Comparison of immune peripheral blood cells in tuberculin reactor cattle that are seropositive or seronegative for Mycobacterium bovis antigens

Abstract

Bovine tuberculosis (bTB) is a major economic problem in animal husbandry and is a public health risk in nonindustrialized countries. It is generally accepted that protection against TB is generated through cell-mediated immunity. Previous investigations have shown that WC1+ γδ, CD4+ and CD8+ T-cell subpopulations are important in the immune response to bTB. It is known that changes in the immune balance from a dominant T helper 1 (Th1)-type response toward a more prominent Th2 response may be observed during disease progression. In this study, we aimed to investigate immune peripheral blood cells in tuberculin reactor cattle that are seropositive or seronegative for Mycobacterium bovis antigens, using flow cytometry and hematological analysis. The evaluation of the T cell subpopulations revealed a decrease in CD8+ T cells of the seropositive and seronegative animals compared with the control animals (p=0.0001). Moreover, the seropositive group exhibited a lower percentage of CD8+ T cells than the seronegative group. The percentage of B cells was significantly increased in the seropositive group compared with the seronegative group and the control group (p=0.0009). No difference was observed in the percentage of WC1+ γδ and CD4+ T cells among the groups. Furthermore, following 24h of peripheral blood culture with bovine purified protein derivative (PPD), both apparently infected groups showed an increase in the levels of cellular activation compared with the control group (p<0.0001). The seropositive group displayed a higher level of cellular activation than the seronegative group. In both apparently infected groups, the hematological analysis showed an increase in total leukocyte (p=0.0012), lymphocyte (p=0.0057), monocyte (p=0.0010) and neutrophil (p=0.0320) counts in comparison with the healthy animals. Our results demonstrated differences in immune peripheral blood cells of tuberculin reactor cattle that are seropositive or seronegative for M. bovis antigens, probably due to different stages of bTB among the groups. The percentages of CD8+ T cells, B cells and the T cell activation levels may represent biomarkers for the progression of the disease. However, general characteristics shared by both apparently infected groups as lymphocytosis and monocytosis may also be indicative of the disease. Further experiments are required to understand the variations between cellular and humoral immunities throughout the course of bTB infection. A detailed knowledge of the peripheral blood cells involved in all stages of the bTB immune response of naturally infected cattle is essential for the optimal exploitation of diagnosis and vaccination models.