Tissue antigens were detected by the avidin-biotinim method. In the formulation of the immunohistochemical reaction, the universal peroxidase DAKO LSAB + kit (Dako Corporation, USA) was used.
Expression of the anti-apoptotic BCL-2 protein and pro-apoptotic VAC protein was assessed by the color intensity in points (0 — no reaction, 1 — weak reaction, 2 — moderate reaction, 3 — strong reaction). Further calculated the values of H-score expression by the formula
H-score=0*n%(0)+ 1*n%(1)+ 2*n%(2)+ 3*n%(3).
Statistical processing of the obtained results was carried out by methods of dispersive and correlation analysis using the software STATISTICA 13.0 (Dell StatSoft Inc., USA).
Results and discussion
In the study of lymphocyte subpopulations, it was found that the most characteristic phenomena of the change in the immunogram were an increase in the content of natural killer cells NK (CD56 +) to 13.2 ± 0.6% in patients of group III and up to 11.3 ± 0.8% - group I and 12 0 ± 0.7% - Group II, despite the fact that in the control this indicator did not exceed 9.4 ± 0.4% (p <0.05). This indicates the likelihood of anti-cytotoxic reactions.
An immunohistochemical study of female CIN II biopsy specimens was established (Table 1), dendritic cells that expressed CD1a (+), CD56 (+) and CD83 (+) antigens showed a high level of expression compared to the control (P <0.05 ).
The definition of the functional state of dendritic cells in patients at risk has a high sensitivity (0.88) and specificity (0.81), which corresponds to positive predictive value (PPV) 0.82, and can be recommended for assessing the prognosis of dysplastic changes in the cervix.
Table 1. Expression of markers of antigen of dendritic cells (H-score) in patients with cervical dysplasia.