RESUMEN
Secretory (S) IgA is the predominant mucosal Ab that protects host epithelial barriers and promotes microbial homeostasis. SIgA production occurs when plasma cells assemble two copies of monomeric IgA and one joining chain (JC) to form dimeric (d) IgA, which is bound by the polymeric Ig receptor (pIgR) on the basolateral surface of epithelial cells and transcytosed to the apical surface. There, pIgR is proteolytically cleaved, releasing SIgA, a complex of the dIgA and the pIgR ectodomain, called the secretory component (SC). The pIgR's five Ig-like domains (D1-D5) undergo a conformational change upon binding dIgA, ultimately contacting four IgA H chains and the JC in SIgA. In this study, we report structure-based mutational analysis combined with surface plasmon resonance binding assays that identify key residues in mouse SC D1 and D3 that mediate SC binding to dIgA. Residues in D1 CDR3 are likely to initiate binding, whereas residues that stabilize the D1-D3 interface are likely to promote the conformational change and stabilize the final SIgA structure. Additionally, we find that the JC's three C-terminal residues play a limited role in dIgA assembly but a significant role in pIgR/SC binding to dIgA. Together, these results inform models for the intricate mechanisms underlying IgA transport across epithelia and functions in the mucosa.
Asunto(s)
Inmunoglobulina A Secretora , Receptores de Inmunoglobulina Polimérica , Componente Secretorio , Animales , Ratones , Inmunoglobulina A Secretora/inmunología , Inmunoglobulina A Secretora/metabolismo , Componente Secretorio/metabolismo , Componente Secretorio/inmunología , Receptores de Inmunoglobulina Polimérica/metabolismo , Receptores de Inmunoglobulina Polimérica/inmunología , Receptores de Inmunoglobulina Polimérica/genética , Unión Proteica , Multimerización de Proteína , Humanos , Inmunoglobulina A/inmunología , Inmunoglobulina A/metabolismo , Conformación ProteicaRESUMEN
Fish rely on mucosal surfaces as their first defence barrier against pathogens. Maintaining mucosal homeostasis is therefore crucial for their overall well-being, and it is likely that secreted immunoglobulins (sIg) play a pivotal role in sustaining this balance. In mammals, the poly-Ig receptor (pIgR) is an essential component responsible for transporting polymeric Igs across mucosal epithelia. In teleost fish, a counterpart of pIgR has been identified and characterized, exhibiting structural differences and broader mRNA expression patterns compared to mammals. Despite supporting evidence for the binding of Igs to recombinant pIgR proteins, the absence of a joining chain (J-chain) in teleosts challenges the conventional understanding of Ig transport mechanisms. The transport of IgM to the intestine via the hepatobiliary route is observed in vertebrates and has been proposed in a few teleosts. Investigations on the stomachless fish, ballan wrasse, revealed a significant role of the hepatobiliary route and interesting possibilities for alternative IgM transport routes that might include pancreatic tissue. These findings highlight the importance of gaining a thorough understanding of the mechanisms behind Ig transport to the gut in various teleosts. This review aims to gather existing information on pIgR-mediated transport across epithelial cells and immunoglobulin transport pathways to the gut lumen in teleost fish. It provides comparative insights into the hepatobiliary transport of Igs to the gut, emphasizing the current understanding in teleost fish while exploring potential alternative pathways for Ig transport to the gut lumen. Despite significant progress in understanding various aspects, there is still much to uncover, especially concerning the diversity of mechanisms across different teleost species.
Asunto(s)
Peces , Inmunoglobulina M , Animales , Inmunoglobulina M/inmunología , Peces/inmunología , Peces/genética , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Receptores de Inmunoglobulina Polimérica/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Proteínas de Peces/química , Tracto Gastrointestinal/inmunologíaRESUMEN
Immunoglobulin A (IgA) plays a crucial role in the human immune system, particularly in mucosal immunity. IgA antibodies that target the mucosal surface are made up of two to five IgA monomers linked together by the joining chain, forming polymeric molecules. These IgA polymers are transported across mucosal epithelial cells by the polymeric immunoglobulin receptor pIgR, resulting in the formation of secretory IgA (SIgA). This review aims to explore recent advancements in our molecular understanding of IgA, with a specific focus on SIgA, and the interaction between IgA and pathogen molecules.
Asunto(s)
Inmunoglobulina A Secretora , Inmunoglobulina A , Humanos , Inmunoglobulina A Secretora/inmunología , Inmunoglobulina A Secretora/metabolismo , Inmunoglobulina A/inmunología , Inmunoglobulina A/química , Inmunoglobulina A/metabolismo , Inmunoglobulina A/genética , Animales , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Receptores de Inmunoglobulina Polimérica/metabolismo , Receptores de Inmunoglobulina Polimérica/química , Inmunidad MucosaRESUMEN
PURPOSE: Transport of secretory immunoglobulin A (SIgA) through the airway epithelial cell barrier into the mucosal lumen by the polymeric immunoglobulin receptor (pIgR) is an important mechanism of respiratory mucosal host defense. Identification of immunomodulating substances that regulate secretory immunity might have therapeutic implications with regard to an improved immune exclusion. Thus, we sought to analyze secretory immunity under homeostatic and immunomodulating conditions in different compartments of the murine upper and lower respiratory tract (URT&LRT). METHODS: Pigr gene expression in lung, trachea, and nasal-associated lymphoid tissue (NALT) of germ-free mice, specific pathogen-free mice, mice with an undefined microbiome, as well as LPS- and IFN-γ-treated mice was determined by quantitative real-time PCR. IgA levels in bronchoalveolar lavage (BAL), nasal lavage (NAL), and serum were determined by ELISA. LPS- and IFN-γ-treated mice were colonized with Streptococcus pneumoniae and bacterial CFUs were determined in URT and LRT. RESULTS: Respiratory Pigr expression and IgA levels were dependent on the degree of exposure to environmental microbial stimuli. While immunostimulation with LPS and IFN-γ differentially impacts respiratory Pigr expression and IgA in URT vs. LRT, only prophylactic IFN-γ treatment reduces nasal colonization with S. pneumoniae. CONCLUSION: Airway-associated secretory immunity can be partly modulated by exposure to microbial ligands and proinflammatory stimuli. Prophylactic IFN-γ-treatment modestly improves antibacterial immunity in the URT, but this does not appear to be mediated by SIgA or pIgR.
Asunto(s)
Inmunoglobulina A Secretora , Receptores de Inmunoglobulina Polimérica , Mucosa Respiratoria , Animales , Antibacterianos/inmunología , Antibacterianos/farmacología , Inmunoglobulina A Secretora/inmunología , Inmunoglobulina A Secretora/metabolismo , Pulmón/efectos de los fármacos , Pulmón/inmunología , Pulmón/metabolismo , Ratones , Receptores de Superficie Celular/inmunología , Receptores de Superficie Celular/metabolismo , Receptores de Inmunoglobulina Polimérica/inmunología , Receptores de Inmunoglobulina Polimérica/metabolismo , Mucosa Respiratoria/efectos de los fármacos , Mucosa Respiratoria/inmunología , Mucosa Respiratoria/metabolismoRESUMEN
The global health crisis and economic tolls of COVID-19 necessitate a panoply of strategies to treat SARS-CoV-2 infection. To date, few treatment options exist, although neutralizing antibodies against the spike glycoprotein have proven to be effective. Because infection is initiated at the mucosa and propagates mainly at this site throughout the course of the disease, blocking the virus at the mucosal milieu should be effective. However, administration of biologics to the mucosa presents a substantial challenge. Here, we describe bifunctional molecules combining single-domain variable regions that bind to the polymeric Ig receptor (pIgR) and to the SARS-CoV-2 spike protein via addition of the ACE2 extracellular domain (ECD). The hypothesis behind this design is that pIgR will transport the molecule from the circulation to the mucosal surface where the ACE ECD would act as a decoy receptor for the nCoV2. The bifunctional molecules bind SARS-Cov-2 spike glycoprotein in vitro and efficiently transcytose across the lung epithelium in human tissue-based analyses. Designs featuring ACE2 tethered to the C-terminus of the Fc do not induce antibody-dependent cytotoxicity against pIgR-expressing cells. These molecules thus represent a potential therapeutic modality for systemic administration of neutralizing anti-SARS-CoV-2 molecules to the mucosa.
Asunto(s)
Anticuerpos Antivirales , Tratamiento Farmacológico de COVID-19 , Receptores de Inmunoglobulina Polimérica , SARS-CoV-2/inmunología , Anticuerpos de Cadena Única , Glicoproteína de la Espiga del Coronavirus/inmunología , Enzima Convertidora de Angiotensina 2/genética , Enzima Convertidora de Angiotensina 2/inmunología , Animales , Anticuerpos Antivirales/genética , Anticuerpos Antivirales/inmunología , Anticuerpos Antivirales/farmacología , Células CHO , COVID-19/genética , COVID-19/inmunología , Cricetulus , Perros , Femenino , Humanos , Células de Riñón Canino Madin Darby , Ratones , Mucosa Bucal/inmunología , Dominios Proteicos , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Receptores de Inmunoglobulina Polimérica/uso terapéutico , SARS-CoV-2/genética , Anticuerpos de Cadena Única/genética , Anticuerpos de Cadena Única/inmunología , Anticuerpos de Cadena Única/farmacocinética , Anticuerpos de Cadena Única/farmacología , Glicoproteína de la Espiga del Coronavirus/antagonistas & inhibidores , Glicoproteína de la Espiga del Coronavirus/genética , PorcinosRESUMEN
The polyimmunoglobulin receptor (pIgR) transcytoses J chain-containing antibodies through mucosal epithelia. In mammals, two cis-duplicates of PIGR, FCMR, and FCAMR, flank the PIGR gene. A PIGR duplication is first found in amphibians, previously annotated as PIGR2 (herein xlFCAMR), and is expressed by APCs. We demonstrate that xlFcamR is the equivalent of mammalian FcamR. It has been assumed that pIgR is the oldest member of this family, yet our data could not distinguish whether PIGR or FCAMR emerged first; however, FCMR was the last family member to emerge. Interestingly, bony fish "pIgR" is not an orthologue of tetrapod pIgR, and possibly acquired its function via convergent evolution. PIGR/FCAMR/FCMR are members of a larger superfamily, including TREM, CD300, and NKp44, which we name the "double-disulfide Ig superfamily" (ddIgSF). Domains related to each ddIgSF family were identified in cartilaginous fish (sharks, chimeras) and encoded in a single gene cluster syntenic to the human pIgR locus. Thus, the ddIgSF families date back to the earliest antibody-based adaptive immunity, but apparently not before. Finally, our data strongly suggest that the J chain arose in evolution only for Ig multimerization. This study provides a framework for further studies of pIgR and the ddIgSF in vertebrates.
Asunto(s)
Antígenos CD/genética , Inmunidad Mucosa/inmunología , Receptores Fc/genética , Receptores Opioides mu/genética , Receptores de Inmunoglobulina Polimérica/genética , Transcitosis/inmunología , Animales , Antígenos CD/inmunología , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Humanos , Inmunoglobulinas/metabolismo , Filogenia , Transporte de Proteínas/fisiología , Receptores Fc/inmunología , Receptores Opioides mu/inmunología , Receptores de Inmunoglobulina Polimérica/inmunología , Transcitosis/genética , Xenopus laevisRESUMEN
The upper respiratory tract is highly exposed to airborne pathogens and serves as an important inductive site for protective antibody responses, including mucosal IgA and systemic IgG. However, it is currently unknown to what extent inhaled environmental toxins, such as a cigarette smoke, affect the ability to induce antibody-mediated immunity at this site. Using a murine model of intranasal lipopolysaccharide and ovalbumin (LPS/OVA) immunization, we show that cigarette smoke exposure compromises the induction of antigen-specific IgA in the upper airways and systemic circulation. Deficits in OVA-IgA were observed in conjunction with a reduced accumulation of OVA-specific IgA antibody-secreting cells (ASCs) in the nasal mucosa, inductive tissues (NALT, cervical lymph nodes, spleen) and the blood. Nasal OVA-IgA from smoke-exposed mice also demonstrated reduced avidity during the acute post-immunization period in association with an enhanced mutational burden in the cognate nasal Igha repertoire. Mechanistically, smoke exposure attenuated the ability of the nasal mucosa to upregulate VCAM-1 and pIgR, suggesting that cigarette smoke may inhibit both nasal ASC homing and IgA transepithelial transport. Overall, these findings demonstrate the immunosuppressive nature of tobacco smoke and illustrate the diversity of mechanisms through which this noxious stimulus can interfere with IgA-mediated immunity in the upper airways.
Asunto(s)
Formación de Anticuerpos/inmunología , Antígenos/inmunología , Inmunidad Mucosa , Inmunoglobulina A Secretora/inmunología , Mucosa Nasal/inmunología , Mucosa Nasal/metabolismo , Fumar Tabaco/efectos adversos , Animales , Biomarcadores , Quimiocinas CC/metabolismo , Inmunización , Inmunofenotipificación , Lipopolisacáridos/inmunología , Tejido Linfoide/inmunología , Tejido Linfoide/metabolismo , Ratones , Ovalbúmina/inmunología , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Hipermutación Somática de Inmunoglobulina , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
The polymeric immunoglobulin receptor (pIgR) transports secretory immunoglobulins across mucosal epithelial cells into external secretions, playing critical roles in mucosal surface defenses, but the regulation mechanism of pIgR expression is not clarified in teleost fish. In this study, the dynamic changes of flounder (Paralichthys olivaceus) pIgR (fpIgR) and pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) mRNA expression in mucosal tissues were first analyzed post inactivated Vibrio anguillarum immunization, and increased production of TNF-α was found to correlate with increased expression of fpIgR. To determine that cytokine TNF-α influenced fpIgR expression, following confirming that natural fpIgR expressed on flounder gill (FG) cells, FG cells were incubated with various concentrations of recombinant TNF-α for different time, the results showed that the expressions of fpIgR were significantly upregulated at gene and protein levels in a dose-dependent and time-dependent manner, and similar change trend was observed for free secretory component (SC) secreted by fpIgR into the culture supernatant. After FG cells were treated with TNF-α, specific phosphoinositide 3-kinase (PI3K) inhibitor wortmannin, nuclear factor kappa-B (NF-κB) inhibitor Bay11-7082, and the mixtures of TNF-α and wortmannin / Bay11-7082 respectively, the fpIgR protein and mRNA levels, together with SC secretion, obviously decreased in wortmannin- and Bay11-7082-treated cells compared with the untreated control, and cotreatment with wortmannin / Bay11-7082 plus TNF-α resulted in lower expression compared with that upon treatment with TNF-α alone, indicating that the inhibition of PI3K and NF-κB both blocked the ability of TNF-α to increase cellular fpIgR and SC levels. Furthermore, the gene expressions of PI3K and NF-κB were upregulated and present a tendency to increase first and then decrease after TNF-α treatment of FG cells; However, the expression of PI3K mRNA was inhibited significantly by wortmannin but not by Bay11-7082, and the expression of NF-κB mRNA was suppressed obviously by Bay11-7082 but not by wortmannin, suggesting that inhibition of PI3K or NF-κB had no influence on each other. All these results collectively revealed that TNF-α could transcriptionally upregulate fpIgR expression and SC production, and this TNF-α-induced pIgR expression was regulated by complex mechanisms that involved PI3K and NF-κB signaling pathways, which provided evidences for pro-inflammatory cytokine TNF-α acting as a regulator in pIgR expression and better understanding of regulation mechanism of pIgR expression in teleost fish.
Asunto(s)
Regulación de la Expresión Génica/inmunología , FN-kappa B/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Receptores de Inmunoglobulina Polimérica/inmunología , Factor de Necrosis Tumoral alfa/inmunología , Animales , Células Cultivadas , Lenguado/inmunología , Branquias/citología , Branquias/inmunología , Inmunización , FN-kappa B/antagonistas & inhibidores , Nitrilos/farmacología , Inhibidores de las Quinasa Fosfoinosítidos-3/farmacología , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Receptores de Inmunoglobulina Polimérica/genética , Transducción de Señal/inmunología , Sulfonas/farmacología , Regulación hacia Arriba/genética , Vibrio/inmunología , Wortmanina/farmacologíaRESUMEN
The polymeric immunoglobulin receptor (pIgR) is an important molecule in the mucosal immunity of teleosts. Previous studies have shown that pIgR can bind and transport polymeric immunoglobulins (pIgs), but few studies have focused on the binding of teleost pIgR to bacteria. In this study, we identified a gene encoding pIgR in largemouth bass (Micropterus salmoides). The pIgR gene contained two Ig-like domains (ILDs), which were homologous to ILD1 and ILD5 of mammalian pIgR. Our results showed that largemouth bass pIgR-ILD could combine with IgM. Moreover, we also found that largemouth bass pIgR-ILD could bind to Aeromonas hydrophila and Micrococcus luteus. Further analysis showed that largemouth bass pIgR-ILD could also combine with lipopolysaccharide (LPS), peptidoglycan (PGN) and various saccharides, and reduced binding to bacteria was observed with LPS and PGN treatment, indicating that largemouth bass pIgR could bind to bacteria to prevent infection and that saccharide binding is an important interaction mechanism between pIgR and bacteria. These results collectively demonstrated that largemouth bass pIgR not only combines with IgM but also binds to bacteria by various saccharides.
Asunto(s)
Aeromonas hydrophila/inmunología , Lubina/inmunología , Inmunoglobulina M/inmunología , Micrococcus luteus/inmunología , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Lubina/genética , Enfermedades de los Peces/inmunología , Inmunidad Mucosa/genética , Inmunidad Mucosa/inmunología , Lipopolisacáridos/inmunología , Peptidoglicano/inmunología , Filogenia , Dominios Proteicos/genética , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
Immunoglobulins are glycoproteins which are produced as membrane-bound receptors on B-cells or in a secreted form, known as antibodies. In teleosts, three immunoglobulin isotypes, IgM, IgT, and IgD, are present, each comprising two identical heavy and two identical light polypeptide chains. The basic mechanisms for generation of immunoglobulin diversity are similar in teleosts and higher vertebrates. The B-cell pre-immune repertoire is diversified by VDJ recombination, junctional flexibility, addition of nucleotides, and combinatorial association of light and heavy chains, while the post-immune repertoire undergoes somatic hypermutation during clonal expansion. Typically, the teleost immunoglobulin heavy chain gene complex has a modified translocon arrangement where the Dτ-Jτ-Cτ cluster of IgT is generally located between the variable heavy chain (VH) region and the Dµ/δ-Jµ/δ-Cµ-Cδ gene segments, or within the set of VH gene segments. However, multiple genome duplication and deletion events and loss of some individual genes through evolution has complicated the IgH gene organization. The IgH gene arrangement allows the expression of either IgT or IgM/IgD. Alternative splicing is responsible for the regulation of IgM/IgD expression and the secreted versus transmembrane forms of IgT, IgD, and IgM. The overall structure of IgM and IgT is usually conserved across species, whereas IgD has a large variety of structures. IgM is the main effector molecule in both systemic and mucosal immunity and shows a broad range of concentrations in different teleost species. Although IgM is usually present in higher concentrations under normal conditions, IgT is considered the main mucosal Ig.
Asunto(s)
Peces/inmunología , Inmunoglobulinas/genética , Inmunoglobulinas/inmunología , Animales , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Peces/genética , Genes de Inmunoglobulinas/genética , Genes de Inmunoglobulinas/inmunología , Inmunidad Mucosa , Inmunoglobulina D/genética , Inmunoglobulina D/inmunología , Inmunoglobulina M/genética , Inmunoglobulina M/inmunología , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Recombinación V(D)JRESUMEN
Porcine deltacoronavirus (PDCoV) is a porcine enteropathogenic coronavirus that causes watery diarrhea, vomiting, and frequently death in piglets, causing serious economic losses to the pig industry. The strain CHN-JS-2017 was isolated and identified by cytopathology, immunofluorescence assays, transmission electron microscopy, and sequence analysis. A nucleotide sequence alignment showed that the whole genome of CHN-JS-2017 is 97.4%-99.6% identical to other PDCoV strains. The pathogenicity of the CHN-JS-2017 strain was investigated in orally inoculated five-day-old piglets; the piglets developed acute, watery diarrhea, but all recovered and survived. CHN-JS-2017 infection-induced microscopic lesions were observed, and viral antigens were detected mainly by immunohistochemical staining in the small intestine. The neonatal Fc receptor (FcRn) and polymeric immunoglobulin receptor (pIgR) are crucial immunoglobulin (Ig) receptors for the transcytosis ofimmunoglobulin G (IgG), IgA, or IgM. Importantly, CHN-JS-2017 infected five-day-old piglets could significantly down-regulate the expression of FcRn, pIgR, and nuclear factor-kappa B (NF-κB)in the intestinal mucosa. Note that the level of FcRn mRNA in the intestinal mucosa of normal piglets is positively correlated with pIgR and NF-κB. At the same time, the expressions of FcRn, pIgR, and NF-κB mRNA are also positively correlated in infected piglets. These results may help explain the immunological and pathological changes associated with porcine deltacorononirus infection.
Asunto(s)
Infecciones por Coronavirus/veterinaria , Coronavirus/clasificación , Antígenos de Histocompatibilidad Clase I/inmunología , Mucosa Intestinal/inmunología , Receptores Fc/inmunología , Receptores de Inmunoglobulina Polimérica/inmunología , Enfermedades de los Porcinos/virología , Animales , Antígenos Virales/análisis , Coronavirus/aislamiento & purificación , Infecciones por Coronavirus/inmunología , Diarrea/veterinaria , Diarrea/virología , Regulación de la Expresión Génica , Mucosa Intestinal/virología , Intestino Delgado/inmunología , Intestino Delgado/virología , FN-kappa B/inmunología , Filogenia , ARN Viral/análisis , Alineación de Secuencia , Análisis de Secuencia de ADN , Porcinos , Enfermedades de los Porcinos/inmunología , Esparcimiento de VirusRESUMEN
The Polymeric Immunoglobulin Receptor (pIgR) gene has been proved to play an important role in transporting polymeric immunoglobulin (Ig) in the mucosal tissues of mammals. pIgR gene also exists in teleost, but the genetic diversity and functions of this gene still need to be further explored. We obtained seven grass carp pIgR splicing transcripts, a full-length pIgR (CipIgR-1) and six truncated variants (CipIgR-2 to CipIgR-7). The full-length pIgR contained two immunoglobulin-like domains (ILD), a transmembrane domain (TMD) and a cytoplasmic domain (CyD). The CipIgR-2 lacked a small part in CyD, and CipIgR-3 lost TMD and CyD. Partial cDNA sequences of the other four grass carp pIgR variants (CipIgR-4 to CipIgR-7) were also cloned. The total expression levels of CipIgR and its variants in different tissues were detected by real-time quantitative PCR. The highest expression was found in the intestine, followed by the spleen and the skin. The function of the two extracellular ILDs of CipIgR was investigated based on its combining capacity with grass carp immunoglobulin M (IgM) and aquatic pathogenic bacteria. The cDNA sequences of two ILDs were cloned and expressed in Escherichia coli BL21 (DE3). Recombinant ILDs protein was purified and incubated with different bacteria respectively. Results of Western blot showed the recombinant protein could combine Bacillus subtilis, Vibrio parahaemolyticus, and Escherichia coli. In addition, binding activity of rILDs with grass carp IgM was detected. Collectively, these results indicated that multiple variants of pIgR gene in grass carp might be involved in the antibacterial immunity.
Asunto(s)
Carpas/genética , Carpas/inmunología , Dominios de Inmunoglobulinas , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Animales , Bacterias/inmunología , Sitios de Unión de Anticuerpos , Clonación Molecular , Escherichia coli/genética , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/microbiología , Variación Genética , Inmunoglobulina M/metabolismo , Filogenia , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunologíaRESUMEN
In the present work, the polymeric immunoglobulin receptor-like (pIgRL) from flounder (Paralichthys olivaceus) was firstly cloned and identified. The full length cDNA of flounder pIgRL was of 1393 bp including an open reading frame of 1053 bp, and the deduced pIgRL sequence encoded 350 amino acids, with a predicted molecular mass of 39â¯kDa. There were two immunoglobulin-like domains in flounder pIgRL. In healthy flounder, the transcriptional level of pIgRL was detected in different tissues by real-time PCR, showing the highest level in the skin and gills, and higher levels in the spleen and hindgut. After flounders were vaccinated with inactivated Vibrio anguillarum via intraperitoneal injection and immersion, the pIgRL mRNA level increased firstly and then declined in all tested tissues during 48â¯h, and the maximum expression levels in the gills, skin, spleen and hindgut in immersion group, or in the spleen, head kidney, skin and gills in injection group, were higher than in other tested tissues. In addition, recombinant protein of the extracellular region of flounder pIgRL was expressed in Escherichia coli BL21 (DE3), and rabbit anti-pIgRL polyclonal antibodies were prepared, which specifically reacted with the recombinant pIgRL, and a 39â¯kDa protein confirmed as natural pIgRL by liquid chromatography-mass spectrometry in skin mucus of flounder. Co-immunoprecipitation assay and western-blotting demonstrated that the pIgRL, together with IgM, could be immunoprecipitated by anti-pIgRL antibody in gut, skin and gill mucus of flounder, suggesting the existence of pIgRL-IgM complexes. These results indicated that the flounder pIgRL was probably involved in the mucosal IgM transportation and played important roles in mucosal immunity.
Asunto(s)
Enfermedades de los Peces/inmunología , Peces Planos/genética , Peces Planos/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Secuencia de Aminoácidos , Animales , Vacunas Bacterianas/inmunología , Secuencia de Bases , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Inyecciones Intraperitoneales/veterinaria , Filogenia , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Receptores de Inmunoglobulina Polimérica/química , Alineación de Secuencia/veterinaria , Vibrio/inmunologíaRESUMEN
Viral entry into the host cell is the first step towards successful infection. Viral entry starts with virion attachment, and binding to receptors. Receptor binding viruses either directly release their genome into the cell, or enter cells through endocytosis. For DNA viruses and a few RNA viruses, the endocytosed viruses will transport from cytoplasm into the nucleus followed by gene expression. Receptors on the cell membrane play a crucial role in viral infection. Although several attachment factors, or candidate receptors, for the infection of white spot syndrome virus (WSSV) were identified in shrimp, the authentic entry receptors for WSSV infection and the intracellular signaling triggering by interaction of WSSV with receptors remain unclear. In the present study, a receptor for WSSV infection in kuruma shrimp, Marsupenaeus japonicus, was identified. It is a member of the immunoglobulin superfamily (IgSF) with a transmembrane region, and is similar to the vertebrate polymeric immunoglobulin receptor (pIgR); therefore, it was designated as a pIgR-like protein (MjpIgR for short). MjpIgR was detected in all tissues tested, and its expression was significantly induced by WSSV infection at the mRNA and protein levels. Knockdown of MjpIgR, and blocking MjpIgR with its antibody inhibited WSSV infection in shrimp and overexpression of MjpIgR facilitated the invasion of WSSV. Further analyses indicated that MjpIgR could independently render non-permissive cells susceptible to WSSV infection. The extracellular domain of MjpIgR interacts with envelope protein VP24 of WSSV and the intracellular domain interacts with calmodulin (MjCaM). MjpIgR was oligomerized and internalized following WSSV infection and the internalization was associated with endocytosis of WSSV. The viral internalization facilitating ability of MjpIgR could be blocked using chlorpromazine, an inhibitor of clathrin dependent endocytosis. Knockdown of Mjclathrin and its adaptor protein AP-2 also inhibited WSSV internalization. All the results indicated that MjpIgR-mediated WSSV endocytosis was clathrin dependent. The results suggested that MjpIgR is a WSSV receptor, and that WSSV enters shrimp cells via the pIgR-CaM-Clathrin endocytosis pathway.
Asunto(s)
Penaeidae/inmunología , Receptores de Inmunoglobulina Polimérica/inmunología , Virus del Síndrome de la Mancha Blanca 1/metabolismo , Animales , Acuicultura/métodos , Virus ADN , Endocitosis , Penaeidae/metabolismo , Penaeidae/patogenicidad , Unión Proteica , Receptores de Inmunoglobulina Polimérica/metabolismo , Proteínas del Envoltorio Viral , Internalización del Virus , Replicación Viral , Virus del Síndrome de la Mancha Blanca 1/patogenicidadRESUMEN
The receptor responsible for maternofetal transmission of immunoglobulin (Igs) in the teleosts is not clear. Polymeric immunoglobulin receptor (pIgR) specifically binds with IgA and IgM and mediates the transcytosis of intracellular polymeric immunoglobulins (pIgs) at the mucosal surface to protect against pathogens. Hence there is a possibility that it may be involved in the transmission of maternal Igs. The aim of the present study was to detect the expression and localization of pIgR during embryonal development in turbot (Scophthalmus maximus). pIgR gene was first cloned from eggs and embryos of turbot with or without parent immunization. The expression and distribution of pIgR in unfertilized egg and in embryos ranging from day 1 to day 5 after fertilization were analyzed using reverse transcriptase quantitative polymerase chain reaction and in situ hybridization. pIgR gene was detected in all eggs and embryos at different stages of development, with the highest level detected on the 5th day. pIgR mRNA was observed to be first located in the whole blastoderm and enveloped the yolk sac. Later, it was located around entoderm including primary digestive tract and pronephric tubule tract, and finally it was located at the joint of abdomen and vitelline membrane. Then, Eukaryotic expression plasmid carrying pIgR gene was constructed and transfected into HEK293T cells. Results showed mature pIgR protein located on the cellular membrane, and could bound IgM in vitro. Our findings provide information for studying the involvement of pIgR in maternal Igs transportation in turbot.
Asunto(s)
Proteínas de Peces/genética , Peces Planos/genética , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Animales , Desarrollo Embrionario/genética , Femenino , Proteínas de Peces/inmunología , Peces Planos/embriología , Peces Planos/metabolismo , Especificidad de ÓrganosRESUMEN
Polymeric immunoglobulin receptor (pIgR) functions in transporting polymeric immunoglobulin across epithelial cells into external secretion in animals. During animal evolution, fish was situated at a transition point on the phylogenetic spectrum between species possessing only innate immunity (i.e., invertebrates) and species depending heavily on adaptive immunity (i.e., mammals). Previous studies reported that fish and mammals significantly differ in pIgR. This review summarized the differences in pIgR structure, function, and transcriptional regulation between fish and mammals. A model of the transcriptional regulation of the pIgR gene was suggested. In this model, microbes could activate Toll-like receptor, trigger the cascade reactions in the signaling pathway, and then activate transcription factors that regulate pIgR expression through combining with the pIgR promoter. This review provides some suggestions for further studies on the function and regulatory mechanism of pIgR in fish and other animals.
Asunto(s)
Peces/inmunología , Mamíferos/inmunología , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Inmunidad Adaptativa , Animales , Células Epiteliales/inmunología , Regulación de la Expresión Génica , Inmunidad Innata , Filogenia , Regiones Promotoras Genéticas , Transducción de Señal , Receptores Toll-Like/inmunología , Factores de Transcripción/genética , Factores de Transcripción/inmunologíaRESUMEN
The gastrointestinal tract houses millions of microbes, and thus has evolved several host defense mechanisms to keep them at bay, and prevent their entry into the host. One such mucosal surface defense is the secretion of secretory immunoglobulins (SIg). Secretion of SIg depends on the polymeric immunoglobulin receptor (pIgR), which transports polymeric Ig (IgA or IgM) from the basolateral surface of the epithelium to the apical side. Upon reaching the luminal side, a portion of pIgR, called secretory component (SC) is cleaved off to release Ig, forming SIg. Through antigen-specific and non-specific binding, SIg can modulate microbial communities and pathogenic microbes via several mechanisms: agglutination and exclusion from the epithelial surface, neutralization, or via host immunity and complement activation. Given the crucial role of SIg as a microbial scavenger, some pathogens also evolved ways to modulate and utilize pIgR and SIg to facilitate infection. This review will cover the regulation of the pIgR/SIg cycle, mechanisms of SIg-mediated mucosal protection as well as pathogen utilization of SIg.
Asunto(s)
Inmunidad Mucosa , Inmunoglobulina A Secretora/inmunología , Inmunoglobulinas/inmunología , Receptores de Inmunoglobulina Polimérica/inmunología , Animales , Transporte Biológico , Tracto Gastrointestinal/inmunología , Humanos , Inmunoglobulina A/inmunología , RatonesRESUMEN
The polymeric immunoglobulin receptor (pIgR) is an essential component of the mucosal immune system in jawed vertebrates including teleost fish, which mediate transepithelial transport of secretory immunoglobulins (sIgs) to protect organisms against environmental pathogens. In this study, we firstly cloned and identified the pIgR from dojo loach (Misgurnus anguillicaudatus). The full-length cDNA of Ma-pIgR was of 1145 bp, containing an open reading frame (ORF) of 1101 bp encoded a predicted protein of 336 amino acids. The structure of Ma-pIgR is comprised of a signal peptide, a transmembrane region, an intracellular region and an extracellular region with two Ig-like domains (ILDs), which are similar to their counterparts described in other teleosts. Multiple sequence alignment and phylogenetic analysis showed the dojo loach is closely related to the fish family Cyprinidae. The transcriptional level of Ma-pIgR was detected by quantitative real-time PCR (qRT-PCR) in different tissues and high expression was found in liver, skin, kidney, eye, fin and gills. Two infection models of the loach with bacteria (Aeromonas hydrophila) and parasite (Ichthyophthirius multifiliis) were constructed for the first time. Histological studies showed the goblet cells in skin significantly increased and the ratio of gill length to width also significantly changed after challenged with A.hydrophila. Both challenge experiments resulted in the significant up-regulated expression of Ma-pIgR not only in kidney and spleen, but also in skin and gills. Our results suggest that pIgR may play an important role in skin and gill mucosal immunity to protect the loach against bacteria and parasite.
Asunto(s)
Cipriniformes/genética , Cipriniformes/inmunología , Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Mucosa/genética , Receptores de Inmunoglobulina Polimérica/genética , Receptores de Inmunoglobulina Polimérica/inmunología , Aeromonas hydrophila/fisiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Infecciones por Cilióforos/inmunología , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica , Infecciones por Bacterias Gramnegativas/inmunología , Hymenostomatida/fisiología , Filogenia , Receptores de Inmunoglobulina Polimérica/química , Alineación de Secuencia/veterinariaRESUMEN
Dengue is a major global disease requiring improved treatment and prevention strategies. The recently licensed Sanofi Pasteur Dengvaxia vaccine does not protect children under the age of nine, and additional vaccine strategies are thus needed to halt this expanding global epidemic. Here, we employed a molecular engineering approach and plant expression to produce a humanized and highly immunogenic poly-immunoglobulin G scaffold (PIGS) fused to the consensus dengue envelope protein III domain (cEDIII). The immunogenicity of this IgG Fc receptor-targeted vaccine candidate was demonstrated in transgenic mice expressing human FcγRI/CD64, by induction of neutralizing antibodies and evidence of cell-mediated immunity. Furthermore, these molecules were able to prime immune cells from human adenoid/tonsillar tissue ex vivo as evidenced by antigen-specific CD4+ and CD8+ T-cell proliferation, IFN-γ and antibody production. The purified polymeric fraction of dengue PIGS (D-PIGS) induced stronger immune activation than the monomeric form, suggesting a more efficient interaction with the low-affinity Fcγ receptors on antigen-presenting cells. These results show that the plant-expressed D-PIGS have the potential for translation towards a safe and easily scalable single antigen-based tetravalent dengue vaccine.
Asunto(s)
Vacunas contra el Dengue/inmunología , Ingeniería Genética , Receptores de Inmunoglobulina Polimérica/genética , Proteínas Recombinantes de Fusión/genética , Tonsila Faríngea/inmunología , Adyuvantes Inmunológicos/genética , Animales , Anticuerpos Neutralizantes/inmunología , Vacunas contra el Dengue/genética , Femenino , Ingeniería Genética/métodos , Humanos , Inmunidad Celular , Inmunoglobulina G/inmunología , Masculino , Ratones , Ratones Transgénicos , Tonsila Palatina/inmunología , Plantas Modificadas Genéticamente , Receptores de IgG/inmunología , Receptores de Inmunoglobulina Polimérica/inmunología , Proteínas Recombinantes de Fusión/inmunología , Nicotiana/genéticaRESUMEN
The polymeric immunoglobulin receptor (pIgR) plays a pivotal role in mucosal immune response by transporting polymeric immunoglobulins onto the surface of mucosal epithelia to protect animals from invading pathogens. In this study, the full-length cDNA of pIgR was firstly cloned in Qihe crucian carp (Carassius auratus), hereafter designated as CapIgR, by using reverse transcription polymerase chain reaction and rapid amplification of cDNA ends. The molecular characterization and expression of CapIgR were investigated. The full-length cDNA sequence of CapIgR was composed of 1409 bp, which included a 112 bp 5'-untranslated region (UTR), a 984 bp ORF, and a 313 bp 3'-UTR, with a putative polyadenylation signal sequence AATAAA located upstream of the poly(A) tail. The deduced amino acid sequence indicated that CapIgR was a single-spanning transmembrane protein with 327 amino acids and possessed a signal peptide, an extracellular region containing two immunoglobulin-like domains, a transmembrane region, and an intracellular region. The mRNA expression levels of CapIgR were detected in different tissues of healthy C. auratus by quantitative real-time PCR, and the highest expression level was found in the liver. After Aeromonas hydrophila challenge, CapIgR expression was upregulated in different tissues at certain time points, and temporal expression changes of CapIgR fluctuated in a time-dependent manner. CapIgR exhibited rapid immune response to A. hydrophila challenge and played an important role in the immune defense of fish. These findings provided insights into the structure, function, and immune defense mechanism of CapIgR in C. auratus. This study can serve as a basis for developing disease control strategies in aquaculture.