[THE IDENTIFICATION OF SUB-POPULATIONS OF B-LYMPHOCYTES OF PERIPHERAL BLOOD USING TECHNIQUE OF FLOW CYTOFLUOROMETRY IN HEALTHY INDIVIDUALS AND PATIENTS WITH RHEUMATOID DISEASES].

Abstract The study was carried out to apply technique of flow cytofluorometry for immunofenotyping of sub-populations of B-cells of peripheral blood in healthy persons and patients with rheumatoid diseases. The samples included 27 healthy donors, 16 patients with rheumatoid arthritis and 9 patients with systemic lupus erythematosus. The peripheral blood of all participants was analyzed for relative number of CD19+B-cells, total population of memory B-cells (CD19+CD27+); unswitched (CD19+IgD+CD27+) and switched (CD19+IgD- CD27+) memory B-cells, naive (CD19+IgD+CD27-) and transitory (CD19+IgD+CD10+CD38++CD27-) B-cells, plasmablasts (CD19+CD38+++IgD-CD27+CD20+) and long-lived plasmatic cells (CD19+CD138+). The sub-populations of B-cells were identified by technique of multicolor flow cytofluorometry using panel of monoclonal antibodies to surface membrane markers of B-lymphocytes. In normal state, relative number (median-Me; interquartile range 25-75 percentiles) of CD19+B-lymphocytes amounted to 9.1 (6.6-11.6)%; CD19+CD27+memory B-cells - 2.2 (1.6-3.3)%; unswitched and switched memory B-cells - 10 (6.4-12.7)% and 17.7 (14.9-27.0)%; naive B-cells - 65.8 (55.1-73.4)%; transitory B-cells - 0.1 (0.1-0.3)%; plasmablasts - 7.0 (5.0-9.4)%. The long-lived plasmatic cells in peripheral blood were absent. In patients with rheumatoid arthritis percentage of content of total population of memory B-cells were lower than in donors (1.6; 1.00-2.3%; p = 0.038). Under systemic lupus erythematosus was detected decreasing of number of naive B-cells (40.2; 19.7-58.2) and increasing of level of switched memory B-cells (34.2; 21.0-52.7) as compared with donors (p = 0.003 in both cases). The study established no reliable differences between healthy donors and patients with rheumatoid arthritis and systemic lupus erythematosus in the rest of sub-populations of B-lymphocytes. The developed technique of multi-parametric flow cytofluorometry can be recommended for studying homeostasis of B-cells of peripheral blood in healthy persons and patients with auto-immune rheumatoid diseases and also for evaluating and forecasting effectiveness of anti-B-cells therapy.