CD40 engagement triggers switching to IgA1 and IgA2 in human B cells through induction of endogenous TGF-beta: evidence for TGF-beta but not IL-10-dependent direct S mu-->S alpha and sequential S mu-->S gamma, S gamma-->S alpha DNA recombination.
IgA are major effectors of antimicrobial defense in the respiratory and digestive tracts. We have analyzed the requirements for and the modalities of switching to IgA using our recently identified monoclonal model of human germinal center differentiation, CL-01 B cells. CL-01 cells bear surface IgM (sIgM) and sIgD and switch to all seven downstream isotypes in response to physiologic stimuli. In these cells, CD40 engagement by CD40 ligand induces production of endogenous TGF-beta and IL-10, expression of germline Ialpha1-Calpha1 and Ialpha2-Calpha2 transcripts, mature VHDJH-Calpha1 and VHDJH-Calpha2 transcripts, and IgA secretion. These events are associated with not only direct Smu-->Salpha, but also sequential Smu-->Sgamma, Sgamma-->Salpha DNA recombination, and are ablated by neutralizing anti-TGF-beta but not IL-10 Ab, and indicating that TGF-beta, not IL-10, is a crucial mediator of the transcriptional activation and recombination of human Calpha1 and Calpha2 genes. Our findings in CL-01 cells were reproduced in freshly isolated naive sIgM+ sIgD+ B lymphocytes. Thus, engagement of CD40, in the absence of other (known) stimuli, is sufficient to effectively induce switching to IgA in human B cells. This is effected by direct and sequential DNA recombination events, which are both dependent upon endogenous TGF-beta secreted by the CD40L-induced B cells.
Zan
H
H
Department of Pathology, Cornell University Medical College, New York 10021, USA.
Cerutti
A
A
Dramitinos
P
P
Schaffer
A
A
Casali
P
P
D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism*; D001483:Base Sequence; D019013:CD40 Antigens / Q000378:metabolism*; D023201:CD40 Ligand; D015322:Gene Rearrangement, B-Lymphocyte; D005803:Genes, Immunoglobulin; D006801:Humans; D007070:Immunoglobulin A / Q000096:biosynthesis* / Q000235:genetics; D017578:Immunoglobulin Class Switching / Q000235:genetics*; D007127:Immunoglobulin Constant Regions / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007137:Immunoglobulin alpha-Chains / Q000235:genetics; D007142:Immunoglobulin gamma-Chains / Q000235:genetics; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D016753:Interleukin-10 / Q000502:physiology*; D008024:Ligands; D008213:Lymphocyte Activation / Q000235:genetics; D008562:Membrane Glycoproteins / Q000502:physiology; D008969:Molecular Sequence Data; D011947:Receptors, Antigen, B-Cell / Q000096:biosynthesis; D014158:Transcription, Genetic / Q000276:immunology; D015533:Transcriptional Activation / Q000276:immunology; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000378:metabolism / Q000502:physiology
D019013:CD40 Antigens; D007070:Immunoglobulin A; D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D007137:Immunoglobulin alpha-Chains; D007142:Immunoglobulin gamma-Chains; D007148:Immunoglobulin mu-Chains; D008024:Ligands; D008562:Membrane Glycoproteins; D011947:Receptors, Antigen, B-Cell; D016212:Transforming Growth Factor beta; D016753:Interleukin-10; D023201:CD40 Ligand