Induction of delayed-type hypersensitivity and interferon-gamma to Candida albicans and anti-hen-egg white lysozyme antibody as phenotypic markers of enhanced bovine immune response

Delayed-type hypersensitivity (DTH) is a protective localized cell-mediated immune response (CMIR), primarily against intracellular pathogens. DTH is widely used in research to assess immune responsiveness and has been a valuable diagnostic test in commercial settings. In pigs and other species both antibody (AMIR) and CMIR have been considered as reliable phenotypic markers of selection programs for disease resistance. Therefore in cattle, it was also considered important to find antigen/adjuvant combinations capable of inducing AMIR and CMIR without interfering with diagnostic tests. The objectives of the present study were to evaluate the combined use of hen-egg white lysozyme (HEWL) and Candida albicans adjuvanted with Quil A in lactacting Holstein cows for the induction of anti-HEWL antibody, as well as DTH and IFN-γ to C. albicans as phenotypic markers of enhanced immune responsiveness. Thirty one lactating Holstein cows were immunized with HEWL to induce antibody responses and C. albicans to sensitize for DTH. Two test antigens, candin and C. albicans whole cell (CaWC), were used to induce the effector phase of DTH. PBS was used as the negative control. In addition, two different skin sites (neck versus tail) were tested to evaluate differences in skin site responsiveness. C. albicans-induced IFN-γ production, as an indicator of a type 1 response, was evaluated by ELISA. Microscopic evaluation of skin samples at DTH sites was performed in five randomly selected cows and these skin biopsies were scored based on inflammation and cell infiltration. Results demonstrated the presence of classical DTH response to C. albicans, in that DTH responses peaked at 24 h post-intradermal injections and cell infiltration was composed largely of mononuclear leukocytes, typical of DTH skin reactions in cattle. The only difference in test antigens was that DTH to candin showed a higher early response (6 h) than CaWC and a rapid decrease in inflammation from 24 to 48 h. The neck was significantly more sensitive than the tail skin-fold as a DTH test site. IFN-γ was detected on days 14 and 21 post-immunization in plasma from blood incubated with candin. Significant primary and secondary anti-HEWL antibodies were also detected, indicating that this combination of test antigens could be used as phenotypic markers of immune responsiveness in cattle.