2 In Vitro Analysis: 2.1 Isolation and Preparation of Lymphocytes from Infected Animals

Abstract

Publisher Summary This chapter describes the isolation and preparation of lymphocytes from infected animals. Lymphocytes are the cells that provide specificity to host defense. Identifying the phenotype and antigen specificity of lymphocytes that have been isolated from animals infected with microbial agents is integral to understanding protective immunity. There are various methods of isolating, purifying, and characterizing lymphocytes from the tissues of infected animals. The unperturbed peritoneal cavity contains a mixed population of mononuclear cells. Neutrophils and eosinophils should be present in very low numbers. Mast cells are a minority population, but are conspicuous because of their large basophilic granules. Lymph node cell suspensions contain B and T lymphocytes, as well as macrophages, dendritic cells, and stromal cells. The numbers of lymphocytes recovered from lymph nodes depend on which nodes are collected and whether there has been recent antigen stimulation. Mononuclear phagocytes obtained from different tissue sites are all adherent, but can differ in how strongly they adhere. Cells activated at the sites of inflammation in vivo are more adherent than resident cells.