DNA Vector-based RNA Interference to Study Gene Function in Cancer

RNA interference (RNAi) inhibits gene expression by specifically degrading target mRNAs. Since the discovery of double-stranded small interference RNA ...

Evaluation of Host-Pathogen Responses and Vaccine Efficacy in Mice

Vaccines are a 20th century medical marvel. They have dramatically reduced the morbidity and mortality caused by infectious diseases and contributed to a ...

Wake Forest University School of Medicine

The Ohio State University

&#x76EE;&#x7684;&#xFF1A; &#x4EE5;&#x524D;&#x7684;&#x5DE5;&#x4F5C;&#x901A;&#x8FC7;&#x6B63;&#x7535;&#x5B50;&#x53D1;&#x5C04;&#x65AD;&#x5C42;&#x626B;&#x63CF; (PET) &#x9886;&#x8FDB;&#x80BF;&#x7624;&#x90E8;&#x4F4D;&#x7684; T &#x7EC6;&#x80DE;&#x8FC1;&#x79FB;&#x7684;&#x5B9A;&#x91CF;&#x6210;&#x50CF;&#x4F7F;&#x7528;&#x7A81;&#x53D8;&#x7684;&#x80F8;&#x82F7;&#x6FC0;&#x9176; (sr39TK) &#x7684;&#x8BB0;&#x8005;&#x57FA;&#x56E0;&#x7684;&#x9006;&#x8F6C;&#x5F55;&#x75C5;&#x6BD2;&#x8F6C;&#x5BFC;&#x514D;&#x75AB; T &#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x3002;&#x7A0B;&#x5E8F;&#x548C;&#x7ED3;&#x679C;&#xFF1A; &#x4E3A;&#x4E86;&#x63D0;&#x9AD8;&#x7075;&#x654F;&#x5EA6;&#x548C;&#x6210;&#x50CF;&#x5206;&#x6790;&#x7684;&#x7075;&#x6D3B;&#x6027;&#xFF0C;&#x6162;&#xFF0C;&#x8868;&#x793A; sr39TK&#xFF0C;&#x88AB;&#x7528;&#x6765;&#x6362;&#x80FD;&#x5668;&#x8FC1;&#x79FB;&#x5230;&#x514D;&#x75AB;&#x539F;&#x6027;&#x8089;&#x7624;&#x7F51;&#x7AD9;&#x7684;&#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x3002;Retrovirally &#x8F6C;&#x5BFC;&#x7684;&#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#xFF0C;&#x4E0E; lentivirally &#x8F6C;&#x5BFC;&#x7684;&#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x663E;&#x793A;&#x589E;&#x5F3A;&#x7684; PET &#x4FE1;&#x53F7;&#x65F6;&#x88AB;&#x8F6C;&#x79FB;&#x7684;&#x8F6C;&#x5BFC;&#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x6570;&#x76EE;&#x76F8;&#x7B49;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x4E3A;&#x4E86;&#x5229;&#x7528;&#x591A;&#x5B66;&#x79D1;&#x7EFC;&#x5408;&#x4F53;&#x5185;&#x6210;&#x50CF;&#x80FD;&#x529B;&#xFF0C;&#x8BB0;&#x8005;&#x4E09;&#x878D;&#x5408;&#x57FA;&#x56E0;&#x5305;&#x542B; sr39TK&#x3001; &#x5408;&#x6210;&#x6D77;&#x80BE;&#x8367;&#x5149;&#x7D20;&#x9176; (hRluc) &#x548C;&#x589E;&#x5F3A;&#x578B;&#x7684;&#x7EFF;&#x8272;&#x8367;&#x5149;&#x86CB;&#x767D; (eGFP) &#x63D2;&#x5165;&#x6162;&#x75C5;&#x6BD2;&#x8F6C;&#x79FB;&#x5230;&#x77E2;&#x91CF;&#x3002;&#x4F7F;&#x7528;&#x8FC7;&#x7EE7;&#x8F6C;&#x8F93;&#x6A21;&#x578B;&#xFF0C;&#x80BF;&#x7624;&#x7279;&#x5F02;&#x6027;&#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x8FC1;&#x79FB; microPET &#x626B;&#x63CF;&#x548C;&#x751F;&#x7269;&#x53D1;&#x5149;&#x6210;&#x50CF;&#x68C0;&#x6D4B;&#x5230;&#x3002;&#x7ED3;&#x8BBA;: &#x5F71;&#x50CF;&#x52A0;&#x4E0A;&#x8FD9;&#x91CC;&#x663E;&#x793A;&#x51FA;&#x7684;&#x6162;&#x75C5;&#x6BD2;&#x8BB0;&#x8005;&#x6784;&#x9020;&#x4EA4;&#x4ED8;&#x7B56;&#x7565;&#x7684;&#x591A;&#x5F0F;&#x8054;&#x8FD0;&#x53EF;&#x4EE5;&#x65B9;&#x4FBF;&#x4ECA;&#x540E;&#x7684;&#x5206;&#x5B50;&#x5F71;&#x50CF;&#x5B66;&#x7814;&#x7A76;&#x3002;

&#x4F7F;&#x7528;&#x57FA;&#x4E8E;&#x6162;&#x75C5;&#x6BD2;&#x7684;&#x4E09;&#x878D;&#x5408;&#x57FA;&#x56E0;&#x8F6C;&#x5BFC;&#x7684;&#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x8FC1;&#x79FB;&#x7684;&#x591A;&#x6A21;&#x6001;&#x6210;&#x50CF;&#x3002;

PURPOSE: Previous work showed quantitative imaging of T-cell migration into a tumor site by positron emission tomography (PET), using retroviral transduction of mutated thymidine kinase (sr39TK) reporter genes into immunized T-lymphocytes. PROCEDURES AND RESULTS: In order to improve the sensitivity and flexibility of the imaging analysis, lentivirus, that expressed sr39TK, was used to transduce the lymphocytes that migrated to an immunogenic sarcoma site. In comparison to retrovirally transduced lymphocytes, the lentivirally transduced lymphocytes showed enhanced PET signal when equal numbers of transduced lymphocytes were transferred. Furthermore, in order to utilize multimodality in vivo imaging capability, a tri-fusion reporter gene containing sr39TK, synthetic Renilla luciferase (hRluc), and enhanced green fluorescent protein (eGFP) was inserted into a lentiviral transfer vector. Using the adoptive transfer model, tumor-specific lymphocytic migration was detected by both microPET scan and bioluminescence imaging. CONCLUSION: The multimodal imaging strategy coupled with lentiviral reporter construct delivery demonstrated here can facilitate future molecular imaging studies.

Multimodality imaging of lymphocytic migration using lentiviral-based transduction of a tri-fusion reporter gene.

BUT : Travaux ant&eacute;rieurs ont montr&eacute; l&#39;imagerie quantitative de la migration des lymphocytes T dans un site de la tumeur par tomographie par &eacute;mission de positrons (PET), &agrave; l&#39;aide de transduction r&eacute;trovirale des g&egrave;nes de rapporteurs mut&eacute; thymidine kinase (sr39TK) en vaccin&eacute;s T-lymphocytes. PROC&Eacute;DURES et r&eacute;sultats : Afin d&#39;am&eacute;liorer la sensibilit&eacute; et la souplesse de l&#39;analyse d&#39;imagerie, lentivirus, qui exprime sr39TK, servait &agrave; transmettre les lymphocytes qui ont migr&eacute; vers un site de sarcome immunog&egrave;ne. Par rapport aux lymphocytes retrovirally transduites, les lymphocytes lentivirally transduites a montr&eacute; signal am&eacute;lior&eacute; de PET lorsqu&#39;un nombre &eacute;gal de lymphocytes transduites ont &eacute;t&eacute; transf&eacute;r&eacute;. En outre, afin de tirer parti de multimodalit&eacute; in vivo imaging capacit&eacute;, un journaliste tri-fusion g&eacute;nique contenant sr39TK et synth&eacute;tique k&ouml;llikeri lucif&eacute;rase (hRluc) am&eacute;lior&eacute; green fluorescent protein (eGFP) a &eacute;t&eacute; ins&eacute;r&eacute; dans un transfert des g&egrave;nes vector. En utilisant le mod&egrave;le de transfert adoptif, tumeur sp&eacute;cifique migration lymphocytaire a &eacute;t&eacute; d&eacute;tect&eacute;e par analyse microPET et imagerie de bioluminescence. CONCLUSION : Le multimodal d&#39;imagerie strat&eacute;gie coupl&eacute; avec des g&egrave;nes reporter construct livraison d&eacute;montr&eacute; ici peut faciliter de futures &eacute;tudes d&#39;imagerie mol&eacute;culaires.

Imagerie multimodalit&eacute; de migration lymphocytaire &agrave; l&#39;aide de transduction ax&eacute;e sur des g&egrave;nes d&#39;un g&egrave;ne rapporteur tri-fusion.

Multimodale Bildgebung von Lymphozyten Migration mit lentiviralen-basierten Transduktion eines Tri-Reportergen-Fusion.

SCOPO: Lavoro precedente ha mostrato quantitativa imaging della migrazione delle cellule T in un sito di tumore di tomografia ad emissione di positroni (PET), utilizzando la trasduzione retrovirale di geni di reporter mutato timidina chinasi (sr39TK) in immunizzati T-linfociti. PROCEDURE e risultati: Al fine di migliorare la sensibilit&agrave; e la flessibilit&agrave; dell&#39;analisi imaging, lentivirus, che ha espresso sr39TK, &egrave; stato utilizzato per trasdurre i linfociti che migrarono in un sito di sarcoma immunogenico. Rispetto a retrovirally trasdotte linfociti, i linfociti lentivirally trasdotte ha mostrato maggiore segnale PET quando vennero trasferito un numero uguale di linfociti trasdotte. Inoltre, al fine di utilizzare la multimodalit&agrave; in vivo imaging capacit&agrave;, un tri-fusione reporter gene contenente sr39TK, sintetico Renilla luciferase (hRluc) e rafforzata green fluorescent protein (eGFP) &egrave; stata inserita in un trasferimento lentivirale vettoriali. Utilizzando il modello di trasferimento adottivo, migrazione linfocitaria tumore-specifico &egrave; stato rilevato da entrambi microPET scansione e imaging bioluminescenza. CONCLUSIONE: Il multimodal imaging strategia accoppiato con reporter lentivirali costrutto consegna dimostrato qui pu&ograve; facilitare futuri studi di imaging molecolari.

Imaging multimodale di migrazione linfocitaria tramite trasduzione lentivirale basato su di un gene reporter tri-fusione.

&#x76EE;&#x7684;: &#x524D;&#x4F5C; T &#x7D30;&#x80DE;&#x904A;&#x8D70;&#x306E;&#x5B9A;&#x91CF;&#x7684;&#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x816B;&#x760D;&#x90E8;&#x4F4D;&#x306B;&#x967D;&#x96FB;&#x5B50;&#x653E;&#x5C04;&#x65AD;&#x5C64;&#x64AE;&#x5F71; (PET) &#x793A;&#x3057;&#x305F;&#x5909;&#x7570;&#x30C1;&#x30DF;&#x30B8;&#x30F3;&#x306E;&#x30AD;&#x30CA;&#x30FC;&#x30BC; (&#x7A81;&#x7136;&#x5909;&#x7570;&#x4F53;&#x5F62;&#x5F0F;) &#x30EC;&#x30DD;&#x30FC;&#x30BF;&#x30FC;&#x907A;&#x4F1D;&#x5B50;&#x3078;&#x306E;&#x30EC;&#x30C8;&#x30ED; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306E;&#x4F1D;&#x9054;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066; T &#x30EA;&#x30F3;&#x30D1;&#x7403;&#x514D;&#x75AB;&#x3002;&#x30D7;&#x30ED;&#x30B7;&#x30FC;&#x30B8;&#x30E3;&#x3068;&#x7D50;&#x679C;: &#x611F;&#x5EA6;&#x3068;&#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x89E3;&#x6790;&#x306E;&#x67D4;&#x8EDF;&#x6027;&#x3092;&#x5411;&#x4E0A;&#x3055;&#x305B;&#x308B;&#x305F;&#x3081;&#x306B;&#x306F;&#x3001;&#x7A81;&#x7136;&#x5909;&#x7570;&#x4F53;&#x5F62;&#x5F0F;&#x8868;&#x73FE;&#x3001;&#x30EC;&#x30F3;&#x30C1; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x3001;&#x514D;&#x75AB;&#x539F;&#x6027;&#x8089;&#x816B;&#x306E;&#x30B5;&#x30A4;&#x30C8;&#x306B;&#x79FB;&#x884C;&#x3059;&#x308B;&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x30D8;&#x30C3;&#x30B8;&#x30DB;&#x30C3;&#x30B0;&#x3057;&#x3066;&#x3044;&#x307E;&#x3057;&#x305F;&#x3002;&#x5F62;&#x8CEA;&#x5C0E;&#x5165;&#x3055;&#x308C;&#x305F;&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x306E;&#x7B49;&#x3057;&#x3044;&#x6570;&#x304C;&#x8EE2;&#x9001;&#x3055;&#x308C;&#x305F;&#x3068;&#x304D;&#x907A;&#x4F1D;&#x5B50;&#x5F62;&#x8CEA;&#x5C0E;&#x5165;&#x3055;&#x308C;&#x305F;&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x3068;&#x6BD4;&#x8F03;&#x3057;&#x3066;&#x5F37;&#x5316;&#x3055;&#x308C;&#x305F;&#x30DA;&#x30C3;&#x30C8; &#x30B7;&#x30B0;&#x30CA;&#x30EB; lentivirally &#x5F62;&#x8CEA;&#x5C0E;&#x5165;&#x3055;&#x308C;&#x305F;&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x3092;&#x793A;&#x3057;&#x305F;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x30DE;&#x30EB;&#x30C1;&#x30E2;&#x30C0;&#x30EA;&#x30C6;&#x30A3;&#x3092;&#x5229;&#x7528;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B; in vivo &#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x6A5F;&#x80FD;&#x3001;&#x30C8;&#x30EA;&#x878D;&#x5408;&#x30EC;&#x30DD;&#x30FC;&#x30BF;&#x30FC;&#x907A;&#x4F1D;&#x5B50;&#x3092;&#x542B;&#x3080;&#x7A81;&#x7136;&#x5909;&#x7570;&#x4F53;&#x5F62;&#x5F0F;&#x3001;&#x5408;&#x6210; Renilla &#x30EB;&#x30B7;&#x30D5;&#x30A7;&#x30E9;&#x30FC;&#x30BC; (hRluc)&#x3001;&#x5F37;&#x5316;&#x7DD1;&#x8272;&#x86CD;&#x5149;&#x30BF;&#x30F3;&#x30D1;&#x30AF;&#x8CEA; (eGFP) &#x30EC;&#x30F3;&#x30C1; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x8EE2;&#x9001;&#x306B;&#x633F;&#x5165;&#x3055;&#x308C;&#x305F;&#x30D9;&#x30AF;&#x30C8;&#x30EB;&#x3057;&#x307E;&#x3059;&#x3002;&#x990A;&#x5B50;&#x8EE2;&#x9001;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x3001;&#x816B;&#x760D;&#x56FA;&#x6709;&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x306E;&#x79FB;&#x884C; microPET &#x30B9;&#x30AD;&#x30E3;&#x30F3;&#x3068;&#x751F;&#x7269;&#x767A;&#x5149;&#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x306B;&#x3088;&#x3063;&#x3066;&#x691C;&#x51FA;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x7D50;&#x8AD6;: &#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x6226;&#x7565;&#x3053;&#x3053;&#x306B;&#x793A;&#x3057;&#x305F;&#x30EC;&#x30F3;&#x30C1; &#x30A6;&#x30A4;&#x30EB;&#x30B9; &#x30EC;&#x30DD;&#x30FC;&#x30BF;&#x30FC; &#x30B3;&#x30F3;&#x30B9;&#x30C8;&#x30E9;&#x30AF;&#x30C8;&#x914D;&#x4FE1;&#x3068;&#x76F8;&#x307E;&#x3063;&#x3066;&#x30DE;&#x30EB;&#x30C1; &#x30E2;&#x30FC;&#x30C0;&#x30EB;&#x5C06;&#x6765;&#x306E;&#x5206;&#x5B50;&#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x7814;&#x7A76;&#x3092;&#x4FC3;&#x9032;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;

&#x30EC;&#x30F3;&#x30C1; &#x30A6;&#x30A4;&#x30EB;&#x30B9; &#x30D9;&#x30FC;&#x30B9;&#x4FE1;&#x53F7;&#x5909;&#x63DB;&#x30C8;&#x30EA;&#x878D;&#x5408;&#x30EC;&#x30DD;&#x30FC;&#x30BF;&#x30FC;&#x907A;&#x4F1D;&#x5B50;&#x3092;&#x7528;&#x3044;&#x305F;&#x30EA;&#x30F3;&#x30D1;&#x7403;&#x306E;&#x79FB;&#x884C;&#x306E;&#x30DE;&#x30EB;&#x30C1;&#x30E2;&#x30C0;&#x30EA;&#x30C6;&#x30A3; &#x30A4;&#x30E1;&#x30FC;&#x30B8;&#x30F3;&#x30B0;&#x3002;

Prostate stem cell antigen (PSCA) is expressed in normal epithelium of various tissues, in embryos and adult animals. PSCA expression is upregulated in up to 70% of prostate tumors and metastases, and a subset of bladder and pancreatic cancers. However, its function is unknown. We studied the effect of targeted gene deletion of PSCA on normal organ development and prostate carcinogenesis.

Deletion of PSCA increases metastasis of TRAMP-induced prostate tumors without altering primary tumor formation.

Antig&#xE8;ne de cellule souche prostatique (PSCA) est exprim&#xE9;e dans l&#x26;#39;&#xE9;pith&#xE9;lium normal des tissus divers, des embryons et des animaux adultes. L&#x26;#39;expression est r&#xE9;gul&#xE9;e positivement dans PSCA jusqu&#x26;#39;&#xE0; 70% des tumeurs de la prostate et les m&#xE9;tastases, et un sous-ensemble de la vessie et du pancr&#xE9;as. Cependant, sa fonction est inconnue. Nous avons &#xE9;tudi&#xE9; l&#x26;#39;effet de la d&#xE9;l&#xE9;tion du g&#xE8;ne cibl&#xE9; de PSCA sur le d&#xE9;veloppement normal des organes et de la prostate canc&#xE9;rogen&#xE8;se.

Suppression de PSCA Augmente la m&#xE9;tastase des tumeurs de la prostate TRAMP-induites Sans modifier Formation tumeur primaire

Prostata Stammzell-Antigen (PSCA) dr&uuml;ckt sich in normalen Epithel der verschiedenen Gewebe, Embryonen und Erwachsene Tiere. PSCA Ausdruck ist herraufreguliert in bis zu 70 % der Prostatatumoren und Metastasen und eine Teilmenge der Blase und Pankreaskrebs. Seine Funktion ist jedoch unbekannt. Wir studierten die Wirkung der gezielten gen L&ouml;schung von PSCA f&uuml;r normale Orgel Entwicklung und Prostata Karzinogenese.

Streichung des PSCA erh&ouml;ht die Metastasierung von TRAMP-induzierte Prostatatumoren ohne Ver&auml;nderung der prim&auml;ren Tumorbildung.

Antigene prostata cellule staminali (PSCA) &egrave; espresso nell&#39;epitelio normale dei vari tessuti, in embrioni e in animali adulti. PSCA espressione &egrave; sovraregolati fino al 70% dei tumori della prostata e metastasi, e un sottoinsieme della vescica e tumori pancreatici. Tuttavia, la sua funzione &egrave; sconosciuta. Abbiamo studiato l&#39;effetto della delezione di un gene mirati di PSCA per lo sviluppo normale organo e carcinogenesi della prostata.

Eliminazione di PSCA aumenta la metastasi dei tumori della prostata TRAMP-indotta senza alterare la formazione del tumore primario.

&#x524D;&#x5217;&#x817A;&#x5E72;&#x7EC6;&#x80DE;&#x6297;&#x539F; (&#x524D;&#x5217;&#x817A;&#x5E72;&#x7EC6;&#x80DE;&#x6297;&#x539F;) &#x8868;&#x793A;&#x6B63;&#x5E38;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x5728;&#x80DA;&#x80CE;&#x548C;&#x6210;&#x5E74;&#x52A8;&#x7269;&#x7684;&#x5404;&#x79CD;&#x7EC4;&#x7EC7;&#x3002;&#x524D;&#x5217;&#x817A;&#x5E72;&#x7EC6;&#x80DE;&#x6297;&#x539F;&#x8868;&#x8FBE;&#x662F;&#x589E;&#x9AD8;&#x8FBE; 70%&#x7684;&#x524D;&#x5217;&#x817A;&#x80BF;&#x7624;&#x548C;&#x8F6C;&#x79FB;&#xFF0C;&#x548C;&#x8180;&#x80F1;&#x548C;&#x80F0;&#x817A;&#x764C;&#x7684;&#x4E00;&#x4E2A;&#x5B50;&#x96C6;&#x3002;&#x4F46;&#x662F;&#xFF0C;&#x5176;&#x529F;&#x80FD;&#x662F;&#x672A;&#x77E5;&#x7684;&#x3002;&#x6211;&#x4EEC;&#x7814;&#x7A76;&#x4E86;&#x6B63;&#x5E38;&#x5668;&#x5B98;&#x53D1;&#x80B2;&#x548C;&#x524D;&#x5217;&#x817A;&#x764C;&#x53D8;&#x524D;&#x5217;&#x817A;&#x5E72;&#x7EC6;&#x80DE;&#x6297;&#x539F;&#x7684;&#x9776;&#x5411;&#x6027;&#x7684;&#x57FA;&#x56E0;&#x5220;&#x9664;&#x7684;&#x5F71;&#x54CD;&#x3002;

&#x524D;&#x5217;&#x817A;&#x5E72;&#x7EC6;&#x80DE;&#x6297;&#x539F;&#x7684;&#x5220;&#x9664;&#x800C;&#x4E0D;&#x4F1A;&#x6539;&#x53D8;&#x539F;&#x53D1;&#x6027;&#x80BF;&#x7624;&#x5F62;&#x6210;&#x589E;&#x52A0;&#x6D41;&#x6D6A;&#x6C49;&#x81F4;&#x524D;&#x5217;&#x817A;&#x80BF;&#x7624;&#x8F6C;&#x79FB;&#x3002;

&#x524D;&#x7ACB;&#x817A;&#x5E79;&#x7D30;&#x80DE;&#x6297;&#x539F;&#xFF08;PSCA&#xFF09;&#x306F;&#x3001;&#x80DA;&#x304A;&#x3088;&#x3073;&#x6210;&#x4F53;&#x52D5;&#x7269;&#x306B;&#x304A;&#x3044;&#x3066;&#x3001;&#x7A2E;&#x3005;&#x306E;&#x7D44;&#x7E54;&#x306E;&#x6B63;&#x5E38;&#x4E0A;&#x76AE;&#x306B;&#x767A;&#x73FE;&#x3057;&#x3066;&#x3044;&#x308B;&#x3002; PSCA&#x306E;&#x767A;&#x73FE;&#x306F;70&#x524D;&#x7ACB;&#x817A;&#x816B;&#x760D;&#x3068;&#x8EE2;&#x79FB;&#x306E;&#xFF05;&#x3067;&#x3042;&#x308A;&#x3001;&#x8180;&#x80F1;&#x3001;&#x81B5;&#x81D3;&#x764C;&#x306E;&#x30B5;&#x30D6;&#x30BB;&#x30C3;&#x30C8;&#x306B;&#x306B;&#x30A2;&#x30C3;&#x30D7;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30C8;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3057;&#x304B;&#x3057;&#x3001;&#x305D;&#x306E;&#x6A5F;&#x80FD;&#x306F;&#x4E0D;&#x660E;&#x3067;&#x3042;&#x308B;&#x3002;&#x6211;&#x3005;&#x306F;&#x6B63;&#x5E38;&#x306A;&#x81D3;&#x5668;&#x306E;&#x958B;&#x767A;&#x3068;&#x524D;&#x7ACB;&#x817A;&#x767A;&#x764C;&#x306B;&#x304A;&#x3051;&#x308B;PSCA&#x306E;&#x6A19;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x6B20;&#x5931;&#x306E;&#x52B9;&#x679C;&#x3092;&#x691C;&#x8A0E;&#x3057;&#x305F;&#x3002;

PSCA&#x306E;&#x524A;&#x9664;&#x306F;&#x3001;&#x539F;&#x767A;&#x816B;&#x760D;&#x306E;&#x5F62;&#x6210;&#x3092;&#x5909;&#x66F4;&#x305B;&#x305A;&#x306B;TRAMP&#x8A98;&#x767A;&#x6027;&#x524D;&#x7ACB;&#x817A;&#x816B;&#x760D;&#x306E;&#x8EE2;&#x79FB;&#x3092;&#x5897;&#x52A0;&#x3055;&#x305B;&#x308B;

&#xC804;&#xB9BD;&#xC120; &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xD56D; &#xC6D0; (PSCA) &#xD0DC;&#xC544; &#xBC0F; &#xC131;&#xC778; &#xB3D9;&#xBB3C;&#xC5D0;&#xC11C; &#xB2E4;&#xC591; &#xD55C; &#xC870;&#xC9C1;&#xC758; &#xC815;&#xC0C1;&#xC801;&#xC778; &#xC0C1;&#xD53C;&#xB85C; &#xD45C;&#xC2DC; &#xB429;&#xB2C8;&#xB2E4;. PSCA &#xC2DD; &#xCD5C;&#xB300; 70% &#xC804;&#xB9BD;&#xC120; &#xC885;&#xC591; &#xBC0F; &#xC804;&#xC774;, &#xADF8;&#xB9AC;&#xACE0; &#xBC29;&#xAD11;&#xACFC; &#xCDCC;&#xC7A5;&#xC554;&#xC758; &#xD558;&#xC704; &#xC9D1;&#xD569;&#xC5D0; &#xC788;&#xB294; upregulated&#xC785;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xADF8; &#xAE30;&#xB2A5;&#xC744; &#xC54C; &#xC218; &#xC788;&#xB2E4;. &#xC6B0;&#xB9AC; &#xC77C;&#xBC18; &#xAE30;&#xAD00; &#xAC1C;&#xBC1C; &#xBC0F; &#xC804;&#xB9BD;&#xC120; &#xBC1C;&#xC554; PSCA&#xC758; &#xD0C0;&#xAC9F;&#xB41C; &#xC720;&#xC804;&#xC790; &#xC0AD;&#xC81C;&#xC758; &#xD6A8;&#xACFC; &#xACF5;&#xBD80; &#xD588;&#xC2B5;&#xB2C8;&#xB2E4;.

PSCA&#xC758; &#xC0AD;&#xC81C; &#xAE30;&#xBCF8; &#xC885;&#xC591;&#xC758; &#xD615;&#xC131;&#xC744; &#xBCC0;&#xACBD; &#xD558;&#xC9C0; &#xC54A;&#xACE0; &#xBC14;&#xB78C;&#xB465;&#xC774; &#xC720;&#xB3C4; &#xC804;&#xB9BD;&#xC120; &#xC885;&#xC591;&#xC758; &#xC804;&#xC774; &#xC99D;&#xAC00; &#xD569;&#xB2C8;&#xB2E4;.

Pr&#xF3;stata ant&#xED;geno de c&#xE9;lulas madre (LCSP) se expresa en el epitelio normal de diversos tejidos, en los embriones y animales adultos. Expresi&#xF3;n de PSCA es upregulated en hasta un 70% de los tumores de pr&#xF3;stata y met&#xE1;stasis, y un subconjunto de la vejiga y el c&#xE1;ncer de p&#xE1;ncreas. Sin embargo, su funci&#xF3;n es desconocida. Se estudi&#xF3; el efecto de la deleci&#xF3;n del gen espec&#xED;fico de la LCSP en el desarrollo normal del organismo y de la carcinog&#xE9;nesis de pr&#xF3;stata.

Supresi&#xF3;n de la LCSP Aumenta la met&#xE1;stasis de los tumores de pr&#xF3;stata TRAMP inducidos Sin alterando la formaci&#xF3;n de tumor primario

Bordetella colonization factor A (BcfA) is an outer membrane immunogenic protein, which is critical for efficient colonization of the murine respiratory tract. These properties of BcfA prompted us to examine its utility in inducing a protective immune response against Bordetella bronchiseptica in a mouse model of intranasal infection. Mice vaccinated with BcfA demonstrated reduced pathology in the lungs and harbored lower bacterial burdens in the respiratory tract. Immunization with BcfA led to the generation of BcfA-specific antibodies in both the sera and lungs, and passive immunization led to the reduction of B. bronchiseptica in the tracheas and lungs. These results suggest that protection after immunization with BcfA is mediated in part by antibodies against BcfA. To further investigate the mechanism of BcfA-induced immune clearance, we examined the role of neutrophils and macrophages. Our results demonstrate that neutrophils are critical for anti-BcfA antibody-mediated clearance and that opsonization with anti-BcfA serum enhances phagocytosis of B. bronchiseptica by murine macrophages. We show that immunization with BcfA results in the production of gamma interferon and subclasses of immunoglobulin G antibodies that are consistent with the induction of a Th1-type immune response. In combination, our findings suggest that the mechanism of BcfA-mediated immunity involves humoral and cellular responses. Expression of BcfA is conserved among multiple clinical isolates of B. bronchiseptica. Our results demonstrate the striking protective efficacy of BcfA-mediated immunization, thereby highlighting its utility as a potential vaccine candidate. These results also provide a model for the development of cell-free vaccines against B. bronchiseptica.

Active and passive immunizations with Bordetella colonization factor A protect mice against respiratory challenge with Bordetella bronchiseptica.

Bordetella pertussis is the causative agent of pertussis, a highly contagious disease of the human respiratory tract. Despite very high vaccine coverage, pertussis has reemerged as a serious threat in the United States and many developing countries. Thus, it is important to pursue research to discover unknown pathogenic mechanisms of B. pertussis. We have investigated a previously uncharacterized locus in B. pertussis, the dra locus, which is homologous to the dlt operons of Gram-positive bacteria. The absence of the dra locus resulted in increased sensitivity to the killing action of antimicrobial peptides (AMPs) and human phagocytes. Compared to the wild-type cells, the mutant cells bound higher levels of cationic proteins and peptides, suggesting that dra contributes to AMP resistance by decreasing the electronegativity of the cell surface. The presence of dra led to the incorporation of d-alanine into an outer membrane component that is susceptible to proteinase K cleavage. We conclude that dra encodes a virulence-associated determinant and contributes to the immune resistance of B. pertussis. With these findings, we have identified a new mechanism of surface modification in B. pertussis which may also be relevant in other Gram-negative pathogens. 

D-alanine modification of a protease-susceptible outer membrane component by the Bordetella pertussis dra locus promotes resistance to antimicrobial peptides and polymorphonuclear leukocyte-mediated killing.

Pertussis, or whooping cough, caused by the obligate human pathogen  is undergoing a worldwide resurgence. The majority of studies of this pathogen are conducted with laboratory-adapted strains which may not be representative of the species as a whole. Biofilm formation by  plays an important role in pathogenesis. We conducted a side-by-side comparison of the biofilm-forming abilities of the prototype laboratory strains and the currently circulating isolates from two countries with different vaccination programs. Compared to the reference strain, all strains examined herein formed biofilms at high levels. Biofilm structural analyses revealed country-specific differences, with strains from the United States forming more structured biofilms. Bacterial hyperaggregation and reciprocal expression of biofilm-promoting and -inhibitory factors were observed in clinical isolates. An association of increased biofilm formation with augmented epithelial cell adhesion and higher levels of bacterial colonization in the mouse nose and trachea was detected. To our knowledge, this work links for the first time increased biofilm formation in bacteria with a colonization advantage in an animal model. We propose that the enhanced biofilm-forming capacity of currently circulating strains contributes to their persistence, transmission, and continued circulation.

Hyperbiofilm Formation by Bordetella pertussis Strains Correlates with Enhanced Virulence Traits.

The reemergence of pertussis or whooping cough in several countries highlights the need for better vaccines. Acellular pertussis vaccines (aPV) contain alum as the adjuvant and elicit Th2 biased immune responses that are less effective in protecting against infection than the reactogenic whole cell pertussis vaccines (wPV), which elicit primarily a Th1/Th17 response. An important goal for the field is to devise aPVs that will induce immune responses similar to wPV. We show that Bordetella Colonization Factor A (BcfA), an outer membrane protein from  has strong adjuvant function and elicits cellular and humoral immune responses to heterologous and  antigens. Addition of BcfA to a commercial aPV resulted in greater reduction of  numbers from the lungs than elicited by aPV alone. More efficient pathogen clearance was accompanied by increased IL-17 and reduced IL-5, and an increased ratio of IgG2/IgG1 antibodies. Thus our results suggest that BcfA improves aPV induced responses by modifying the alum-induced Th2-biased aPV response towards Th1/Th17. A re-designed aPV containing BcfA may allow better control of pertussis reemergence by reshaping immune responses to resemble those elicited by wPV immunization.

The adjuvant Bordetella Colonization Factor A attenuates alum-induced Th2 responses and enhances  clearance from mouse lungs.

Reporter gene-based molecular imaging is a powerful means to detect the movement and function of diverse cell populations in vivo. Reconstitution of an immune system marked with molecular imaging reporter genes permits tracking of primary immune responses to pathogens and cancer in experimental systems. This chapter describes the methods to isolate bone marrow stem/progenitors, transduce them with imaging reporter genes, and track the reconstitution of the peripheral immune system by bioluminescent imaging.

Purnima Dubey

Department of Pathology and Comprehensive Cancer Center

Wake Forest University School of Medicine

DNA Vector-based RNA Interference to Study Gene Function in Cancer

Evaluation of Host-Pathogen Responses and Vaccine Efficacy in Mice

Purnima Dubey

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Pathology and Comprehensive Cancer Center

Wake Forest University School of Medicine

DNA Vector-based RNA Interference to Study Gene Function in Cancer

Evaluation of Host-Pathogen Responses and Vaccine Efficacy in Mice

Department of Pathology and Comprehensive Cancer Center