Detection of equine and bovine T- and B-lymphocytes in formalin-fixed paraffin-embedded tissues.
Formalin-fixed paraffin-embedded sections of equine and bovine lymph nodes, spleen, thymus, and Peyer's patches were incubated with monoclonal antibodies to B-lymphocyte markers BLA.36, B29, and mb-1 and T-lymphocyte markers CD3 and CD5. The monoclonal antibody BLA.36 reacted with 80-90% of lymphocytes in the germinal centers and mantle zones of follicles in lymph nodes, spleen, and Peyer's patches. In addition, 90% of lymphocytes in the marginal zone of the spleen, and variable numbers of lymphocytes within lymph node medullary cords were immunopositive for BLA.36. Antibodies to B29 and mb-1 produced similar staining patterns as BLA.36 with fewer positive cells in the germinal centers and medullary cords. BLA.36, B29, and mb-1 reacted with 30-50% of lymphocytes in the medulla of the thymus and with 5-10% of lymphocytes in the cortex. CD3 and CD5 reacted with 90% of lymphocytes in the paracortex and parafollicular zones of lymph nodes, spleen, and Peyer's patches; 40-50% of lymphocytes in the medullary cords of lymph nodes, and scattered positive cells within follicles. Anti-CD3 antibody reacted with 95% of lymphocytes in the splenic red pulp, but antibodies directed against CD5 reacted only faintly with approximately 5-10% of lymphocytes in the red pulp. CD3 and CD5 reacted with 50-60% of cells in the medulla of the thymus and with 40-80% of lymphocytes in the thymic cortex. The biochemical characterization of the antibodies by Western blotting against lysates of equine and bovine peripheral blood mononuclear cells confirmed that antibodies to BLA.36, mb-1, B29, CD3, and CD5 detected molecules of the same approximate molecular mass as found on lymphoid cells of human beings and rats.
Kelley
L C
LC
Pathology Division, Food Safety and Inspection Service, United States Department of Agriculture, Athens, GA 30604, USA.
Mahaffey
E A
EA
Bounous
D I
DI
Antczak
D F
DF
Brooks
R L
RL
D000818:Animals; D001402:B-Lymphocytes / Q000737:chemistry*; D002417:Cattle / Q000276:immunology*; D005557:Formaldehyde; D006736:Horses / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D008198:Lymph Nodes / Q000737:chemistry; D008221:Lymphoid Tissue / Q000737:chemistry* / Q000166:cytology; D008970:Molecular Weight; D016612:Paraffin Embedding / Q000662:veterinary*; D010581:Peyer's Patches / Q000737:chemistry; D013154:Spleen / Q000737:chemistry; D013601:T-Lymphocytes / Q000737:chemistry*; D013950:Thymus Gland / Q000737:chemistry; D016707:Tissue Fixation / Q000662:veterinary*
D005557:Formaldehyde