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Application of Multiparametric Flow Cytometry Panels to Study Lymphocyte Subpopulations in Tuberculin-Positive Cattle
Anabelle Manzo-Sandoval1, Laura Jaramillo-Meza1, Roxana Olguín-Alor2
1Laboratorio de Inmunología, Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto de Investigaciones Forestales, Agrícolas y Pecuarias, Carretera Federal México-Toluca Km 15.5, Ciudad de México 05110, Mexico.
Insights
Flow cytometry (FC) is a valuable tool in veterinary immunology. New multicolor panels were developed to identify T cell subpopulations in dairy heifers, aiding in bovine tuberculosis research and vaccine development.
Area of Science:
- Veterinary Immunology
- Cellular Immunology
- Flow Cytometry
Background:
- Flow cytometry (FC) is crucial in veterinary research for studying immune responses and vaccine efficacy in cattle.
- Limited availability of fluorochrome-conjugated antibodies for bovine antigens restricts multiparametric analysis.
- This study addresses the need for advanced FC tools in bovine immunology.
Purpose of the Study:
- To design and implement two five-color flow cytometry panels for T cell analysis in dairy heifers.
- To identify T cell populations and subpopulations in peripheral blood mononuclear cells.
- To investigate differences in T cell profiles related to bovine tuberculosis status.
Main Methods:
- Development of two novel five-color flow cytometry panels.
- Analysis of peripheral blood mononuclear cells from dairy heifers.
- Comparison of T cell subpopulations between tuberculin-positive and negative heifers.
Main Results:
- The developed panels successfully identified T cell subpopulations.
- Significant differences in T cell subpopulations were observed between tuberculin-positive and negative heifers.
- Overexpression of CD25+ and CD45RO+ T cells was noted in tuberculin-positive heifers post-stimulation with Mycobacterium bovis CFPE.
Conclusions:
- The implemented multicolor flow cytometry panels are effective for analyzing bovine T cell subpopulations.
- These panels can be valuable tools for immunopathogenesis studies and vaccine development in cattle.
- The strategy holds potential for application in other veterinary species.
Abstract:
Flow cytometry (FC) is widely used in microbiology, immunology, hematology, and oncology. In the veterinary field, FC enabled the study of the immune response in cattle infected with different pathogens, as well as vaccine testing. However, few fluorochrome-conjugated antibodies recognize bovine antigens, limiting the possible benefits of FC and the implementation of multiparametric analysis for more complex studies. Two cytometry panels with five colors each were designed and implemented for the study and identification of populations and subpopulations of T cells derived from the peripheral blood mononuclear cells of dairy heifers. Both panels detected differences in T cell subpopulations between heifers positively and negatively tested for tuberculin; they detected overexpression of CD25+ and CD45RO+ in tuberculin-positive heifers after stimulation with a culture filtrate protein extract (CFPE) from Mycobacterium bovis (M. bovis). We identified subpopulations of T cells from peripheral blood mononuclear cells using two multicolor panels. These panels could be used to analyze total bovine blood in immunopathogenic studies and vaccine development. The same strategy could be implemented in other species of veterinary interest.
