a-choroid-plexus-epithelial-cell-based-model-human-blood

A Choroid Plexus Epithelial Cell-based Model of the Human Blood-Cerebrospinal Fluid Barrier to Study Bacterial Infection from the Basolateral Side

The epithelial cells of the choroid plexus (CP), located in the ventricular system of the brain, form the blood-cerebrospinal fluid barrier (BCSFB). The ...

Heidelberg University

Knowing the inherent stimulatory properties of the lipid moiety of bacterial lipoproteins, we first hypothesized that Brucella abortus outer membrane protein (Omp)16 lipoprotein would be able to elicit a protective immune response without the need of external adjuvants. In this study, we demonstrate that Omp16 administered by the i.p. route confers significant protection against B. abortus infection and that the protective response evoked is independent of the protein lipidation. To date, Omp16 is the first Brucella protein that without the requirement of external adjuvants is able to induce similar protection levels to the control live vaccine S19. Moreover, the protein portion of Omp16 (unlipidated Omp16 [U-Omp16]) elicits a protective response when administered by the oral route. Either systemic or oral immunization with U-Omp16 elicits a Th1-specific response. These abilities of U-Omp16 indicate that it is endowed with self-adjuvanting properties. The adjuvanticity of U-Omp16 could be explained, at least in part, by its capacity to activate dendritic cells in vivo. U-Omp16 is also able to stimulate dendritic cells and macrophages in vitro. The latter property and its ability to induce a protective Th1 immune response against B. abortus infection have been found to be TLR4 dependent. The facts that U-Omp16 is an oral protective Ag and possesses a mucosal self-adjuvanting property led us to develop a plant-made vaccine expressing U-Omp16. Our results indicate that plant-expressed recombinant U-Omp16 is able to confer protective immunity, when given orally, indicating that a plant-based oral vaccine expressing U-Omp16 could be a valuable approach to controlling this disease.

The protein moiety of Brucella abortus outer membrane protein 16 is a new bacterial pathogen-associated molecular pattern that activates dendritic cells in vivo, induces a Th1 immune response, and is a promising self-adjuvanting vaccine against systemic and oral acquired brucellosis.

The Gram-positive zoonotic bacterium Streptococcus suis (S. suis) is responsible for a wide range of diseases including meningitis in pigs and humans. The blood-cerebrospinal fluid (CSF) barrier is constituted by the epithelial cells of the choroid plexus, which execute barrier function also after bacteria have entered the central nervous system (CNS). We show that the bacterial capsule, a major virulence factor, strongly attenuates adhesion of S. suis to the apical side of porcine choroid plexus epithelial cells (PCPEC). Oligonucleotide microarray analysis and quantitative PCR surprisingly demonstrated that adherent wild-type and capsule-deficient S. suis influenced expression of a pronounced similar pattern of genes in PCPEC. Investigation of purified capsular material provided no evidence for a significant role of the capsule. Enriched among the regulated genes were those involved in "inflammatory response", "defense response" and "cytokine activity". These comprised several cytokines and chemokines including the interleukins 6 and 8, which could be detected on protein level. We show that after infection with S. suis the choroid plexus contributes to the immune response by actively producing cytokines and chemokines. Other virulence factors than the bacterial capsule may be relevant in inducing a strong inflammatory response in the CNS during S. suis meningitis.

In vitro transcriptome analysis of porcine choroid plexus epithelial cells in response to Streptococcus suis: release of pro-inflammatory cytokines and chemokines.

Acute bacterial meningitis is a life-threatening disease in humans. Discussed as entry sites for pathogens into the brain are the blood-brain and the blood-cerebrospinal fluid barrier (BCSFB). Although human brain microvascular endothelial cells (HBMEC) constitute a well established human in vitro model for the blood-brain barrier, until now no reliable human system presenting the BCSFB has been developed. Here, we describe for the first time a functional human BCSFB model based on human choroid plexus papilloma cells (HIBCPP), which display typical hallmarks of a BCSFB as the expression of junctional proteins and formation of tight junctions, a high electrical resistance and minimal levels of macromolecular flux when grown on transwell filters. Importantly, when challenged with the zoonotic pathogen Streptococcus suis or the human pathogenic bacterium Neisseria meningitidis the HIBCPP show polar bacterial invasion only from the physiologically relevant basolateral side. Meningococcal invasion is attenuated by the presence of a capsule and translocated N. meningitidis form microcolonies on the apical side of HIBCPP opposite of sites of entry. As a functionally relevant human model of the BCSFB the HIBCPP offer a wide range of options for analysis of disease-related mechanisms at the choroid plexus epithelium, especially involving human pathogens.

Polar invasion and translocation of Neisseria meningitidis and Streptococcus suis in a novel human model of the blood-cerebrospinal fluid barrier.

&#x80A0;&#x75C5;&#x6BD2;&#x662F;&#x6700;&#x5E38;&#x89C1;&#x75C5;&#x539F;&#x83CC;&#x75C5;&#x6BD2;&#x6027;&#x8111;&#x819C;&#x708E;&#x5C24;&#x5176;&#x662F;&#x5728;&#x513F;&#x7AE5;&#x4E2D;&#x3002;&#x9664;&#x4E86;&#x8840;&#x8111;&#x5C4F;&#x969C; (BBB) &#x7684;&#x8109;&#x7EDC;&#x4E1B;&#xFF0C;&#x5F62;&#x6210;&#x8840;&#x6DB2;&#x8111;&#x810A;&#x6DB2; (CSF) &#x969C;&#x788D; (BCSFB)&#xFF0C;&#x8868;&#x660E;&#x662F;&#x53C2;&#x4E0E;&#x7684;&#x80A0;&#x75C5;&#x6BD2;&#x8111;&#x819C;&#x708E;&#x53D1;&#x75C5;&#x673A;&#x5236;&#x3002;&#x5728;&#x4EBA;&#x7C7B;&#x7684;&#x4F53;&#x5916;&#x6A21;&#x578B;&#x7EC4;&#x6210;&#x7684;&#x4EBA;&#x4F53;&#x8109;&#x7EDC;&#x4E1B;&#x4E73;&#x5934;&#x72B6;&#x7624;&#x7EC6;&#x80DE; (HIBCPP) &#x7684; BCSFB &#x4E2D;&#xFF0C;&#x653E;&#x7EB5; Echovirus 30 (EV30) &#x4E1B;&#x4E0A;&#x76AE;&#x7EC6;&#x80DE;&#x662F;&#x901A;&#x8FC7;&#x514D;&#x75AB;&#x5370;&#x8FF9;&#x6CD5;&#x548C;&#x5B9A;&#x91CF;&#x5B9E;&#x65F6;&#x805A;&#x5408;&#x9176;&#x94FE;&#x53CD;&#x5E94; (PCR) &#x8FDB;&#x884C;&#x5206;&#x6790;&#x3002;&#x4ECE;&#x6839;&#x5C16;&#x548C;&#x751F;&#x7406;&#x6709;&#x5173;&#x57FA;&#x5E95;&#x5916;&#x4FA7;&#x7AEF;&#xFF0C;&#x53EF;&#x4EE5;&#x76F4;&#x63A5;&#x611F;&#x67D3;&#x7684; EV30 HIBCPP&#x3002;5 H &#x611F;&#x67D3;&#x671F;&#x95F4;&#x53EF;&#x80FD;&#x89C2;&#x5BDF;&#x5230;&#x7684;&#x5C4F;&#x969C;&#x529F;&#x80FD;&#x548C;&#x7EC6;&#x80DE;&#x6D3B;&#x529B;&#x6CA1;&#x6709;&#x53D8;&#x5316;&#x3002;&#x4E4B;&#x540E;&#x5F0F;&#x5FAE; (CBA) &#x548C; PCR &#x7684;&#x80A0;&#x75C5;&#x6BD2;&#x611F;&#x67D3;&#x7684;&#x7EC6;&#x80DE;&#x56E0;&#x5B50;/&#x8D8B;&#x5316;&#x56E0;&#x5B50;-&#x914D;&#x7F6E;&#x7684;&#x5206;&#x6790;&#x8868;&#x660E;&#x589E;&#x5F3A;&#x7684;&#x5206;&#x6CCC;&#x7684; PanGRO &#xFF08;CXCL1&#x3001; CXCL2 &#x548C; CXCL3&#xFF09;&#xFF0C;IL8 &#x548C; CCL5&#x3002;Q-&#x805A;&#x5408;&#x9176;&#x94FE;&#x53CD;&#x5E94;&#x65F6;&#x95F4;&#x53D7;&#x517B;&#x4EBA;&#x7684;&#x65B9;&#x5F0F;&#x663E;&#x793A; CXCL1&#x3001; CXCL2 &#x548C; CXCL3 &#x7684;&#x663E;&#x8457;&#x7684;&#x4E0A;&#x8C03;&#x3002;&#x4E0D;&#x8FC7;&#xFF0C;&#x6709;&#x53EA;&#x8F83;&#x5C0F;&#x7684;&#x5F71;&#x54CD;&#x7684; EV30 HIBCPP &#x611F;&#x67D3; &mdash; T &#x6DCB;&#x5DF4;&#x7EC6;&#x80DE;&#x8FC1;&#x79FB;&#x6216;&#x4E0D;&#x8D8B; CXCL12 &#x8DE8;&#x4E0A;&#x76AE;&#x4E0A;&#x3002;&#x6B64;&#x5916;&#xFF0C;CXCL3 &#x4E0D;&#x663E;&#x8457;&#x63D0;&#x9AD8; T &#x7EC6;&#x80DE;&#x7684;&#x8FC1;&#x79FB;&#x3002;&#x56E0;&#x6B64;&#x5FC5;&#x987B;&#x4F53;&#x5185;&#x62A5;&#x544A;&#x589E;&#x5F3A;&#x7684; T &#x7EC6;&#x80DE;&#x8FC1;&#x79FB;&#x5230;&#x4E2D;&#x67A2;&#x795E;&#x7ECF;&#x7CFB;&#x7EDF;&#x4E0A;&#x4E0B;&#x6587;&#x4E2D;&#x7684;&#x80A0;&#x75C5;&#x6BD2;&#x8111;&#x819C;&#x708E;&#x6D89;&#x53CA;&#x5176;&#x4ED6;&#x56E0;&#x7D20;&#x3002;&#x6B63;&#x5982; HIBCPP &#x6240;&#x7EB5;&#x5BB9; EV30 &#x611F;&#x67D3;&#xFF0C;&#x5B83;&#x4EEC;&#x6784;&#x6210;&#x5B9D;&#x8D35;&#x7684;&#x4EBA;&#x7C7B;&#x4F53;&#x5916;&#x6A21;&#x578B;&#xFF0C;&#x7814;&#x7A76;&#x5728; BCSFB &#x7684;&#x75C5;&#x6BD2;&#x6027;&#x611F;&#x67D3;&#x3002;

T &#x7EC6;&#x80DE;&#x8D8B;&#x5316;&#x540E;&#x8840;-&#x8111;&#x810A;&#x6DB2;&#x5C4F;&#x969C;&#x4F53;&#x5916;&#x6A21;&#x578B;&#x4E0E; Echovirus 30 &#x4EBA;&#x4F53;&#x8109;&#x7EDC;&#x4E1B;&#x4E73;&#x5934;&#x72B6;&#x7624;&#x7EC6;&#x80DE;&#x7684;&#x611F;&#x67D3;&#x3002;

Enterovirus is the most common pathogen causing viral meningitis especially in children. Besides the blood-brain barrier (BBB) the choroid plexus, which forms the blood-cerebrospinal-fluid (CSF) barrier (BCSFB), was shown to be involved in the pathogenesis of enteroviral meningitis. In a human in vitro model of the BCSFB consisting of human choroid plexus papilloma cells (HIBCPP), the permissiveness of plexus epithelial cells for Echovirus 30 (EV30) was analyzed by immunoblotting and quantitative real-time PCR (Q-PCR). HIBCPP could be directly infected by EV30 from the apical as well as from the physiological relevant basolateral side. During an infection period of 5h no alterations of barrier function and cell viability could be observed. Analysis of the cytokine/chemokine-profile following enteroviral infection with a cytometric bead array (CBA) and Q-PCR revealed an enhanced secretion of PanGRO (CXCL1, CXCL2 and CXCL3), IL8 and CCL5. Q-PCR showed a significant upregulation of CXCL1, CXCL2 and CXCL3 in a time dependant manner. However, there was only a minor effect of HIBCPP-infection with EV30 on transepithelial T lymphocyte migration with or without the chemoattractant CXCL12. Moreover, CXCL3 did not significantly enhance T cell migrations. Therefore additional factors must be involved for the in vivo reported enhanced T cell migration into the CNS in the context of enteroviral meningitis. As HIBCPP are permissive for infection with EV30, they constitute a valuable human in vitro model to study viral infection at the BCSFB.

Chemotaxis of T-cells after infection of human choroid plexus papilloma cells with Echovirus 30 in an in vitro model of the blood-cerebrospinal fluid barrier.

Ent&eacute;rovirus est le plus commun pathog&egrave;ne causant une m&eacute;ningite virale en particulier chez les enfants. Sans compter que la sang barri&egrave;re - enc&eacute;phalique (BHE) les plexus choro&iuml;des, qui forme le mur du sang-fluide c&eacute;phalo-rachidien (CSF) (BCSFB), s&#39;est av&eacute;r&eacute; &ecirc;tre impliqu&eacute;s dans la pathogen&egrave;se de la m&eacute;ningite &agrave; ent&eacute;rovirus. Dans un mod&egrave;le in vitro humain de la BCSFB compos&eacute;e de cellules de papillome des plexus choro&iuml;des humaine (HIBCPP), la permissivit&eacute; des cellules &eacute;pith&eacute;liales du plexus pour &eacute;chovirus 30 (EV30) a &eacute;t&eacute; analys&eacute;e par immunotransfert et quantitative PCR en temps r&eacute;el (Q-PCR). HIBCPP pourrait s&#39;infecter directement par EV30 de l&#39;apicale, ainsi que du c&ocirc;t&eacute; basolat&eacute;ral pertinentes physiologique. Pendant une p&eacute;riode d&#39;infection de 5 h aucune alt&eacute;ration de viabilit&eacute; cellulaire et de la fonction de barri&egrave;re pourrait &ecirc;tre observ&eacute;e. Analyse de la cytokine/chemokine-profil suite &agrave; une infection ent&eacute;rovirale avec un tableau de Perle en cytom&eacute;trie de flux (CBA) et Q-PCR a r&eacute;v&eacute;l&eacute; une s&eacute;cr&eacute;tion accrue de PanGRO (CXCL1, CXCL2 et CXCL3), IL8 et CCL5. Q-PCR a montr&eacute; une importante upregulation of CXCL1, CXCL2 et CXCL3 d&#39;une mani&egrave;re d&eacute;pendante de temps. Toutefois, il n&#39;y n&#39;avait qu&#39;un effet mineur de HIBCPP-Infection&agrave; EV30 sur trans&eacute;pith&eacute;lial migration des lymphocytes T avec ou sans le facteur chimiotactique CXCL12. En outre, CXCL3 n&#39;augmente pas significativement les migrations des lymphocytes T. C&#39;est pourquoi les facteurs suppl&eacute;mentaires doivent &ecirc;tre impliqu&eacute;es pour la migration in vivo des lymphocytes T am&eacute;lior&eacute; signal&eacute;s dans le SNC dans le contexte de m&eacute;ningite &agrave; ent&eacute;rovirus. HIBCPP sont permissifs pour Infection&agrave; EV30, elles constituent un pr&eacute;cieux mod&egrave;le in vitro humain pour &eacute;tudier une infection virale &agrave; le BCSFB.

Chimiotactisme des cellules T apr&egrave;s l&#39;infection de cellules de papillome des plexus choro&iuml;des humaine avec &eacute;chovirus 30 dans un mod&egrave;le in vitro de la barri&egrave;re sang-c&eacute;phalorachidienne de fluide.

Enterovirus es el pat&oacute;geno m&aacute;s com&uacute;n que causa meningitis viral especialmente en los ni&ntilde;os. Adem&aacute;s de la barrera blood - brain (BBB) el plexo coroideo, que forma la barrera (CSF) de sangre-cerebrospinal fluid (BCSFB), mostr&oacute; ser implicados en la patogenia de la meningitis enteroviral. En un modelo humano in vitro de la BCSFB consiste en c&eacute;lulas de papiloma del plexo coroideo humana (HIBCPP), la permisividad de las c&eacute;lulas epiteliales del plexo por Echovirus 30 (EV30) fue analizada por immunoblotting y PCR cuantitativa a tiempo real (Q-PCR). HIBCPP podr&iacute;a estar infectado directamente por EV30 de la apical, as&iacute; como de la parte fisiol&oacute;gica basolateral pertinentes. Durante un per&iacute;odo de infecci&oacute;n de 5 h no pudieran observarse alteraciones de la viabilidad de c&eacute;lula y la funci&oacute;n de barrera. An&aacute;lisis del citoquina/chemokine-perfil despu&eacute;s de la infecci&oacute;n enteroviral con una matriz de grano cytometric (CBA) y Q-PCR revel&oacute; una mayor secreci&oacute;n de PanGRO (CXCL1, CXCL2 y CXCL3), IL8 y CCL5. Q-PCR mostr&oacute; una upregulation significativo de CXCL1, CXCL2 y CXCL3 de una manera dependiente de tiempo. Sin embargo, hubo s&oacute;lo un efecto menor de HIBCPP-infecci&oacute;n con EV30 sobre transepitelial migraci&oacute;n de linfocitos T con o sin el quimioatrayentes CXCL12. Por otra parte, CXCL3 no mejorar considerablemente las migraciones T cell. Por lo tanto, deben estar involucrados factores adicionales para la migraci&oacute;n de c&eacute;lulas T de mayor divulgado in vivo en el SNC en el contexto de meningitis enteroviral. Como HIBCPP son permisivas para infecci&oacute;n con EV30, constituyen un modelo in vitro humano valioso para el estudio de la infecci&oacute;n viral en el BCSFB.

Quimiotaxis de c&eacute;lulas T despu&eacute;s de la infecci&oacute;n de c&eacute;lulas de papiloma del plexo coroides humano con 30 ecovirus en un modelo in vitro de la barrera de fluido cerebroespinal de sangre.

Enteroviren ist der h&auml;ufigste Erreger, virale Meningitis verursachen, besonders bei Kindern. Neben die Blut - Hirn Schranke (BBB) des Plexus choroideus bildet wurde die Blut-cerebrospinal-Fl&uuml;ssigkeit (CSF) Schranke (BCSFB), gezeigt, in der Pathogenese des enteroviraler Meningitis einbezogen werden. In einem menschlichen in-vitro-Modell von der BCSFB, bestehend aus menschlichen Plexus choroideus Papilloma Zellen (HIBCPP) wurde die Zul&auml;ssigkeit der Plexus Epithelzellen f&uuml;r Echovirus 30 (EV30) von Immunoblotting und quantitative Echtzeit-PCR (Q-PCR) analysiert. HIBCPP k&ouml;nnten direkt von EV30 aus dem apikalen sowie aus der physiologischen relevanten Basolateraler Seite infiziert. Zeitraum eine Infektion von 5 h konnte keine Ver&auml;nderungen der Barriere-Funktion und Zelle Entwicklungsf&auml;higkeit beobachtet werden. Analyse des Cytokine/Chemokin-Profils nach enteroviraler Infektionen mit Cytometric Perlen Array (CBA) und Q-PCR ergab eine erh&ouml;hte Sekretion von PanGRO (CXCL1, CXCL2 und CXCL3), IL8 und CCL5. Q-PCR zeigte eine signifikante Hochregulation von CXCL1, CXCL2 und CXCL3 in einer Zeit-abh&auml;ngigen Weise. Allerdings gab es nur eine geringe Auswirkung der HIBCPP-Infektion mit EV30 auf T-Lymphozyten-Migration mit oder ohne Chemoattractant CXCL12 transepithelialen. Dar&uuml;ber hinaus CXCL3 T-Zell-Migrationen nicht erheblich verbessern. Daher m&uuml;ssen weitere Faktoren f&uuml;r die in-vivo gemeldeten erh&ouml;hte T-Zell-Migration in der CNS im Kontext von enteroviraler Meningitis einbezogen werden. Wie HIBCPP sind permissive Infektion mit EV30, bilden sie eine wertvolle menschliche in vitro Modell Virusinfektion bei der BCSFB zu studieren.

Chemotaxis von T-Zellen nach Infektion der menschlichen Plexus choroideus Papilloma Zellen mit Echovirus 30 in einem in-vitro-Modell der Blut-cerebro-spinale Fl&uuml;ssigkeit Barriere.

Enterovirus &egrave; il pi&ugrave; comune agente patogeno causando meningite virale, soprattutto nei bambini. Oltre la barriera emato - encefalica (BBB) il plesso coroideo, che forma la barriera di sangue fluido cerebrospinale (CSF) (BCSFB), ha dimostrato di essere coinvolti nella patogenesi della meningite classica. In un modello umano in vitro di BCSFB costituiti da cellule di papilloma umano plesso coroideo (HIBCPP), la permissivit&agrave; delle cellule epiteliali del plesso per 30 Echovirus (EV30) &egrave; stato analizzato da immunoblotting e PCR quantitativa in tempo reale (Q-PCR). HIBCPP potrebbe essere infettato direttamente da EV30 dall&#39;apicale, come pure dal lato basolaterale fisiologici rilevanti. Durante un periodo di infezione di h 5 non potrebbero essere osservati alterazioni della vitalit&agrave; delle cellule e la funzione di barriera. Analisi delle citochine/chemochine-profilo seguendo la classica infezione con un citometro perlina array (CBA) e Q-PCR ha rivelato una maggiore secrezione di PanGRO (CXCL1, CXCL2 e CXCL3), IL8 e CCL5. Q-PCR ha mostrato una significativa sovraregolazione di CXCL1, CXCL2 e CXCL3 in modo tempo dipendente. Tuttavia, c&#39;era solo un effetto minore di HIBCPP-infezione con EV30 sulla migrazione dei linfociti T con o senza chemioattrattanti CXCL12 transepiteliale. Inoltre, CXCL3 non ha fatto migliorare in modo significativo le migrazioni delle cellule T. Quindi fattori supplementari devono essere coinvolte per la migrazione di cellule T in vivo enhanced segnalati nel sistema nervoso centrale nel contesto della meningite classica. Come HIBCPP sono permissivi per infezione con EV30, essi costituiscono un prezioso modello umano in vitro per lo studio di infezione virale presso il BCSFB.

Chemiotassi di cellule T dopo l&#39;infezione delle cellule di papilloma umano plesso coroideo con 30 Echovirus in un modello in vitro della barriera sangue-cerebrospinale fluido.

&#x30A8;&#x30F3;&#x30C6;&#x30ED; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306F;&#x3001;&#x7279;&#x306B;&#x5B50;&#x4F9B;&#x306B;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x6027;&#x9AC4;&#x819C;&#x708E;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059;&#x6700;&#x3082;&#x4E00;&#x822C;&#x7684;&#x306A;&#x75C5;&#x539F;&#x4F53;&#x3067;&#x3059;&#x3002;&#x8840;&#x6DB2;&#x8133;&#x5F62;&#x6210;&#x95A2;&#x9580;&#x307B;&#x304B; (BBB) &#x8108;&#x7D61;&#x53E2;&#x8840;&#x6DB2;&#x8133;&#x810A;&#x9AC4;&#x6DB2; &#xFF08;CSF&#xFF09; &#x30D0;&#x30EA;&#x30A2; (&#x30E9;&#x30AF;&#x30C8;&#x30D5;&#x30A8;&#x30EA;&#x30F3;), &#x30A8;&#x30F3;&#x30C6;&#x30ED; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306E;&#x9AC4;&#x819C;&#x708E;&#x306E;&#x75C5;&#x56E0;&#x306B;&#x95A2;&#x4E0E;&#x3059;&#x308B;&#x3053;&#x3068;&#x793A;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x4EBA;&#x9593;&#x306E;&#x8108;&#x7D61;&#x53E2;&#x4E73;&#x982D;&#x816B;&#x7D30;&#x80DE; (HIBCPP) &#x304B;&#x3089;&#x6210;&#x308B;&#x30E9;&#x30AF;&#x30C8;&#x30D5;&#x30A8;&#x30EA;&#x30F3;&#x306E;&#x4EBA;&#x9593; in vitro &#x30E2;&#x30C7;&#x30EB; &#x30A8;&#x30B3;&#x30FC; &#x30A6;&#x30A4;&#x30EB;&#x30B9; 30 (EV30) &#x306E;&#x795E;&#x7D4C;&#x53E2;&#x306E;&#x4E0A;&#x76AE;&#x7D30;&#x80DE;&#x306E;&#x7518;&#x3044;&#x30A4;&#x30E0;&#x30CE;&#x30D6;&#x30ED;&#x30C3;&#x30C8;&#x6CD5;&#x304A;&#x3088;&#x3073;&#x5B9A;&#x91CF;&#x7684;&#x30EA;&#x30A2;&#x30EB;&#x30BF;&#x30A4;&#x30E0; PCR (Q PCR) &#x306B;&#x3088;&#x3063;&#x3066;&#x89E3;&#x6790;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;HIBCPP &#x76F4;&#x63A5; EV30 &#x306B;&#x3088;&#x3063;&#x3066;&#x9802;&#x7AEF;&#x304B;&#x3089;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x3001;&#x751F;&#x7406;&#x5B66;&#x7684;&#x95A2;&#x9023;&#x6027;&#x306E;&#x9AD8;&#x3044;&#x57FA;&#x5E95;&#x5074;&#x306B;&#x3082;&#x611F;&#x67D3;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;5 H &#x306E;&#x611F;&#x67D3;&#x671F;&#x9593;&#x4E2D;&#x306B;&#x30D0;&#x30EA;&#x30A2;&#x306E;&#x6A5F;&#x80FD;&#x3068;&#x7D30;&#x80DE;&#x306E;&#x751F;&#x5B58;&#x7387;&#x306E;&#x5909;&#x5316;&#x304C;&#x8A8D;&#x3081;&#x3089;&#x308C;&#x306A;&#x304B;&#x3063;&#x305F;&#x3002;&#x6B21;&#x306E;&#x30A8;&#x30F3;&#x30C6;&#x30ED; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x611F;&#x67D3;&#x75C7; Q PCR &#x3068;&#x30D5;&#x30ED;&#x30FC;&#x30B5;&#x30A4;&#x30C8;&#x30E1;&#x30C8;&#x30EA;&#x30FC;&#x306B;&#x3088;&#x308B;&#x30D3;&#x30FC;&#x30BA; &#x30A2;&#x30EC;&#x30A4; &#xFF08;CBA&#xFF09; &#x30B5;&#x30A4;&#x30C8;&#x30AB;&#x30A4;&#x30F3;/&#x30B1;&#x30E2;&#x30AB;&#x30A4;&#x30F3;-&#x30D7;&#x30ED;&#x30D5;&#x30A1;&#x30A4;&#x30EB;&#x5206;&#x6790;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3001;PanGRO (CXCL1&#x3001;CXCL2 &#x304A;&#x3088;&#x3073; CXCL3) &#x306E;&#x5F37;&#x5316;&#x3055;&#x308C;&#x305F;&#x5206;&#x6CCC; IL8 &#x3068; CCL5&#x3002;Q PCR &#x6709;&#x610F;&#x306E;&#x30A2;&#x30C3;&#x30D7;&#x30EC;&#x30AE;&#x30E5;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306E; CXCL1&#x3001;CXCL2&#x3001;CXCL3&#x3001;&#x6642;&#x9593;&#x4F9D;&#x5B58;&#x306E;&#x65B9;&#x6CD5;&#x3067;&#x793A;&#x3057;&#x305F;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;EV30 &#x3068; HIBCPP &#x611F;&#x67D3;&#x75C7;&#x306E;&#x30DE;&#x30A4;&#x30CA;&#x30FC;&#x306E;&#x52B9;&#x679C;&#x4E0A;&#x76AE; T &#x30EA;&#x30F3;&#x30D1;&#x7403;&#x306E;&#x79FB;&#x884C;&#x904A;&#x8D70; cxcl 12 &#x306E;&#x6709;&#x7121;&#x306B;&#x3060;&#x3051;&#x3060;&#x3063;&#x305F;&#x3002;&#x307E;&#x305F;&#x3001;CXCL3 &#x304C; T &#x7D30;&#x80DE;&#x306E;&#x79FB;&#x884C;&#x3092;&#x5927;&#x5E45;&#x306B;&#x5411;&#x4E0A;&#x3057;&#x305F;&#x306A;&#x3044;&#x3067;&#x3059;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x8FFD;&#x52A0;&#x306E;&#x8981;&#x56E0;&#x30A8;&#x30F3;&#x30C6;&#x30ED; &#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x306E;&#x9AC4;&#x819C;&#x708E;&#x306E;&#x30B3;&#x30F3;&#x30C6;&#x30AD;&#x30B9;&#x30C8;&#x3067; CNS &#x306B; in vivo &#x3067;&#x306E;&#x5F37;&#x5316;&#x3055;&#x308C;&#x305F; T &#x7D30;&#x80DE;&#x3067;&#x5831;&#x544A;&#x3055;&#x308C;&#x305F;&#x79FB;&#x884C;&#x306E;&#x95A2;&#x4E0E;&#x3059;&#x308B;&#x5FC5;&#x8981;&#x304C;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;HIBCPP &#x5BDB;&#x5BB9; EV30 &#x3068;&#x611F;&#x67D3;&#x75C7;&#x306E;&#x305F;&#x3081;&#x3068;&#x3057;&#x3066;&#x3001;&#x5F7C;&#x3089;&#x306F;&#x30E9;&#x30AF;&#x30C8;&#x30D5;&#x30A8;&#x30EA;&#x30F3;&#x3067;&#x306E;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#x611F;&#x67D3;&#x3092;&#x52C9;&#x5F37;&#x3059;&#x308B;&#x8CB4;&#x91CD;&#x306A;&#x4EBA;&#x9593;&#x306E; in vitro &#x30E2;&#x30C7;&#x30EB;&#x3092;&#x69CB;&#x6210;&#x3057;&#x307E;&#x3059;&#x3002;

T &#x7D30;&#x80DE;&#x306E;&#x8D70;&#x5316;&#x6027;&#x611F;&#x67D3;&#x8840;&#x6DB2;&#x8133;&#x810A;&#x9AC4;&#x6DB2;&#x95A2;&#x9580;&#x306E; in vitro &#x30E2;&#x30C7;&#x30EB; &#x30A8;&#x30B3;&#x30FC; &#x30A6;&#x30A4;&#x30EB;&#x30B9; 30 &#x3068;&#x4EBA;&#x9593;&#x306E;&#x8108;&#x7D61;&#x53E2;&#x4E73;&#x982D;&#x816B;&#x7D30;&#x80DE;&#x306E;&#x5F8C;&#x3002;

&#xC5D4;&#xD14C;&#xB85C;&#xBC14;&#xC774;&#xB7EC;&#xC2A4;&#xC5D0; &#xD2B9;&#xD788; &#xC5B4;&#xB9B0;&#xC774; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xC131; &#xC218; &#xB9C9; &#xC5FC;&#xC744; &#xC77C;&#xC73C;&#xD0A4;&#xB294; &#xAC00;&#xC7A5; &#xD754;&#xD55C; &#xBCD1;&#xC6D0; &#xCCB4; &#xC774;&#xB2E4;. &#xC678;&#xC5D0; &#xD608;&#xC561;-&#xB1CC; &#xC7A5;&#xBCBD; (BBB) &#xB9E5;&#xB77D; &#xB9C9; &#xC2E0;&#xACBD; &#xCD1D;&#xC744; &#xD615;&#xC131; &#xD558;&#xB294; &#xD608;&#xC561; cerebrospinal &#xC561;&#xCCB4; (CSF) &#xC7A5;&#xBCBD; (BCSFB), enteroviral &#xC218; &#xB9C9; &#xC5FC;&#xC758; &#xBCD1; &#xC778;&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xD45C;&#xC2DC; &#xD588;&#xB2E4;. &#xC778;&#xAC04;&#xC758; &#xB9E5;&#xB77D; &#xB9C9; &#xC2E0;&#xACBD; &#xCD1D; &#xC720; &#xB450; &#xC885; &#xC140; (HIBCPP)&#xB85C; &#xAD6C;&#xC131; &#xB41C; BCSFB&#xC758; &#xC778;&#xAC04;&#xC758; &#xCCB4; &#xC678; &#xBAA8;&#xB378;, Echovirus 30 (EV30)&#xC5D0; &#xB300; &#xD55C; &#xCD1D; &#xC0C1;&#xD53C; &#xC138;&#xD3EC; &#xD5C8;&#xC6A9;&#xD568;&#xC740; immunoblotting &#xBC0F; &#xC591;&#xC801; &#xC2E4;&#xC2DC;&#xAC04; PCR (Q-PCR) &#xBD84;&#xC11D; &#xD588;&#xB2E4;. HIBCPP ev30&#xC5D0; &#xC758;&#xD574; &#xC0DD;&#xB9AC; &#xAD00;&#xB828; basolateral &#xCE21;&#xBA74;&#xC5D0;&#xC11C; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C;&#xB97C; &#xD600;&#xB05D;&#xC5D0;&#xC11C; &#xC9C1;&#xC811; &#xAC10;&#xC5FC; &#xB420; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. 5 H&#xC758; &#xAC10;&#xC5FC; &#xAE30;&#xAC04; &#xB3D9;&#xC548; &#xC7A5;&#xBCBD; &#xAE30;&#xB2A5; &#xBC0F; &#xC138;&#xD3EC; &#xC0DD;&#xC874; &#xB2A5;&#xB825;&#xC758; &#xC544;&#xBB34; &#xBCC0;&#xACBD; &#xAD00;&#xCC30; &#xB420; &#xC218; &#xC788;&#xC5C8;&#xB2E4;. Cytokine/chemokine-&#xD504;&#xB85C;&#xD544; cytometric &#xBE44;&#xB4DC; &#xC5B4;&#xB808;&#xC774; (CBA)&#xC640; Q PCR enteroviral &#xAC10;&#xC5FC;&#xC5D0; &#xB530;&#xB77C; &#xBD84;&#xC11D; &#xACF5;&#xAC1C; PanGRO (CXCL1, CXCL2 &#xBC0F; CXCL3)&#xC758; &#xD5A5;&#xC0C1; &#xB41C; &#xBD84; &#xBE44; IL8 &#xBC0F; CCL5. Q PCR &#xC2DC;&#xAC04; &#xC758;&#xC874; &#xBC29;&#xC2DD;&#xC73C;&#xB85C; CXCL1, CXCL2 &#xBC0F; cxcl3&#xC758; &#xC0C1;&#xB2F9;&#xD55C; upregulation&#xC744; &#xBCF4;&#xC600;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, transepithelial T &#xB9BC;&#xD504; &#xAD6C; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158; chemoattractant CXCL12 &#xC720;&#xBB34;&#xC5D0; ev30&#xC640; HIBCPP-&#xAC10;&#xC5FC;&#xC758; &#xB2E8; &#xBD80; &#xD6A8;&#xACFC;. &#xB610;&#xD55C;, cxcl3&#xB294; &#xD06C;&#xAC8C; T &#xC138;&#xD3EC; &#xC774;&#xC8FC;&#xB97C; &#xAC15;&#xD654; &#xD558;&#xC9C0; &#xC54A;&#xC558;&#xB2E4;. &#xB530;&#xB77C;&#xC11C; &#xCD94;&#xAC00; &#xC694;&#xC18C; enteroviral &#xC218; &#xB9C9; &#xC5FC;&#xC758; &#xB9E5;&#xB77D;&#xC5D0;&#xC11C; CNS&#xC5D0; vivo&#xC5D0;&#xC11C; &#xBCF4;&#xACE0; &#xB41C; &#xD5A5;&#xC0C1; &#xB41C; T &#xC138;&#xD3EC; &#xB9C8;&#xC774;&#xADF8;&#xB808;&#xC774;&#xC158;&#xC5D0; &#xCC38;&#xC5EC; &#xD574;&#xC57C; &#xD569;&#xB2C8;&#xB2E4;. HIBCPP&#xB294; ev30&#xACFC; &#xAC10;&#xC5FC;&#xC5D0; &#xB300; &#xD55C; &#xAD00;&#xB300; &#xD55C;, &#xADF8;&#xB4E4;&#xC740; &#xADC0;&#xC911; &#xD55C; &#xC778;&#xAC04;&#xC758; &#xCCB4; &#xC678; &#xBAA8;&#xB378; &#xC5F0;&#xAD6C;&#xB294; BCSFB&#xC5D0;&#xC11C; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; &#xAC10;&#xC5FC;&#xC744; &#xAD6C;&#xC131;.

&#xC778;&#xAC04;&#xC758; &#xB9E5;&#xB77D; &#xB9C9; &#xC2E0;&#xACBD; &#xCD1D; &#xC720; &#xB450; &#xC885; &#xC140; Echovirus 30 &#xD608;&#xC561; cerebrospinal &#xC720;&#xCCB4; &#xBC30;&#xB9AC;&#xC5B4;&#xC758; &#xCCB4; &#xC678; &#xBAA8;&#xB378;&#xC5D0;&#xC11C;&#xC758; &#xAC10;&#xC5FC; &#xD6C4; T-&#xC138;&#xD3EC;&#xC758; chemotaxis.

Bacterial invasion through the blood-cerebrospinal fluid barrier (BCSFB) during bacterial meningitis causes secretion of proinflammatory cytokines/chemokines followed by the recruitment of leukocytes into the CNS. In this study, we analyzed the cellular and molecular mechanisms of polymorphonuclear neutrophil (PMN) and monocyte transepithelial transmigration (TM) across the BCSFB after bacterial infection.

Transmigration of polymorphnuclear neutrophils and monocytes through the human blood-cerebrospinal fluid barrier after bacterial infection in vitro.

Human milk oligosaccharides help to prevent infectious diseases in breastfed infants. Larger scale testing, particularly in animal models and human clinical studies, is still limited due to shortened availability of more complex oligosaccharides. The purpose of this study was to evaluate 2'-fucosyllactose (2'-FL) and 3-fucosyllactose (3-FL) synthesized by whole-cell biocatalysis for their biological activity in vitro. Therefore, we have tested these oligosaccharides for their inhibitory potential of pathogen adhesion in two different human epithelial cell lines. 2'-FL could inhibit adhesion of Campylobacter jejuni, enteropathogenic Escherichia coli, Salmonella enterica serovar fyris, and Pseudomonas aeruginosa to the intestinal human cell line Caco-2 (reduction of 26%, 18%, 12%, and 17%, respectively), as could be shown for 3-FL (enteropathogenic E coli 29%, P aeruginosa 26%). Furthermore, adherence of P aeruginosa to the human respiratory epithelial cell line A549 was significantly inhibited by 2'-FL and 3-FL (reduction of 24% and 23%, respectively). These results confirm the biological and functional activity of biotechnologically synthesized human milk oligosaccharides. Mass-tailored human milk oligosaccharides could be used in the future to supplement infant formula ingredients or as preventatives to reduce the impact of infectious diseases.

Bioengineered 2'-fucosyllactose and 3-fucosyllactose inhibit the adhesion of Pseudomonas aeruginosa and enteric pathogens to human intestinal and respiratory cell lines.

The human-specific, Gram-negative bacterium Neisseria meningitidis (Nm) is a leading cause of bacterial meningitis worldwide. The blood-cerebrospinal fluid barrier (BCSFB), which is constituted by the epithelial cells of the choroid plexus (CP), has been suggested as one of the potential entry sites of Nm into the CSF and can contribute to the inflammatory response during infectious diseases of the brain. Toll-like receptors (TLRs) are involved in mediating signal transduction caused by the pathogens.

Article	Year
The protein moiety of Brucella abortus outer membrane protein 16 is a new bacterial pathogen-associated molecular pattern that activates dendritic cells in vivo, induces a Th1 immune respons... Journal of immunology (Baltimore, Md. : 1950)\| PubMed ID: 20351187	2010
In vitro transcriptome analysis of porcine choroid plexus epithelial cells in response to Streptococcus suis: release of pro-inflammatory cytokines and chemokines. Microbes and infection / Institut Pasteur\| PubMed ID: 21683799	2011
Polar invasion and translocation of Neisseria meningitidis and Streptococcus suis in a novel human model of the blood-cerebrospinal fluid barrier. PloS one\| PubMed ID: 22253884	2012
Chemotaxis of T-cells after infection of human choroid plexus papilloma cells with Echovirus 30 in an in vitro model of the blood-cerebrospinal fluid barrier. Virus research\| PubMed ID: 23000117	2012
Transmigration of polymorphnuclear neutrophils and monocytes through the human blood-cerebrospinal fluid barrier after bacterial infection in vitro. Journal of neuroinflammation\| PubMed ID: 23448224	2013
Bioengineered 2'-fucosyllactose and 3-fucosyllactose inhibit the adhesion of Pseudomonas aeruginosa and enteric pathogens to human intestinal and respiratory cell lines. Nutrition research (New York, N.Y.)\| PubMed ID: 24074741	2013
Neisseria meningitidis elicits a pro-inflammatory response involving IκBζ in a human blood-cerebrospinal fluid barrier model. Journal of neuroinflammation\| PubMed ID: 25347003	2014

Julia Borkowski

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