Mild heat stress induced adaptive immune response in blood mononuclear cells and leukocytes from mesenteric lymph nodes of primiparous lactating Holstein cows - Supplemental Material

Heat stress negatively affects the metabolism and physiology of the bovine gut. However, it is not known whether heat stress induces an inflammatory response in mesenteric lymph nodes (MLN), the primary origin of gut immune cells, thus contributes to inflammatory processes in the circulation. Therefore, our objective was to elucidate the effects of chronic heat stress on the systemic activation of acute-phase response in blood, pro-inflammatory cytokine production in peripheral blood mononuclear cells (PBMC) and the activation of the Toll-like receptor signaling (TLR) 2/4 pathway in MLN leucocytes and their chemokines and chemokine receptors profiles in Holstein cows. Primiparous Holstein cows (n = 30; 169 ± 48 DIM) were exposed to a temperature-humidity-index (THI) of 60 (16°C, 63% relative humidity (RH)) for 6 days. Thereafter, cows were evenly assigned to 3 groups: heat-stressed (HS; 28°C, 50% RH, THI = 76), control (CON; 16°C, 69% RH, THI = 60) or pair-feeding (PF; 16°C, 69% RH, THI = 60) group for 7 days. On day 6, PBMC were isolated and on day 7 MLN. Plasma haptoglobin, TNFα and IFNγ concentrations increased more in HS than CON cows. Concomitantly, <em>TNFA</em> mRNA abundance was higher in PBMC and MLN leucocytes of HS than PF cows, whilst <em>IFNG</em> mRNA abundance tended to be higher in MLN leucocytes of HS than PF cows, but not for chemokines (<em>CCL20</em>, <em>CCL25</em>) or chemokine receptors (<em>ITGB7</em>, <em>CCR6</em>, <em>CCR7</em>, <em>CCR9</em>). Furthermore, the TLR2 protein expression tended to be higher abundant in MLN leucocytes of HS than PF cows. These results suggest that heat stress induced an adaptive immune response in blood, PBMC and MLN leukocytes involving the acute-phase protein haptoglobin, pro-inflammatory cytokine production, and TLR2 signaling in MLN leucocytes. However, chemokines regulating the leucocyte trafficking between MLN and gut seem not to be involved in the adaptive immune response to heat stress.