Shreffler:Notebook/R03 Baso/2010/03/08

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*Wayne G. Shreffler 18:41, 8 March 2010 (EST):

Mechanisms of IgE-dependent immune modulation by basophils Hypothesis: Basophils can sequester small amounts of antigen via the presence of specific IgE bound to FcεRI and present that antigen directly to B cells and directly (CD8) and/or indirectly (CD4) to T cells to illicit Th2-biased de novo immune responses.

Aim 1. To determine the fate of FcεRI-Ag complexes following cross-linking by confocal microscopy using fluorochrome tagged Ag and mAb (Derp2)

  • Reagents from Kaare Lund at ALK

Aim 2. To characterize the capacity of human basophils to elicit Derp2-specific Ab from syngeneic naïve B cells.

  • Co-culture basophils +/- Derp2-specific IgE mAb, +/- Derp2 Ag with naïve B cells
  • Measure total and Derp2-specific Ab IgM, IgA, IgG, IgE in supernatant

Aim 3. To characterize the capacity of human basophils to elicit class I and class II restricted Derp2-specific T cell responses (proliferation and cytokine)

  • Co-culture basophils +/- Derp2-specific IgE mAb, +/- Derp2 Ag with CD8 and CD4 Derp2-specific HLA matched T cell clones
  • Same with the addition of syngeneic DCs
  • Same with naïve T cells



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