Shreffler:Notebook/R03 Baso/2010/03/08: Difference between revisions
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Mechanisms of IgE-dependent immune modulation by basophils | 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. | 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) | 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 | *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. | 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 | *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 | *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) | 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 | *Co-culture basophils +/- Derp2-specific IgE mAb, +/- Derp2 Ag with CD8 and CD4 Derp2-specific HLA matched T cell clones | ||
Revision as of 16:42, 8 March 2010
 Basophil Project
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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)
Aim 2. To characterize the capacity of human basophils to elicit Derp2-specific Ab from syngeneic naïve B cells.
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)
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