RESUMO
Immunoglobulin D (IgD) is an enigmatic antibody isotype that mature B cells express together with IgM through alternative RNA splicing. Here we report active T cell-dependent and T cell-independent IgM-to-IgD class switching in B cells of the human upper respiratory mucosa. This process required activation-induced cytidine deaminase (AID) and generated local and circulating IgD-producing plasmablasts reactive to respiratory bacteria. Circulating IgD bound to basophils through a calcium-mobilizing receptor that induced antimicrobial, opsonizing, inflammatory and B cell-stimulating factors, including cathelicidin, interleukin 1 (IL-1), IL-4 and B cell-activating factor (BAFF), after IgD crosslinking. By showing dysregulation of IgD class-switched B cells and 'IgD-armed' basophils in autoinflammatory syndromes with periodic fever, our data indicate that IgD orchestrates an ancestral surveillance system at the interface between immunity and inflammation.
Assuntos
Linfócitos B/imunologia , Basófilos/imunologia , Imunoglobulina D/imunologia , Imunoglobulina M/imunologia , Mucosa Respiratória/imunologia , Fator Ativador de Células B/metabolismo , Basófilos/metabolismo , Catelicidinas/metabolismo , Linhagem Celular , Citidina Desaminase/metabolismo , Febre Familiar do Mediterrâneo/imunologia , Haemophilus influenzae/crescimento & desenvolvimento , Haemophilus influenzae/fisiologia , Humanos , Switching de Imunoglobulina , Imunoglobulina D/biossíntese , Interleucina-1/metabolismo , Interleucina-4/metabolismo , Deficiência de Mevalonato Quinase/imunologia , Moraxella catarrhalis/crescimento & desenvolvimento , Moraxella catarrhalis/fisiologia , Ligação Proteica , Mucosa Respiratória/metabolismo , Mucosa Respiratória/microbiologiaRESUMO
Contact-dependent communication between immune cells generates protection but also facilitates viral spread. Here we found that macrophages formed long-range actin-propelled conduits in response to negative factor (Nef), a human immunodeficiency virus type 1 (HIV-1) protein with immunosuppressive functions. Conduits attenuated immunoglobulin G2 (IgG2) and IgA class switching in systemic and intestinal lymphoid follicles by shuttling Nef from infected macrophages to B cells through a guanine-exchange factor-dependent pathway involving the amino-terminal anchor, central core and carboxy-terminal flexible loop of Nef. By showing stronger virus-specific IgG2 and IgA responses in patients with Nef-deficient virions, our data suggest that HIV-1 exploits intercellular 'highways' as a 'Trojan horse' to deliver Nef to B cells and evade humoral immunity systemically and at mucosal sites of entry.
Assuntos
Linfócitos B/metabolismo , Comunicação Celular , Anticorpos Anti-HIV/imunologia , HIV-1/imunologia , Imunoglobulina A/imunologia , Imunoglobulina G/imunologia , Produtos do Gene nef do Vírus da Imunodeficiência Humana/fisiologia , Actinas/metabolismo , Antígenos CD40/fisiologia , Centro Germinativo/fisiologia , Proteína do Núcleo p24 do HIV/fisiologia , Humanos , Switching de Imunoglobulina , Macrófagos/virologia , Células U937RESUMO
Class switch DNA recombination (CSR) from IgM to IgG and IgA is crucial for antiviral immunity. Follicular B cells undergo CSR upon engagement of CD40 by CD40 ligand on CD4+ T cells. This T cell-dependent pathway requires 5-7 days, which is too much of a delay to block quickly replicating pathogens. To compensate for this limitation, extrafollicular B cells rapidly undergo CSR through a T cell-independent pathway that involves innate Ag receptors of the TLR family. We found that a subset of upper respiratory mucosa B cells expressed TLR3 and responded to viral dsRNA, a cognate TLR3 ligand. In the presence of dsRNA, mucosal B cells activated NF-kappaB, a transcription factor critical for CSR. Activation of NF-kappaB required TRIF (Toll/IL-1R domain-containing protein inducing IFN-beta), a canonical TLR3 adapter protein, and caused germline transcription of downstream CH genes as well as expression of AID (activation-induced cytidine deaminase), a DNA-editing enzyme essential for CSR. Subsequent IgG and IgA production was enhanced by BAFF (B cell-activating factor of the TNF family), an innate mediator released by TLR3-expressing mucosal dendritic cells. Indeed, these innate immune cells triggered IgG and IgA responses upon exposure to dsRNA. By showing active TLR3 signaling and ongoing CSR in upper respiratory mucosa B cells from patients with CD40 signaling defects, our findings indicate that viral dsRNA may initiate frontline IgG and IgA responses through an innate TLR3-dependent pathway involving BAFF.
Assuntos
Fator Ativador de Células B/metabolismo , Linfócitos B/imunologia , Switching de Imunoglobulina/imunologia , Imunoglobulinas/imunologia , Transdução de Sinais/imunologia , Receptor 3 Toll-Like/metabolismo , Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Linfócitos B/citologia , Linfócitos B/metabolismo , Antígenos CD40/imunologia , Proliferação de Células , Células Cultivadas , Citidina Desaminase/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Humanos , Switching de Imunoglobulina/genética , Imunoglobulinas/genética , Imunoglobulinas/metabolismo , Mucosa/citologia , Mucosa/imunologia , RNA de Cadeia Dupla/genética , Regulação para CimaRESUMO
Epithelial cells (ECs) transport class-switched immunoglobulin G (IgG) and IgA antibodies across mucous membranes. Whether ECs initiate class switching remains unknown. Here we found that ECs lining tonsillar crypts formed pockets populated by B cells expressing activation-induced cytidine deaminase (AID), an enzyme associated with ongoing class switching. ECs released B cell-activating AID-inducing factors after sensing microbial products through Toll-like receptors. The resulting class switching was amplified by thymic stromal lymphopoietin, an epithelial interleukin 7-like cytokine that enhanced the B cell 'licensing' function of dendritic cells, and was restrained by secretory leukocyte protease inhibitor, an epithelial homeostatic protein that inhibited AID induction in B cells. Thus, ECs may function as mucosal 'guardians' orchestrating frontline IgG and IgA class switching through a Toll-like receptor-inducible signaling program regulated by secretory leukocyte protease inhibitor.NOTE: In the version of this article initially published online, the middle label above Figure 6c is incorrect. The correct label should be 'BAFF'. The error has been corrected for all versions of the article.
Assuntos
Linfócitos B/imunologia , Células Epiteliais/imunologia , Switching de Imunoglobulina , Inibidor Secretado de Peptidases Leucocitárias/imunologia , Transdução de Sinais/imunologia , Fator Ativador de Células B/imunologia , Fator Ativador de Células B/metabolismo , Linfócitos B/metabolismo , Células Cultivadas , Citocinas/imunologia , Citocinas/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/metabolismo , Citometria de Fluxo , Humanos , Imunidade nas Mucosas , Immunoblotting , Imunoglobulina A/imunologia , Imunoglobulina G/imunologia , Imuno-Histoquímica , Hibridização In Situ , Interleucina-10/imunologia , Interleucina-10/metabolismo , Tonsila Palatina/citologia , Tonsila Palatina/imunologia , RNA Viral/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Inibidor Secretado de Peptidases Leucocitárias/metabolismo , Receptores Toll-Like/imunologia , Receptores Toll-Like/metabolismo , Linfopoietina do Estroma do TimoRESUMO
Bacteria colonize the intestine shortly after birth and thereafter exert several beneficial functions, including induction of protective immunoglobulin A (IgA) antibodies. The distal intestine contains IgA(2), which is more resistant to bacterial proteases than is IgA(1). The mechanism by which B cells switch from IgM to IgA(2) remains unknown. We found that human intestinal epithelial cells (IECs) triggered IgA(2) class switching in B cells, including IgA(1)-expressing B cells arriving from mucosal follicles, through a CD4(+) T cell-independent pathway involving a proliferation-inducing ligand (APRIL). IECs released APRIL after sensing bacteria through Toll-like receptors (TLRs) and further increased APRIL production by activating dendritic cells via thymic stromal lymphopoietin. Our data indicate that bacteria elicit IgA(2) class switching by linking lamina propria B cells with IECs through a TLR-inducible signaling program requiring APRIL. Thus, mucosal vaccines should activate IECs to induce more effective IgA(2) responses.
Assuntos
Linfócitos B/imunologia , Bactérias/imunologia , Imunoglobulina A/genética , Switching de Imunoglobulina , Mucosa Intestinal/imunologia , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/metabolismo , Linfócitos B/microbiologia , Vacinas Bacterianas/imunologia , Sequência de Bases , Linfócitos T CD4-Positivos/imunologia , Antígenos CD40/análise , Citocinas/metabolismo , Células Dendríticas/imunologia , Humanos , Mucosa Intestinal/microbiologia , Intestinos/citologia , Intestinos/imunologia , Intestinos/microbiologia , Dados de Sequência Molecular , Mucosa/imunologia , Receptores Toll-Like/imunologiaRESUMO
Meiosis is a specialized cell division that halves the genome complement, producing haploid gametes/spores from diploid cells. Proper separation of homologous chromosomes at the first meiotic division requires the production of physical connections (chiasmata) between homologs through recombinational exchange of chromosome arms after sister-chromatid cohesion is established but before chromosome segregation takes place. The events of meiotic prophase must thus occur in a strictly temporal order, but the molecular controls coordinating these events have not been well elucidated. Here, we demonstrate that the budding yeast cyclin-dependent kinase Cdc28 directly regulates the formation of the DNA double-strand breaks that initiate recombination by phosphorylating the Mer2/Rec107 protein and thereby modulating interactions of Mer2 with other proteins required for break formation. We propose that this function of Cdc28 helps to coordinate the events of meiotic prophase with each other and with progression through prophase.
Assuntos
Proteína Quinase CDC28 de Saccharomyces cerevisiae/metabolismo , Meiose/genética , Meiose/fisiologia , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , Sequência de Aminoácidos , Sítios de Ligação/genética , Cromatina/metabolismo , Cromossomos Fúngicos/metabolismo , Replicação do DNA , DNA Fúngico/genética , DNA Fúngico/metabolismo , Modelos Biológicos , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Fosforilação , Prófase , Recombinação Genética , Saccharomyces cerevisiae/citologia , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismoRESUMO
Most of what we know about proteins reflects their native folded structure. Much less is understood about the structure of unfolded proteins, which tends to be referred to as "random coil", lacking extended alpha-helix or beta-strand structure. Recent work suggests that unfolded proteins might adopt significant population of PII structure, an extended left-handed helix found in collagen and proline-rich peptides. A series of short peptides AcGGXGGNH2 has been adopted as a model for studying unfolded protein structure because of the minimal steric effect imposed by flanking glycines. Peptide AcGGAGGNH2 makes possible a host-guest conformation analysis of the middle residue alanine. NMR experiments reveal that the Phi and Psi dihedral angles of the central alanine are -73 degrees and 125 degrees , respectively, placing the alanine in the PII region of the Ramachandran plot. Circular dichroism shows a typical PII spectrum with a strong negative absorbance at 190 nm. Temperature experiments show the alanine structure shifts to increasing beta-strand at high temperature. Because the alanine side chain most closely represents unsubstituted peptide backbone, these results have significant implications for the conformational entropy of unfolded polypeptide chains.