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Published on: February 8, 2016
Development of an advanced flow cytometry based high-resolution immunophenotyping method to benchmark early immune
Sabine Farschtschi1, Martin Mattes2, Alex Hildebrandt2
1Division of Animal Physiology and Immunology, TUM School of Life Sciences, Technical University of Munich, Freising, Germany. sabine.farschtschi@wzw.tum.de.
Insights
High-resolution differential cell counts (HRDCC) in cow blood and milk reveal immune responses to vaccination. This advanced method offers diagnostic benefits over standard tests for monitoring udder health and systemic diagnostics.
Area of Science:
- Veterinary immunology
- Dairy cow health monitoring
- Flow cytometry applications
Background:
- Somatic cell count is standard for udder health but lacks cell type specificity.
- Immune cell populations in milk are diverse and not fully characterized by current methods.
Purpose of the Study:
- To develop a high-resolution differential cell count (HRDCC) for bovine blood and milk using flow cytometry.
- To assess the diagnostic utility of HRDCC in monitoring immune responses to vaccination in dairy cows.
Main Methods:
- Flow cytometry was used to analyze immune cell subpopulations and viability in blood and milk.
- Blood samples were also analyzed for standard biomarkers (differential counts, haptoglobin, metabolic parameters).
- Three vaccines were administered to groups of healthy cows, with samples collected pre-, inter-, and post-vaccination.
Main Results:
- HRDCC identified ten immune cell subpopulations in both blood and milk.
- Inclusion of HRDCC provided diagnostic benefits compared to standard parameters alone.
- Vaccinations induced visible immune system impacts in both blood and milk HRDCCs, with CD8+ T cells, B cells, and monocytes/macrophages being key indicators.
Conclusions:
- HRDCC in blood and milk is highly relevant for early systemic diagnostics in dairy cows.
- Further research is needed to validate these subpopulations across different breeds, lactation stages, and health statuses.
Abstract:
The determination of the somatic cell count of a milk sample is one of the most common methods to monitor udder health of a dairy cow. However, this procedure does not take into account the fact that cells in milk present a great variety of different cell types. The objective of our study was to establish a high-resolution differential cell count (HRDCC) by means of flow cytometry in blood and milk. We were able to detect ten subpopulations among the three main populations of immune cells and to determine their viability. Additionally, blood samples were analyzed for common laboratory biomarkers, i.e. differential blood counts, haptoglobin levels and several metabolic parameters. In this first feasibility study, we used three different vaccines to stimulate the immune system of five healthy cows each. Samples were collected shortly before, in between and after the vaccinations. Using multivariate statistical methods we saw a diagnostic benefit when HRDCCs were included compared to only the standard laboratory parameters. The impacts of all three vaccinations on the immune system were visible in blood HRDCCs as well as in milk HRDCCs. Cluster of Differentiation 8+ (CD8+) T cells, B cells and monocyte/macrophage subpopulations were among the most important and statistically relevant parameters for all treatments in both biofluids. Moreover, in one of the treatment groups intermediate monocytes showed a significant increase after both vaccinations. Although the use of HRDCC in blood or milk was shown to be highly relevant for early systemic diagnostic, to confirm these subpopulations further investigations in cows of different breed, lactation stage or health status are required.
