CD28 functions as an adhesion molecule and is involved in the regulation of human IgE synthesis.
Activated T cells induce IgE switching in B cells via a combination of lymphokines and direct T:B cell contact. As CD28-deficient mice have reduced basal levels of IgG1 and IgG2a and diminished Ig class switching, we investigated whether the CD28/B7.1 (CD80) ligand pairing might also be involved in human IgE regulation. Co-incubation of an allergen-specific, human T cell clone with tonsillar B cells caused a marked up-regulation of CD28 expression, whereas, in contrast, CD45 RB expression was unaffected. To test whether blocking the CD28: B7.1 interaction affected IgE synthesis, a dialyzed anti-CD28 monoclonal antibody (mAb) was added to cultures containing tonsillar B cells, pre-activated T cell clones and interleukin-4. Anti-CD28 treatment caused a reproducible, dose-dependent inhibition of IgE, but not IgG synthesis that was accompanied by a visible decrease in cell aggregate formation. Conversely, an anti-B7.1 mAb had no effect in this system. The effect of blocking CD28-ligand interactions on lymphocyte adhesion was formally assessed on human T cell clones and B cell lines using dual intracellular staining and flow cytometry. Co-incubation with an anti-CD28 mAb, but not control IgG or anti-B7.1 mAb, resulted in a marked impairment of conjugate formation that correlated well with T cell surface expression of CD28. Using this system we found that an anti-CTLA-4 mAb but not an anti-B7.2 mAb inhibited T:B cell conjugate formation. Lastly, in addition to a direct effect of anti-CD28 mAb on conjugate formation, 14-day culture of T and B cells in the presence of anti-CD28 caused a marked decrease of ICAM-1 (CD54) expression on aggregated lymphocytes. In contrast, LFA-1 (CD18) expression was unaffected. We, therefore, conclude that the T cell co-stimulatory molecule CD28 is involved in the regulation of IgE synthesis in vitro. CD28 may act to a limited extent as an adhesion molecule, though apparently not by pairing with B7.1 or B7.2. It is more likely that ligation of CD28 under certain conditions modulates the expression of other T and B cell surface molecules.
Life
P
P
Glaxo Institute for Molecular Biology, Geneva.
Aubry
J P
JP
Estoppey
S
S
Schnuriger
V
V
Bonnefoy
J Y
JY
D000069594:Abatacept; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000502:physiology; D018122:B7-1 Antigen / Q000502:physiology; D018821:CD18 Antigens / Q000032:analysis; D018106:CD28 Antigens / Q000502:physiology*; D060908:CTLA-4 Antigen; D015815:Cell Adhesion Molecules / Q000502:physiology*; D002449:Cell Aggregation; D002478:Cells, Cultured; D006801:Humans; D018796:Immunoconjugates; D007073:Immunoglobulin E / Q000096:biosynthesis*; D018799:Intercellular Adhesion Molecule-1 / Q000032:analysis
D015703:Antigens, CD; D000943:Antigens, Differentiation; D018122:B7-1 Antigen; D018821:CD18 Antigens; D018106:CD28 Antigens; D060908:CTLA-4 Antigen; C556706:CTLA4 protein, human; D015815:Cell Adhesion Molecules; C556707:Ctla4 protein, mouse; D018796:Immunoconjugates; D018799:Intercellular Adhesion Molecule-1; D007073:Immunoglobulin E; D000069594:Abatacept