IMMUNOCOMPETENT CELLS AND LOCAL IMMUNOREGULATORY INDEX IN TISSUE LESIONS AT PULMONARY TUBERCULOMA WITH ACTIVITY OF A SPECIFIC PROCESS
DOI:
https://doi.org/10.11603/1811-2471.2019.v0.i1.10055Keywords:
pulmonary tuberculoma, antigen presenting cells, mycobacterial antigens, immunohistochemistryAbstract
Currently, tuberculomas remain a common form of pulmonary tuberculosis. The progression of disease may indicate about inadequate control of tuberculosis infection in the affected organ at the local level with the possible absence of any systemic changes in the immune status. That's why the study of local immune responses in lesions, in particular in the lung tissue, is of principal importance.
The aim – to establish local patterns of localization and a relative number of antigen-presenting cells, cells with M. tuberculosis antigens and the value of the local CD4/CD8 ratio in the lung tissue with active tuberculomas.
Material and Methods. An immunohistochemical study of serial sections of lung tissue on the material of 19 lung resectates from patients with pulmonary tuberculoma and morphological signs of the activity of a specific tuberculous process was conducted.
Fragments of lung tissue with mycobacteria and/or their antigens, namely, the granulation layer of the capsule of tuberculoma, tuberculosis foci and granulomas were investigated. The relative amount of CD4 + and CD8 + lymphocytes, CD68 + cells and cells with the presence of M. Tuberculosis antigens was determined.
An immunohistochemical study was performed on AUTOSTAINER 360-2D manufactured by Thermo Fisher Scientific (USA), and the Ultra Vision Quanto HRP DAB system was used to visualize of cells. The following monoclonal antibodies by Thermo Fisher Scientific were used: mouse CD4 Clon 4B12, rabbit CD8 Clon SP16, mouse CD68 Clon KR1 and polyclonal rabbit Mycobacterium tuberculosis antibody PA1-7231 (Pierce Biotechnology, USA).
Statistical data processing was carried out using licensed software products that are included in the package of Microsoft Office Professional 2000 (Excel).
Results. In the lung tissue samples the tuberculoma capsule was presented in all cases, granulomas outside of the tuberculoma were defined in 13 (68.4 %) cases, and tuberculous foci – in 10 (52.6 %) cases. CD68 + cells and macrophages with the presence of MBT antigens (infected) were detected in 100 % of cases, but the number of these cells was very varied in different parts of the tissue.
CD8 + lymphocytes were identified in 100.0 % of cases in the granulation layer of tuberculoma and tuberculosis foci, and in 90.9 % in granulomas. CD4 + cells were observed in 100.0 % of cases in tuberculous foci, in 89.5 % of cases in the granulation layer of TB, and in 72.7 % of cases in granulomas. The relative number of these cells showed significant distinctions in both individual histological structures and in individual observations.
Calculated values of local CD4/CD8 ratio at active pulmonary tuberculomas, were within the limits of their physiological variations (1.1 – 1.61).
Conclusions. In zones of lung tissue with tuberculoma, where there is a contact of antigen-presenting cells with mycobacteria of tuberculosis, the greatest number of macrophages with different morpho-functional properties are localized in the granulation layer of tuberculoma, while CD4 + and CD8 + cells show significant heterogeneity of quantitative distribution regardless of the number of infected macrophages.
Outside of the tuberculoma in lung parenchyma granulomas of various cell types are determined – mainly epithelioid-lymphoid, but there are also macrophage and giant cell domination. Most of them have multifunctional macrophages, and the number of CD4 + and CD8 + lymphocytes varies greatly, which leads to the different values of the CD4/CD8 ratio.
The smallest relative number of CD68 + cells that are simultaneously infected with MBT is observed in tuberculosis foci, and the number of CD4 + and CD8 + cells varies. The CD4/CD8 ratio in these areas of the tissue indicates the prepotency of a cytotoxic immune response.
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