Immunophenotypical characterization in Andalusian horse: Variations with age and gender

Assessment of lymphocyte subsets is an effective method for characterizing disorders such as leukemia, lymphomas, autoimmune and infectious diseases. In order to clinically interpret these parameters, normal reference values should be set, estimating age- and gender-related variations. This research aimed to: (1) characterize lymphocyte subpopulations in Andalusian horse, and (2) evaluate age and gender-related variations of lymphocyte subsets. Jugular blood samples were obtained from 159 animals, 77 males and 82 females, belonging to four age groups—1: 1–2 years ( N = 39; 21 males and 18 females), 2: 2–3 years ( N = 38; 16 males and 22 females), 3: 3–4 years ( N = 41; 19 males and 22 females) and 4: 4–7 years ( N = 41; 21 males and 20 females). T lymphocytes subsets were quantified by flow cytometry with monoclonal antibodies specific for CD2, CD4 and CD8 cell markers. B and NK cell counts were estimated by using a mathematical formula. No variations were found in T, B lymphocytes and NK cells between males and females. Animals of group 1 and 2 had a higher number of CD2, T, CD4+, CD8+, B lymphocytes and NK cells than animals of groups 3 and 4. The percentage of CD2 in group 1 was significantly lower than in group 4. The percentage of T and CD4+ lymphocytes in the group 1 were significantly higher than groups 2 and 3, respectively. Whereas the percentage of B cells calculated by flow cytometry was significantly lower in group 2 compared to group 4, the percentage of B cells calculated by a mathematical formula was higher in group 1. NK cells percentage was significantly lower in group 3 and 4 than in younger animals. In conclusion, in Andalusian horse, gender does not influence absolute numbers and percentages of T, B and NK. There is an age-related decline in absolute number of CD2, T, CD4+ and CD8+ lymphocytes, B lymphocytes and NK cells, with increasing percentage of CD2, T, CD4+ and B lymphocytes, and a decrease in NK with no differences in CD4/CD8 ratio. The decline of lymphocyte population numbers with age is a natural process in many animal species, and could be the origin for immune dysfunction observed in geriatric individuals.