Инфекция и иммунитет. 2013; 3: 235-242
ПОИСК ОПТИМАЛЬНОЙ КОМБИНАЦИИ АНТИГЕНОВ ДЛЯ СЕРОДИАГНОСТИКИ ТУБЕРКУЛЕЗА
https://doi.org/10.15789/2220-7619-2013-3-235-242Аннотация
Резюме. Методами генной инженерии получено четыре химерных рекомбинантных антигена: CBD-CFP10, CBD-ESAT6, ESAT6-CFP10 и CBD-P38, содержащих аминокислотные последовательности полноразмерных белков ESAT6 и CFP10 и зрелого белка P38 M. tuberculosis, состыкованные с аминокислотной последовательностью целлюлозосвязывающего домена эндоглюконазы А (CBD) из Cellumonas fimi. Рекомбинантные белки были очищены методом аффинной хроматографии на колонке с Ni-NTA-сефарозе 6В-CL и, наряду с PPDN-3, были использованы для определения их антигенной активности в непрямом варианте ИФА при серодиагностике ТБ. В работе исследовались сыворотки, полученные от больных туберкулезом легких (n = 321), от лиц, имеющих профессиональный контакт с больными ТБ (n = 42), от здоровых доноров (n = 366) и от больных заболеваниями легких нетуберкулезной этиологии (n = 68). Было выявлено наличие достоверной положительной корреляции между уровнями антител против всех исследованных антигенов, сравниваемых попарно. Показано, что антигены существенно отличаются по своим антигенным и иммунобиологическим свойствам, но дополняют друг друга в составе антигенных диагностических композиций и могут найти применение в тест-системах для серодиагностики ТБ. Использование данных антигенов позволит выявлять лиц, инфицированных ТБ, а также выявлять больных с активным ТБ.
Список литературы
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Russian Journal of Infection and Immunity. 2013; 3: 235-242
THE SEARCH OF OPTIMAL COMBINATION OF ANTIGENS FOR SEROLOGICAL DIAGNOSTICS OF TUBERCULOSIS
https://doi.org/10.15789/2220-7619-2013-3-235-242Abstract
Abstract. The four chimeric recombinant antigens CBD-CFP10, CBD-ESAT6, ESAT6-CFP10 and CBD-P38 contained aminoacid sequences of full-size proteins ESAT6, CFP10 and matured protein P38 of M. tuberculosis, joined with aminoacid sequences of cellulose bind domain of endogluconase A (CBD) from Cellumonas fimi have been obtained by gene engineering methods. Recombinant proteins were purified by affine chromatography in column with Ni-NTA-sepharose 6В-CL and as PPDN-3 were used for detection of their antigenic activity in indirect ELISA for TB serological diagnostics. The sera from patients with lung tuberculosis (n = 321), from persons who had professional contacts with TB patients (n = 42), from healthy blood donors (n = 366) and from patients with lung diseases of non-TB etiology were tested. It was detected that there was positive correlation between antibodies level for all studied antigens compared by pair. It has been demonstrated that although antigens were different by antigenic and immunobiological characteristics they add each other in the content of antigenic diagnostics compositions. Thus, all these antigens can be used in the test kits for serological diagnostics of TB. Using of these antigens will allow to detect persons infected by TB and patients with active tuberculosis.
References
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6. Chegou N.N., Hoek K.G., Kriel M., Warren R.M., Victor T.C., Walzl G. Tuberculosis assays: past, present and future. Expert Rev. Anti Infect. Ther., 2011, vol. 9, no. 4, pp. 457–469.
7. Cole S.T., Brosch R., Parkhill J. Deciphering the biology of Mycobacterium Tuberculosis from the complete genome sequence. Nature, 1998, vol. 393, no. 6685, pp. 537–544.
8. Davidow A., Kanaujia G.V., Shi L., Kaviar J., Guo X., Sung N., Kaplan G., Menzies D., Gennaro M.L. Antibody profiles characteristic of Mycobacterium tuberculosis infection state. Infect. Immun., 2005, vol. 73, no. 10, pp. 6846–6851.
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10. Hatfield G. W., Roth D.A. Optimizing scaleup yield for protein production: Computationally Optimized DNA Assembly [CODA] and Translation Engineering. Biotechnology Annual Review, 2007, vol. 13, pp. 27–42.
11. He X.Y., Li J., Hao J., Chen H.B., Zhao Y.Z., Huang X.Y., He K., Xiao L., Ye L.P., Qu Y.M., Ge L.H. Assessment of five antigens from Mycobacterium tuberculosis for serodiagnosis of tuberculosis. Clin. Vaccine Immunol., 2011, vol. 18, no. 4, pp. 565–570.
12. Manca C., Lyashchenko K., Wiker H.G., Usai D., Colangeli R., Gennaro M.L. Molecular cloning, purification, and serological characterization of MPT63, a novel antigen secreted by Mycobacterium tuberculosis. Infect. Immun., 1997, vol. 65, no. 1, pp. 16–23.
13. Min F., Zhang Y., Huang R., Li W., Wu Y., Pan J., Zhao W., Liu X. Serum antibody responses to 10 Mycobacterium tuberculosis proteins, purified protein derivative, and old tuberculin in natural and experimental tuberculosis in rhesus monkeys. Clin. Vaccine Immunol., 2011, vol. 18, no. 12, pp. 2154–2160.
14. Pinto L.M., Grenier J., Schumacher S.G., Denkinger C.M., Steingart K.R., Pai M. Immunodiagnosis of tuberculosis: state of the art. Med. Princ. Pract., 2012, vol. 21, no. 1, pp. 4–13.
15. Randall L.L., Hardy S. J. S. Correlation of competence for export with lack of tertiary structure of the mature species: a study in vivo of maltose-binding protein in E. coli. Cell, 1986, vol. 46, pp. 921–928.
16. Singh S., Singh J., Kumar S., Gopinath K., Balooni V., Singh N., Mani K. Poor performance of serological tests in the diagnosis of pulmonary tuberculosis: evidence from a contact tracing field study. PLoS One, 2012, vol. 7, no. 7. Available at: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3393741, free access (accessed: 14.03.13).
17. Steingart K.R., Dendukuri N., Henry M., Schiller I., Nahid P., Hopewell P.C., Ramsay A., Pai M., Laal S. Performance of purified antigens for serodiagnosis of pulmonary tuberculosis: a meta-analysis. Clin. Vaccine Immunol., 2009, vol. 16, no. 2, pp. 260–276.
18. Steingart K.R., Flores L.L., Dendukuri N., Schiller I., Laal S., Ramsay A., Hopewell P.C., Pai M. Commercial serological tests for the diagnosis of active pulmonary and extrapulmonary tuberculosis: an updated systematic review and meta-analysis. PLoS Med., 2011, vol. 8, no. 8. Available at: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3153457, free access (accessed: 14.03.13).
19. Wallis R.S., Pai M., Menzies D., Doherty T.M., Walzl G., Perkins M.D., Zumla A. Biomarkers and diagnostics for tuberculosis: progress, needs, and translation into practice. Lancet, 2010, vol. 375, iss. 9729, pp. 1920–1937.
20. Walzl G., Ronacher K., Hanekom W., Scriba T.J., Zumla A. Immunological biomarkers of tuberculosis. Nat. Rev. Immunol., 2011, vol. 11, no. 5, pp. 343–354.
21. Weldingh K., Rosenkrands I., Okkels L.M., Doherty T.M., Andersen P. Assessing the serodiagnostic potential of 35 Mycobacterium tuberculosis proteins and identification of four novel serological antigens. J. Clin. Microbiol., 2005, vol. 43, no. 1, pp. 57–65.
22. Wu X., Yang Y., Zhang J., Li B., Liang Y., Zhang C., Dong M. Comparision of antibody responses to seventeen antigens from Mycobacterium tuberculosis. Clinica Shim. Acta., 2010, vol. 411, iss. 19–20, pp. 1520–1528.
23. Wu X.,Yang Y., Zhang J., Li B., Liang Y., Zhang C., Dong M., Cheng H., He J. Humoral immune responses against the Mycobacterium tuberculosis 38-kilodalton, MTB48, and CFP-10/ESAT-6 antigens in tuberculosis. Clin. Vaccine Immunol., 2010, vol. 17, no. 3, pp. 372–375.
24. Xu J.N., Chen J.P., Chen D.L.Serodiagnosis efficacy and immunogenicity of the fusion protein of Mycobacterium tuberculosis composed of the 10-kilodalton culture filtrate protein, ESAT-6, and the extracellular domain fragment of PPE68. Clin. Vaccine Immunol., 2012, vol. 19, no. 4, pp. 536–544.
25. Zhu C., Liu J., Ling Y., Yang H., Liu Z., Zheng R., Qin L., Hu Z. Evaluation of the clinical value of ELISA based on MPT64 antibody aptamer for serological diagnosis of pulmonary tuberculosis. BMC Infect. Dis., 2012, vol. 12: 96. Available at: http://www.biomedcentral.com/1471–2334/12/96, free access (accessed: 14.03.13).
