Quantitative and quantitative analysis of spleen and bone marrow CD8+ and CD4+ T cell populations in lactating mice

Abstract

Purpose: to investigate the phenotype of adaptive immunity cells in the spleen and bone marrow of lactating mice.Materials and methods. The studies were carried out on lactating mice. For the experiment, the spleen and bone marrow were taken from animals. A suspension of individual splenocytes was prepared by grinding the spleen. Bone marrow was obtained by rinsing the medullary cavity with a syringe with 1–2 ml of a balanced salt solution. The phenotype of adaptive immunity cells was determined using a set of fluorochrome-conjugated antibodies: CD4-PerCP-Cy5.5, CD8-PE/Cy7, CD62L-APC/Cy7, CD44-BV510 (Biolegend, USA), in the presence of True Stain reagent containing antibodies to CD16/CD32 (Biolegend, USA) to block nonspecific antibody binding. Data collection was carried out on a CytoFlex flow cytometer (Beckman Coulter, USA). The results were analyzed using the Kaluza Analysis 2.1 program (Beckman Coulter, USA).Results. As a result of the study, it was found that there are 2.2 times more CD8+ T-cells of effector (TEM) and central memory (TCM) in the red bone marrow, while all subpopulations of CD4+ T-memory cells (TCM; TEM; TNV) predominate in the spleen . The content of the subpopulation of CD8+ T-cells of naive memory (TNV) in the bone marrow and spleen was almost the same and amounted to 52.57±1.58 % and 57.40±2.63 %, respectively. A significantly low content (p<0.001) of populations of CD8+ T-cells of effector memory (TEM; CD44+CD62L-) in the spleen was found to be 4.9±1.39 % compared with T-cells (TEM; CD44+CD62L-) of red bone marrow brain 11.04±2.58 %.Conclusion. The population of CD4+ T-cells of effector memory (TEM) accumulates in large numbers in the spleen in order to further respond with a cellular or humoral mechanism in response to the penetration of an antigen into the body. The bone marrow quantitatively and qualitatively surpasses the spleen in the accumulation and placement of effector (TEM) and central memory (TCM) CD8+ T cells, which are responsible for immunological memory and adaptive immune response.