Dendritic Cell Dependent Expression of IgA By Clones in T/B Microcultures
We sought to develop a T-dependent, clonal B cell microculture in order to assess changes in freguencies and Ig isotype potential of antigen (Ag)-specific B cells, associated with oral/gut mucosal exposure to Ag. Particularly, we were concerned with the development of IgA-memory B cells (1) as predictive of an in vivo, secondary mucosal IgA antibody (Ab) response (2) and with the role of germinal centers (GC) in Peyer’s patches (PP) in the generation of these IgA-memory cells as well as IgA pre-plasmablasts. Our original T/B microculture was based on clonal culturing of B cells responsive to thymus-independent Ags, as practiced in the Nossal laboratory (3) using Ag-specific B cells enriched by panning on haptenated gelatin (4), except that we used cloned, Ag (conalbumin)-specific D10, TH 2 cells (5) and haptenated-Ag as stimuli (6). This system exhibited H-2 haplotype restriction and a requirement for linked recognition of hapten and carrier. An Ag-independent version of this T/B microculture utilized the alloreactivity of the D1O cells versus I-Ab molecules and purified F1, k — b, B cells (6). Both systems promoted a high frequency of productive B cell clones (1-50%) that exhibited intraclonal isotype switching.
KeywordsConditioned Medium Cell Clone Clonal Culture Productive Clone Culture Dendritic Cell
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