Subpopulation Composition of Peripheral Blood Lymphocytes in Uveal Melanoma

Abstract

Uveal melanoma is a malignant neuroectodermal tumor. Despite of radical primary treatment with the use of enucleation of the eye or radiation therapy, almost in 50% of patients develop metastatic disease. The effectiveness of various methods of treatment of malignant tumors, the prognosis of the disease largely depend on the state of the patient's immune system. Data on the ratio of lymphocytes of different populations, their relationship with the stages of uveal melanoma are currently not available. 84 patients with uveal melanoma were examined (44 women and 40 men; mean age 53.7 ± 12.2 years). In group 1 (small tumors) included 36 patients, in the 2nd (medium-sized tumors) - 26 patients, 22 patients made up the 3rd group (large tumors). The control group included 33 practically healthy donors. The material of the study was samples of whole blood taken from the ulnar vein on an empty stomach in the morning with the help of vacuum systems in test tubes Vacuette® with anticoagulant K3EDTA. Immunophenotyping was performed using flow laser cytofluorimetry using the Multitest 6-Color TBNK Reagent monoclonal antibody system in BD TruСount test tubes (Becton Dickinson, USA), BD FACSCanto II cytometer (Becton Dickinson, USA). Erythrocyte lysis and leukocyte fixation were performed using a BD FACS ™ Lysing Solution lysing solution (Becton Dickinson, USA). The relative and absolute levels of lymphocyte populations and subpopulations were determined in the Canto program (Becton Dickinson, USA), with the selection of the analyzed region in the general population expressing CD45 + antigen and cell granularity (CD45 + PerCP-Cy5,5 * / SSC); fluorochromes labeled antibodies to CD3 + (FITC), CD4 + (PE-Cy7 *), CD8 + (APC-CY7 *), CD16 + / 56 + (PE), CD19 + (APC *) were used to differentiate: T-lymphocytes (CD3 +), T-helper cells (CD3 + CD4 + CD8-), T-cytotoxic (CD3 + CD4-CD8 +), T-double positive (CD3 + CD4 + CD8 +), NK-cells (CD16 + CD56 +), B-lymphocytes (CD19 +) , calculate the ratio of CD3 + CD4 + / CD3 + CD8 + subpopulations - the index reflecting the balance of T-helper cells and cytotoxic T-cells (CD4 + / CD8 +). Our individual analysis of the composition of lymphocytes allows us to conclude that the growth of uveal melanoma is accompanied by systemic multidirectional changes in the qualitative and quantitative composition of immunocompetent cells, affecting both innate (NK cells) and adaptive (T-lymphocytes) links of antitumor immune defense. The results are important for the development of personalized approaches to the prognosis and treatment of patients with uveal melanoma. The dynamics of changes in the quantitative composition of lymphocyte subpopulations in oncological diseases may be of value for monitoring immunotherapeutic effects and supplement the clinical assessment of the course of the underlying disease.