SURFACE ANTIGENS OF C. DIPHTHERIAE OBTAINED BY PHYSICAL FACTORS AS A BIOLOGICAL PLATFORM FOR THE DEVELOPMENT OF COMBINED DIPHTHERIAL CANDIDATE-VACCINE

Yelyseyeva I. V., Babych E. M., Zhdamarova L. A., Belozersky V. I., Kolpak S. A.

MICROBIOLOGY

Scentific article

Diphtheria infection is currently recorded mainly in countries endemic in diphtheria. According to the WHO, in 2017, Nigeria (7616), India (5293), Indonesia (954), Venezuela (786), Nepal (728), Pakistan (560) reported the highest incidence of diphtheria. In all of these countries, there is a tendency to increase the incidence compared to 2016, especially in Pakistan – 46.7 times and in Venezuela – 25.4 times. Within them, diphtheria remains a serious health problem and is, as a result of intensive migration processes in the modern world, a potential risk for countries epidemiologically well-off with diphtheria. The last epidemic rise of diphtheria in the countries of Eastern Europe in the 90s of the last century was on the background of significant immunization rates and a high proportion of vaccinated among the sick. Clinically expressed and toxic forms of diphtheria have been recorded even in fully vaccinated people. Among the causes of the incidence of diphtheria in vaccinated children, scientists call the possible use of drugs with insufficient antigenic load, insufficient immunogenicity of drugs used, violation of immunization schemes, impairment of the immune status of children and a certain percentage of truly refractory children. The indicated epidemiological features of the rise of morbidity, as well as the existence of endemic diphtheria territories raised a number of questions about the effectiveness of vaccine prophylaxis of this dangerous infection and forcing scientists to continue to seek ways to improve it. Practical aspects of the development of a combined diphtheria vaccine based on bacterial diphtheria antigen obtained by physical factors were studied. The aim of the study was development of approaches to obtaining surface antigens of C. diphtheriae bacterial cells as a biological platform for the creation of a combined vaccine against diphtheria with a bacterial component by physical factors. The efficiency of various physical factors (ultrasound, narrow-band electromagnetic radiation of ultrahigh frequency, laser radiation) was evaluated for C. diphtheriae microbial cells according to the biochemical composition of the preparations obtained. Optimization of ultrasonic irradiation of biomass pathogen for obtaining surface bacterial antigens was carried out. Antigenic and adjuvant properties of experimental preparations were studied. Particular attention is paid to ensuring their specific safety, which is achieved by sequential purification of the microbial disintegration by physical methods (centrifugation, filtration, preparative liquid gel chromatography) or by adequate dilution. It is shown the principle opportunity of improving the diphtheria vaccine by combining the native purified diphtheria toxoid and adjuvant of bacterial origin made from the microbial mass of the toxigenic strain C. diphtheriae by various physical factors. Vaccination of animals with investigated diphtheria bacterial antigenic preparations of different degree of purification (disintegrate, soluble antigenic complexes, filtrate, gel-chromatographic fractions) together with native purified diphtheria toxoid leads to stimulation of humoral antitoxic immunity. Sequential purification of the microbial disintegrator by physical methods (centrifugation, filtration, preparative liquid gel chromatography), or adequate dilution of antigenic preparations, effectively eliminate the skin reactions to vaccination with experimental C. diphtheriae antigen preparations and preservation of adjuvant effect. Prepared C. diphtheriae surface antigen agents obtained by different physical factors are biological platforms for the further development of advanced combined diphtheria vaccines.

diphtheria vaccine, bacterial diphtheria antigen, physical factors, ultrasound, electromagnetic radiation of ultrahigh frequency, laser irradiation

1. WHO/Diphtheria reported cases. Last update: 15-July-2018 (data received as of 09-Jul-18).
2. Global Vaccine Action Plan 2011 – 2020. Immunization, Vaccines and Biologicals.
3. WHO data. WHO vaccine-preventable diseases: monitoring system. 2018 global summary. Coverage time series for Ukraine (UKR). Last updated 15-Jul-2018 (data as of 15-Jul-2018).
4. Chudnaya LM, Oksiyuk VG, Krasyuk LS, Moroz LV, Bryzhata SI, Skuratovskaya IM, i dr. Ehpidemiologicheskaya situaciya po difterii na Ukraine. Ehpidemiologiya i infekcionnye bolezni. 1999;1:10-2. [in Russian].
5. Kolesnikov MM, Petrusevich TV. Vivchennya mozhlivostі viniknennya timchasvoi sprijnyatlivostі do difterіjnogo toksinu u shcheplenih tvarin. Suchasnі іnfekcіi. 2002;4:69-71. [in Ukrainian].
6. Demіhovs’ka OV, Chudna LM. Epіdemіya difterіi v Ukraynі: pіdsumki і uzagal’nennya. Ukrayns’kij medichnij chasopis. 1999;3(12):56-8. [in Ukrainian].
7. Narkevich NI, Tymchakovskaya IM. Osobennosti raspostraneniya difterii na fone immunizacii detej. Zhurnal mikrobiologii, ehpidemiologi, immunologii. 1996;2:25-9. [in Russian].
8. Kharseeva GG, Alieva AA. Adhesion of Corynebacterium diphtheriae: the role of surface structures and formation mechanism. Zh. Mikrobiol. (Moscow). 2014;4:109-17.
9. Kawai T, Akira S. Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. Immunity. 2011;34(5):637-50.
10. Rusek P, Wala M, Druszczyńska M, Fol M. Infectious Agents as Stimuli of Trained Innate Immunity. Int J Mol Sci. 2018 Feb 3;19(2). pii: E456. DOI: 10.3390/ijms19020456
11. Netea MG, Joosten LA, Latz E, Mills KH, Natoli G, Stunnenberg HG, et al. Trained immunity: A program of innate immune memory in health and disease. Science. 2016 Apr;22(352):6284.
12. Locht C, Mielcarek N. Live attenuated vaccines against pertussis. Expert Rev Vaccines. 2014 Sep;13:1147.
13. Shmeleva EO, avtor; Shmeleva EO, patentoobladatel’. Sposob povysheniya nespecificheskoj rezistentnosti organizma i sposob polucheniya preparata dlya povysheniya nespecificheskoj rezistentnosti organizma. № 2019181 RU; 1994 Sent 15. [in Russian].
14. Isaenko EYu. Primenenie ul’trazvuka dlya dezitegracii mikrobnіh kletok. Annals of Mechnicov Institute. 2008;1. Dostupno: www.imiamn.org/ journal.htm 5 [in Russian].
15. Yelyseieva ІV, Doroshenko AO, Babich EM, Zhdamarova LA, Bіlozers’kij VІ, Gorbach TV. Viznachennya osoblivostej spektra fluorescencіi zrazkіv nativnogo ochishchenogo difterіjnogo anatoksinu. Zhurnal klinicheskih i ehksperimental’nyh medicinskih issledovanij. 2017;5(1):680-9. [in Ukrainian].
16. Babych YeM, Yelyseieva IV, Bilozerskyi VI, Zhdamarova LA, Isaienko OYu, Bobyrieva IV, Horbach TV, vynakhidnyky; Derzhavna ustanova «Instytut mikrobiolohiyi ta imunolohiyi im. I. I. Mechnykova NAMN Ukrayiny», patentovlasnyk. Sposib otrymannia bakteriinoho dyfteriinoho antyhenu. № 86891 UA. 2014 Sich 10. [in Ukrainian].
17. Ada G. The importance of vaccination. Front Biosci. 2007Jan;1(12):1278-90.

«Bulletin of problems biology and medicine» Issue 3 (145), 2018 year, 251-256 pages, index UDK 615.371:579.871.1:534.321.9:616-097

10.29254/2077-4214-2018-3-145-251-256