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Related Experiment Videos

Differential Thy-1 expression by splenic fibroblasts defines functionally distinct subsets

M A Borrello1, R P Phipps

  • 1Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, New York 14642, USA.

Cellular Immunology
|November 1, 1996
PubMed
Summary
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Splenic fibroblasts are diverse, with Thy-1-positive cells producing more IL-6 and Thy-1-negative cells presenting antigens after IFN-gamma treatment, suggesting roles in immune responses.

Area of Science:

  • Immunology
  • Cell Biology
  • Spleen Microenvironment

Background:

  • Fibroblasts form the spleen's structural extracellular matrix scaffold for immune cells.
  • Splenic fibroblasts, often termed 'stromal' or 'reticular' components, remain poorly characterized.
  • Understanding fibroblast heterogeneity is crucial for comprehending spleen immune regulation.

Purpose of the Study:

  • To establish and phenotypically characterize normal splenic fibroblast lines from mice.
  • To investigate the heterogeneity of splenic fibroblasts based on Thy-1 antigen expression.
  • To explore the distinct functional capabilities of Thy-1-positive and Thy-1-negative splenic fibroblast subsets.

Main Methods:

  • Establishment of normal fibroblast lines from mouse spleen.

Related Experiment Videos

  • Phenotypic characterization using morphology, marker expression (vimentin, CD45, epithelial/endothelial markers), and phagocytosis assays.
  • Flow cytometry (FACS) and limiting dilution cloning for isolating Thy-1-positive and Thy-1-negative splenic fibroblasts.
  • Analysis of cytokine production (IL-6) and cell surface molecule expression (MHC class I/II, ICAM-1, VCAM-1, CD44).
  • Antigen presentation assays using an I-A(b)-restricted T cell line after IFN-gamma stimulation.

    • Main Results:

    • Established stable fibroblast lines confirmed by morphology and vimentin expression, lacking hematopoietic, epithelial, and endothelial markers.
    • Identified two distinct splenic fibroblast subpopulations: Thy-1-positive (50-65%) and Thy-1-negative.
    • Both subsets constitutively synthesized collagen and expressed MHC class I, ICAM-1, VCAM-1, and CD44.
    • Thy-1-positive fibroblasts produced significantly higher levels of IL-6 compared to Thy-1-negative fibroblasts.
    • Thy-1-negative fibroblasts, but not Thy-1-positive, expressed MHC class II and presented antigens after IFN-gamma treatment, suggesting a role in T cell interaction.

    Conclusions:

    • Splenic fibroblasts are a heterogeneous and dynamic cell population.
    • Thy-1 expression defines functionally distinct splenic fibroblast subsets with differential immune-modulatory capacities.
    • Thy-1-negative splenic fibroblasts may present antigens to T lymphocytes under inflammatory conditions, highlighting their immunological relevance.
    • These findings contribute to understanding the spleen's microenvironment and fibroblast roles in immune regulation and fibrotic conditions.