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Recombinant soluble trimeric CD40 ligand is biologically active

G J Mazzei1, M D Edgerton, C Losberger

  • 1Glaxo Institute for Molecular Biology, Geneva, Switzerland.

The Journal of Biological Chemistry
|March 31, 1995
PubMed
Summary
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The 18-kDa fragment of CD40 ligand (CD40L) is the sole biologically active component, stimulating B cell proliferation and differentiation. This soluble fragment functions identically to the membrane-bound form, offering insights into immune cell interactions.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Biology

Background:

  • CD40 ligand (CD40L) on activated immune cells binds CD40 on B cells, promoting B cell activation.
  • Previous studies utilized a soluble recombinant CD40L preparation containing multiple fragments.

Purpose of the Study:

  • To identify the specific biologically active fragment of soluble CD40L.
  • To characterize the function and structure of the active CD40L fragment.

Main Methods:

  • Purification of individual CD40L fragments.
  • Recombinant expression and functional assays (B cell proliferation, differentiation, apoptosis rescue, CD40 binding).
  • Sucrose gradient sedimentation to determine protein structure.

Main Results:

Related Experiment Videos

  • Only the 18-kDa CD40L fragment (Glu-108 to Leu-261) demonstrated biological activity.
  • The 18-kDa fragment induced B cell proliferation, differentiation, and rescued B cells from apoptosis.
  • The 18-kDa fragment bound CD40 and formed an apparent homotrimer, consistent with CD40L structure.

Conclusions:

  • The 18-kDa fragment is the essential functional unit of soluble CD40L.
  • Soluble CD40L can activate CD40-expressing cells similarly to membrane-bound CD40L.
  • This finding has implications for understanding B cell regulation and immune responses.