Liskina I. V., Kuzovkova S. D., Zagaba L. M., Melnik O. A.

CLINICAL AND EXPERIMENTAL MEDICINE

Scentific article

Despite the high efficiency of microbiological and immunological methods for the diagnosis of tuberculosis in patients with co-infection TB-HIV, morphological diagnosis remains relevant in clinical practice. In recent decades, immunohistochemical methods for detecting M. tuberculosis in affected body tissues have been actively developed and applied mainly for scientific research. The aim of the study was to analyze the features of the immunomorphological detection of antigens and whole structures of the M. tuberculosis by different antibodies in the lung tissue during co-infection with TB-HIV. Twenty eight specimens of biopsy (20 cases) and autopsy (8 cases) material were selected. In all cases diagnosis of HIV infection in association with various forms of pulmonary tuberculosis was previously verified. An immunohistochemical study of lung tissue was performed to detect mycobacterial antigens using two polyclonal antibodies: M. tuberculosis antibody PA1-7231 and M. tuberculosis antibody with a fluorescent label PA1- 73134 (Thermo Fisher Scientific, USA). Immunohistochemical treatment was performed automatically using AUTOSTAINER 360-2D system and reagents manufactured by Thermo Fisher Scientific (USA) with UltraVision Quanto imaging system. Microscopy of samples was performed on an Olympus BX51 microscope with an Olympus U-RFL-T fluorescence attachment at a wavelength range of exciting light of 461-489 nm and emitting light of 497-532.5 nm, working magnification x400. It was found that an antibody with a fluorescent label allows for making a relative quantitative assessment of mycobacteria in lung tissue and to assess the presence of mycobacterial polymorphism. Another polyclonal antibody allows the maximum detection of immunopositive material in the form of granules in the cells of various structures of the lung tissue and extracellularly. The simultaneous use of two antibodies expands the possibilities of immunohistochemical analysis.

mycobacterium tuberculosis, polyclonal antibody to MBT, immunohistochemistry, HIV infection, pulmonary tuberculosis.

1. Lawn SD, Zumla AI. Tuberculosis. Lancet. [Internet] 2011 Mar 18 [cited 2020 Jun 3];378:57-72. DOI: 10.1016/S0140-6736(10)62173-3
2. Parhomenko YuG, Zyuzya YuR, Mazus AI. Morfologicheskie aspekty VICH-infekcii. M.: Litterra; 2016. 168 s. [in Russian].
3. Zimina VN, Kravchenko AV, Viktorov IB. Epidemiologiya, techenie i osobennosti lecheniya tuberkul’oza u bol’nyh VICH-infekciej. Medicina v Kuzbasse. 2011 Okt;10(3):6-13. [in Russian].
4. Cinzerling VA, Svistunov VV. Tuberkulez v sochetanii s VICH-infekciej: kliniko-morfologicheskie aspekty. Tuberkulez i bolezni legkih. 2014 Jun;6:56-60. [in Russian].
5. Gupta S, Shenoy VP, Bairy I, Muralidharan S. Comparison among three cold staining methods in the primary diagnosis of tuberculosis: a pilot study. J. Bras. Pneumol. 2010 Sep-Oct;36(5):612-6. DOI: 10.1590/s1806-37132010000500013
6. Ulrichs T, Lefmann M, Reich M, Morawietz L, Roth A, Brinkmann V, et al. Modified immunohistological staining allows detection of ZiehlNeelsen negative Mycobacterium tuberculosis organisms and their precise localization in human tissue. J. Pathol. 2005 Mar;205:633-40. doi: 10.1002/path.1728
7. Ellinidi VN, Ariel’ BM, Samusenko IA, Tugolukova LV. Immunogistohimicheskij metod v diagnostike tuberkuleza. Arh. patologii. 2007 Maj;5:36-7. [in Russian].
8. Dvorakovskaya IV, Majskaya MYu, Nasyrov RA, Baranova OP, Ariel’ BM. Morfologicheskoe issledovanie v differencial’noj diagnostike tuberkuleza i sarkoidoza. Arh. patologii. 2014 Yanv;1:27-31. [in Russian].
9. Karimi S, Shamaei M, Pourabdollah M, Sadr M, Karbasi M, Kiani A, et al. Histopathological findings in immunohistological staining of the granulomatous tissue reaction associated with tuberculosis [Internet]. Tuberculosis Research and Treatment. 2014 Jan. Available from: http://dx.doi.org/
10. 1155/2014/858396 10.Ihama Y, Hokama A, Hibiya K, Kishimoto K. Diagnosis of intestinal tuberculosis using a monoclonal antibody to Mycobacterium tuberculosis. World J. Gastroenterol. 2012 Dec 21;18(47):6974-80. DOI: 10.3748/wjg.v18.i47.6974
11. Cinzerling VA, Agapov MM. Sovremennye podhody k morfologicheskoj diagnostike tuberkuleza. Tuberkulez i bolezni legkih. 2017 Fevr;95(2):7-12. [in Russian].
12. Dorozhkova IR. Vozbuditel’ tuberkuleza: istoriya otkrytiya i izucheniya. Tuberkulez i bolezni legkih. 2012 Maj 03;3:3-14. [in Russian].
13. Kuzovkova SD, Liskina IV, Zagaba LM, Mel`ny`k OO. Diagnosty`chni mozhly`vosti gistologichnogo vy`yavlennya mikobakterial`noyi infekciyi v legenyakh pry` VIL-infekciyi. V materiali mizhnarodnoyi nauk.-prakt. konf. Suchasny`j vy`mir medy`chnoyi nauky` ta prakty`ky`; 2020 Maj 8-9; Dnipro. Organizaciya naukovy`kh medy`chny`kh doslidzhen` Salutem. 2020. s. 68-71. [in Ukrainian].

«Bulletin of problems biology and medicine» Issue 3 (157), 2020 year, 113-117 pages, index UDK 616.24-002.5:576.858:612.017.1:576.852.211:616-097-07

10.29254/2077-4214-2020-3-157-113-117