Advertisement

Dendritic Cell Dependent Expression of IgA By Clones in T/B Microcultures

  • Carol E. Schrader
  • John J. Cebra
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 329)

Abstract

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.

Keywords

Conditioned Medium Cell Clone Clonal Culture Productive Clone Culture Dendritic Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    D. A. Lebman, P. M. Griffin, and J. J. Cebra, Relationship between expression of IgA by Peyer’s patch cells and functional IgA memory cells, J. Exp. Med. 166: 1405 (1987).PubMedCrossRefGoogle Scholar
  2. 2.
    J. A. Fuhrman and J. J. Cebra, Special features of the priming process for a secretory IgA response. B cell priming with Cholera toxin, J. Exp. Med. 153: 534 (1981).PubMedCrossRefGoogle Scholar
  3. 3.
    G. J. V. Nossal, B. L. Pike, and F. L. Battye, Sequential use of hapten-gelatin fractionation and fluorescence activated cell sorting in the enrichment of haptenspecific B lymphocytes, Eur. J. Immunol. 8: 151 (1978).PubMedCrossRefGoogle Scholar
  4. 4.
    P. A. Schweitzer and J. J. Cebra, Quality of antibodies secreted by clones in microcultures from B cells enriched on haptenated gelatin: isotypes and avidities, Mol. Immunol. 25: 231 (1988).PubMedCrossRefGoogle Scholar
  5. 5.
    J. Kaye, S. Porcelli, J. Tite, B. Jones, and C. A. Janeway, Jr., Both a monoclonal antibody and antisera specific for determinants unique to individual cloned helper T cell lines can substitute for antigen and antigen-presenting cells in the activation of T cells. J. Exp. Med. 158: 836 (1983).PubMedCrossRefGoogle Scholar
  6. 6.
    C. E. Schrader, A. George, R. L. Kerlin, and J. J. Cebra, Dendritic cells support production of IgA and other non-IgM isotypes in clonal microculture, Int. Immunol. 2: 563 (1990).PubMedCrossRefGoogle Scholar
  7. 7.
    D. B. Kotloff and J. J. Cebra, Effect of TH-lines and clones on the growth and differentiation of B cells clones in microculture, Mol. Immunol. 25: 147 (1988).PubMedCrossRefGoogle Scholar
  8. 8.
    R. M. Steinman, G. Kaplan, M. D. Witmer, and Z. A. Cohn, Identification of a novel cell type in peripheral lymphoid organs of mice. V. Purification of spleen dendritic cells, new surface markers, and maintenance in vitro. J. Exp. Med. 149: 1 (1979).PubMedCrossRefGoogle Scholar
  9. 9.
    D. M. Spaulding, W. J. Koopman, J. H. Eldridge, J. R. McGhee, and R. M. Steinman, Accessory cells in murine Peyer’s patch. I. Identification and enrichment of a functional dendritic cell. J. Exp. Med. 157: 1646 (1983).CrossRefGoogle Scholar
  10. 10.
    A. George and J. J. Cebra, Responses of single germinalcenter B cells in T-cell-dependent microculture, Proc. Natl. Acad. Sci. USA 88: 11 (1991).PubMedCrossRefGoogle Scholar
  11. 11.
    R. L. Coffman, D. A. Lebman, and B. Shrader, Transforming growth factor-β specifically enhances IgA production by lipopolysaccharide-stimulated murine B lymphocytes, J. Exp. Med. 170: 1039 (1989).PubMedCrossRefGoogle Scholar
  12. 12.
    J. F. Elliott, Y. Lin, S. B. Mizel, R. C. Bleackley, D. G. Harnish, and V. Paetkau, Induction of Interleukin 2 messenger RNA inhibited by cyclosporin A, Science, 226: 1439 (1984).PubMedCrossRefGoogle Scholar
  13. 13.
    C. B. Thompson, T. Lindsten, J. A. Ledbetter, S. L. Kunkel, H. A. Young, S. G. Emerson, J. M. Leiden, and C. H. June, CD28 activation pathway regulates the production of multiple T-cell-derived lymphokines/cytokines, Proc. Natl. Acad. Sci. USA 86: 1333 (1989).PubMedCrossRefGoogle Scholar
  14. 14.
    A. Vallé, J-P. Aubry, I. Durand, and J. Banchereau, IL-4 and IL-2 upregulate the expression of antigen B7, the B cell counterstructure to T cell CD28: an amplification mechanism for T-B interactions, Int. Immunol. 3: 229 (1991).PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1993

Authors and Affiliations

  • Carol E. Schrader
    • 1
  • John J. Cebra
    • 1
  1. 1.Department of BiologyUniversity of PennsylvaniaPhiladelphiaUSA

Personalised recommendations