the blood. In this regard, the use of immunological tests can be effective to confirm the diagnosis of STEC infection. In our studies, enzyme immunoassay allowed us to detect antibodies to Stx2A in 63.3%, and to Stx2B in 43.3% of patients. Using immunoblotting, antibodies to Stx2A were detected in all sera obtained from patients, and in 66.7% of cases to Stx2B. Immunoblot analysis was characterized by higher sensitivity for detecting antibodies to Stx2, however, due to the presence of an immunological layer among healthy people, it is preferable to use ELISA analysis. In healthy donors with antibodies to Stx2, the antibody titer was significantly lower than in patients. Thus, laboratory confirmation of the diagnosis of STEC infection is difficult when conducting microbiological and molecular genetic studies, which is confirmed in this work. The effectiveness of laboratory diagnostics can be expanded by performing an ELISA aimed at detecting antibodies to Stx2A.

About the authors

Maria A. Shkuratova

STATE RESEARCH CENTER FOR APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

Author for correspondence.
Email: shkuratova@obolensk.org
ORCID iD: 0000-0001-8021-8453
SPIN-code: 7112-9372

Junior Researcher, Laboratory of Molecular Biology

Russian Federation, SRCAMB Bld. 24, “Quarter A” Territory, 142279, Obolensk, City District Serpukhov, Moscow Region, Russian Federation

Anna E. Khlyntseva

STATE RESEARCH CENTER FOR APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

Email: khlyntseva_anna@mail.ru
ORCID iD: 0000-0003-0052-2602
SPIN-code: 3279-6161

PhD (Biological Science), Researcher, Laboratory of Molecular Biology

Russian Federation, SRCAMB Bld. 24, “Quarter A” Territory, 142279, Obolensk, City District Serpukhov, Moscow Region, Russian Federation

Olga V. Kalmantaeva

STATE RESEARCH CENTER FOR APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

Email: kalmantaevaov@yandex.ru
ORCID iD: 0000-0002-2838-6879
SPIN-code: 4657-5192

PhD (Biological Science), Researcher, Laboratory of Molecular Biology

Russian Federation, SRCAMB Bld. 24, “Quarter A” Territory, 142279, Obolensk, City District Serpukhov, Moscow Region, Russian Federation

Nikolay N. Kartsev

STATE RESEARCH CENTER FOR APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

Email: kartsev@obolensk.org
ORCID iD: 0000-0002-2006-9131
SPIN-code: 5463-3016

MD, PhD, Senior Researcher of Antimicrobial Agents Laboratory, Molecular Microbiology Department, 

Russian Federation, SRCAMB Bld. 24, “Quarter A” Territory, 142279, Obolensk, City District Serpukhov, Moscow Region, Russian Federation

Alexandr L. Muzurov Alexandr Lvovich

Moscow Clinical Municipal Children Hospital St. Vladimir;
Russian Medical Academy of Continuous Professional Education Ministry of Healthcare of Russia

Federal State Budgetary Educational Institution of Further Professional Education "Russian Medical Academy of Continuous Professional Education" of the Ministry of Healthcare of the Russian Federation

Email: al_muz@mail.ru
ORCID iD: 0000-0003-4131-9440
SPIN-code: 8489-9991

Ph. D. Med., Head of Department of Center of Gravitational Blood Surgery and Hemodialysis

Associate Professor, Department of Pediatric Anesthesiology, Critical Care Medicine and Toxicology

Russian Federation, 1/3 k1, Rubtsovsko-Dvortsovaya Street, Moscow, Russia, zipcode 107014 2/1, Barrikadnaya st., Moscow, 125993, Russia

Victoria V. Firstova

STATE RESEARCH CENTER FOR APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

Email: firstova@obolensk.org
ORCID iD: 0000-0002-9898-9894
SPIN-code: 9166-9151

PhD, DSc (Biological Science), Chief Researcher, Laboratory of Molecular Biology

Russian Federation, SRCAMB Bld. 24, “Quarter A” Territory, 142279, Obolensk, City District Serpukhov, Moscow Region, Russian Federation

References

  1. Малов В.А., Малеев В.В., Козловская Н.Л., Цветкова Н.А., Сметанина С.В., Горобченко А.Н., Серова В.В., Ченцов В.Б., Волков А.Г., Фаллер А.П. Трудности диагностики гемолитико-уремического синдрома, ассоциированного с диареей, у взрослых // Терапевтический архив. - 2017. Т. 89, №11, C. 69-78. [Malov V.A., Maleev V.V., Kozlovskaya N.L., Tsvetkova N.A., Smetanina S.V., Gorobchenko A.N., Serova V.V., Chentsov V.B., Volkov A.G., Faller A.P. Difficulties in the diagnosis of diarrhea-associated hemolytic uremic syndrome in adults // Terapevticheskii arkhiv. - 2017. - Vol. 89, no. 11, pp. 69-78.]
  2. Шкуратова М.А., Марьин М.А., Рогозин М.М., Сурин А.К., Коломбет Л.В., Фирстова В.В. Получение и масс-спектрометрическая характеристика рекомбинантных субъединиц шига-токсинов первого и второго типов (Stx1 и Stx2) Escherichia сoli. Бактериология. 2021. Т. 6, №4, С. 30-38, [Shkuratova M.A., Maryin M.A., Rogozin M.M., Surin A.K., Kolombet L.V., Firstova V.V. Preparation and mass-spectrometric characteristics of recombinant shiga-toxin units of the first and second types (Stx1 and Stx2) Escherichia coli. Bacteriology. 2021. Vol. 6, no. 4, pp. 30-38 (In Russ.)]
  3. Эмирова Х.М., Толстова Е.М., Каган М.Ю. и др. Гемолитико-уремический синдром, ассоциированный с шига-токсин-продуцирующей Escherichia coli. Нефрология. 2016. Т.20, №2, С. 18-32. [Emirova Kh., Tolstova E.M., Kagan O.M., Orlova M.Yu., Abaseeva T., Pancratenko T.E., Shpikalova I.Yu. Hemolytic uremic syndrome associated with shiga-toxin-producing Esherichia coli. Nephrology (Saint-Petersburg). 2016. Vol. 20, no. 2, pp. 18-32. (In Russ.)]
  4. Ariceta G. Hemolytic Uremic Syndrome. Current Treatment Options in Pediatrics. 2020. Vol. 6, no. 4, pp.252-262.
  5. Barrett TJ, Green JH, Griffin PM, Pavia AT, Ostroff SM, Wachsmuth LK: Enzyme linked immunosorbent assays for detecting antibodies to Shiga-like toxin I, Shiga-like toxin II, and Escherichia coli O157:H7 lipopolysaccharide in human serum. Curr Microbiol. 1991. Vol. 23, pp.189-195.
  6. Bruyand M., Mariani-Kurkdjian P., Le Hello S., King L.A., Van Cauteren D., Lefevre S., Gouali M., Jourdan-da Silva N., Mailles A., Donguy M.P., Loukiadis E., Sergentet-Thevenot D., Loirat C., Bonacorsi S., Weill F.X., De Valk H.; Réseau Français Hospitalier de Surveillance du Shu Pédiatrique. Paediatric haemolytic uraemic syndrome related to Shiga toxin-producing Escherichia coli, an overview of 10 years of surveillance in France, 2007 to 2016. Euro Surveill. 2019. Vol. 24, no.8, pp.1800068.
  7. Cai K, Gao X, Li T, Wang Q, Hou X, Tu W, Xiao L, Tian M, Liu Y, Wang H: Enhanced immunogenicity of a novel Stx2Am-Stx1B fusion protein in a mice model of enterohemorrhagic Escherichia coli O157:H7 infection. Vaccine. 2010. Vol. 29, no. 5, pp. 946-952.
  8. Joseph A, Cointe A, Mariani Kurkdjian P, Rafat C, Hertig A. Shiga Toxin-Associated Hemolytic Uremic Syndrome: A Narrative Review. Toxins (Basel). 2020. Vol. 12, no.2, pp. 67.
  9. Karmali M.A., Mascarenhas M., Petric M., Dutil L., Rahn K., Ludwig K., Arbus G.S., Michel P., Sherman P.M., Wilson J., Johnson R., Kaper J.B. Age-specific frequencies of antibodies to Escherichia coli verocytotoxins (Shiga toxins) 1 and 2 among urban and rural populations in southern Ontario. J Infect Dis. 2003, Vol. 188, no. 11, pp. 1724-1729.
  10. Majowicz S.E., Scallan E., Jones-Bitton A., Sargeant J.M., Stapleton J., Angulo F.J., Yeung D.H., Kirk M.D. Global incidence of human Shiga toxin-producing Escherichia coli infections and deaths: a systematic review and knowledge synthesis. Foodborne Pathog Dis. 2014. Vol. 11, no. 6, pp. 447-55.
  11. O'Brien A.D., Tesh V.L., Donohue-Rolfe A., Jackson M.P., Olsnes S., Sandvig K., Lindberg A.A., Keusch G.T. Shiga toxin: biochemistry, genetics, mode of action, and role in pathogenesis. Curr Top Microbiol Immunol. 1992. Vol. 180, pp. 65-94.

Supplementary files

Supplementary Files
Action
1. JATS XML

Copyright (c) Shkuratova M.A., Khlyntseva A.E., Kalmantaeva O.V., Kartsev N.N., Muzurov Alexandr Lvovich A.L., Firstova V.V.

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

СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: серия ПИ № 77 - 11525 от 04.01.2002.


This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies