Selective adherence of IgA to murine Peyer's patch M cells: evidence for a novel IgA receptor.

M cells represent the primary route by which mucosal Ags are transported across the intestinal epithelium and delivered to underlying gut-associated lymphoid tissues. In rodents and rabbits, Peyer's patch M cells selectively bind and endocytose secretory IgA (SIgA) Abs. Neither the nature of the M cell IgR nor the domains of SIgA involved in this interaction are known. Using a mouse ligated ileal loop assay, we found that monoclonal IgA Abs with or without secretory component, but not IgG or IgM Abs, bound to the apical surfaces of Peyer's patch M cells, indicating that the receptor is specific for the IgA isotype. Human serum IgA and colostral SIgA also bound to mouse M cells. The asialoglycoprotein receptor or other lectin-like receptors were not detected on the apical surfaces of M cells. We used recombinant human IgA1 and human IgA2 Abs and domain swapped IgA/IgG chimeras to determine that both domains Calpha1 and Calpha2 are required for IgA adherence to mouse Peyer's patch M cells. This distinguishes the M cell IgA receptor from CD89 (FcalphaI), which binds domains Calpha2-Calpha3. Finally, we observed by immunofluorescence microscopy that some M cells in the human ileum are coated with IgA. Together these data suggest that mouse, and possibly human, M cells express an IgA-specific receptor on their apical surfaces that mediates the transepithelial transport of SIgA from the intestinal lumen to underlying gut-associated organized lymphoid tissues

Four promoters subject to regulation by ExoR and PhoB direct transcription of the Sinorhizobium meliloti exoYFQ operon involved in the biosynthesis of succinoglycan

Quester I, Becker A. Four promoters subject to regulation by ExoR and PhoB direct transcription of the Sinorhizobium meliloti exoYFQ operon involved in the biosynthesis of succinoglycan. JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY. 2004;7(3):115-132.Succinoglycan (EPS I), the main acidic exopolysaccharide of Sinorhizobium meliloti, is required for the initiation and elongation of infection threads during nodulation of the host plant alfalfa. The gene products of the exoYFQ operon are involved in the first step of succinoglycan biosynthesis as well as in the polymerisation of subunits to the high-molecular-mass form of this exopolysaccharide. One promoter region that directs transcription of exoX and two promoter regions that drive transcription of exoY were mapped in the exoX-exoY intergenic region. The distal exoY promoter region containing three putative -10 promoter elements was active under standard growth conditions and was subject to ExoR-dependent regulation. Although this promoter region was stimulated in a phoB mutant, no PHO box-like sequences were found, suggesting an indirect regulatory effect of PhoB. The proximal promoter contains a PHO box-like sequence in the putative - 35 region and was affected by low and high phosphate concentrations dependent on PhoB. In the case of deleted upstream regions, this promoter was also controlled by ExoR. An additional promoter displaying activity in exoR, mucR and phoB mutants under standard conditions was identified upstream of exoF. The putative - 35 promoter element of this promoter is covered by a second PHO boxlike sequence. Copyright (C) 2004 S. Karger AG, Basel

A two-component regulatory system playing a critical role in plant pathogens and endosymbionts is present in Brucella abortus and controls cell invasion and virulence

Two mutants showing increased sensitivity to polycations and surfactants were obtained by transposon mutagenesis of virulent Brucella abortus 2308 Nal(r). These mutants showed no obvious in vitro growth defects and produced smooth-type lipopolysaccharides. However, they hardly multiplied or persisted in mouse spleens, displayed reduced invasiveness in macrophages and HeLa cells, lost the ability to inhibit lysosome fusion and were unable to replicate intracellularly. Subsequent DNA analyses identified a two-component regulatory system [Brucella virulence related (Bvr)] with a regulatory (BvrR) and sensory (BvrS) protein. Cloning on bvrR in the BvrR-deficient mutant restored the resistance to polycations and, in part, the invasiveness to polycations and, in part, the invasiveness and the ability to multiply intracellularly. BvrR and BvrS were highly similar (87-89% and 70-80% respectively) to the regulatory and sensory proteins of the chromosomally encoded Rhizobium meliloti Chvl-ExoS and Agrobacterium tumefaciens Chvl-ChvG systems previously shown to be critical for endosymbiosis and pathogenicity in plants. Divergence among the three sensory proteins was located mostly within a periplasmic domain probably involved in stimulus sensing. As B. abortus, R. meliloti and A. tumefaciens are phylogenetically related, these observations suggest that these systems have a common ancestor that has evolved to sense stimuli in plant and animal microbial environments.Dos mutantes que muestran una mayor sensibilidad a los policationes y a los tensioactivos se obtuvieron por mutagénesis de transposones de la Brucella abortus 2308 Nal(r) virulenta. Estos mutantes no mostraron defectos evidentes de crecimiento in vitro y produjeron lipopolisacáridos de tipo suave. Sin embargo, apenas se multiplicaron o persistieron en el bazo de los ratones, mostraron una menor capacidad de invasión en macrófagos y células HeLa, perdieron la capacidad de inhibir la fusión de lisosomas y fueron incapaces de replicarse intracelularmente. Los análisis posteriores del ADN identificaron un sistema regulador de dos componentes [Brucella virulence related (Bvr)] con una proteína reguladora (BvrR) y otra sensorial (BvrS). La clonación en bvrR en el mutante deficiente en BvrR restauró la resistencia a los policationes y, en parte, la capacidad de invasión a los policationes y, en parte, la capacidad de invasión y de multiplicación intracelular. BvrR y BvrS eran muy similares (87-89% y 70-80% respectivamente) a las proteínas reguladoras y sensoriales de los sistemas Rhizobium meliloti Chvl-ExoS y Agrobacterium tumefaciens Chvl-ChvG codificados cromosómicamente, que habían demostrado ser críticos para la endosimbiosis y la patogenicidad en las plantas. La divergencia entre las tres proteínas sensoriales se localizó sobre todo en un dominio periplásmico probablemente implicado en la detección de estímulos. Dado que B. abortus, R. meliloti y A. tumefaciens están filogenéticamente relacionados, estas observaciones sugieren que estos sistemas tienen un ancestro común que ha evolucionado para percibir estímulos en entornos microbianos vegetales y animales.Universidad Nacional, Costa RicaEscuela de Medicina Veterinari