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Enumeration of single IFN-gamma-producing cells in mice during viral and bacterial infection
A Gessner1, D Moskophidis, F Lehmann-Grube
1Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie, Universität Hamburg, Germany.
Abstract:
A solid phase immunoenzymatic technique was employed for detecting single IFN-gamma-producing cells (IFN-gamma PC) in the mouse. After infection with lymphocytic choriomeningitis virus or Listeria monocytogenes, the numbers of IFN-gamma PC in spleens began to rise on day 4, attained maxima on days 7 and 8, and declined thereafter. Negative selection in vitro by use of mAb and C allowed phenotypic identification of the producer cells; most, if not all, carried Thy-1, and approximately one half expressed CD4, the other half, CD8. Depletion of cells in vivo by treatment of mice with mAb led to somewhat different results; again, anti-Thy-1 antibody eliminated essentially all IFN-gamma PC, but considerably more than 50% were either CD4+ or CD8+, suggesting regulatory interactions between these T lymphocyte subsets with regard to generation of the lymphokine.
Insights
This study identifies interferon-gamma-producing cells (IFN-gamma PC) in mice post-infection. Results show these cells peak around day 7-8 and are primarily Thy-1 positive, with roughly half being CD4+ and half CD8+ T cells.
Area of Science:
- Immunology
- Cell Biology
- Virology
Background:
- Interferon-gamma (IFN-gamma) is a critical cytokine in immune responses.
- Identifying the cellular sources of IFN-gamma is crucial for understanding immune regulation.
- Mouse models are essential for studying cellular immunity against viral and bacterial infections.
Purpose of the Study:
- To develop and apply a method for detecting single IFN-gamma-producing cells (IFN-gamma PC) in mice.
- To characterize the phenotype of IFN-gamma PC following viral and bacterial infections.
- To investigate the roles of different T lymphocyte subsets in IFN-gamma production.
Main Methods:
- Solid phase immunoenzymatic technique for single-cell detection of IFN-gamma PC.
- Infection of mice with lymphocytic choriomeningitis virus and Listeria monocytogenes.
- In vitro negative selection using monoclonal antibodies (mAb) and in vivo cell depletion.
Main Results:
- IFN-gamma PC numbers in mouse spleens increased post-infection, peaking around days 7-8.
- The majority of IFN-gamma PC expressed Thy-1.
- Approximately 50% of IFN-gamma PC were CD4+ and 50% were CD8+ T cells.
- In vivo depletion studies suggested complex regulatory interactions between CD4+ and CD8+ T cells in IFN-gamma generation.
Conclusions:
- The study successfully identified and phenotyped IFN-gamma PC in mice.
- Thy-1+ cells are the primary source of IFN-gamma.
- Both CD4+ and CD8+ T cells contribute to IFN-gamma production, with evidence of regulatory interactions between these subsets.