Cattle vary in their susceptibility to infection and immunopathology, but our ability to measure and longitudinally profile immune response variation is limited by the lack of standardized immune phenotyping assays for high-throughput analysis. Here we report longitudinal innate immune response profiles in cattle using a low-blood volume, whole blood stimulation system—the ImmunoChek (IChek) assay. By minimizing cell manipulation, our standardized system minimizes the potential for artefactual results and enables repeatable temporal comparative analysis in cattle. IChek successfully captured biological variation in innate cytokine (IL-1β and IL-6) and chemokine (IL-8) responses to 24-hr stimulation with either Gram-negative (LPS), Gram-positive (PamCSK4) bacterial or viral (R848) pathogen-associated molecular patterns (PAMPs) across a 4-month time window. Significant and repeatable patterns of inter-individual variation in cytokine and chemokine responses, as well as consistent high innate immune responder individuals were identified at both baseline and induced levels. Correlation coefficients between immune response read-outs (IL-1β, IL-6 and IL-8) varied according to PAMP. Strong significant positive correlations were observed between circulating monocytes and IL-6 levels for null and induced responses (0.49–0.61) and between neutrophils and cytokine responses to R848 (0.38–0.47). The standardized assay facilitates high-throughput bovine innate immune response profiling to identify phenotypes associated with disease susceptibility and responses to vaccination.

Introduction: Early detection of vascular alterations associated with disease can play a key role in diagnosis and evaluation of treatment strategies. Ultrasound is commonly used for real-time assessment of human blood vessels but the efficacy of preclinical ultrasound at providing a quantitative assessment of mouse models of vascular disease is relatively unknown. In this study, preclinical ultrasound was used in combination with a semi-automatic image processing method to track arterial distension alterations in mouse models of abdominal aortic aneurysm (AAA) and atherosclerosis. Methods: Longitudinal B-mode ultrasound images of the abdominal aorta were acquired using a preclinical ultrasound scanner. Arterial distension was assessed using a semi-automatic image processing algorithm to track vessel wall motion over the cardiac cycle and a standard, manual analysis method was applied for comparison. Results: Mean arterial distension was significantly lower in AAA mice between day 0 and day 7 post-onset of disease (p<0.01) and between day 0 and day 14 (p<0.001), while no difference was observed in sham control mice. The manual method detected a significant decrease (p<0.05) between day 0 and day 14 only. Atherosclerotic mice showed alterations in arterial distension relating to genetic modification and diet. Arterial distension was significantly lower (p<0.05) in Ldlr-/-(++/-) mice fed high fat western diet when compared with both wild type (++/++) mice and Ldlr-/-(++/-) mice fed chow diet. The manual method did not detect a significant difference between these groups. Conclusions: Arterial distension can be used as an early marker for the detection of arterial disease in small animal models. The semi-automatic analysis method provided increased sensitivity to differences between experimental groups when compared to the manual analysis method. Furthermore, a retrospective power calculation revealed that use of the semi-automatic method reduces the number of animals required per experiment.

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