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Updated: Sep 27, 2025
Enumeration of Major Peripheral Blood Leukocyte Populations for Multicenter Clinical Trials Using a Whole Blood Phenotyping Assay
Published on: September 16, 2012
Flow cytometry panels for immunophenotyping dairy cattle peripheral blood leukocytes
S Casaro1, M G Marrero2, D M C Madrid2
1Department of Large Animal Clinical Sciences, University of Florida, Gainesville, FL, USA.
Insights
Bovine leukocyte subsets differ between calves and cows, impacting dairy cattle health. Understanding these immune differences is key to addressing cattle diseases.
Area of Science:
- Veterinary Immunology
- Dairy Cattle Health
- Leukocyte Phenotyping
Background:
- The bovine immune system's complexity hinders dairy cow health management.
- Characterizing leukocyte subpopulations is crucial for understanding immune responses in cattle.
Purpose of the Study:
- To phenotype leukocyte subpopulations in Holstein calves and cows using flow cytometry.
- To identify differences in immune cell profiles between young and mature dairy cattle.
Main Methods:
- Development and application of two flow cytometry panels with specific antibody markers (CD8α, CD4, TCR-δ, CD172α, CD14, MHCII, CD21, CD62L, CD11b).
- Analysis of peripheral blood leukocytes from 30-day-old calves and cows at 260 days of gestation and calving.
Main Results:
- Calf leukocytes showed higher proportions of CD3+ lymphocytes, γδ T-cells, CD8+ γδ T-cells, and monocytes compared to cows.
- Cow lymphocytes had a greater proportion of CD4+ and CD8+ T-cells, and B-cells than calves.
- Differences in CD62L, CD11b, and MHCII expression were observed between calf and cow leukocyte subsets.
Conclusions:
- Significant differences exist in peripheral blood leukocyte profiles between Holstein calves and cows.
- These findings provide a foundational resource for understanding bovine immunity and developing strategies against dairy cattle diseases.
Abstract:
Many aspects of the bovine immune system remain poorly characterized, which poses an obstacle to improving dairy cow health. Herein, we describe two flow cytometry panels that included antibodies against CD8α, CD4, TCR-δ, CD172α, CD14, MHCII, CD21, CD62L, and CD11b. These panels were used to characterize the phenotype of leukocyte subpopulations from the peripheral blood of 30-day old Holstein calves and Holstein cows at 260 d of gestation and calving. No leukocyte subset differences were found between the pre- and post-partum cows. However, calf leukocytes presented a higher proportion of CD3+ lymphocytes, γδ T-cells, CD8+ γδ T-cells, and monocytes when compared with mature cows. Conversely, cow lymphocytes had a higher proportion of CD4+ and CD8+ T-cells, and B-cells than calf lymphocytes. The proportion of CD4+ T-cells and B-cells expressing CD62L was greater in calves than in cows, while cow B-cells expressed greater levels of CD11b than calf B-cells. In contrast, calf polymorphonuclear cells (PMN) and monocytes expressed greater levels of CD11b compared to cows. Moreover, calf monocytes expressed higher levels of MHCII compared with those of cows. Collectively, our data provides a resource to better understand the bovine immune system as well as immune-related diseases that affect dairy cattle.
