Isolation and phenotypic characterization of abomasal mucosal lymphocytes in the course of a primary Ostertagia ostertagi infection in calves

Isolation and characterization of surface marker phenotypes of abomasal intraepithelial (IEL), lamina propria (LPL) and abomasal lymph node lymphocytes (ABLN) from uninfected calves were conducted, and the dynamics of change in these populations during the course of a primary Ostertagia ostertagi infection were defined. To obtain viable IEL and LPL from the abomasal mucosa of cattle, a modified isolation method was developed. The phenotypic characterization of abomasal lymphocytes was accomplished by indirect immunofluorescence staining. In uninfected animals, numbers of T cells exceeded the number of immunoglobulin-bearing cells in IEL, LPL and ABLN. The predominant T cell type in IEL and LPL was CD8+ cells, while the CD4+ T cell predominated in ABLN. Levels of activated cells and T cell receptor-1 γδ T cells were higher in IEL and LPL compared to ABLN. Within 3 weeks of infection, the number of lymphocytes recovered from the abomasal lamina propria and the mass of the ABLN was dramatically increased when compared to uninfected animals. Laser flow cytometric analysis demonstrated increased levels of immunoglobulin-bearing cells, γδ T cells, and activated T cells in IEL, LPL and ABLN in the infected animals. The greatest changes in LPL and ABLN took place during the first days of infection, and these changes were apparent throughout the 28 days covered by the experiment.