PECULIARITIES OF IMMUNOGISTOCHEMICAL DETECTION OF MYCOBACTERIA ANTIGENS WITH DIFFERENT ANTIBODIES IN PULMONARY TISSUE OF HIV-INFECTED PATIENTS

Abstract

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.

Keywords

• mycobacterium tuberculosis
• polyclonal antibody to mbt
• immunohistochemistry
• hiv infection
• pulmonary tuberculosis.