B cell activation and Ig, especially IgE, production is inhibited by high CD23 levels in vivo and in vitro.
The capacity of CD23 to regulate IgE production was evaluated in both an in vitro and an in vivo system. The decreased IgE response seen in CD23 transgenic mice was confirmed and observed to occur at all antigen doses used. In addition, purified B cells from the Tg animals in general exhibited lower IgE production when stimulated with CD40L and IL-4. To examine this down-regulating activity of CD23 an in vitro model system was developed. CHO cells were transfected with CD23, ICAM-1, or both CD23 and ICAM-1. ICAM-1 was chosen to enhance B cell-B cell interaction. Purified resting B cells were placed into culture with the mitomycin C-treated transfected or control CHO cells and activated with IL-4, IL-5, and CD40L-CHO. A dose-dependent decrease in IgE production was observed with increasing cell numbers of the CHO transfectants that expressed CD23. The effect lasted up to Day 3 of culture. B cell proliferation was also inhibited in a dose-dependent fashion by increasing numbers of CD23-expressing cells suggesting a potential effect of CD23 on B cell apoptosis. In contrast, ICAM-1-transfected or CHO control cells had minimal effects on either Ig production or B cell proliferation. While IgE production was inhibited up to 95% by high numbers of CD23-transfected CHO cells, some inhibition of IgG and IgM production was also seen. Finally, the mechanism of CD23-mediated inhibition of IgE production was compared with the inhibition in IgE production seen when B cell were coactivated with multivalent anti-IgD in conjunction with CD40L plus optimal IL-4. To this end we used RT-PCR to compare the relative levels of epsilon-germline transcripts in control cultures and cultures coactivated by anti-IgD, CD40L, and IL-5 or activated in the presence of high levels of CD23-expressing cells. CD22 was used as an internal standard since levels change little with B cell activation. Coactivation strongly inhibited epsilon-germline transcript levels but the presence of CD23-expressing cells did not. Thus, coactivation potentially operates prior to isotype switching, while high CD23 coculture blocks either recombination or more likely B cell differentiation to high Ig producers stage. Our data support the hypothesis that IL-4 induces both IgE and a controlling agent for IgE, namely, CD23.
Cho
S W
SW
Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond 23298, USA.
Kilmon
M A
MA
Studer
E J
EJ
van der Putten
H
H
Conrad
D H
DH
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D016466:CHO Cells; D006224:Cricetinae; D015536:Down-Regulation; D006433:Hemocyanins / Q000276:immunology; D017578:Immunoglobulin Class Switching; D007073:Immunoglobulin E / Q000096:biosynthesis*; D007136:Immunoglobulins / Q000096:biosynthesis; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D015847:Interleukin-4 / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D017455:Receptors, IgE / Q000235:genetics / Q000276:immunology*; D014162:Transfection
C042158:2,4-dinitrophenyl keyhole limpet hemocyanin; D007136:Immunoglobulins; D017455:Receptors, IgE; D018799:Intercellular Adhesion Molecule-1; D015847:Interleukin-4; D007073:Immunoglobulin E; D006433:Hemocyanins