Stress-Related Immunomodulation of Canine Lymphocyte Responses and Hematologic Profiles

The immune status of dogs is shaped by continuous exposure to antigenic and various environmental stimuli, which together influence the development, regulation, and effectiveness of immune responses. Stress-related immune alterations may not be evident at the systemic level but can emerge at cellular and molecular scales. Therefore, this study aimed to comprehensively characterize the hematological and immunological profiles of dogs in different environments. We evaluated lymphocyte responses under basal conditions and following CD3/CD28-mediated in vitro activation, with subsequent long-term culture. Gene expression analyses targeted markers of early T cell activation, cytotoxic effector function, cytokine signaling, and inhibitory immune regulation. The memory phenotype of T lymphocytes was evaluated after blood collection and prolonged in vitro culture. In addition, hematological and biochemical profiles were assessed, including basic parameters, cortisol, and C-reactive protein. Our results revealed that client-owned dogs exhibited lower baseline expression of activation markers, especially in comparison with the short-term stay group, indicating an early immune activation state upon entry to the shelter environment. Furthermore, T lymphocytes from short- and long-term shelter dogs exhibited marked differences in the distribution of naïve and effector-memory subsets as well as different expansion capacity. These alterations persisted during prolonged in vitro culture, indicating that stress duration and environmental antigen exposure differentially shape immune responsiveness. In summary, chronic stress modulates canine immune status in a time-dependent manner, highlighting the importance of integrated cellular and molecular approaches in assessing the impact of environmental stressors on dogs' health and welfare.