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Detection and quantitation of cells secreting IL-6 under physiologic conditions in BALB/c mice

A Shirai1, K Holmes, D Klinman

  • 1Laboratory of Retrovirus Research, Food and Drug Administration, Bethesda, MD 20892.

Journal of Immunology (Baltimore, Md. : 1950)
|February 1, 1993
PubMed

Insights

A new ELIspot assay quantifies Interleukin-6 (IL-6) producing cells in mice. Macrophages and specific B-cell subsets are key IL-6 sources in bone marrow and spleen.

Area of Science:

  • Immunology
  • Cell Biology

Background:

  • Interleukin-6 (IL-6) is a critical cytokine regulating hematopoiesis and lymphocyte activation.
  • Understanding IL-6 production at the single-cell level is crucial for immunological research.

Purpose of the Study:

  • To develop and validate a highly sensitive ELIspot assay for detecting and enumerating IL-6 secreting cells.
  • To characterize the frequency and cellular sources of IL-6 production in normal BALB/c mice.

Main Methods:

  • Development of an extremely sensitive ELIspot assay.
  • Flow cytometry (FACS) analysis for cell identification.
  • Enumeration of IL-6 secreting cells in bone marrow, spleen, and mesenteric lymph nodes.

Main Results:

  • The ELIspot assay detected IL-6 production in normal mice at frequencies of approximately 0.5% (bone marrow), 0.1% (spleen), and 0.01% (mesenteric lymph nodes).
  • Macrophages and Mac1- B220- cells were identified as major IL-6 producers in the bone marrow.
  • T lymphocytes (4%) and B lymphocytes (38%) were significant contributors to IL-6 production in the spleen.
  • An average activated cell was estimated to produce ~85 molecules of IL-6 per second.

Conclusions:

  • The developed ELIspot assay provides a sensitive tool for quantifying IL-6 producing cells.
  • This study elucidates the cellular distribution of IL-6 production in key lymphoid organs of normal mice.
  • Macrophages and specific lymphocyte subsets are identified as primary sources of IL-6 under physiological conditions.

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