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An antigen-specific DTH-initiating cell clone. Functional, phenotypical, and partial molecular characterization.
W R Herzog1, I Millet, N R Ferreri
1Department of Internal Medicine, Yale University School of Medicine, New Haven, CT 06510-8056.
Journal of Immunology (Baltimore, Md. : 1950)
|May 15, 1990
Summary
Delayed-type hypersensitivity (DTH) involves two Ag-specific Thy-1+ cells. A novel DTH-initiating cell clone was generated, producing Ag-specific factors that initiate DTH reactions with a unique cell surface phenotype.
Area of Science:
- Immunology
- Cell Biology
- Hypersensitivity Reactions
Background:
- Delayed-type hypersensitivity (DTH) reactions are mediated by sequential actions of distinct Ag-specific Thy-1+ cells.
- An early-acting DTH-initiating cell produces a circulating, Ag-specific factor, functionally analogous to IgE, which sensitizes local tissues for serotonin release.
- Previous studies identified a unique, unusual surface phenotype for polyclonal DTH-initiating cells, including Thy-1+, Lyt-1+ (CD5), triple negative (CD4-, CD8-, CD3-), B220+, IL-3R+, and IL-2R-.
Purpose of the Study:
- To generate and characterize an Ag-specific DTH-initiating cell clone.
- To confirm the unique surface phenotype and functional abilities of this cloned cell.
- To elucidate the molecular characteristics of Ag-specific DTH initiation.
Main Methods:
- Generation of an Ag-specific DTH-initiating cell clone from nude mice immunized with oxazolone (OX).
- Flow microfluorometry analysis to determine the surface phenotype of the cloned cells.
- In vivo assays to assess the functional abilities of the cloned cells in initiating DTH.
- Specific cDNA probes and Northern blot analysis to examine gene expression.
Main Results:
- An Ag-specific DTH-initiating cell clone (OX-F) was successfully generated, exhibiting similar unique surface phenotype and functional abilities to polyclonal DTH-initiating cells.
- The cloned cells produce Ag-specific OX-F, which initiates DTH in an Ag-specific manner.
- Northern blot analysis confirmed that these Ag-specific DTH-initiating cells express Thy-1+, CD3-, and IL-3R+ markers.
Conclusions:
- DTH initiation is mediated by an Ag-specific lymphoid cell with a distinct surface phenotype: Thy-1+, CD5+, sIg-, CD4-, CD8-, CD3-, CD45RA+, IL-2R-, and IL-3R+.
- This cell produces an Ag-specific factor (e.g., OX-F) crucial for initiating the DTH response.
- The findings provide a deeper understanding of the cellular and molecular mechanisms underlying DTH.