following-real-time-impact-pneumococcal-virulence-factors-an-acute

Following in Real Time the Impact of Pneumococcal Virulence Factors in an Acute Mouse Pneumonia Model Using Bioluminescent Bacteria

Pneumonia is one of the major health care problems in developing and industrialized countries and is associated with considerable morbidity and mortality. ...

University of Greifswald

Increased golli MBP (golli) expression has been observed in the peripheral immune system of mice in the relapsing phase of EAE, raising the possibility that golli MBP expression in the periphery may contribute to relapses. Here we describe the generation of golli MBP-deficient mice and a comparison of the clinical course of EAE between heterozygous (golli(+/-)) and wild-type (golli(+/+)) mice. There was no difference between the two groups in incidence of disease, severity of the first episode of disease, or remission after the first episode. However, there was a significant reduction in relapses in golli(+/-) mice vs. controls, suggesting a role for golli proteins in the relapses in EAE.

Experimental autoimmune encephalomyelitis relapses are reduced in heterozygous golli MBP knockout mice.

The myelin basic protein (MBP) gene encodes two families of proteins, the classic MBP constituents of myelin and the golli-MBPs, the function of which is less well understood. In this study, targeted ablation of the golli-MBPs, but not the classic MBPs, resulted in a distinct phenotype unlike that of knock-outs (KOs) of the classic MBPs or other myelin proteins. Although the golli KO animals did not display an overt dysmyelinating phenotype, they did exhibit delayed and/or hypomyelination in selected areas of the brain, such as the visual cortex and the optic nerve, as determined by Northern and Western blots and immunohistochemical analysis with myelin protein markers. Hypomyelination in some areas, such as the visual cortex, persisted into adulthood. Ultrastructural analysis of the KOs confirmed both the delay and hypomyelination and revealed abnormalities in myelin structure and in some oligodendrocytes. Abnormal visual-evoked potentials indicated that the hypomyelination in the visual cortex had functional consequences in the golli KO brain. Evidence that the abnormal myelination in these animals was a consequence of intrinsic problems with the oligodendrocyte was indicated by an impaired ability of oligodendrocytes to form myelin sheets in culture and by the presence of abnormal Ca2+ transients in purified cortical oligodendrocytes studied in vitro. The Ca2+ results reported in this study complement previous results implicating golli proteins in modulating intracellular signaling in T-cells. Together, all these findings suggest a role for golli proteins in oligodendrocyte differentiation, migration, and/or myelin elaboration in the brain.

Region-specific myelin pathology in mice lacking the golli products of the myelin basic protein gene.

Mitochondrial F(1)-ATPase contains a hexamer of alternating alpha and beta subunits. The assembly of this structure requires two specialized chaperones, Atp11p and Atp12p, that bind transiently to beta and alpha. In the absence of Atp11p and Atp12p, the hexamer is not formed, and alpha and beta precipitate as large insoluble aggregates. An early model for the mechanism of chaperone-mediated F(1) assembly (Wang, Z. G., Sheluho, D., Gatti, D. L., and Ackerman, S. H. (2000) EMBO J. 19, 1486-1493) hypothesized that the chaperones themselves look very much like the alpha and beta subunits, and proposed an exchange of Atp11p for alpha and of Atp12p for beta; the driving force for the exchange was expected to be a higher affinity of alpha and beta for each other than for the respective chaperone partners. One important feature of this model was the prediction that as long as Atp11p is bound to beta and Atp12p is bound to alpha, the two F(1) subunits cannot interact at either the catalytic site or the noncatalytic site interface. Here we present the structures of Atp11p from Candida glabrata and Atp12p from Paracoccus denitrificans, and we show that some features of the Wang model are correct, namely that binding of the chaperones to alpha and beta prevents further interactions between these F(1) subunits. However, Atp11p and Atp12p do not resemble alpha or beta, and it is instead the F(1) gamma subunit that initiates the release of the chaperones from alpha and beta and their further assembly into the mature complex.

Chaperones of F1-ATPase.

Studies in yeast have shown that a deficiency in Atp12p prevents assembly of the extrinsic domain (F(1)) of complex V and renders cells unable to make ATP through oxidative phosphorylation. De Meirleir et al. (De Meirleir, L., Seneca, S., Lissens, W., De Clercq, I., Eyskens, F., Gerlo, E., Smet, J., and Van Coster, R. (2004) J. Med. Genet. 41, 120-124) have reported that a homozygous missense mutation in the gene for human Atp12p (HuAtp12p), which replaces Trp-94 with Arg, was linked to the death of a 14-month-old patient. We have investigated the impact of the pathogenic W94R mutation on Atp12p structure/function. Plasmid-borne wild type human Atp12p rescues the respiratory defect of a yeast ATP12 deletion mutant (Deltaatp12). The W94R mutation alters the protein at the most highly conserved position in the Pfam sequence and renders HuAtp12p insoluble in the background of Deltaatp12. In contrast, the yeast protein harboring the corresponding mutation, ScAtp12p(W103R), is soluble in the background of Deltaatp12 but not in the background of Deltaatp12Deltafmc1, a strain that also lacks Fmc1p. Fmc1p is a yeast mitochondrial protein not found in higher eukaryotes. Tryptophan 94 (human) or 103 (yeast) is located in a positively charged region of Atp12p, and hence its mutation to arginine does not alter significantly the electrostatic properties of the protein. Instead, we provide evidence that the primary effect of the substitution is on the dynamic properties of Atp12p.

Defining the pathogenesis of the human Atp12p W94R mutation using a Saccharomyces cerevisiae yeast model.

&#x80BA;&#x708E;&#x94FE;&#x7403;&#x83CC;&#x662F;&#x793E;&#x533A;&#x83B7;&#x5F97;&#x6027;&#x80BA;&#x708E;&#x7684;&#x4E3B;&#x8981;&#x539F;&#x56E0;&#x3002;&#x5728;&#x6B64;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x5BA1;&#x67E5;&#x611F;&#x5B98;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x611F;&#x67D3;&#x7684;&#x5148;&#x5929;&#x514D;&#x75AB;&#x8BC6;&#x522B;&#x901A;&#x8DEF;&#xFF0C;&#x89E6;&#x53D1;&#x5668; I &#x578B;&#x5E72;&#x6270;&#x7D20;&#x751F;&#x4EA7;&#xFF0C;&#x5E76;&#x89C4;&#x5B9A; RANTES &#x751F;&#x4EA7;&#x3002;&#x6211;&#x4EEC;&#x53D1;&#x73B0;&#x4EBA;&#x7C7B;&#x548C;&#x5C0F;&#x9F20;&#x80BA;&#x6CE1;&#x5DE8;&#x566C;&#x7EC6;&#x80DE;&#x4EE5;&#x53CA;&#x5C0F;&#x9F20;&#x9AA8;&#x9AD3;&#x5DE8;&#x566C;&#x7EC6;&#x80DE;&#xFF0C;&#x4F46;&#x4E0D;&#x662F;&#x80BA;&#x6CE1;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#xFF0C;&#x4EA7;&#x751F;&#x4E00;&#x7C7B; IFNs S.&#x80BA;&#x708E;&#x611F;&#x67D3;&#x540E;&#x3002;&#x8FD9;&#x79CD;&#x53CD;&#x5E94;&#x53D6;&#x51B3;&#x4E8E;&#x5B54;&#x9699;&#x5F62;&#x6210;&#x7684;&#x6BD2;&#x7D20;&#x6EB6;&#x8840;&#xFF0C;&#x4F3C;&#x4E4E;&#x4ECB;&#x5BFC;&#x6E38;&#x79BB; DNA &#x9065;&#x611F;&#x9014;&#x5F84;&#xFF0C;&#x6D89;&#x53CA;&#x5230;&#x9002;&#x914D;&#x5668;&#x5206;&#x5B50;&#x9489;&#x523A;&#x548C;&#x8F6C;&#x5F55;&#x56E0;&#x5B50;&#x5E72;&#x6270;&#x7D20;&#x8C03;&#x8282;&#x56E0;&#x5B50; 3&#x3002;&#x4E8B;&#x5B9E;&#x4E0A;&#xFF0C;DNA &#x662F;&#x672C;&#x671F;&#x95F4;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x611F;&#x67D3;&#x7EC6;&#x80DE;&#x6EB6;&#x8D28;&#x4E2D;&#x7684; AIM2 inflammasome&#xFF0C;&#x8FD9;&#x5DF2;&#x77E5;&#x611F;&#x89C9;&#x5FAE;&#x751F;&#x7269; DNA &#x6FC0;&#x6D3B;&#x6240;&#x793A;&#x3002;&#x7C7B;&#x578B;&#x6211; IFNs S.&#x611F;&#x67D3;&#x80BA;&#x708E;&#x7684;&#x5DE8;&#x566C;&#x7EC6;&#x80DE;&#x4EA7;&#x751F;&#x79EF;&#x6781;&#x8C03;&#x8282;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x548C; RANTES &#x751F;&#x4EA7;&#x4F53;&#x5916;&#x590D;&#x5408;&#x57F9;&#x517B;&#x80BA;&#x6CE1;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x548C;&#x5DE8;&#x566C;&#x7EC6;&#x80DE;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x952E;&#x5165;&#x6211; IFNs &#x63A7;&#x5236; RANTES &#x751F;&#x4EA7;&#x671F;&#x95F4;&#x4F53;&#x5185;&#x7684;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x80BA;&#x708E;&#x3002;&#x6700;&#x540E;&#xFF0C;&#x6211;&#x4EEC;&#x786E;&#x5B9A;&#x4E86;&#x9065;&#x611F;&#x63A2;&#x6D4B;&#x89E6;&#x53D1;&#x7C7B;&#x578B;&#x7684; S.&#x80BA;&#x708E;&#x7684;&#x9014;&#x5F84;&#x514D;&#x75AB;&#x6211;&#x5E72;&#x6270;&#x7D20;&#x53CD;&#x5E94;&#xFF0C;&#x5E76;&#x79EF;&#x6781;&#x8C03;&#x8282; RANTES &#x751F;&#x4EA7;&#x3002;

&#x80BA;&#x708E;&#x94FE;&#x7403;&#x83CC;&#x523A;&#x6FC0;&#x523A;-&#x548C;&#x5E72;&#x6270;&#x7D20;&#x8C03;&#x8282;&#x56E0;&#x5B50; 3 &#x4F9D;&#x8D56;&#x7684; I &#x578B;&#x5E72;&#x6270;&#x7D20;&#x751F;&#x4EA7;&#x5DE8;&#x566C;&#x7EC6;&#x80DE;&#xFF0C;&#x8BE5;&#x6CD5;&#x89C4;&#x5B9A;&#x5728;&#x5DE8;&#x566C;&#x7EC6;&#x80DE;&#x3001; &#x590D;&#x5408;&#x57F9;&#x517B;&#x7684;&#x80BA;&#x6CE1;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x548C;&#x9F20;&#x6807;&#x80BA;&#x90E8; RANTES &#x751F;&#x4EA7;&#x3002;

&#xC5F0;&#xC1C4; &#xC0C1; &#xAD6C; &#xADE0; &#xADE0; &#xC9C0;&#xC5ED; &#xC0AC;&#xD68C; &#xD68D;&#xB4DD; &#xD3D0; &#xB834;&#xC758; &#xC8FC;&#xC694; &#xC6D0;&#xC778;&#xC785;&#xB2C8;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC;&#xB294; &#xD0C0;&#xACE0; &#xB09C; &#xBA74;&#xC5ED; &#xC778;&#xC2DD; &#xD1B5;&#xB85C; &#xAC10;&#xC9C0; &#xD558;&#xB294; &#xD3D0; &#xB834; &#xAC10;&#xC5FC; &#xAC80;&#xC0AC;, &#xD2B8;&#xB9AC;&#xAC70; &#xC720;&#xD615; I IFN &#xC0DD;&#xC0B0; RANTES &#xC0DD;&#xC0B0;&#xC744; &#xADDC;&#xC81C; &#xD558; &#xACE0;. &#xC6B0;&#xB9AC;&#xB294; &#xC778;&#xAC04;&#xACFC; murine &#xCE58;&#xACBD; &#xB300; &#xC2DD; &#xC138;&#xD3EC; murine &#xACE8; &#xB300;&#xB9CC; &#xC0DD;&#xC0B0; &#xD558;&#xC9C0; &#xCE58;&#xACBD; &#xC0C1;&#xD53C; &#xC138;&#xD3EC; &#xC720;&#xD615; I IFNs &#xBBF8; &#xADE0; &#xAC10;&#xC5FC; &#xC2DC; &#xBC1C;&#xACAC;. &#xC774; &#xC751;&#xB2F5; &#xAE30; &#xACF5; &#xD615;&#xC131; &#xB3C5; &#xC18C; pneumolysin&#xC5D0; &#xC885;&#xC18D; &#xB418;&#xC5C8;&#xACE0; &#xC5B4;&#xB311;&#xD130; &#xBD84;&#xC790; &#xC2A4; &#xD305;&#xACFC; &#xB179;&#xC74C; &#xBC29;&#xC1A1; &#xC694;&#xC778; IFN &#xADDC;&#xC81C; &#xC694;&#xC18C; 3 cytosolic DNA &#xAC10;&#xC9C0; &#xD1B5;&#xB85C; &#xC758;&#xD574; &#xC911;&#xC7AC; &#xB420; &#xAC83;&#xC73C;&#xB85C; &#xBCF4;&#xC778;&#xB2E4;. &#xC0AC;&#xC2E4;, DNA &#xBBF8;&#xC0DD;&#xBB3C; DNA&#xC744; &#xAC10;&#xC9C0; &#xC54C;&#xB824;&#xC838; &#xC788;&#xB2E4; AIM2 inflammasome &#xD65C;&#xC131;&#xD654; &#xD558; &#xC5EC; &#xD45C;&#xC2DC; &#xB41C; &#xB300;&#xB85C; &#xD3D0; &#xB834; &#xAC10;&#xC5FC; &#xC2DC; cytosol&#xC5D0; &#xC874;&#xC7AC; &#xD588;&#xB2E4;. &#xC720;&#xD615; &#xB098; IFNs &#xAE0D;&#xC815;&#xC801;&#xC73C;&#xB85C; &#xBBF8; &#xADE0;&#xC5D0; &#xAC10;&#xC5FC; &#xB41C; &#xB300; &#xC2DD; &#xC138;&#xD3EC;&#xC5D0; &#xC758;&#xD574; &#xC0DD;&#xC0B0; &#xC720;&#xC804;&#xC790; &#xBC1C;&#xD604; &#xBC0F; &#xB300; &#xC2DD; &#xC138;&#xD3EC;&#xC640; cocultured &#xD3D0; &#xD3EC; &#xC0C1;&#xD53C; &#xC138;&#xD3EC; &#xCCB4; &#xC678;&#xC5D0;&#xC11C; RANTES &#xC0DD;&#xC0B0; &#xADDC;&#xC81C;. &#xB610;&#xD55C;, &#xB0B4;&#xAC00; IFNs &#xC81C;&#xC5B4; RANTES &#xC0DD;&#xC0B0; vivo&#xC5D0;&#xC11C; &#xD3D0; &#xB834; &#xADE0; &#xC131; &#xD3D0; &#xB834;&#xC744; &#xC785;&#xB825; &#xD569;&#xB2C8;&#xB2E4;. &#xB05D;&#xC73C;&#xB85C;, &#xD1B5;&#xB85C;&#xAC00; &#xD2B8;&#xB9AC;&#xAC70; &#xD615;&#xC2DD; &#xBBF8; &#xADE0; &#xAC10;&#xC9C0; &#xAC10;&#xC9C0;&#xB294; &#xBA74;&#xC5ED; &#xC2DD;&#xBCC4; &#xB098; IFN &#xC751;&#xB2F5; RANTES &#xC0DD;&#xC0B0;&#xC744; &#xC801;&#xADF9;&#xC801;&#xC73C;&#xB85C; &#xADDC;&#xC81C; &#xD558; &#xACE0; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xC5F0;&#xC1C4; &#xC0C1; &#xAD6C; &#xADE0; &#xADE0; &#xC790;&#xADF9; &#xC2A4; &#xD305;-&#xD558; &#xACE0; IFN &#xADDC;&#xC81C; &#xC694;&#xC18C; 3 &#xC885;&#xC18D; &#xC720;&#xD615; I IFN &#xC0DD;&#xC0B0; &#xB300; &#xC2DD; &#xC138;&#xD3EC;, &#xB300; &#xC2DD; &#xC138;&#xD3EC;, cocultured &#xD3D0; &#xD3EC; &#xC0C1;&#xD53C; &#xC138;&#xD3EC; &#xBC0F; &#xB9C8;&#xC6B0;&#xC2A4; &#xD3D0; RANTES &#xC0DD;&#xC0B0; &#xC870;&#xC815;&#xC5D0;.

Streptococcus pneumoniae is the leading cause of community-acquired pneumonia. In this study, we examine an innate immune recognition pathway that senses pneumococcal infection, triggers type I IFN production, and regulates RANTES production. We found that human and murine alveolar macrophages as well as murine bone marrow macrophages, but not alveolar epithelial cells, produced type I IFNs upon infection with S. pneumoniae. This response was dependent on the pore-forming toxin pneumolysin and appeared to be mediated by a cytosolic DNA-sensing pathway involving the adapter molecule STING and the transcription factor IFN regulatory factor 3. Indeed, DNA was present in the cytosol during pneumococcal infection as indicated by the activation of the AIM2 inflammasome, which is known to sense microbial DNA. Type I IFNs produced by S. pneumoniae-infected macrophages positively regulated gene expression and RANTES production in macrophages and cocultured alveolar epithelial cells in vitro. Moreover, type I IFNs controlled RANTES production during pneumococcal pneumonia in vivo. In conclusion, we identified an immune sensing pathway detecting S. pneumoniae that triggers a type I IFN response and positively regulates RANTES production.

Streptococcus pneumoniae stimulates a STING- and IFN regulatory factor 3-dependent type I IFN production in macrophages, which regulates RANTES production in macrophages, cocultured alveolar epithelial cells, and mouse lungs.

Streptococcus pneumoniae est la cause de la pneumonie extra-hospitali&egrave;re. Dans cette &eacute;tude, nous examinons une voie de reconnaissance immunitaire inn&eacute;e qui d&eacute;tecte l&#39;infection pneumococcique, d&eacute;clencheurs de type I production IFN et r&eacute;gule la production de RANTES. Nous avons trouv&eacute; que les macrophages alv&eacute;olaires humains et murins ainsi que macrophages murins de la moelle osseuse, mais les cellules &eacute;pith&eacute;liales alv&eacute;olaires pas, produits type I IFNs apr&egrave;s infection avec S. pneumoniae. Cette r&eacute;ponse d&eacute;pend de la pneumolysin de toxines formant des pores et semble &ecirc;tre m&eacute;di&eacute; par une voie de d&eacute;tection ADN cytosolique impliquant la mol&eacute;cule adaptateur STING et le facteur r&eacute;glementaire de l&#39;IFN transcription factor 3. En effet, l&#39;ADN a &eacute;t&eacute; pr&eacute;sente dans le cytosol au cours de l&#39;infection &agrave; pneumocoques, comme indiqu&eacute; par l&#39;activation de l&#39;inflammasome AIM2, qui est connu pour d&eacute;tecter l&#39;ADN microbien. Type qu&#39;i IFNs produite par les macrophages infect&eacute;s par pneumoniae S. positivement r&eacute;glement&eacute; l&#39;expression des g&egrave;nes et production de RANTES dans les macrophages et les cellules &eacute;pith&eacute;liales alv&eacute;olaires cocultured in vitro. En outre, le type I IFNs production contr&ocirc;l&eacute;e de RANTES au cours de la pneumonie &agrave; pneumocoque in vivo. En conclusion, nous avons identifi&eacute; un immunitaire d&eacute;tection d&eacute;tection de S. pneumoniae qui d&eacute;clenche un type de voie j&#39;ai IFN r&eacute;ponse et r&eacute;gule positivement la production RANTES.

Streptococcus pneumoni&aelig; stimule un STING - et le facteur de normalisation IFN 3-d&eacute;pendant de type I production IFN dans les macrophages, qui r&eacute;gule la production de RANTES dans les macrophages, les cellules &eacute;pith&eacute;liales alv&eacute;olaires cocultured et les poumons de souris.

Streptococcus Pneumoniae ist die f&uuml;hrende Ursache der Gemeinschaft-erworbenen Pneumonie. In dieser Studie untersuchen wir eine angeborene immune Anerkennung-Weg, die Pneumokokken-Infektion sp&uuml;rt, Trigger Typ I IFN-Produktion und RANTES reguliert. Wir fanden, dass Mensch und murinen alveol&auml;ren Makrophagen sowie Knochenmark murinen Makrophagen, aber nicht alveolaren Epithelzellen produziert Typ I IFNs auf Infektion mit S. Pneumoniae. Diese Antwort war die Pore-forming Toxin Pneumolysin abh&auml;ngig und schien sich ein cytosolisches DNA-Sensorik-Option unter Beteiligung des Adapter-Molek&uuml;ls STING und Faktor IFN regulatorischen Transkriptionsfaktors 3 vermittelt zu werden. Tats&auml;chlich war die DNA in das Zytosol w&auml;hrend Pneumokokken-Infektion vorhanden, wie durch die Aktivierung von den AIM2 Inflammasom, das bekannt ist, mikrobiellen DNA zu sp&uuml;ren. Typ, den ich IFNs durch S. Pneumoniae infizierte Makrophagen positiv produziert reguliert Genexpression und RANTES Produktion in Makrophagen und cocultured alveolaren Epithelzellen in vitro. Dar&uuml;ber hinaus geben Sie I IFNs kontrollierten RANTES Produktion w&auml;hrend der in-vivo Pneumokokken Lungenentz&uuml;ndung. Abschlie&szlig;end ermittelten wir eine Immune Sensorik Weg erkennen von S. Pneumoniae, der eine Art ausl&ouml;st ich IFN Antwort und RANTES positiv reguliert.

Pneumokokken stimuliert ein STING - und IFN ordnungspolitischen Faktor 3-abh&auml;ngige Typ I IFN-Produktion in Makrophagen, die RANTES Produktion in Makrophagen, cocultured alveolaren Epithelzellen und Maus-Lunge regelt.

Lo Streptococcus pneumoniae &egrave; la causa principale di polmonite acquisita in comunit&agrave;. In questo studio, esaminiamo un percorso di riconoscimento immunitaria innata che rileva l&#39;infezione da pneumococco, trigger produzione di IFN di tipo I e regola la produzione di RANTES. Abbiamo trovato che umani e murini macrofagi alveolari nonch&eacute; i macrofagi di midollo osseo murino, ma cellule epiteliali non alveolari, prodotte tipo I IFN all&#39;infezione con S. pneumoniae. Questa risposta dipendeva la pneumolysin tossina formando dei pori e sembrava essere mediata da un percorso di rilevazione del DNA citosolico che coinvolgono la molecola adattatore STING e il trascrizione IFN normativo fattore 3. Infatti, il DNA era presente nel citosol durante l&#39;infezione da pneumococco, come indicato dall&#39;attivazione di inflammasome di AIM2, che &egrave; noto a senso del DNA microbico. Tipo che i IFN prodotta dai macrofagi S. pneumoniae infetta positivamente disciplinata espressione genica e la produzione di RANTES in macrofagi e cellule epiteliali alveolari cocultured in vitro. Inoltre, tipo I IFN produzione RANTES controllata durante la polmonite pneumococcica in vivo. In conclusione, abbiamo identificato un immunitario pathway rilevazione S. pneumoniae che innesca un tipo di rilevamento che IFN risposta e regola positivamente la produzione di RANTES.

Streptococcus pneumoniae stimola un PUNGIGLIONE - e fattore regolatore IFN 3-dipendente di tipo produzione di IFN in macrofagi, che regola la produzione di RANTES in macrofagi, cellule epiteliali alveolari cocultured e polmoni del mouse.

&#x80BA;&#x708E;&#x7403;&#x83CC;&#x30B3;&#x30DF;&#x30E5;&#x30CB;&#x30C6;&#x30A3;-&#x53D6;&#x5F97;&#x80BA;&#x708E;&#x306E;&#x4E3B;&#x8981;&#x306A;&#x539F;&#x56E0;&#x3067;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x6211;&#x3005; &#x306F;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x611F;&#x67D3;&#x75C7;&#x306E;&#x611F;&#x899A;&#x751F;&#x5F97;&#x306E;&#x514D;&#x75AB;&#x8A8D;&#x8B58;&#x7D4C;&#x8DEF;&#x3092;&#x8ABF;&#x3079;&#x308B;&#x3001;&#x30C8;&#x30EA;&#x30AC;&#x30FC; I &#x578B;&#x30A4;&#x30F3;&#x30BF;&#x30FC;&#x30D5;&#x30A7;&#x30ED;&#x30F3;&#x7523; RANTES &#x751F;&#x7523;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306F;&#x4EBA;&#x9593;&#x304A;&#x3088;&#x3073;&#x30DE;&#x30A6;&#x30B9;&#x80BA;&#x80DE;&#x30DE;&#x30AF;&#x30ED;&#x30D5;&#x30A1;&#x30FC;&#x30B8;&#x3068;&#x540C;&#x69D8;&#x306B;&#x30DE;&#x30A6;&#x30B9;&#x9AA8;&#x9AC4;&#x30DE;&#x30AF;&#x30ED;&#x30D5;&#x30A1;&#x30FC;&#x30B8;&#x304C;&#x751F;&#x7523;&#x3001;&#x306A;&#x3044;&#x80BA;&#x80DE;&#x4E0A;&#x76AE;&#x7D30;&#x80DE;&#x3068; I &#x578B; Ifn &#x3068;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x611F;&#x67D3;&#x75C7;&#x306E;&#x6642;&#x306B;&#x767A;&#x898B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3053;&#x306E;&#x5FDC;&#x7B54;&#x306F;&#x3001;&#x7D30;&#x5B54;&#x5F62;&#x6210;&#x6BD2;&#x7D20;&#x30CB;&#x30E5;&#x30FC;&#x30E2;&#x30EA;&#x30B7;&#x30F3;&#x306B;&#x4F9D;&#x5B58;&#x3057;&#x3066;&#x3044;&#x305F;&#x3001;&#x30A2;&#x30C0;&#x30D7;&#x30BF;&#x30FC;&#x5206;&#x5B50;&#x30B9;&#x30C6;&#x30A3;&#x30F3;&#x30B0;&#x3068;&#x8EE2;&#x5199;&#x56E0;&#x5B50; IFN &#x898F;&#x5236;&#x56E0;&#x5B50; 3 &#x3092;&#x542B;&#x3080;&#x3001;&#x30BE;&#x30EB;&#x6027;&#x7D30;&#x80DE;&#x8CEA; DNA &#x30BB;&#x30F3;&#x30B7;&#x30F3;&#x30B0;&#x306E;&#x7D4C;&#x8DEF;&#x3092;&#x4ECB;&#x3059;&#x308B;&#x3088;&#x3046;&#x306B;&#x898B;&#x3048;&#x305F;&#x3002;&#x78BA;&#x304B;&#x306B;&#x3001;DNA &#x5FAE;&#x751F;&#x7269;&#x306E; DNA &#x3092;&#x611F;&#x77E5;&#x3059;&#x308B;&#x77E5;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B; AIM2 &#x30A4;&#x30F3;&#x30D5;&#x30E9;&#x30DE;&#x30BD;&#x30FC;&#x30E0;&#x306E;&#x6D3B;&#x6027;&#x5316;&#x306B;&#x3088;&#x3063;&#x3066;&#x793A;&#x3055;&#x308C;&#x308B;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x611F;&#x67D3;&#x6642;&#x306B;&#x7D30;&#x80DE;&#x8CEA;&#x306B;&#x5B58;&#x5728;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;&#x79C1;&#x306F; Ifn S. &#x80BA;&#x708E;&#x306B;&#x611F;&#x67D3;&#x3057;&#x305F;&#x30DE;&#x30AF;&#x30ED;&#x30D5;&#x30A1;&#x30FC;&#x30B8;&#x306B;&#x3088;&#x3063;&#x3066;&#x7A4D;&#x6975;&#x7684;&#x306B;&#x751F;&#x7523;&#x578B;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x3068;&#x30DE;&#x30AF;&#x30ED;&#x30D5;&#x30A1;&#x30FC;&#x30B8;&#x304A;&#x3088;&#x3073;&#x4F53;&#x5916;&#x57F9;&#x990A;&#x306E;&#x80BA;&#x80DE;&#x4E0A;&#x76AE;&#x7D30;&#x80DE;&#x306E; RANTES &#x751F;&#x7523;&#x898F;&#x5236;&#x3002;&#x307E;&#x305F;&#x3001;&#x79C1;&#x306F; Ifn &#x5236;&#x5FA1; RANTES &#x751F;&#x7523;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x6027;&#x80BA;&#x708E; in vivo &#x3067;&#x306E;&#x4E2D;&#x306B;&#x5165;&#x529B;&#x3057;&#x307E;&#x3059;&#x3002;&#x7D50;&#x8AD6;&#x3068;&#x3057;&#x3066;&#x3001;&#x6211;&#x3005; &#x306F;&#x7D4C;&#x8DEF;&#x306E;&#x30BF;&#x30A4;&#x30D7;&#x3092;&#x30C8;&#x30EA;&#x30AC;&#x30FC;&#x80BA;&#x708E;&#x7403;&#x83CC;&#x3092;&#x691C;&#x51FA;&#x3059;&#x308B;&#x30BB;&#x30F3;&#x30B7;&#x30F3;&#x30B0;&#x514D;&#x75AB;&#x8B58;&#x5225;&#x79C1;&#x306F; IFN &#x5FDC;&#x7B54;&#x3068;&#x7A4D;&#x6975;&#x7684;&#x306B; RANTES &#x751F;&#x7523;&#x3092;&#x8ABF;&#x7BC0;&#x3057;&#x307E;&#x3059;&#x3002;

&#x80BA;&#x708E;&#x7403;&#x83CC;&#x30B9;&#x30C6;&#x30A3;&#x30F3;&#x30B0;-&#x3092;&#x523A;&#x6FC0;&#x3057;&#x3001;&#x30A4;&#x30F3;&#x30BF;&#x30FC;&#x30D5;&#x30A7;&#x30ED;&#x30F3;&#x5236;&#x5FA1;&#x56E0;&#x5B50; 3 &#x4F9D;&#x5B58; I &#x578B;&#x30A4;&#x30F3;&#x30BF;&#x30FC;&#x30D5;&#x30A7;&#x30ED;&#x30F3;&#x7523; RANTES &#x751F;&#x7523;&#x30DE;&#x30AF;&#x30ED;&#x30D5;&#x30A1;&#x30FC;&#x30B8;, &#x57F9;&#x990A;&#x306E;&#x80BA;&#x80DE;&#x4E0A;&#x76AE;&#x7D30;&#x80DE;&#x3068;&#x30DE;&#x30A6;&#x30B9;&#x80BA;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x30DE;&#x30AF;&#x30ED;&#x30D5;&#x30A1;&#x30FC;&#x30B8;&#x3002;

Streptococcus pneumoniae es la principal causa de neumon&iacute;a adquirida en la comunidad. En este estudio examinamos un camino de reconocimiento inmune innata que detecta la infecci&oacute;n neumoc&oacute;cica, tipo I producci&oacute;n de IFN y regula la producci&oacute;n de RANTES. Encontramos que los macr&oacute;fagos alveolares murinos y humanos as&iacute; como macr&oacute;fagos murinos de m&eacute;dula &oacute;sea, pero las c&eacute;lulas epiteliales no alveolares, producida IFN tipo I sobre la infecci&oacute;n por S. pneumoniae. Esta respuesta depende de la formaci&oacute;n de poros toxina inmunoprotectores y parece estar mediado por una v&iacute;a citos&oacute;lica de detecci&oacute;n de ADN la mol&eacute;cula adaptador STING y el factor regulador IFN 3 de transcripci&oacute;n factor. De hecho, ADN estuvo presente en el citosol durante la infecci&oacute;n neumoc&oacute;cica seg&uacute;n lo indicado por la activaci&oacute;n de la AIM2 inflammasome, que se conoce para detectar ADN microbiano. Tipo que i IFN producido por macr&oacute;fagos de S. pneumoniae-infectado positivamente regula la expresi&oacute;n g&eacute;nica y la producci&oacute;n de RANTES en macr&oacute;fagos y c&eacute;lulas epiteliales alveolares morfolog&iacute;a in vitro. Por otra parte, el tipo I IFN producci&oacute;n controlada de RANTES durante la neumon&iacute;a neumoc&oacute;cica en vivo. En conclusi&oacute;n, identificamos un inmune detecci&oacute;n v&iacute;a de detecci&oacute;n de S. pneumoniae que desencadena un tipo me IFN respuesta y regula positivamente la producci&oacute;n de RANTES.

Streptococcus pneumoniae estimula una picadura - y factor regulador de IFN 3-dependiente tipo I producci&oacute;n de IFN en macr&oacute;fagos, que regula la producci&oacute;n de RANTES en macr&oacute;fagos, c&eacute;lulas epiteliales alveolares morfolog&iacute;a y pulmones de rat&oacute;n.

Adherence of Streptococcus pneumoniae is directly mediated by interactions of adhesins with eukaryotic cellular receptors or indirectly by exploiting matrix and serum proteins as molecular bridges. Pneumococci engage vitronectin, the human adhesive glycoprotein and complement inhibitor, to facilitate attachment to epithelial cells of the mucosal cavity, thereby modulating host cell signaling. In this study, we identified PspC as a vitronectin-binding protein interacting with the C-terminal heparin-binding domain of vitronectin. PspC is a multifunctional surface-exposed choline-binding protein displaying various adhesive properties. Vitronectin binding required the R domains in the mature PspC protein, which are also essential for the interaction with the ectodomain of the polymeric immunoglobulin receptor and secretory IgA. Consequently, secretory IgA competitively inhibited binding of vitronectin to purified PspC and to PspC-expressing pneumococci. In contrast, Factor H, which binds to the N-terminal part of mature PspC molecules, did not interfere with the PspC-vitronectin interaction. Using a series of vitronectin peptides, the C-terminal heparin-binding domain was shown to be essential for the interaction of soluble vitronectin with PspC. Binding experiments with immobilized vitronectin suggested a region N-terminal to the identified heparin-binding domain as an additional binding region for PspC, suggesting that soluble, immobilized, as well as cellularly bound vitronectin possesses different conformations. Finally, vitronectin bound to PspC was functionally active and inhibited the deposition of the terminal complement complex. In conclusion, this study identifies and characterizes (on the molecular level) the interaction between the pneumococcal adhesin PspC and the human glycoprotein vitronectin.

The choline-binding protein PspC of Streptococcus pneumoniae interacts with the C-terminal heparin-binding domain of vitronectin.

Streptococcus pneumoniae is a Gram-positive human pathogen with a complex lipoteichoic acid (pnLTA) structure. Because the current structural model for pnLTA shows substantial inconsistencies, we reinvestigated purified and, more importantly, O-deacylated pnLTA, which is most suitable for NMR spectroscopy and electrospray ionization-MS spectrometry. We analyzed pnLTA of nonencapsulated pneumococcal strains D39Δcps and TIGR4Δcps, respectively. The data obtained allowed us to (re)define (i) the position and linkage of the repeating unit, (ii) the putative α-GalpNAc substitution at the ribitiol 5-phosphate (Rib-ol-5-P), and (iii) the length of (i.e. the number of repeating units in) the pnLTA chain. We here also describe for the first time that the terminal sugar residues in the pnLTA (Forssman disaccharide; α-D-GalpNAc-(1→3)-β-D-GalpNAc-(1→)), responsible for the cross-reactivity with anti-Forssman antigen antibodies, can be heterogeneous with respect to its degree of phosphorylcholine substitution in both O-6-positions. To assess the proinflammatory potency of pnLTA, we generated a (lipopeptide-free) Δlgt mutant of strain D39Δcps, isolated its pnLTA, and showed that it is capable of inducing IL-6 release in human mononuclear cells, independent of TLR2 activation. This finding was quite in contrast to LTA of the Staphylococcus aureus SA113Δlgt mutant, which did not activate human mononuclear cells in our experiments. Remarkably, this is also contrary to various other reports showing a proinflammatory potency of S. aureus LTA. Taken together, our study refines the structure of pnLTA and indicates that pneumococcal and S. aureus LTAs differ not only in their structure but also in their bioactivity.

Structural reevaluation of Streptococcus pneumoniae Lipoteichoic acid and new insights into its immunostimulatory potency.

The respiratory pathogen Streptococcus pneumoniae has evolved efficient mechanisms to resist oxidative stress conditions and to displace other bacteria in the nasopharynx. Here we characterize at physiological, functional and structural levels two novel surface-exposed thioredoxin-family lipoproteins, Etrx1 and Etrx2. The impact of both Etrx proteins and their redox partner methionine sulfoxide reductase SpMsrAB2 on pneumococcal pathogenesis was assessed in mouse virulence studies and phagocytosis assays. The results demonstrate that loss of function of either both Etrx proteins or SpMsrAB2 dramatically attenuated pneumococcal virulence in the acute mouse pneumonia model and that Etrx proteins compensate each other. The deficiency of Etrx proteins or SpMsrAB2 further enhanced bacterial uptake by macrophages, and accelerated pneumococcal killing by H2 O2 or free methionine sulfoxides (MetSO). Moreover, the absence of both Etrx redox pathways provokes an accumulation of oxidized SpMsrAB2 in vivo. Taken together our results reveal insights into the role of two extracellular electron pathways required for reduction of SpMsrAB2 and surface-exposed MetSO. Identification of this system and its target proteins paves the way for the design of novel antimicrobials.

Molecular architecture of Streptococcus pneumoniae surface thioredoxin-fold lipoproteins crucial for extracellular oxidative stress resistance and maintenance of virulence.

Surface proteins are important for the fitness and virulence of the Gram-positive pathogen Streptococcus pneumoniae. They are crucial for interaction of the pathogen with its human host during infection. Therefore, the analysis of the pneumococcal surface proteome is an important task that requires powerful tools. In this study, two different methods, an optimized biotinylation approach and shaving with trypsin beads, were applied to study the pneumococcal surface proteome and to identify surface-exposed protein domains, respectively. The identification of nearly 95% of the predicted lipoproteins and 75% of the predicted sortase substrates reflects the high coverage of the two classical surface protein classes accomplished in this study. Furthermore, the biotinylation approach was applied to study the impact of an impaired lipoprotein maturation pathway on the cell envelope proteome and exoproteome. Loss of the lipoprotein diacylglyceryl transferase Lgt leads to striking changes in the lipoprotein distribution. Many lipoproteins disappear from the surface proteome and accumulate in the exoproteome. Further insights into lipoprotein processing in pneumococci are provided by immunoblot analyses of bacterial lysates and corresponding supernatant fractions. Taken together, the first comprehensive overview of the pneumococcal surface and exoproteome is presented, and a model for lipoprotein processing in S. pneumoniae is proposed.

Article	Year
Experimental autoimmune encephalomyelitis relapses are reduced in heterozygous golli MBP knockout mice. Journal of neuroimmunology\| PubMed ID: 12799019	2003
Region-specific myelin pathology in mice lacking the golli products of the myelin basic protein gene. The Journal of neuroscience : the official journal of the Society for Neuroscience\| PubMed ID: 16049176	2005
Chaperones of F1-ATPase. The Journal of biological chemistry\| PubMed ID: 19383603	2009
Defining the pathogenesis of the human Atp12p W94R mutation using a Saccharomyces cerevisiae yeast model. The Journal of biological chemistry\| PubMed ID: 19933271	2010
Streptococcus pneumoniae stimulates a STING- and IFN regulatory factor 3-dependent type I IFN production in macrophages, which regulates RANTES production in macrophages, cocultured alveolar... Journal of immunology (Baltimore, Md. : 1950)\| PubMed ID: 22156592	2012
The choline-binding protein PspC of Streptococcus pneumoniae interacts with the C-terminal heparin-binding domain of vitronectin. The Journal of biological chemistry\| PubMed ID: 23603906	2013
Structural reevaluation of Streptococcus pneumoniae Lipoteichoic acid and new insights into its immunostimulatory potency. The Journal of biological chemistry\| PubMed ID: 23603911	2013
Molecular architecture of Streptococcus pneumoniae surface thioredoxin-fold lipoproteins crucial for extracellular oxidative stress resistance and maintenance of virulence. EMBO molecular medicine\| PubMed ID: 24136784	2013
Influence of Impaired Lipoprotein Biogenesis on Surface and Exoproteome of Streptococcus pneumoniae. Journal of proteome research\| PubMed ID: 24387739	2014

Thomas Pribyl

Biography

JoVE Journal Publications

Other Publications