determination-biofilm-initiation-on-virus-infected-cells-bacteria

Determination of Biofilm Initiation on Virus-infected Cells by Bacteria and Fungi

The study of polymicrobial interactions across the taxonomic kingdoms that include fungi, bacteria and virus have not been previously examined with ...

anaerobic-growth-and-maintenance-of-mammalian-cell-lines

Anaerobic Growth and Maintenance of Mammalian Cell Lines

Most mucosal surfaces along with the midpoints in tumors and stem cell niches are geographic areas of the body that are anoxic. Previous studies show that ...

Midwestern University

&#x6027;&#x75C5;&#x6C99;&#x773C;&#x8863;&#x539F;&#x4F53;&#x611F;&#x67D3;&#x7684;&#x540E;&#x9057;&#x75C7;&#x5305;&#x62EC;&#x8F93;&#x5375;&#x7BA1;&#x7684;&#x75A4;&#x75D5;&#xFF0C;&#x8FD9;&#x610F;&#x5473;&#x7740;&#x7EC6;&#x80DE;&#x5916;&#x57FA;&#x8D28;&#x7684;&#x6539;&#x6027;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x76EE;&#x6807;&#x662F;&#x63CF;&#x8FF0;&#x4E24;&#x4E2A;&#x57FA;&#x8D28;&#x91D1;&#x5C5E;&#x86CB;&#x767D;&#x9176;&#x5728;&#x4F53;&#x5916;&#x7684;&#x8863;&#x539F;&#x4F53;&#x611F;&#x67D3;&#x53CD;&#x5E94;&#x7684;&#x751F;&#x4EA7;&#x3002;

&#x6C99;&#x773C;&#x8863;&#x539F;&#x4F53;&#x589E;&#x5F3A;&#x4EBA;&#x7C7B;&#x8F93;&#x5375;&#x7BA1;&#x611F;&#x67D3;&#x4F53;&#x5916;&#x6A21;&#x578B;&#x4E2D;&#x57FA;&#x8D28;&#x91D1;&#x5C5E;&#x86CB;&#x767D;&#x9176;&#x7684;&#x8868;&#x8FBE;&#x3002;

The sequelae of sexually transmitted Chlamydia trachomatis infection include fallopian tube scarring, which implies modification of the extracellular matrix. Our objective was to describe the production of two matrix metalloproteinases in response to chlamydial infection in vitro.

Chlamydia trachomatis enhances the expression of matrix metalloproteinases in an in vitro model of the human fallopian tube infection.

Les s&eacute;quelles d&#39;infections sexuellement transmissibles Chlamydia trachomatis incluent une cicatrisation des trompes de Fallope, qui implique la modification de la matrice extracellulaire. Notre objectif &eacute;tait de d&eacute;crire la production de deux m&eacute;talloprot&eacute;inases matricielles en r&eacute;ponse &agrave; l&#39;infection &agrave; Chlamydia in vitro.

Chlamydia trachomatis favorise l&#39;expression des prot&eacute;inases m&eacute;talliques de matrice dans un mod&egrave;le in vitro de l&#39;infection des trompes de Fallope.

Die Folgeerscheinungen von sexuell &uuml;bertragbaren Chlamydia-Trachomatis-Infektion z&auml;hlen Eileiter Narben, was bedeutet &Auml;nderung der extrazellul&auml;ren Matrix. Unser Ziel war es, die Produktion von zwei Matrix Metalloproteinasen als Reaktion auf in-vitro Chlamydieninfektion zu beschreiben.

Chlamydia Trachomatis erh&ouml;ht den Ausdruck der Matrix Metalloproteinasen in einem in-vitro-Modell der menschlichen Eileiter Infektion.

Le sequele di infezioni a trasmissione sessuale Chlamydia trachomatis includono cicatrici tuba di Falloppio, che implica la modifica della matrice extracellulare. Il nostro obiettivo era quello di descrivere la produzione di due matrix metalloproteinases in risposta all&#39;infezione da clamidia in vitro.

Chlamydia trachomatis migliora l&#39;espressione delle metalloproteinasi di matrice in un modello in vitro l&#39;infezione umana tuba di Falloppio.

&#x6027;&#x611F;&#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2;&#x611F;&#x67D3;&#x75C7;&#x306E;&#x5F8C;&#x907A;&#x75C7;&#x5375;&#x7BA1;&#x7622;&#x75D5;&#x3001;&#x7D30;&#x80DE;&#x5916;&#x306E;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x306E;&#x6539;&#x8CEA;&#x3092;&#x610F;&#x5473;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306E;&#x76EE;&#x7684;&#x306F;&#x3001;&#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2;&#x611F;&#x67D3;&#x4F53;&#x5916;&#x306B;&#x5FDC;&#x7B54;&#x3057;&#x3066;&#x305D;&#x306E; 2 &#x3064;&#x306E;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x30E1;&#x30BF;&#x30ED;&#x30D7;&#x30ED;&#x30C6;&#x30A2;&#x30FC;&#x30BC;&#x306E;&#x751F;&#x7523;&#x3092;&#x8A18;&#x8FF0;&#x3059;&#x308B;&#x305F;&#x3081;&#x3060;&#x3063;&#x305F;&#x3002;

&#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2; &#x30FB; &#x30C8;&#x30E9;&#x30B3;&#x30DE;&#x30C1;&#x30B9;&#x4EBA;&#x5375;&#x7BA1;&#x611F;&#x67D3;&#x306E; in vitro &#x30E2;&#x30C7;&#x30EB;&#x306B;&#x304A;&#x3051;&#x308B;&#x30DE;&#x30C8;&#x30EA;&#x30C3;&#x30AF;&#x30B9;&#x30E1;&#x30BF;&#x30ED;&#x30D7;&#x30ED;&#x30C6;&#x30A2;&#x30FC;&#x30BC;&#x306E;&#x767A;&#x73FE;&#x3092;&#x5F37;&#x5316;&#x3057;&#x307E;&#x3059;&#x3002;

&#xC131;&#xBCD1;&#xB41C; &#xD074; &#xB77C; &#xBBF8;&#xB514; &#xC544;&#xC758; trachomatis &#xAC10;&#xC5FC; sequelae &#xD3EC;&#xD568; &#xB098; &#xD314; &#xAD00;, &#xD749;&#xD130;&#xB294; &#xC138;&#xD3EC; &#xC678; &#xB9E4;&#xD2B8;&#xB9AD;&#xC2A4;&#xC758; &#xC218;&#xC815;&#xC744; &#xC758;&#xBBF8; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xBAA9;&#xD45C;&#xB294; &#xB450; &#xD589;&#xB82C; metalloproteinases chlamydial &#xAC10;&#xC5FC; &#xCCB4; &#xC678;&#xC5D0; &#xB300; &#xD55C; &#xC751;&#xB2F5;&#xC73C;&#xB85C; &#xC0DD;&#xC0B0;&#xC744; &#xC124;&#xBA85; &#xD588;&#xB2E4;.

&#xD074; &#xB77C; &#xBBF8;&#xB514; &#xC544;&#xC758; trachomatis &#xC778;&#xAC04;&#xC758; &#xB098; &#xD314; &#xAD00; &#xAC10;&#xC5FC;&#xC758; &#xCCB4; &#xC678; &#xBAA8;&#xB378;&#xC5D0; &#xB9E4;&#xD2B8;&#xB9AD;&#xC2A4; metalloproteinases&#xC758; &#xC2DD;&#xC744; &#xD5A5;&#xC0C1; &#xC2DC;&#xD0B5;&#xB2C8;&#xB2E4;.

Las secuelas de la infecci&oacute;n de transmisi&oacute;n sexual Clamidia trachomatis son cicatrices de la trompa de Falopio, que implica la modificaci&oacute;n de la matriz extracelular. Nuestro objetivo era describir la producci&oacute;n de metaloproteinasas de matriz dos en respuesta a la infecci&oacute;n por clamidia in vitro.

Chlamydia trachomatis aumenta la expresi&oacute;n de metaloproteinasas de matriz en un modelo in vitro de la infecci&oacute;n de la trompa de Falopio humana.

We sought to determine if intraluminal occluding fibrosis of the oviduct occurs after urogenital Chlamydia muridarum infection in mice.

Histopathologic changes related to fibrotic oviduct occlusion after genital tract infection of mice with Chlamydia muridarum.

Matrix metalloproteinases (MMP) are a family of host-derived enzymes involved in the turnover of extracellular matrix molecules. We have previously reported enhanced expression of matrix metalloproteinases in Chlamydia muridarum urogenital tract infection of female mice. Kinetics and patterns of MMP expression as well as enhanced expression in susceptible strains of mice in the prior study implied a role for MMP in pathogenesis. To explore this further, we infected a susceptible strain of mice (C3H/HeN) with C. muridarum and treated two groups of mice with either one of two chemical inhibitors of MMP (MMPi; captopril and a chemically modified tetracycline) and reserved infected sham-treated mice as controls. Neither of the treatments affected shedding of viable chlamydiae from the lower urogenital tract, but the administration of either MMPi protected mice from the formation of hydrosalpinx-a surrogate marker of oviduct occlusion and infertility. Interestingly, the mechanism of protection for mice treated with chemically modified tetracycline 3, appeared to be related to prevention of ascending upper genital tract infection. These results imply that MMP are involved in pathogenesis of chlamydial infection in this model by mediating ascension of the infection into the upper genital tract.

Inhibition of matrix metalloproteinases protects mice from ascending infection and chronic disease manifestations resulting from urogenital Chlamydia muridarum infection.

CD14 has been postulated to play a role in chlamydial immunity and immunopathology. There is evidence to support this role in human infections but its function in a mouse model has not been investigated.

Outcome of urogenital infection with Chlamydia muridarum in CD14 gene knockout mice.

Matrix metalloproteinases (MMPs) are a family of host-derived enzymes involved in the turnover of extracellular matrix (ECM) molecules and the processing of cytokines, chemokines and growth factors. We have previously reported that global inhibition of MMP in Chlamydia muridarum urogenital tract infection of susceptible strains of female mice impeded ascension of C. muridarum into the upper genital tract, blunted acute inflammatory responses and reduced the rate of formation of chronic disease. Because we have also observed that MMP-9 (also known as gelatinase B) is expressed in relatively large quantities in susceptible strains of mice in response to infection during acute phases of infection, we explored this further in a more selected fashion. We infected MMP-9 gene knockout mice and wild type controls intravaginally with C. muridarum. Both groups of mice had similar isolation rates from the lower urogenital tract but the absence of MMP-9 resulted in a slightly lower isolation rate in the upper genital tract, blunted acute inflammatory indices in the affected tissues and a reduced rate of formation of hydrosalpinx-a surrogate marker of infertility. These results imply that MMP-9 is involved in pathogenesis of chlamydial infection in this model possibly by amplifying inflammatory responses.

A role for matrix metalloproteinase-9 in pathogenesis of urogenital Chlamydia muridarum infection in mice.

&#x9F20;&#x6807;&#x8863;&#x539F;&#x4F53;&#x75C5;&#x539F;&#x4F53;&#x8863;&#x539F;&#x4F53; muridarum &#x5DF2;&#x88AB;&#x7528;&#x4F5C;&#x4EBA;&#x7C7B;&#x8863;&#x539F;&#x4F53;&#x6C99;&#x773C;&#x6CCC;&#x5C3F;&#x751F;&#x6B96;&#x9053;&#x548C;&#x547C;&#x5438;&#x9053;&#x547C;&#x5438;&#x9053;&#x611F;&#x67D3;&#x7684;&#x7814;&#x7A76;&#x7684;&#x6A21;&#x5F0F;&#x751F;&#x7269;&#x3002;&#x5230;&#x76EE;&#x524D;&#x4E3A;&#x6B62;&#xFF0C;&#x4E24;&#x4E2A;&#x5E38;&#x7528; C.muridarum &#x682A;&#x5DF2;&#x88AB;&#x4EA4;&#x66FF;&#x4F7F;&#x7528;&#x548C;&#x57FA;&#x672C;&#x4E0A;&#x662F;&#x91C7;&#x53D6;&#x5B8C;&#x5168;&#x76F8;&#x540C;&#x3002;&#x672C;&#x6587;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x63D0;&#x51FA;&#x8868;&#x660E;&#x8FD9;&#x4E0D;&#x662F;&#x8FD9;&#x79CD;&#x60C5;&#x51B5;&#x7684;&#x6570;&#x636E;&#x3002;C.muridarum Weiss &#x9694;&#x79BB;&#x548C; C.muridarum &#x5C3C;&#x683C;&#x5206;&#x79BB;&#x5DEE;&#x522B;&#x4E5F;&#x5F88;&#x5927;&#x4E2D;&#x5176;&#x6BD2;&#x5728;&#x4F53;&#x5185;&#x548C;&#x62E5;&#x6709;&#x4E0D;&#x540C;&#x751F;&#x957F;&#x7279;&#x6027;&#x4F53;&#x5916;&#x3002;&#x4E0D;&#x540C;&#x7684;&#x5DEE;&#x5F02;&#x53D1;&#x73B0;&#x5728;&#x9634;&#x9053; 50%&#x4F20;&#x67D3;&#x6027;&#x5242;&#x91CF;&#x548C;&#x6311;&#x6218;&#x611F;&#x67D3; Weiss &#x5206;&#x79BB;&#x663E;&#x793A;&#x66F4;&#x5927;&#x6BD2;&#x529B;&#x3002;&#x547C;&#x5438;&#x9053;&#x611F;&#x67D3;&#x9F3B;&#x8DEF;&#x7EBF;&#x8FD8;&#x8868;&#x793A; Weiss &#x5206;&#x79BB;&#x66F4;&#x6BD2;&#x3002;&#x4F53;&#x5916;&#xFF0C;&#x5F62;&#x6001;&#x8BA1;&#x91CF;&#x5B66;&#x5206;&#x6790;&#x63ED;&#x793A; Weiss &#x5206;&#x79BB;&#x4EA7;&#x751F;&#x4E00;&#x8D2F;&#x5C0F;&#x5305;&#x88F9;&#x4F53;&#x5B50;&#x5BAB;&#x9888;&#x817A;&#x764C;&#x7EC6;&#x80DE; (HeLa 229&#xFF09; &#x53CA;&#x5C0F;&#x6591;&#x5757;&#x7684;&#x5355;&#x5206;&#x5B50;&#x819C;&#x7684;&#x5C0F;&#x9F20;&#x6210;&#x7EA4;&#x7EF4;&#x7EC6;&#x80DE; (L929) &#x6BD4;&#x5C3C;&#x683C;&#x9694;&#x79BB;&#x3002;&#x6B64;&#x5916;&#xFF0C;Weiss &#x5206;&#x79BB;&#x62E5;&#x6709;&#x663E;&#x8457;&#x63D0;&#x9AD8;&#x7E41;&#x6B96;&#x4EA7;&#x91CF;&#x4F53;&#x5916;&#x6BD4;&#x5C3C;&#x683C;&#x5206;&#x79BB;&#x3002;&#x6591;&#x5757;&#x51C0;&#x5316;&#x5206;&#x79BB;&#x6765;&#x81EA;&#x6211;&#x4EEC;&#x6240;&#x50A8;&#x5B58;&#x7684;&#x8FD9;&#x4E9B;&#x4E24;&#x682A;&#xFF0C;&#x5168;&#x57FA;&#x56E0;&#x7EC4;&#x6D4B;&#x5E8F;&#x786E;&#x5B9A;&#x51E0;&#x4E2A;&#x72EC;&#x7279;&#x7684;&#x7A81;&#x53D8;&#x5355;&#x6838;&#x82F7;&#x9178;&#x591A;&#x6001;&#x6027;&#x548C;&#x63D2;&#x5165;/&#x7F3A;&#x5931;&#x7A81;&#x53D8;&#x65F6;&#x6211;&#x4EEC; Weiss &#xFF08;n = 4) &#x548C;&#x5C3C;&#x683C; (n = 5) &#x5206;&#x79BB;&#x4E0E;&#x5DF2;&#x53D1;&#x5E03;&#x7684;&#x5C3C;&#x683C;&#x5E8F;&#x5217;&#x4F5C;&#x4E86;&#x6BD4;&#x8F83;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x8FD9;&#x4E24;&#x4E2A;&#x83CC;&#x682A;&#x76F8;&#x6BD4;&#x5DF2;&#x53D1;&#x5E03;&#x5C3C;&#x683C;&#x5E8F;&#x5217;&#x7684;&#x5171;&#x4EAB; 11 &#x7A81;&#x53D8;&#x3002;&#x8FD9;&#x4E9B;&#x7ED3;&#x679C;&#x8BC1;&#x660E;&#x6709;&#x57FA;&#x56E0;&#x578B;&#x548C;&#x6BD2;&#x529B;&#x591A;&#x6837;&#x6027;&#x4E4B;&#x95F4; C.muridarum &#x682A;&#x3002;&#x53EF;&#x4EE5;&#x5229;&#x7528;&#x8FD9;&#x4E9B;&#x7814;&#x7A76;&#x7ED3;&#x679C;&#x6765;&#x786E;&#x5B9A;&#x8863;&#x539F;&#x4F53;&#x81F4;&#x75C5;&#x4E0E;&#x514D;&#x75AB;&#x6709;&#x5173;&#x7684;&#x56E0;&#x7D20;&#x3002;

&#x5728;&#x9F20;&#x6807;&#x75C5;&#x539F;&#x4F53;&#x8863;&#x539F;&#x4F53; muridarum &#x5E94;&#x53D8;&#x548C;&#x6BD2;&#x529B;&#x7684;&#x591A;&#x6837;&#x6027;&#x3002;

The mouse chlamydial pathogen Chlamydia muridarum has been used as a model organism for the study of human Chlamydia trachomatis urogenital and respiratory tract infections. To date, two commonly used C. muridarum isolates have been used interchangeably and are essentially taken to be identical. Herein, we present data that indicate that this is not the case. The C. muridarum Weiss isolate and C. muridarum Nigg isolate varied significantly in their virulences in vivo and possessed different growth characteristics in vitro. Distinct differences were observed in intravaginal 50% infectious doses and in challenge infections, with the Weiss isolate displaying greater virulence. Respiratory infection by the intranasal route also indicated a greater virulence of the Weiss isolate. In vitro, morphometric analysis revealed that the Weiss isolate produced consistently smaller inclusions in human cervical adenocarcinoma cells (HeLa 229) and smaller plaques in monolayers of mouse fibroblasts (L929) than did the Nigg isolate. In addition, the Weiss isolate possessed significantly higher replicative yields in vitro than did the Nigg isolate. In plaque-purified isolates derived from our stocks of these two strains, total genomic sequencing identified several unique nonsynonymous single nucleotide polymorphisms and insertion/deletion mutations when our Weiss (n = 4) and Nigg (n = 5) isolates were compared with the published Nigg sequence. In addition, the two isolates shared 11 mutations compared to the published Nigg sequence. These results prove that there is genotypic and virulence diversity among C. muridarum isolates. These findings can be exploited to determine factors related to chlamydial virulence and immunity.

Strain and virulence diversity in the mouse pathogen Chlamydia muridarum.

Le pathog&egrave;ne Chlamydia souris Chlamydia muridarum a &eacute;t&eacute; utilis&eacute; comme un organisme mod&egrave;le pour l&#39;&eacute;tude des humaines infections des voies respiratoires et urog&eacute;nitales &agrave; Chlamydia trachomatis. A ce jour, deux couramment utilis&eacute; c. muridarum isolats ont &eacute;t&eacute; utilis&eacute;s de fa&ccedil;on interchangeable et sont essentiellement consid&eacute;r&eacute;s comme identiques. Ici, nous pr&eacute;sentons des donn&eacute;es qui indiquent que ce n&#39;est pas le cas. Le c. muridarum Weiss isoler et c. muridarum Nigg isolat variait significativement leurs virulences in vivo et poss&eacute;d&eacute;e de croissance diff&eacute;rents caract&eacute;ristiques in vitro. Des diff&eacute;rences marqu&eacute;es ont &eacute;t&eacute; observ&eacute;es en intravaginal doses infectieuses de 50 % et dans les infections de d&eacute;fi, avec la Weiss isolat affiche une plus grande virulence. Infection des voies respiratoires par voie intranasale a &eacute;galement indiqu&eacute; une plus grande virulence de l&#39;isolat de Weiss. In vitro, analyse morphom&eacute;trique r&eacute;v&egrave;le que l&#39;isolat Weiss produit toujours plus petites inclusions dans les cellules humaines ad&eacute;nocarcinome du col ut&eacute;rin (HeLa 229) et isoler les plaques plus petites en monocouches de fibroblastes de souris (L929) que fait le Nigg. En outre, l&#39;isolat de Weiss poss&eacute;dait significativement plus &eacute;lev&eacute;es r&eacute;plicatifs rendements in vitro que ne l&#39;isolat Nigg. Dans la purifi&eacute; plaque isolats provenant de nos stocks de ces deux souches, s&eacute;quen&ccedil;age g&eacute;nomique total identifi&eacute; plusieurs polymorphismes unique non-synonymes et des mutations de l&#39;insertion/d&eacute;l&eacute;tion quand notre Weiss (n = 4) et Nigg (n = 5) isolats ont &eacute;t&eacute; compar&eacute;s avec la s&eacute;quence Nigg publi&eacute;e. En outre, les deux isolats des 11 mutations partag&eacute;es par rapport &agrave; la s&eacute;quence de Nigg publi&eacute;e. Ces r&eacute;sultats prouvent qu&#39;il existe g&eacute;notypique et diversit&eacute; de la virulence chez c. muridarum isole. Ces r&eacute;sultats peuvent &ecirc;tre exploit&eacute;es afin de d&eacute;terminer les facteurs li&eacute;s &agrave; l&#39;immunit&eacute; et la virulence de Chlamydia trachomati.

Diversit&eacute; de souche et de la virulence dans le souris pathog&egrave;ne Chlamydia muridarum.

Der Maus-Chlamydien-Erreger Chlamydia Muridarum wurde als Modellorganismus zur Erforschung der menschlichen Chlamydia Trachomatis Urogenitale und respiratorischen Trakt Infektionen eingesetzt. Bis dato verwendete zwei h&auml;ufig C. Muridarum Isolate Synonym verwendet wurden und werden im Wesentlichen identisch sein. Hier pr&auml;sentieren wir Daten, die zeigen, dass dies nicht der Fall ist. C. Muridarum Weiss isolieren und C. Muridarum Nigg Isolat deutlich in ihrer Virulences in Vivo und besa&szlig; verschiedene Wachstum Merkmale in-vitro-variiert. Deutliche Unterschiede wurden in intrauterinelle 50 % infekti&ouml;se Dosis und Herausforderung-Infektionen, mit Weiss Eingreiftruppe anzeigen gr&ouml;&szlig;ere Virulenz beobachtet. Infektion der Atemwege durch die intranasale Route angegeben auch eine st&auml;rkere Virulenz von Weiss Isolat. In vitro morphometrischer Analysen ergab, dass das Weiss-Isolat konsequent kleinere Einschl&uuml;sse in menschlichen zervikalen Adenokarzinom Zellen (HeLa 229 produziert) und kleinere Plaketten in Monolayers der Maus-Fibroblasten (L929) als die Nigg Tat isolieren. Dar&uuml;ber hinaus besa&szlig; das Weiss-Isolat deutlich h&ouml;here replicative Ertr&auml;ge in-vitro-als Isolat Nigg Tat. In Tafel-gereinigt-Isolate, die von unserer Best&auml;nde diese zwei St&auml;mme abgeleitet insgesamt genomische Sequenzierung identifiziert mehrere einzigartige nonsynonymous Einzel-Nukleotid-Polymorphismen und Einf&uuml;gung/Auslassung Mutationen bei unserer Weiss (n = 4) und Nigg (n = 5) Isolate wurden verglichen mit der ver&ouml;ffentlichten Nigg-Sequenz. Dar&uuml;ber hinaus die beiden isoliert freigegebene 11 Mutationen im Vergleich zu den ver&ouml;ffentlichten Nigg-Sequenz. Diese Ergebnisse beweisen, dass es genotypische und Virulenz Vielfalt unter C. Muridarum isoliert. Diese Erkenntnisse k&ouml;nnen ausgenutzt werden, um zu bestimmen, im Zusammenhang mit Chlamydien Virulenz und Immunit&auml;t.

Belastung und Virulenz Vielfalt in der Maus Erreger Chlamydia Muridarum.

L&#39;agente patogeno di mouse da clamidia Chlamydia muridarum &egrave; stato utilizzato come organismo modello per lo studio delle infezioni del tratto urogenitale e respiratorio nell&#39;uomo Chlamydia trachomatis. Ad oggi, due comunemente utilizzati c. muridarum isolati sono stati usati in modo intercambiabile e sono essenzialmente prese in modo identico. Vi presentiamo qui, dati che indicano che questo non &egrave; il caso. Il c. muridarum Weiss isolare e c. muridarum Nigg isolato varia sensibilmente in loro virulences in vivo e crescita posseduto diversa caratteristiche in vitro. Distinte differenze sono state osservate in intravaginale 50% dosi infettive e infezioni di sfida, con Weiss isolata visualizzazione maggiore virulenza. Infezione delle vie respiratorie per via intranasale indicata anche una maggiore virulenza dell&#39;isolato Weiss. In vitro, analisi morfometriche ha rivelato che l&#39;isolato Weiss prodotta costantemente piccole inclusioni in cellule di adenocarcinoma cervicale umano (HeLa 229) e piccole placche in monostrati di fibroblasti (L929) che ha fatto la Nigg isolare. Inoltre, l&#39;isolato Weiss ha posseduto significativamente replicativa rendimenti pi&ugrave; alti in vitro di fatto isolato Nigg. Negli isolati di placca-purificato derivati dal nostro stock di questi due ceppi, sequenziamento genomico totale identificati diversi unico nonsynonymous single nucleotide polymorphisms e mutazioni di inserimento/cancellazione quando nostro Weiss (n = 4) e Nigg (n = 5) isolati sono stati confrontati con la sequenza di Nigg pubblicata. Inoltre, i due isolati condivise 11 mutazioni rispetto alla sequenza Nigg pubblicata. Questi risultati dimostrano che c&#39;&egrave; genotipica e diversit&agrave; di virulenza tra c. muridarum isolati. Questi risultati possono essere sfruttati per determinare i fattori legati all&#39;immunit&agrave; e virulenza da clamidia.

Ceppo e virulenza la diversit&agrave; del mouse patogeno Chlamydia muridarum.

&#x30DE;&#x30A6;&#x30B9; &#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2;&#x75C5;&#x539F;&#x4F53;&#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2; muridarum &#x30E2;&#x30C7;&#x30EB;&#x751F;&#x7269;&#x3068;&#x3057;&#x3066;&#x4EBA;&#x9593;&#x306E;&#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2; &#x2010; &#x30C8;&#x30E9;&#x30B3;&#x30DE;&#x30C6;&#x30A3;&#x30B9;&#x6CCC;&#x5C3F;&#x751F;&#x6B96;&#x5668;&#x30FB;&#x547C;&#x5438;&#x5668;&#x306E;&#x5C3F;&#x8DEF;&#x611F;&#x67D3;&#x75C7;&#x306E;&#x7814;&#x7A76;&#x306E;&#x305F;&#x3081;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x65E5;&#x4ED8;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;2 &#x3064;&#x306E;&#x5206;&#x96E2;&#x682A;&#x306F;&#x540C;&#x3058;&#x610F;&#x5473;&#x3067;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x3057;&#x3001;&#x306F;&#x672C;&#x8CEA;&#x7684;&#x306B;&#x540C;&#x4E00;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x53D6;&#x3089;&#x308C;&#x308B; C. muridarum &#x4E00;&#x822C;&#x7684;&#x3002;&#x672C;&#x660E;&#x7D30;&#x66F8;&#x3067;&#x306F;&#x30B1;&#x30FC;&#x30B9;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3059;&#x30C7;&#x30FC;&#x30BF;&#x3092;&#x8868;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;C. muridarum &#x30EF;&#x30A4;&#x30B9;&#x5206;&#x96E2;&#x3057;&#x3001;C. muridarum &#x30CB;&#x30B0;&#x5206;&#x96E2;&#x5927;&#x5E45;&#x306B;&#x5F7C;&#x3089;&#x306E;&#x75C5;&#x6BD2;&#x6027;&#x751F;&#x4F53;&#x5185;&#x3068;&#x4FDD;&#x6709;&#x306E;&#x7570;&#x306A;&#x308B;&#x6210;&#x9577;&#x306E;&#x7279;&#x6027;&#x3092; in vitro &#x3067;&#x69D8;&#x3005; &#x306A;&#x3002;&#x660E;&#x78BA;&#x306A;&#x9055;&#x3044;&#x30D7;&#x30ED;&#x30B8;&#x30A7;&#x30B9;&#x30C6;&#x30ED;&#x30F3; 50% &#x611F;&#x67D3;&#x7528;&#x91CF;&#x3068;&#x6311;&#x6226;&#x306E;&#x611F;&#x67D3;&#x75C7;&#x3001;&#x30EF;&#x30A4;&#x30B9;&#x306E;&#x5206;&#x96E2;&#x8868;&#x793A;&#x3059;&#x308B;&#x5927;&#x304D;&#x3044;&#x75C5;&#x539F;&#x3068;&#x89B3;&#x5BDF;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x547C;&#x5438;&#x5668;&#x611F;&#x67D3;&#x75C7;&#x3001;&#x9F3B;&#x8154;&#x5185;&#x306E;&#x30EB;&#x30FC;&#x30C8;&#x306F;&#x307E;&#x305F;&#x30F4;&#x30A1;&#x30A4;&#x30B9;&#x30B7;&#x30E5;&#x30F4;&#x30A1;&#x30EB;&#x30C4;&#x5206;&#x96E2;&#x306E;&#x5927;&#x304D;&#x3044;&#x75C5;&#x539F;&#x6027;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;In vitro &#x306B;&#x304A;&#x3051;&#x308B;&#x5F62;&#x614B;&#x8A08;&#x6E2C;&#x7684;&#x89E3;&#x6790;&#x3067;&#x306F;&#x3001;&#x30EF;&#x30A4;&#x30B9;&#x5206;&#x96E2;&#x3072;&#x3068;&#x5B50;&#x5BAE;&#x981A;&#x90E8;&#x817A;&#x764C;&#x7D30;&#x80DE; &#xFF08;HeLa 229) &#x3067;&#x4E00;&#x8CAB;&#x3057;&#x3066;&#x3088;&#x308A;&#x5C0F;&#x3055;&#x3044;&#x4ECB;&#x5728;&#x7269;&#x751F;&#x6210;&#x3057;&#x3001;&#x30CB;&#x30B0;&#x304C;&#x3084;&#x3063;&#x305F;&#x3088;&#x308A;&#x3082;&#x5C0F;&#x578B;&#x306E;&#x30D7;&#x30E9;&#x30FC;&#x30AF; &#x30DE;&#x30A6;&#x30B9;&#x7E4A;&#x7DAD;&#x82BD;&#x7D30;&#x80DE; (L929) &#x306E;&#x5358;&#x5206;&#x5B50;&#x819C;&#x3067;&#x5206;&#x96E2;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x307E;&#x305F;&#x3001;&#x30EF;&#x30A4;&#x30B9;&#x5206;&#x96E2;&#x30CB;&#x30B0;&#x5206;&#x96E2;&#x3088;&#x308A;&#x3082;&#x5927;&#x5E45;&#x306B;&#x9AD8;&#x3044;&#x8907;&#x88FD;&#x5229;&#x56DE;&#x308A; in vitro &#x3067;&#x6240;&#x6709;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;&#x6211;&#x3005; &#x3053;&#x308C;&#x3089; 2 &#x7CFB;&#x7D71;&#x306E;&#x682A;&#x304B;&#x3089;&#x6D3E;&#x751F;&#x3057;&#x305F;&#x30D7;&#x30E9;&#x30FC;&#x30AF;&#x7CBE;&#x88FD;&#x306E;&#x83CC;&#x682A;&#x3067;&#x5408;&#x8A08;&#x30B2;&#x30CE;&#x30E0; &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E;&#x30E6;&#x30CB;&#x30FC;&#x30AF;&#x306A;&#x55C5;&#x899A;&#x4E00;&#x5869;&#x57FA;&#x591A;&#x578B;&#x3068;&#x633F;&#x5165;/&#x524A;&#x9664;&#x5909;&#x7570;&#x8B58;&#x5225;&#x3068;&#x304D;&#x79C1;&#x305F;&#x3061;&#x30EF;&#x30A4;&#x30B9; &#xFF08;n = 4) &#x3068;&#x30CB;&#x30B0; (n = 5) &#x5206;&#x96E2;&#x682A;&#x516C;&#x958B;&#x30CB;&#x30B0; &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;2 &#x3064;&#x516C;&#x958B;&#x30CB;&#x30B0; &#x30B7;&#x30FC;&#x30B1;&#x30F3;&#x30B9;&#x306B;&#x6BD4;&#x8F03;&#x3057;&#x3066;&#x5171;&#x6709; 11 &#x5909;&#x7570;&#x3092;&#x5206;&#x96E2;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D50;&#x679C;&#x907A;&#x4F1D;&#x5B50;&#x578B;&#x3060;&#x3057;&#x3001;&#x5206;&#x96E2;&#x682A;&#x306E;&#x75C5;&#x539F;&#x6027;&#x306E;&#x591A;&#x69D8;&#x6027; C. muridarum &#x306E;&#x9593;&#x3067;&#x8A3C;&#x660E;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x6240;&#x898B;&#x306F;&#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2;&#x306E;&#x75C5;&#x539F;&#x6027;&#x3068;&#x514D;&#x75AB;&#x306B;&#x95A2;&#x9023;&#x3059;&#x308B;&#x8981;&#x56E0;&#x3092;&#x6C7A;&#x5B9A;&#x3059;&#x308B;&#x60AA;&#x7528;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;

&#x3072;&#x305A;&#x307F;&#x3068;&#x75C5;&#x539F;&#x6027;&#x306E;&#x591A;&#x69D8;&#x6027;&#x30DE;&#x30A6;&#x30B9;&#x75C5;&#x539F;&#x4F53;&#x30AF;&#x30E9;&#x30DF;&#x30B8;&#x30A2; muridarum &#x306B;&#x3002;

&#xB9C8;&#xC6B0;&#xC2A4; chlamydial &#xBCD1;&#xC6D0; &#xCCB4; &#xD074; &#xB77C; &#xBBF8;&#xB514; &#xC544; muridarum &#xC778;&#xAC04;&#xC758; &#xD074; &#xB77C; &#xBBF8;&#xB514; &#xC544;&#xC758; trachomatis &#xBE44;&#xB1E8; &#xC0DD;&#xC2DD; &#xAE30; &#xBC0F; &#xD638;&#xD761;&#xAE30;&#xB85C; &#xAC10;&#xC5FC;&#xC758; &#xC5F0;&#xAD6C;&#xC5D0; &#xB300; &#xD55C; &#xBAA8;&#xB378; &#xC0DD;&#xBB3C;&#xC73C;&#xB85C; &#xC0AC;&#xC6A9; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC9C0;&#xAE08;&#xAE4C;&#xC9C0; &#xB450;&#xB294; &#xC77C;&#xBC18;&#xC801;&#xC73C;&#xB85C; &#xACA9;&#xB9AC; &#xAC19;&#xC740; &#xC758;&#xBBF8;&#xB85C; &#xC0AC;&#xC6A9; &#xB418; &#xACE0; &#xAE30;&#xBCF8;&#xC801;&#xC73C;&#xB85C; &#xB3D9;&#xC77C;&#xD55C; &#xAC83;&#xC73C;&#xB85C; &#xCD2C;&#xC601; &#xD558;&#xB294; C. muridarum &#xC0AC;&#xC6A9;. &#xC5EC;&#xAE30;&#xC5D0;,&#xC774; &#xC0AC;&#xC2E4;&#xC774; &#xB418;&#xC9C0; &#xB098;&#xD0C0;&#xB0B4;&#xB294; &#xB370;&#xC774;&#xD130; &#xC120;&#xBB3C;&#xC774;. C. muridarum&#xC640; &#xC774;&#xC988; &#xACA9;&#xB9AC; &#xBC0F; C. muridarum Nigg &#xBD84;&#xB9AC; &#xBCC0;&#xD654;&#xB97C; &#xD06C;&#xAC8C; vivo &#xBC0F; &#xBCF4;&#xC720; &#xB2E4;&#xB978; &#xC131;&#xC7A5;&#xC5D0; &#xADF8;&#xB4E4;&#xC758; virulences&#xC5D0; &#xD2B9;&#xC131; &#xC0DD;&#xCCB4; &#xC678;&#xC5D0;&#xC11C;. &#xC640; &#xC774;&#xC988; &#xACA9;&#xB9AC; &#xD45C;&#xC2DC; &#xD070; &#xB3C5;&#xC131;&#xACFC; &#xB69C;&#xB837;&#xD55C; &#xCC28;&#xC774;&#xAC00; &#x8FBC; 50% &#xAC10;&#xC5FC; &#xBCF5;&#xC6A9;&#xB7C9; &#xBC0F; &#xB3C4;&#xC804; &#xAC10;&#xC5FC; &#xAD00;&#xCC30; &#xB418;&#xC9C0; &#xC54A;&#xC558;&#xB2E4;. Intranasal &#xACBD;&#xB85C; &#xC758;&#xD574; &#xD638;&#xD761;&#xAE30; &#xAC10;&#xC5FC; &#xB610;&#xD55C;&#xC640; &#xC774;&#xC988; &#xBD84;&#xB9AC;&#xC758; &#xD070; &#xB3C5;&#xC131;&#xC744; &#xAC00;&#xB9AC;&#xD0A8;&#xB2E4;. &#xCCB4; &#xC678;, &#xD615;&#xD0DC;&#xD559; &#xBD84;&#xC11D;&#xC774; &#xBC1D;&#xD614;&#xB2E4;&#xC640; &#xC774;&#xC988; &#xBD84;&#xB9AC; &#xC778;&#xAC04;&#xC758; &#xC790; &#xAD81; &#xACBD;&#xBD80; &#xC120; &#xC554; &#xC138;&#xD3EC; (HeLa 229)&#xC5D0;&#xC11C; &#xC77C;&#xAD00; &#xB418; &#xAC8C; &#xC791;&#xC740; &#xD760;&#xB3C4; &#xC0DD;&#xC0B0; &#xD558; &#xACE0; &#xB354; &#xC791;&#xC740; &#xD50C; &#xB77C;&#xD06C; &#xB9C8;&#xC6B0;&#xC2A4; &#xAC8C;&#xC7AC; (L929)&#xC758; monolayers&#xC5D0;&#xC11C; &#xBCF4;&#xB2E4;&#xB294; Nigg &#xACA9;&#xB9AC;. &#xB610;&#xD55C;,&#xC640; &#xC774;&#xC988; &#xBD84;&#xB9AC; &#xC0C1;&#xB2F9;&#xD788; &#xB192;&#xC740; &#xC77C;&#xCC28; &#xC0DD;&#xC0B0;&#xB7C9; &#xCCB4; &#xC678; &#xBCF4;&#xB2E4; Nigg &#xBD84;&#xB9AC;&#xB97C; &#xD640;&#xB9AC; &#xACE0;. &#xC774; &#xB450; &#xC885;&#xC790;&#xC758; &#xC6B0;&#xB9AC;&#xC758; &#xC8FC;&#xC2DD;&#xC5D0;&#xC11C; &#xD30C;&#xC0DD; &#xD558;&#xB294; &#xD50C; &#xB77C;&#xD06C; &#xC815;&#xD654; &#xACA9;&#xB9AC; &#xCD1D; &#xAC8C;&#xB188; &#xC2DC;&#xD000;&#xC2F1; &#xC2DD;&#xBCC4; &#xBA87; &#xAC00;&#xC9C0; &#xACE0;&#xC720;&#xC758; nonsynonymous &#xB2E8;&#xC77C; &#xB274;&#xD074;&#xB808;&#xC624;&#xD2F0;&#xB4DC; polymorphisms &#xBC0F; &#xC0BD;&#xC785;/&#xC0AD;&#xC81C; &#xBCC0;&#xC774; &#xB54C; &#xC800;&#xD76C;&#xC640; &#xC774;&#xC988; (n = 4)&#xACFC; Nigg (n = 5) &#xACA9;&#xB9AC; &#xAC8C;&#xC2DC; Nigg &#xC2DC;&#xD000;&#xC2A4;&#xC640; &#xBE44;&#xAD50; &#xD588;&#xB2E4;. &#xB610;&#xD55C;, &#xB450; &#xAC8C;&#xC2DC; Nigg &#xC2DC;&#xD000;&#xC2A4;&#xC5D0; &#xBE44;&#xD574; &#xACF5;&#xC720; 11 &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xBD84;&#xB9AC; &#xD569;&#xB2C8;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xACB0;&#xACFC; genotypic C. muridarum &#xC911; &#xB3C5;&#xC131; &#xB2E4;&#xC591;&#xC131; &#xACA9;&#xB9AC; &#xC99D;&#xBA85;. &#xC774;&#xB7EC;&#xD55C; &#xC5F0;&#xAD6C; &#xACB0;&#xACFC; chlamydial &#xB3C5;&#xC131; &#xBC0F; &#xBA74;&#xC5ED;&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xC694;&#xC18C;&#xB97C; &#xD655;&#xC778; &#xD558;&#xAE30; &#xC704;&#xD574; &#xC545;&#xC6A9; &#xB420; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xB9C8;&#xC6B0;&#xC2A4; &#xBCD1;&#xC6D0; &#xCCB4; &#xD074; &#xB77C; &#xBBF8;&#xB514; &#xC544; muridarum&#xC5D0;&#xC11C; &#xBCC0;&#xD615; &#xBC0F; &#xB3C5;&#xC131; &#xB2E4;&#xC591;&#xC131;.

El pat&oacute;geno por clamidia Chlamydia muridarum de rat&oacute;n se ha utilizado como organismo modelo para el estudio de las infecciones humanas Chlamydia trachomatis tracto urogenital y respiratoria. Hasta la fecha, dos de uso general C. muridarum aislados han sido utilizados indistintamente y se toman esencialmente id&eacute;ntico. Adjunto, presentamos datos que indican que este no es el caso. El C. muridarum Weiss aislar y C. muridarum Nigg aislante vari&oacute; significativamente en sus virulences en vivo y crecimiento pose&iacute;do diferente caracter&iacute;sticas in vitro. Se observaron diferencias distintas en intravaginal dosis infecciosa de 50% y en infecciones de desaf&iacute;o, con el aislante Weiss exhiben mayor virulencia. Infecci&oacute;n respiratoria por v&iacute;a intranasal tambi&eacute;n indica una mayor virulencia de la cepa de Weiss. In vitro, an&aacute;lisis morfom&eacute;trico revelaron que el aislante de Weiss hab&iacute;a producido consistentemente m&aacute;s peque&ntilde;as inclusiones en c&eacute;lulas de adenocarcinoma cervical humano (HeLa 229) y peque&ntilde;as placas en monocapas de fibroblastos de rat&oacute;n (eluci&oacute;n de L929) que hicieron la Nigg aislar. Adem&aacute;s, el aislante de Weiss posey&oacute; significativamente mayores rendimientos replicativos in vitro que hicieron el aislante Nigg. En aislamientos de placa purificado derivados de nuestra acci&oacute;n de estas dos cepas, secuenciaci&oacute;n total identific&oacute; varios polimorfismos de nucle&oacute;tido &uacute;nico nonsynonymous y mutaciones de inserci&oacute;n/deleci&oacute;n cuando nuestro Weiss (n = 4) y Nigg (n = 5) aislantes fueron comparados con la secuencia de Nigg publicada. Adem&aacute;s, los dos aislados de 11 mutaciones compartidas frente a la secuencia de Nigg publicada. Estos resultados demuestran que hay genot&iacute;pico y diversidad de virulencia entre C. muridarum aislantes. Estos resultados pueden explotarse para determinar factores relacionados con la inmunidad y virulencia por clamidia.

Diversidad de cepa y virulencia en el pat&oacute;geno de rat&oacute;n Chlamydia muridarum.

Vigorous acute inflammatory responses accompany Chlamydia muridarum infections in mice and are positively correlated with adverse urogenital and respiratory tract infection outcomes in the mouse model. Thus, we tested the hypothesis that neutrophils induce an acute inflammatory insult that, in the repair phase, leads to the chronic sequelae of hydrosalpinx - a surrogate marker of infertility in the mouse model. To this end, we induced neutropenia in mice using a neutrophil-depleting monoclonal antibody during acute phases of C. muridarum urogenital infection only (days 2-21 postinfection). To prove induced neutropenia, peripheral blood was monitored for neutrophils during the treatment regimen. Neutropenic mice had a similar infection course as control mice, but had significantly reduced levels of certain histopathological parameters, reduced production of matrix metalloproteinase-9 (MMP-9) and reduced rates of hydrosalpinx following resolution of the infection. We conclude that neutrophils are a major source of MMP-9, a previously proved pathological factor in this model. Further, we conclude that acute inflammation in the form of neutrophils and neutrophil activation products are at least partially responsible for inducing the histological changes that ultimately result in fibrosis and infertility in the mouse model of chlamydial upper genital tract disease.

A link between neutrophils and chronic disease manifestations of Chlamydia muridarum urogenital infection of mice.

We tested the hypothesis that a specific chemokine receptor, CXC chemokine receptor-2 (CXCR2), mediates acute inflammatory damage during chlamydial urogenital infection, which ultimately leads to the chronic sequelae of hydrosalpinx - a surrogate marker of infertility. Homozygous CXCR2 genetic knockouts (CXCR2-/-), heterozygous littermates (CXCR2+/-) or homozygous wild-type (wt) controls (CXCR2+/+) were infected intravaginally with Chlamydia muridarum. Although no change was observed in the infection in the lower genital tract based on CXCR zygosity, a delay in the ascension of infection into the upper genital tract was seen in CXCR2-/- mice. Significantly elevated peripheral blood neutrophil counts were observed in CXCR2-/- mice when compared with controls. Reduced rates of acute inflammatory indices were observed in the affected tissue, indicating reduced neutrophil extravasation capacity in the absence of CXCR2. Of note was a reduction in the postinfection development of hydrosalpinx that correlated with CXCR2 zygosity, with both CXCR2-/- (13%) and their CXCR2+/- (35%) littermates displaying significantly lower rates of hydrosalpinx formation than the wt CXCR2-sufficient mice (93%). We conclude that CXCR2 ligands are a major chemotactic signal that induces damaging acute inflammation and the resulting chronic pathology during the repair phase of the host response, but are dispensable for the resolution of infection.

A role for CXC chemokine receptor-2 in the pathogenesis of urogenital Chlamydia muridarum infection in mice.

Cell surface heparan sulfate (HS), a polysaccharide composed of alternating uronic acid and glucosamine residues, represents a common link that many sexually transmitted infections (STIs) require for infection. Variable modifications within the monomeric units of HS chains together with their unique structural conformations generate heterogeneity, which expands the ability of HS to bind a diverse array of host and microbial proteins. Recent advances made in the field of glycobiology have critically enhanced our understanding of HS and its interactions with microbes and their significance in important human diseases. The role of HS has been elaborated for several STIs to include those caused by herpes simplex virus, human immunodeficiency virus, human papillomavirus, and Chlamydia. In addition, gonorrhea, syphilis, and yeast infections are also dependent on the presence of HS on human target cells. Critical steps such as pathogen adhesion or binding to host cells followed by internalization to enhance intracellular survival and possible spread to other cells are mediated by HS. In addition, HS guided cell signaling plays a role in the development of angiogenesis and inflammation associated with many STIs. Past and ongoing investigations are providing new push for the development of HS-mimetics and analogs as novel prevention strategies against many different STIs. This review article summarizes the significance of HS in STIs and describes how emerging new products that target HS can be used to control the spread of STIs.

Role of heparan sulfate in sexually transmitted infections.

We hypothesized that the plasmid of urogenital isolates of Chlamydia trachomatis would modulate infectivity and virulence in a mouse model. To test this hypothesis, we infected female mice in the respiratory or urogenital tract with graded doses of a human urogenital isolate of C. trachomatis, serovar F, possessing the cognate plasmid. For comparison, we inoculated mice with a plasmid-free serovar F isolate. Following urogenital inoculation, the plasmid-free isolate displayed significantly reduced infectivity compared with the wild-type strain with the latter yielding a 17-fold lower infectious dose to yield 50% infection. When inoculated via the respiratory tract, the plasmid-free isolate exhibited reduced infectivity and virulence (as measured by weight change) when compared to the wild-type isolate. Further, differences in infectivity, but not in virulence were observed in a C. trachomatis, serovar E isolate with a deletion within the plasmid coding sequence 1 when compared to a serovar E isolate with no mutations in the plasmid. We conclude that plasmid loss reduces virulence and infectivity in this mouse model. These findings further support a role for the chlamydial plasmid in infectivity and virulence in vivo.

Plasmid deficiency in urogenital isolates of Chlamydia trachomatis reduces infectivity and virulence in a mouse model.

We have previously shown that Chlamydia muridarum has multiple genomic variants that concomitantly vary in their in vitro and in vivo phenotype. Herein, we used real-time polymerase chain reaction-based genotyping assays to query plaque-cloned isolates of C. muridarum for the frequency of eight selected polymorphisms. These strains had no history of passage in vivo since their original isolation from laboratory mice. There was significant variance in the frequency of two of the eight polymorphisms assessed with the remaining exhibiting a low rate of variance. To determine if any of these polymorphisms were more favorable for in vivo conditions, we blindly passaged non-clonal C. muridarum three times at 7-day intervals through the urogenital tract of mice. Seven of the eight polymorphisms varied in frequency following in vivo passage and four of these varied between C. muridarum strains. Selected isolates displayed variable growth rates and cytopathic effect in vitro. We conclude that multiple genotypic variants are present within the existing known C. muridarum strains and that the frequency of these variants changes upon introduction into the mouse host. These findings lend support to the concept that genotypic proportional representation in a chlamydial population is dynamic and adaptive.

Genomic variant representation in a Chlamydia population is dynamic and adaptive with dependence on in vitro and in vivo passage.

To determine if Chlamydia muridarum, or other chlamydiae, are enzootic in rodents, we probed a serum bank of wild Peromyscus spp. mice for immunoglobulin G-antibody reactivity to ultraviolet light-inactivated C. muridarum elementary bodies (EBs) using an enzyme-linked immunoassay. Applying a cut-off for a positive reaction of OD(405) nm = 0.1 at a 1:20 dilution, we found titratable antibody reactivity in 190 of 247 specimens surveyed (77%, mean OD(405) = 0.33 ± 0.26, range = 0.11-1.81, median = 0.25). In addition, serum samples were obtained from a colony of specific pathogen-free Peromyscus spp. maintained at the University of South Carolina and six of 12 samples were reactive (50%, mean OD(405) = 0.19 +/- 0.08, range = 0.1-0.32, median = 0.18). Lastly, 40 additional wild Peromyscus spp. were captured in a disparate region of Midwestern USA and 22 serum specimens were reactive (55%, mean OD(405) = 0.22 +/- 0.11, range = 0.1-0.48, median = 0.2). Specificity of selected reactive sera for chlamydial antigen was confirmed on Western blot using resolved purified EBs as the detecting antigen. From tissues removed from several mice at necropsy, the gene for chlamydial 16S ribosomal ribonucleic acid (rRNA) was amplified by polymerase chain reaction (PCR). Positive samples of 16S rRNA were subjected to additional PCR for the major outer membrane protein gene (ompA). The amplicons of three select ompA positive samples were sequenced with ≥99% homology with C. muridarum. Our findings indicate that chlamydial infection is enzootic for Peromyscus spp., and that C. muridarum, or a closely related species or strain, is likely the agent in the tested rodent species.

Detection of Chlamydia infection in Peromyscus species rodents from sylvatic and laboratory sources.

Although herpes simplex virus type-1 (HSV-1), and type-2 (HSV-2), Staphylococcus aureus and Candida albicans co-habit the oral and genital mucosa, their interaction is poorly understood. We determined the effect HSV has on bacterial and/or fungal adherence, the initial step in biofilm formation. HeLa229 cells were infected with HSV-1 (KOS) gL86 or HSV-2 (KOS) 333gJ (-) at a multiplicity of infection (MOI) of 50 and 10. S. aureus (ATCC 25923) and/or C. albicans (yeast forms or germ tube forms) were co-incubated for 30 min (37 °C; 5 % CO2; 5:1 organism: HeLa cell ratio; n = 16) with virus-infected HeLa cells or uninfected HeLa cell controls. Post-incubation, the monolayers were washed (3x; PBS), lysed (RIPA), and the lysate plated onto Fungisel and/or mannitol salts agar for standard colony count. The level of HeLa-associated S. aureus was significantly decreased (P < 0.05) for both HSV-1- and HSV-2-infected cells, as compared to virus-free HeLa cell controls (38 and 59 % of control, respectively). In contrast, HSV-1 and HSV-2 significantly (P < 0.05) enhanced HeLa cell association of C. albicans yeast forms and germ tube approximately two-fold, respectively. The effect of S. aureus on germ tube and yeast form adherence to HSV-1- and HSV-2-infected cells was specific for the Candida phenotype tested. Our study suggests that HSV, while antagonist towards S. aureus adherence enhances Candida adherence. Furthermore, the combination of the three pathogens results in S. aureus adherence that is either unaffected, or partially restored depending on both the herpes viral species and the fungal phenotype present.

Ira M. Sigar

Department of Microbiology and Immunology

Midwestern University

Determination of Biofilm Initiation on Virus-infected Cells by Bacteria and Fungi

Anaerobic Growth and Maintenance of Mammalian Cell Lines