A pilot study evaluating the utility of commercially available antibodies for flow cytometric analysis of Panthera species lymphocytes

• Published in: BMC veterinary research Vol. 14; no. 1; p. 410
• Main Authors: Sylvester, Tashnica Taime, Parsons, Sven David Charles, van Helden, Paul David, Miller, Michele Ann, Loxton, Andre Gareth
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 19-12-2018; BioMed Central; BMC
• Subjects: African lion; Animals; Antibodies; Antibodies - metabolism; Antigens; Blood; CD14 antigen; CD25 antigen; CD4 antigen; CD44 antigen; CD45 antigen; CD5 antigen; CD8 antigen; Cell surface; Centrifugation; Cloning; Cross-reactivity; Effector cells; Flow cytometery; Flow cytometry; Flow Cytometry - methods; Health aspects; Immune response; Immunoglobulins; Immunology; Immunophenotype; Infections; Laboratories; Lion; Lymphocytes; Lymphocytes - cytology; Lymphocytes T; Lysis; Medical research; Panthera; Panthera leo; Phenotypes; Pilot Projects; Reagents; Surface markers; T cell receptors; T-Lymphocytes - cytology; Time lag; Tuberculosis; Veterinary medicine; United States > US; Flow cytometery; Panthera leo; African lion; Immunophenotype
• ISSN: 1746-6148; 1746-6148
• DOI: 10.1186/s12917-018-1717-4

The immune response against tuberculosis in lions is still poorly defined and our understanding is hampered by the lack of lion specific reagents. The process for producing antibodies against a specific antigen is laborious and not available to many research laboratories. As the search for antibody cross-reactivity is an important strategy for immunological studies in veterinary medicine, we have investigated the use of commercially available antibodies to characterize T cell subsets in African lions (Panthera leo).
Commercially available antibodies were screened and investigated the influence of two different sample processing methods, as well as the effect of time delay on cell surface marker expression on lion lymphocytes. Using commercially available antibodies, we were able to identify CD4
, CD5
, CD8
, CD14
, CD25
, CD44
and CD45
T lymphocytes in samples obtained by density gradient centrifugation as well as red cell lysis of lion whole blood. Two distinct lymphocyte populations, which differed in size and phenotype, were observed in the samples processed by density gradient centrifugation.
Commercially available antibodies are able to differentiate between T lymphocyte subsets including immune effector cells in African lion whole blood, and possibly give insight into unique specie phenotypes.