The ADP-ribosylating CTA1-DD adjuvant enhances T cell-dependent and independent responses by direct action on B cells involving anti-apoptotic Bcl-2- and germinal center-promoting effects.
We recently developed a novel immunomodulating gene fusion protein, CTA1-DD, that combines the ADP-ribosylating ability of cholera toxin (CT) with a dimer of an Ig-binding fragment, D, of Staphylococcus aureus protein A. The CTA1-DD adjuvant was found to be nontoxic and greatly augmented T cell-dependent responses to soluble protein Ags after systemic as well as mucosal immunizations. Here we show that CTA1-DD does not appear to form immune complexes or bind to soluble Ig following injections, but, rather, it binds directly to B cells of all isotypes, including naive IgD+ cells. No binding was observed to macrophages or dendritic cells. Immunizations in FcepsilonR (common FcRgamma-chain)- and FcgammaRII-deficient mice demonstrated that CTA1-DD exerted unaltered enhancing effects, indicating that FcgammaR-expressing cells are not required for the adjuvant function. Whereas CT failed to augment Ab responses to high m.w. dextran B512 in athymic mice, CTA1-DD was highly efficient, demonstrating that T cell-independent responses were also enhanced by this adjuvant. In normal mice both CT and CTA1-DD, but not the enzymatically inactive CTA1-R7K-DD mutant, were efficient enhancers of T cell-dependent as well as T cell-independent responses, and both promoted germinal center formation following immunizations. Although CT augmented apoptosis in Ag receptor-activated B cells, CTA1-DD strongly counteracted apoptosis by inducing Bcl-2 in a dose-dependent manner, a mechanism that was independent of the CD19 coreceptor. However, in the presence of CD40 stimulation, apoptosis was low and unaffected by CT, suggesting that the adjuvant effect of CT is dependent on the presence of activated CD40 ligand-expressing T cells.
Agren
L
L
Department of Medical Microbiology and Immunology, University of Göteborg, Sweden.
Sverremark
E
E
Ekman
L
L
Schön
K
K
Löwenadler
B
B
Fernandez
C
C
Lycke
N
N
D000276:Adjuvants, Immunologic / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*; D000818:Animals; D000917:Antibody Formation / Q000276:immunology; D000955:Antigens, T-Independent / Q000502:physiology*; D017209:Apoptosis / Q000276:immunology*; D016175:B-Lymphocyte Subsets / Q000166:cytology / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism; D002772:Cholera Toxin / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*; D003911:Dextrans / Q000276:immunology; D005260:Female; D018858:Germinal Center / Q000166:cytology / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism; D006241:Haptens / Q000276:immunology; D007072:Immunoglobulin D / Q000096:biosynthesis / Q000378:metabolism; D007132:Immunoglobulin Isotypes / Q000096:biosynthesis / Q000378:metabolism; D007275:Injections, Intravenous; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008819:Mice, Nude; D011065:Poly(ADP-ribose) Polymerases / Q000502:physiology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis / Q000502:physiology*; D011961:Receptors, Fc / Q000096:biosynthesis; D013205:Staphylococcal Protein A / Q000008:administration & dosage / Q000235:genetics / Q000494:pharmacology*
D000276:Adjuvants, Immunologic; D000955:Antigens, T-Independent; D003911:Dextrans; D006241:Haptens; D007072:Immunoglobulin D; D007132:Immunoglobulin Isotypes; D019253:Proto-Oncogene Proteins c-bcl-2; D011961:Receptors, Fc; D013205:Staphylococcal Protein A; D002772:Cholera Toxin; C042157:2,4-dinitrophenyl dextran; D011065:Poly(ADP-ribose) Polymerases