Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

manufacturing-devices-instruments-for-easier-rat-liver

Manufacturing Devices and Instruments for Easier Rat Liver Transplantation

Orthotopic rat liver transplantation is a popular model, which has been shown in a recent JoVE paper with the use of the "quick-linker" device. This ...

University of Geneva

Carbon nanotubes are emerging as innovative tools in nanobiotechnology. However, their toxic effects on environment and health have become an issue of strong concern. In the present study, we address the impact of functionalized carbon nanotubes (f-CNTs) on cells of the immune system. We have prepared two types of f-CNTs, following the 1,3-dipolar cycloaddition reaction (f-CNTs 1 and 2) and the oxidation/amidation treatment (f-CNTs 3 and 4), respectively. We have found that both types of f-CNTs are uptaken by B and T lymphocytes as well as macrophages in vitro, without affecting cell viability. Subsequently, the functionality of the different cells was analyzed carefully. We discovered that f-CNT 1, which is highly water soluble, did not influence the functional activity of immunoregulatory cells. f-CNT 3, which instead possesses reduced solubility and forms mainly stable water suspensions, preserved lymphocytes' functionality while provoking secretion of proinflammatory cytokines by macrophages.

Functionalized carbon nanotubes are non-cytotoxic and preserve the functionality of primary immune cells.

Nanotubes de carbone se pr&eacute;sentent comme des outils innovants en nanobiotechnologie. Cependant, leurs effets toxiques sur l&#39;environnement et la sant&eacute; sont devenues un sujet de forte pr&eacute;occupation. Dans la pr&eacute;sente &eacute;tude, nous abordons l&#39;impact des nanotubes de carbone fonctionnalis&eacute;s (f-CNTs) sur les cellules du syst&egrave;me immunitaire. Nous avons pr&eacute;par&eacute; deux types de f-CNTs, suite &agrave; la r&eacute;action de cycloaddition 1, 3-dipolaire (f-CNTs 1 et 2) et le traitement d&#39;oxydation/amidation (f-CNTs 3 et 4), respectivement. Nous avons trouv&eacute; que les deux types de f-CNTs sont adopt&eacute;s par les lymphocytes B et T ainsi que les macrophages in vitro, sans affecter la viabilit&eacute; des cellules. Par la suite, les fonctionnalit&eacute;s des diff&eacute;rentes cellules a &eacute;t&eacute; analys&eacute;e avec soin. Nous avons d&eacute;couvert ce CNT-f 1, qui est tr&egrave;s soluble dans l&#39;eau, n&#39;a pas influenc&eacute; l&#39;activit&eacute; fonctionnelle des cellules immunor&eacute;gulatrices. f-CNT 3, qui au contraire poss&egrave;de une solubilit&eacute; r&eacute;duite et forme des suspensions principalement stable de l&#39;eau, conserve la fonctionnalit&eacute; des lymphocytes tout en provoquant la s&eacute;cr&eacute;tion de cytokines pro-inflammatoires par les macrophages.

Nanotubes de carbone fonctionnalis&eacute;s sont non cytotoxiques et conserver la fonctionnalit&eacute; des cellules immunitaires primaires.

Nanotubi di carbonio stanno emergendo come strumenti innovativi in nanobiotecnologie. Tuttavia, loro effetti tossici su ambiente e salute sono diventati un problema di forte preoccupazione. Nel presente studio, affrontiamo l&#39;impatto dei nanotubi di carbonio funzionalizzate (f-CNTs) sulle cellule del sistema immunitario. Abbiamo preparato due tipi di f-CNTs, seguendo la reazione di cicloaddizione 1,3-dipolare (f-CNTs 1 e 2) e il trattamento di ossidazione/amidazione (f-CNTs 3 e 4), rispettivamente. Abbiamo trovato che entrambi i tipi di f-CNTs sono uptaken da linfociti B e T, come pure i macrofagi in vitro, senza compromettere la vitalit&agrave; cellulare. Successivamente, &egrave; stata analizzata con attenzione la funzionalit&agrave; delle cellule differenti. Abbiamo scoperto che CNT-f 1, che &egrave; altamente solubile in acqua, non ha influenzato l&#39;attivit&agrave; funzionale delle cellule immunoregulatory. f-CNT 3, che invece possiede ridotta solubilit&agrave; e forma sospensioni stabili principalmente acqua, conservata funzionalit&agrave; dei linfociti mentre provocando la secrezione di citochine proinfiammatorie da macrofagi.

Nanotubi di carbonio funzionalizzate sono non citotossici e preservare la funzionalit&agrave; delle cellule del sistema immunitario primarie.

CXCR3 is a G protein-coupled, seven-transmembrane receptor that binds and is activated by the three IFN-gamma-inducible chemokines of the CXC family named CXCL9, CXCL10, and CXCL11. These chemokines are not constitutively expressed but are up-regulated in a proinflammatory cytokine milieu. Consequently, their major function is to selectively recruit immune cells at inflammation sites, but they also play a role in angiogenesis mechanisms. In the last few years, strong experimental and clinical evidence has been obtained supporting the idea that the CXCR3 pathway is involved in the development of autoimmune diseases, especially by creating local amplification loops of inflammation in target organs, thereby inducing worsening of clinical manifestations. This article briefly reviews what we know today about the nature and functions of CXCR3, with special emphasis on its involvement in two main rheumatic systemic autoimmune diseases, namely rheumatoid arthritis and systemic lupus erythematosus.

CXCR3, inflammation, and autoimmune diseases.

CXCR3 est un r&eacute;cepteur de sept domaines transmembranaire, coupl&eacute;s aux prot&eacute;ines G qui lie et est activ&eacute; par les ch&eacute;mokines IFN-gamma-inducible trois de la famille CXC nomm&eacute;s CXCL9, CXCL10 et CXCL11. Ces chimiokines ne sont pas exprim&eacute;s de fa&ccedil;on constitutive mais sont r&eacute;gul&eacute;e dans un milieu de cytokines pro-inflammatoires. Par cons&eacute;quent, leur fonction principale est de recruter s&eacute;lectivement des cellules immunitaires dans les sites de l&#39;inflammation, mais ils &eacute;galement jouent un r&ocirc;le dans les m&eacute;canismes d&#39;angiogen&egrave;se. Dans ces derni&egrave;res ann&eacute;es, des preuves exp&eacute;rimentales et cliniques solides a &eacute;t&eacute; obtenu en soutenant l&#39;id&eacute;e que la voie CXCR3 est impliqu&eacute;e dans le d&eacute;veloppement de maladies auto-immunes, en particulier en cr&eacute;ant des boucles locales de l&#39;amplification de l&#39;inflammation dans les organes cibles, induisant ainsi une aggravation des manifestations cliniques. Cet article examine bri&egrave;vement ce que nous savons aujourd&#39;hui sur la nature et les fonctions de CXCR3, avec un accent particulier sur sa participation aux deux principaux syst&eacute;miques auto-immunes maladies rhumatismales, nomm&eacute;ment la polyarthrite rhumato&iuml;de et le lupus &eacute;ryth&eacute;mateux diss&eacute;min&eacute;.

CXCR3, inflammation et maladies auto-immunes.

CXCR3 &egrave; un recettore G protein-coupled, 7-transmembrana che si lega e viene attivato mediante le chemiochine IFN-gamma-inducible tre della famiglia CXC denominato CXCL9, CXCL10 e CXCL11. Queste chemochine non sono costitutivamente espressi ma sono up-regolate in un milieu di citochine proinfiammatorie. Di conseguenza, la loro funzione principale &egrave; quella di reclutare selettivamente le cellule immunitarie nei siti di infiammazione, ma svolgono anche un ruolo fondamentale nei meccanismi di angiogenesi. Negli ultimi anni, sono stata ottenuta forti prove sperimentali e cliniche sostenendo l&#39;idea che il percorso CXCR3 &egrave; coinvolto nello sviluppo di malattie autoimmuni, soprattutto mediante la creazione di cicli di amplificazione locale dell&#39;infiammazione in organi bersaglio, quindi indurre peggioramento delle manifestazioni cliniche. Questo articolo recensisce brevemente che cosa sappiamo oggi sulla natura e le funzioni di CXCR3, con speciale enfasi sul suo coinvolgimento in due principali malattie reumatiche malattie autoimmunitarie sistemiche, vale a dire l&#39;artrite reumatoide e lupus eritematoso sistemico.

CXCR3, infiammazione e malattie autoimmuni.

An important hallmark of systemic lupus erythematosus is the production of autoantibodies specific for nuclear Ags, among which nucleosomes and their constituents, DNA and histones. It is widely admitted that some of these autoantibodies contribute largely in lupus pathogenesis because of their nephritogenic potential. However, the underlying mechanisms are still debated. In this study, we analyzed the autoimmune response against histone H2B during the course of the disease in lupus-prone (NZBxNZW)F1 mice, both in lymphoid organs and kidneys, and we assessed its potential involvement in lupus pathogenicity. We found that the N-terminal region of histone H2B represents a preferential target for circulating autoantibodies, which kinetics of appearance positively correlates with disease development. Furthermore, immunization of preautoimmune (NZBxNZW)F1 mice with H2B peptide 1-25 accelerates the disease. Kidney eluates from diseased (NZBxNZW)F1 mice do contain IgG Abs reacting with this peptide, and this H2B sequence was found to be accessible to specific Ab probes in Ag-containing deposits detected in nephritic kidneys. Finally, compared with control normal mice and to young preautoimmune (NZBxNZW)F1 animals, the frequency of cells secreting autoantibodies reacting with peptide 1-25 was significantly raised in the spleen and bone marrow and most importantly on a pathophysiological point of view, locally, in nephritic kidneys of diseased (NZBxNZW)F1 mice. Altogether our results demonstrate the existence in (NZBxNZW)F1 mice of both a systemic and local B cell response targeting the N-terminal region of histone H2B, and highlight the potential implication of this nuclear domain in lupus pathology.

Identification of new pathogenic players in lupus: autoantibody-secreting cells are present in nephritic kidneys of (NZBxNZW)F1 mice.

Une caract&eacute;ristique importante du lupus &eacute;ryth&eacute;mateux diss&eacute;min&eacute; est la production d&#39;auto-anticorps sp&eacute;cifiques pour Ags nucl&eacute;aires, entre les nucl&eacute;osomes et leurs constituants, ADN et les histones. Il est largement admis que certains de ces autoanticorps contribuent en grande partie dans la pathog&eacute;nie du lupus en raison de leur potentiel de n&eacute;phritog&egrave;ne. Cependant, les m&eacute;canismes sous-jacents sont encore d&eacute;battues. Dans cette &eacute;tude, nous avons analys&eacute; la r&eacute;action auto-immune contre histone H2B au cours de la maladie chez les souris F1 (NZBxNZW) lupus sujettes, aussi bien dans les organes lympho&iuml;des et les reins, et nous avons &eacute;valu&eacute; son implication potentielle dans la pathog&eacute;nie du lupus. Nous avons constat&eacute; que la r&eacute;gion N-terminale de l&#39;histone H2B repr&eacute;sente une cible pr&eacute;f&eacute;rentielle pour faire circuler des auto-anticorps, qui cin&eacute;tique d&#39;apparition est corr&eacute;l&eacute;e positivement avec le d&eacute;veloppement de la maladie. Par ailleurs, l&#39;immunisation des souris F1 (NZBxNZW) preautoimmune peptide H2B 1-25 acc&eacute;l&egrave;re la maladie. &Eacute;luats de rein de souris malades de F1 (NZBxNZW) contiennent Abs IgG r&eacute;agissant avec ce peptide, et cette s&eacute;quence H2B s&#39;est av&eacute;r&eacute;e pour &ecirc;tre accessible &agrave; des sondes sp&eacute;cifiques de Ab dans les d&eacute;p&ocirc;ts contenant Ag d&eacute;tect&eacute;s dans les reins. Enfin, en comparaison avec des souris normales de contr&ocirc;le et pour les jeunes animaux de F1 (NZBxNZW) preautoimmune, la fr&eacute;quence des cellules s&eacute;cr&eacute;tant des autoanticorps r&eacute;agissant avec le peptide 1-25 consid&eacute;rablement grandit dans la rate et la moelle osseuse et surtout sur un physiopathologique point de vue, localement, &agrave; n&eacute;phrotique reins des souris malades de F1 (NZBxNZW). Au total, nos r&eacute;sultats d&eacute;montrent l&#39;existence chez les souris F1 (NZBxNZW) des deux une r&eacute;ponse syst&eacute;mique et locale des lymphocytes B ciblant la r&eacute;gion N-terminale de l&#39;histone H2B et mettre en &eacute;vidence l&#39;implication potentielle de ce domaine nucl&eacute;aire en pathologie de lupus.

Identification de nouveaux joueurs de pathog&egrave;nes dans le lupus : cellules s&eacute;cr&eacute;tant des auto-anticorps sont pr&eacute;sents dans les reins n&eacute;phrotique de souris F1 (NZBxNZW).

Un&#39;importante caratteristica del lupus eritematoso sistemico &egrave; la produzione di autoanticorpi specifici per Ags nucleari, tra cui nucleosomi e loro costituenti, il DNA e gli istoni. Esso &egrave; largamente ammesso che alcuni di questi autoanticorpi contribuiscono in gran parte nella patogenesi di lupus a causa del loro potenziale nephritogenic. Tuttavia, i meccanismi sottostanti sono ancora dibattuti. In questo studio, abbiamo analizzato la risposta autoimmune contro istone H2B durante il decorso della malattia nei topi lupus-inclini di F1 (NZBxNZW), sia negli organi linfoidi e reni, e abbiamo valutato il potenziale coinvolgimento nella patogenicit&agrave; di lupus. Abbiamo trovato che la regione N-terminale dell&#39;istone H2B rappresenta un bersaglio preferenziale per circolazione di autoanticorpi, quali cinetica di aspetto correla positivamente con lo sviluppo della malattia. Inoltre, l&#39;immunizzazione dei topi di F1 (NZBxNZW) preautoimmune con H2B peptide 1-25 accelera la malattia. Eluati di rene da topi malati di F1 (NZBxNZW) contengono IgG Abs reagendo con questo peptide, e questa sequenza H2B &egrave; stata trovata per essere accessibili alle sonde specifiche Ab Ag-contenenti depositi rilevati nei reni nefritica. Infine, confrontato con topi normali di controllo e per gli animali giovani F1 (NZBxNZW) preautoimmune, la frequenza di cellule che secernono gli autoanticorpi che reagiscono con peptide 1-25 &egrave; significativamente cresciuta nella milza e midollo osseo e soprattutto su fisiopatologico punto di vista, localmente, a nefritica reni dei topi malati di F1 (NZBxNZW). Complessivamente i nostri risultati dimostrano l&#39;esistenza in F1 (NZBxNZW) topi di entrambi una risposta sistemica e locale delle cellule B targeting per regione N-terminale dell&#39;istone H2B ed evidenziano il potenziale implicazione di questo dominio nucleare nella patologia di lupus.

Identificazione di nuovi giocatori patogeni nel lupus: cellule secernenti degli autoanticorpi sono presenti in nefritica reni dei topi F1 (NZBxNZW).

Improved posttransplant monitoring and on-time detection of rejection could improve islet transplantation outcome. The present study explored the possibility of detecting harmful events after mouse islet transplantation measuring the immune responsiveness against islet extracts. Mouse islet transplantations were performed using various donor/recipient combinations, exploring autoimmune (NOD/SCID to NOD, n = 6) and alloimmune events (C57BL/6 to BALB/c, n = 20), a combination of both (C57BL/6 to NOD, n = 8), the absence of both (BALB/c to BALB/c, n = 21), or naive, nontransplanted control mice (n = 14). The immune reactivity was measured by ELISPOT, looking at the ex vivo release of IFN-Î³ from splenocytes stimulated by islet donor extracts (sonicated islets). The immune reactivity was not altered in the syngeneic and autoimmune models, demonstrating similar levels as nontransplanted controls (p = 0.46 and p = 0.6). Conversely, the occurrence of an allogeneic rejection alone or in combination to autoimmunity was associated to an increase in the level of immune reactivity (p = 0.023 and p = 0.003 vs. respective controls). The observed increase was transient and lost in the postrejection period or after treatment with CTLA4-Ig. Overall, allogeneic rejection was associated to a transient increase in the reactivity of splenocytes against islet proteins. Such a strategy has the potential to improve islet graft monitoring in human and should be further explored.

Detecting rejection after mouse Islet transplantation utilizing islet protein-stimulated ELISPOT.

Surveillance posttransplantation am&eacute;lior&eacute;e et les d&eacute;lais d&eacute;tection du rejet pourraient am&eacute;liorer l&#39;&icirc;lot issue de transplantation. La pr&eacute;sente &eacute;tude explore la possibilit&eacute; de d&eacute;tecter des &eacute;v&eacute;nements n&eacute;fastes apr&egrave;s que transplantation d&#39;&icirc;lots pancr&eacute;atiques de souris mesurant la r&eacute;ponse immunitaire contre l&#39;&icirc;lot extraits. Transplantation d&#39;&icirc;lots pancr&eacute;atiques souris ont &eacute;t&eacute; effectu&eacute;e &agrave; l&#39;aide de diff&eacute;rentes combinaisons de donateurs, explorant auto-immune (NOD/SCID &agrave; ADI, n = 6) et &eacute;v&eacute;nements de l&#39;IG (C57BL/6 &agrave; BALB/c, n = 20), une combinaison des deux (C57BL/6 &agrave; ADI, n = 8), l&#39;absence des deux (BALB/c et BALB/c, n = 21), ou na&iuml;f, les souris t&eacute;moins nontransplanted (n = 14). La r&eacute;activit&eacute; immunitaire a &eacute;t&eacute; mesur&eacute;e par ELISPOT, en regardant l&#39;ex vivo lib&eacute;ration d&#39;IFN-&gamma; de spl&eacute;nocytes stimul&eacute;s par les donateurs de l&#39;&icirc;lot extraits (&icirc;lots trait&eacute; aux ultrasons). La r&eacute;activit&eacute; immunitaire n&#39;a pas &eacute;t&eacute; modifi&eacute;e dans les mod&egrave;les syng&eacute;niques et auto-immunes, d&eacute;montrant des niveaux similaires comme nontransplanted contr&ocirc;les (p = 0,46 et p = 0,6). &Agrave; l&#39;inverse, la pr&eacute;sence d&#39;un rejet allog&eacute;nique seul ou en combinaison &agrave; l&#39;auto-immunit&eacute; &eacute;tait associ&eacute;e &agrave; une augmentation du niveau de la r&eacute;activit&eacute; immunitaire (p = 0,023 et p = 0,003 vs t&eacute;moins respectifs). L&#39;augmentation observ&eacute;e &eacute;tait transitoire et perdus dans la p&eacute;riode postrejection ou apr&egrave;s le traitement avec CTLA4-Ig. Dans l&#39;ensemble, allog&eacute;nique rejet a &eacute;t&eacute; associ&eacute;e &agrave; une augmentation transitoire de la r&eacute;activit&eacute; des spl&eacute;nocytes contre les prot&eacute;ines de l&#39;&icirc;lot. Une telle strat&eacute;gie a le potentiel d&#39;am&eacute;liorer la greffe d&#39;&icirc;lots surveillance chez l&#39;homme et devrait &ecirc;tre &eacute;tudi&eacute;e plus avant.

D&eacute;tection de rejet apr&egrave;s que utilisant &icirc;lot de souris Islet transplantation ELISPOT prot&eacute;ine stimul&eacute;e.

Migliore monitoraggio infettivo e puntuale rilevazione del rifiuto potrebbe migliorare islet trapianto esito. Il presente studio ha esplorato la possibilit&agrave; di rilevare eventi dannosi dopo trapianto di isolotto del mouse misura la capacit&agrave; di risposta immunitaria contro isolotto estratti. Trapianti isolotto del mouse sono stati eseguiti utilizzando varie combinazioni di donatore/ricevente, esplorando autoimmuni (NOD/SCID a NOD, n = 6) e alloimmune eventi (C57BL/6 a BALB/c, n = 20), una combinazione di entrambi (C57BL/6 a NOD, n = 8), l&#39;assenza di entrambi (BALB/c a BALB/c, n = 21), o ingenui, topi di controllo nontransplanted (n = 14). La reattivit&agrave; immunitaria &egrave; stata misurata mediante ELISPOT, guardando l&#39;ex vivo rilascio di IFN-&gamma; da splenocytes stimolata da donatore isolotto estratti (lisati mediante isolotti). La reattivit&agrave; del sistema immunitaria non &egrave; stata alterata nei modelli Carlo e autoimmuni, dimostrando livelli simili come nontransplanted controlli (p = 0,46 e p = 0,6). Al contrario, la presenza di un rifiuto allogenico da solo o in combinazione per autoimmunit&agrave; era associata ad un aumento del livello di reattivit&agrave; immunitaria (p = 0,023 e p = 0.003 vs rispettivi controlli). L&#39;incremento osservato &egrave; stato perduto e transitori nel periodo postrejection o dopo il trattamento con CTLA4-Ig. Rifiuto globale, allogenico &egrave; stato associato a un aumento transitorio della reattivit&agrave; degli splenocytes contro le proteine isolotto. Tale strategia ha il potenziale per migliorare l&#39;innesto isolotto di monitoraggio in essere umano e dovr&agrave; essere ulteriormente esplorata.

Rilevamento di rifiuto dopo l&#39;isolotto utilizzando mouse Islet transplantation proteina-stimolato ELISPOT.

Long-term clinical outcomes of islet transplantation are hampered by rejection and recurrence of autoimmunity, which lead to a gradual decrease in islet function usually taking place over the first five years after transplantation. An accurate monitoring strategy could allow for the detection and treatment of harmful immune events, potentially resulting in higher rates of insulin-independence.

Immune monitoring of pancreatic islet graft: towards a better understanding, detection and treatment of harmful events.

Les r&eacute;sultats cliniques &agrave; long terme de la transplantation d&#39;&icirc;lots pancr&eacute;atiques sont entrav&eacute;s par le rejet et la r&eacute;currence de l&#39;auto-immunit&eacute;, conduisant &agrave; une diminution progressive de la fonction d&#39;&icirc;lots ayant g&eacute;n&eacute;ralement lieu au cours des cinq premi&egrave;res ann&eacute;es apr&egrave;s la transplantation. Une strat&eacute;gie de surveillance pr&eacute;cise pourrait permettre la d&eacute;tection et le traitement des nuisibles &eacute;v&eacute;nements immunitaires, pouvant provoquer des taux plus &eacute;lev&eacute;s d&#39;insuline-ind&eacute;pendance.

Surveillance immunitaire de greffe d&#39;&icirc;lots pancr&eacute;atiques : vers une meilleure compr&eacute;hension, la d&eacute;tection et traitement des &eacute;v&eacute;nements n&eacute;fastes.

I risultati clinici a lungo termine del trapianto isolotto sono ostacolati dal rifiuto e reiterazione di autoimmunit&agrave;, che portano ad una graduale diminuzione della funzione isolotto che solitamente si svolgono sopra i primi cinque anni dopo il trapianto. Un&#39;accurata strategia di monitoraggio potrebbe consentire la rilevazione e il trattamento di eventi dannosi immuni, potenzialmente con conseguente tassi pi&ugrave; elevati di insulina-indipendenza.

Sorveglianza immunitaria del trapianto di isole pancreatiche: verso una migliore comprensione, rilevamento e trattamento di eventi dannosi.

The aim of the present study was to assess the efficiency of cell-based immune assays in the detection of alloreactivity after islet transplantation and to correlate these results with clinical outcome. Mixed lymphocyte cultures were performed with peripheral blood mononuclear cells from recipients (n = 14), donors, or third party. The immune reactivity was assessed by the release of IFN-Î³ (ELISpot), cell proliferation (FACS analysis for Ki67), and cytokine quantification (Bioplex). Islet function correlated with the number of IFN-Î³-secreting cells following incubation with donor cells (p = 0.007, r = -0.50), but not with third party cells (p = 0.61). Similarly, a high number of donor-specific proliferating cells was associated with a low islet function (p = 0.006, r = -0.51). Proliferating cells were mainly CD3(+)CD4(+) lymphocytes and CD3(-)CD56(+) natural killer cells (with low levels of CD3(+)CD8(+) lymphocytes). Patients with low islet function had increased levels of CD4(+)Ki67(+)cells (p â‰¤ 0.0001), while no difference was observed in CD8(+)Ki67(+) and CD56(+)Ki67(+) cells. IFN-Î³, IL-5, and IL-17 levels were increased in patients with low islet function, but IL-10 levels tended to be lower. IFN-Î³-ELISpot, proliferation, and cytokines were similarly accurate in predicting clinical outcome (AUC = 0.77 Â± 0.088, 0.85 Â± 0.084, and 0.88 Â± 0.074, respectively). Cellular immune reactivity against donor cells correlates with posttransplant islet function. The tested assays have the potential to be of substantial help in the management of islet graft recipients and deserve prospective validation.

Posttransplant cellular immune reactivity against donor antigen correlates with clinical islet transplantation outcome: towards a better posttransplant monitoring.

Le but de cette &eacute;tude &eacute;tait d&#39;&eacute;valuer l&#39;efficacit&eacute; des analyses cellulaires immunitaires dans la d&eacute;tection des alloreactivity apr&egrave;s la transplantation d&#39;&icirc;lots et de corr&eacute;ler ces r&eacute;sultats avec des r&eacute;sultats cliniques. Lymphocytaire mixte cultures ont &eacute;t&eacute; r&eacute;alis&eacute;es avec des cellules mononucl&eacute;aires de sang p&eacute;riph&eacute;rique des b&eacute;n&eacute;ficiaires (n = 14), donateurs, ou tierce partie. La r&eacute;activit&eacute; immunitaire a &eacute;t&eacute; &eacute;valu&eacute;e par la lib&eacute;ration d&#39;IFN-&gamma; (ELISpot), la prolif&eacute;ration des cellules (analyse de FACS pour Ki67) et quantification de cytokine (Bioplex). Fonction de l&#39;&icirc;lot en corr&eacute;lation avec le nombre de cellules s&eacute;cr&eacute;trices d&#39;IFN-&gamma; apr&egrave;s incubation avec les cellules du donneur (p = 0, 007, r = -0.50), mais pas avec des cellules de tierce partie (p = 0,61). De m&ecirc;me, un nombre &eacute;lev&eacute; de cellules en prolif&eacute;ration donneur sp&eacute;cifique a &eacute;t&eacute; associ&eacute; &agrave; une fonction d&#39;&icirc;lot faible (p = 0,006, r = -0,51). Les cellules en prolif&eacute;ration ont &eacute;t&eacute; principalement lymphocytes CD3(+)CD4(+) et CD3(-)CD56(+) des cellules tueuses naturelles (ayant un faible niveau de lymphocytes CD3(+)CD8(+)). Patients dont la fonction faible &icirc;lot avaient augment&eacute; le taux de CD4 (+) Ki67 (+) des cellules (p &le; 0,0001), alors qu&#39;aucune diff&eacute;rence a &eacute;t&eacute; observ&eacute;e dans les cellules CD8(+)Ki67(+) et CD56(+)Ki67(+). IFN-&gamma;, IL-niveaux 5 et IL-17 ont augment&eacute; chez les patients dont la fonction &icirc;lot faible, mais IL-10 niveaux avaient tendance &agrave; &ecirc;tre plus faibles. IFN-&gamma;-ELISpot, prolif&eacute;ration et cytokines &eacute;taient de m&ecirc;me pr&eacute;cis pour pr&eacute;dire les r&eacute;sultats cliniques (AUC = 0,77 &plusmn; 0,088, 0,85 &plusmn; 0,084 et &plusmn; 0,88 0,074, respectivement). Cellulaire r&eacute;activit&eacute; immunitaire contre les cellules du donneur est corr&eacute;l&eacute;e avec la fonction &icirc;lot posttransplantation. Les dosages test&eacute;s ont le potentiel pour &ecirc;tre d&#39;une aide substantielle dans la gestion des destinataires de greffe &icirc;lot et m&eacute;ritent la validation prospective.

Posttransplantation r&eacute;activit&eacute; immunitaire cellulaire contre l&#39;antig&egrave;ne du donneur est corr&eacute;l&eacute;e avec le r&eacute;sultat de transplantation clinique &icirc;lot : vers une meilleure surveillance posttransplantation.

Lo scopo del presente studio era di valutare l&#39;efficienza dell&#39;analisi immune cell-based nella rilevazione di alloreactivity dopo trapianto isolotto e correlare questi risultati con esito clinico. Colture miste linfocitarie sono state eseguite con cellule mononucleate di sangue periferico da destinatari (n = 14), donatori, o terze parti. La reattivit&agrave; immunitaria &egrave; stata valutata dal rilascio di IFN-&gamma; (ELISpot), la proliferazione cellulare (analisi FACS per Ki67) e quantificazione delle citochine (Bioplex). Funzione isolotto correlato con il numero di cellule che secernono IFN-&gamma; dopo incubazione con cellule del donatore (p = 0.007, r = -0,50), ma non con le cellule di terze parti (p = 0,61). Allo stesso modo, un elevato numero di cellule proliferanti specifiche del donatore &egrave; stato associato con una funzione di basso isolotto (p = 0.006, r = 060x). Le cellule in proliferazione sono stati principalmente linfociti CD3(+)CD4(+) e CD3(-)CD56(+) le cellule natural killer (con bassi livelli di linfociti CD3(+)CD8(+)). Pazienti con funzione di basso isolotto avevano aumentato i livelli di CD4 (+) Ki67 (+) cellule (p &le; 0,0001), mentre nessuna differenza &egrave; stata osservata nelle cellule CD8(+)Ki67(+) e CD56(+)Ki67(+). IFN-&gamma;, IL-5 e IL-17 livelli sono stati aumentati nei pazienti con funzione di basso isolotto, ma livelli di IL-10 tendevano ad essere pi&ugrave; basso. IFN-&gamma;-ELISpot, proliferazione e citochine erano similmente accurata nel predire l&#39;esito clinico (AUC = 0.77 &plusmn; 0.088, 0.85 &plusmn; 0.084 e 0.88 &plusmn; 0.074, rispettivamente). Cellulare reattivit&agrave; immunitaria contro le cellule del donatore correla con funzione isolotto infettivo. I dosaggi testati hanno il potenziale per essere di notevole aiuto nella gestione dei destinatari dell&#39;innesto isolotto e meritano convalida futuro.

Infettivo reattivit&agrave; immunitaria cellulare contro l&#39;antigene donatore correla con esito trapianto clinico isolotto: verso un migliore monitoraggio infettivo.

Humoral responses are central to the development of chronic autoimmune diseases such as systemic lupus erythematosus. Indeed, autoantibody deposition is responsible for tissue damage, the kidneys being one of the main target organs. As the source of pathogenic antibodies, plasma cells are therefore critical players in this harmful scenario, both at systemic and local levels. The aim of the present study was to analyze plasma cells in NZB/W lupus mice and to get a better understanding of the mechanisms underlying their involvement in the renal inflammation process. Using various techniques (i.e. flow cytometry, quantitative PCR, ELISpot), we identified and extensively characterized three plasma cell intermediates, according to their B220/CD138/MHCII expression levels. Each of these cell subsets displays specific proliferation and antibody secretion capacities. Moreover, we evidenced that the inflammation-related CXCR3 chemokine receptor is uniquely expressed by CD138(high)MHCII(+) plasma cells, which encompass both short- and long-lived cells and mostly produce IgG (auto)antibodies. Expression of CXCR3 allows efficient chemotactic responsiveness of these cells to cognate chemokines, which production is up-regulated in the kidneys of diseased NZB/W mice. Finally, using fluorescence and electron microscopy, we demonstrated the presence of CD138(+)CXCR3(+)IgG(+) cells in inflammatory areas in the kidneys, where they are very likely involved in the injury process. Thus, early differentiated CD138(high)MHCII(+) rather than terminally differentiated CD138(high)MHCII(low) plasma cells may be involved in the renal inflammatory injury in lupus, due to CXCR3 expression and IgG secretion.

Early differentiated CD138(high) MHCII+ IgG+ plasma cells express CXCR3 and localize into inflamed kidneys of lupus mice.

R&eacute;ponses humorales sont essentiels au d&eacute;veloppement de la chroniques maladies auto-immunes comme le lupus &eacute;ryth&eacute;mateux diss&eacute;min&eacute;. En effet, d&eacute;p&ocirc;ts d&#39;auto-anticorps sont responsable de l&eacute;sions tissulaires, les reins &eacute;tant l&#39;un des organes cibles principaux. Comme source d&#39;anticorps pathog&egrave;nes, cellules plasmatiques sont donc critiques joueurs dans ce sc&eacute;nario dangereux, tant au niveau syst&eacute;mique et locale. Le but de cette &eacute;tude &eacute;tait d&#39;analyser les cellules plasmatiques chez des souris NZB/W lupus et d&#39;obtenir une meilleure compr&eacute;hension des m&eacute;canismes qui sous-tendent leur implication dans le processus d&#39;inflammation r&eacute;nale. En utilisant diverses techniques (c.-&agrave;-d. cytom&eacute;trie, PCR quantitative, ELISpot), nous avons identifi&eacute; et largement caract&eacute;ris&eacute; trois plasmocyte interm&eacute;diaires, selon leurs niveaux d&#39;expression B220/CD138/MHCII. Chacun de ces sous-ensembles de cellule affiche prolif&eacute;ration sp&eacute;cifique et des capacit&eacute;s de s&eacute;cr&eacute;tion d&#39;anticorps. En outre, nous avons mis en &eacute;vidence que le r&eacute;cepteur de chimiokine CXCR3 li&eacute;es &agrave; l&#39;inflammation est particuli&egrave;rement bien exprim&eacute; par plasmocytes de CD138(High)mhcii(+), qui englobent les deux cellules &agrave; court et long - lived et surtout produire des anticorps IgG (auto). Expression de CXCR3 permet une r&eacute;activit&eacute; chimiotactique efficace de ces cellules &agrave; chimiokines apparent&eacute;s, dont la production est r&eacute;gul&eacute;e dans les reins des malades souris NZB/W. Enfin, &agrave; l&#39;aide de fluorescence et la microscopie &eacute;lectronique, nous avons d&eacute;montr&eacute; la pr&eacute;sence de cellules CD138(+)CXCR3(+)IgG(+) dans les zones inflammatoires dans les reins, o&ugrave; ils sont tr&egrave;s probablement impliqu&eacute;s dans le processus de la blessure. Ainsi, au d&eacute;but diff&eacute;renci&eacute;s CD138(high)MHCII(+) plut&ocirc;t que des cellules de plasma CD138(high)MHCII(low) terminalement diff&eacute;renci&eacute;es pourraient &ecirc;tre impliqu&eacute;s dans la l&eacute;sion inflammatoire r&eacute;nale dans le lupus, en raison de l&#39;expression CXCR3 et IgG de s&eacute;cr&eacute;tion.

D&eacute;but CD138(high) MHCII + IgG + plasma cellules diff&eacute;renci&eacute;es express CXCR3 et localiser dans les reins enflamm&eacute;s des souris de lupus.

Le risposte umorali sono centrali allo sviluppo di malattie croniche autoimmuni come il lupus eritematoso sistemico. Infatti, deposizione di autoanticorpi &egrave; responsabile per danni ai tessuti, i reni, essendo uno degli organi bersaglio principale. Come fonte di anticorpi patogeni, cellule del plasma sono quindi critici giocatori in questo scenario &egrave; nocivo, sia a livello sistemico e locale. Lo scopo del presente studio era di analizzare cellule del plasma in topi lupus NZB/W e per ottenere una migliore comprensione dei meccanismi sottostanti il loro coinvolgimento nel processo di infiammazione renale. Utilizzando varie tecniche (cio&egrave; flusso cytometry, PCR quantitativa, ELISpot), abbiamo individuato e ampiamente caratterizzata tre prodotti intermedi delle cellule di plasma, secondo i loro livelli di espressione B220/CD138/MHCII. Ognuno di questi sottoinsiemi di cella Visualizza proliferazione specifiche e capacit&agrave; di secrezione di anticorpi. Inoltre, abbiamo evidenziato che il recettore delle chemochine CXCR3 correlati a infiammazione &egrave; espressa in modo univoco da cellule del plasma CD138(high)MHCII(+), che comprendono sia cellule a breve e a lungo lived e soprattutto producono anticorpi IgG (auto). Espressione di CXCR3 permette efficiente risposta chemiotattica di queste cellule a cognate chemiochine, cui la produzione &egrave; up-regolata nei reni dei topi NZB/W malati. Infine, mediante fluorescenza e microscopia elettronica, ha dimostrato la presenza di cellule CD138(+)CXCR3(+)IgG(+) in aree infiammatorie nei reni, dove sono molto probabilmente coinvolti nel processo di lesioni. Cos&igrave;, presto differenziato CD138(high)MHCII(+) piuttosto che terminalmente differenziate CD138(high)MHCII(low) plasmacellule possono essere coinvolti nella lesione infiammatoria renale nel lupus, a causa di espressione CXCR3 e secrezione di IgG.

ArticleTotal : 2	Year
Publication title Cited by 31	2012
Publication title Cited by 1	2013

Article	Year
Functionalized carbon nanotubes are non-cytotoxic and preserve the functionality of primary immune cells. Nano letters\| PubMed ID: 16834443	2006
CXCR3, inflammation, and autoimmune diseases. Annals of the New York Academy of Sciences\| PubMed ID: 19758167	2009
Identification of new pathogenic players in lupus: autoantibody-secreting cells are present in nephritic kidneys of (NZBxNZW)F1 mice. Journal of immunology (Baltimore, Md. : 1950)\| PubMed ID: 20181885	2010
Detecting rejection after mouse Islet transplantation utilizing islet protein-stimulated ELISPOT. Cell transplantation\| PubMed ID: 21054945	2011
Immune monitoring of pancreatic islet graft: towards a better understanding, detection and treatment of harmful events. Expert opinion on biological therapy\| PubMed ID: 21073277	2011
Posttransplant cellular immune reactivity against donor antigen correlates with clinical islet transplantation outcome: towards a better posttransplant monitoring. Cell transplantation\| PubMed ID: 22963841	2012
Early differentiated CD138(high) MHCII+ IgG+ plasma cells express CXCR3 and localize into inflamed kidneys of lupus mice. PloS one\| PubMed ID: 23520491	2013

Stephanie Lacotte

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