Detection of equine and bovine T- and B-lymphocytes in formalin-fixed paraffin-embedded tissues

• Published in: Veterinary immunology and immunopathology Vol. 57; no. 3; pp. 187 - 200
• Main Authors: Collins Kelley, L., Mahaffey, E.A., Bounous, D.I., Antczak, D.F., Brooks, R.L.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-07-1997
• Subjects: animal health; Animals; B-Lymphocytes - chemistry; Bovine; Cattle - immunology; Cluster of differentiation; Equine; Formaldehyde; Horses - immunology; Humans; Immunohistochemistry; Leukocyte antigens; Lymph Nodes - chemistry; Lymphoid Tissue - chemistry; Lymphoid Tissue - cytology; Molecular Weight; Paraffin Embedding - veterinary; Peyer's Patches - chemistry; Spleen - chemistry; T-Lymphocytes - chemistry; Thymus Gland - chemistry; Tissue Fixation - veterinary; veterinary medicine; Western blotting; Immunohistochemistry; PBS; CD; NBT; Cluster of differentiation; SDS; Bovine; TBS; PAGE; Leukocyte antigens; EDTA; Western blotting; CHAPS; PMSF; BCIP; Equine; PBL
• ISSN: 0165-2427; 1873-2534
• DOI: 10.1016/S0165-2427(97)00006-8

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.