Name: expanding cytotoxic t lymphocytes from umbilical cord blood that target cytomegalovirus, epstein-barr virus, and adenovirus
Uploaded: 2012-05-07T13:00:00
Description: Watch this Scientific Journal Video about Expanding Cytotoxic T Lymphocytes from Umbilical Cord Blood that Target Cytomegalovirus, Epstein-Barr Virus, and Adenovirus at JoVE.com

Ce travail a été soutenu par un Dan L. Duncan subvention de recherche concertée (CMB et EJS), le National Heart, Lung, and Blood Institute (US4HL081007), une Société de leucémie et lymphome prix de recherche clinique Scholar (CMB) et le National Cancer Institute (RO1 CA06150816; EJS).

1. Isolement de cellules mononucléaires (jour 0) <ol><li> Insérer pointe de raccord luer femelle au port de sortie de la fraction de 20% de l&#39;unité de sang de cordon, fixer une seringue et retirer le sang. Transférer le sang décongelé à 20 mL de RPMI réchauffé dans 50 ml tube à centrifuger. Rincer sac de sang de cordon ombilical avec 5 ml de RPMI et transférer dans le tube à centrifugation. </li><li> Centrifuger les cellules pendant 10 minutes à 400 x g. Aspirer le surnageant. </li><li> Remettre en su...

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<html> Les stratégies actuelles visant à lutter contre les infections virales après CBT peuvent être efficaces, mais ils sont associés à des toxicités importantes, sont coûteux, et ne confèrent pas une protection à long terme contre l&#39;infection plus tard. En fait, l&#39;utilisation de certains médicaments antiviraux peuvent limiter l&#39;expansion des cellules T spécifiques du virus qui seraient autrement protection. 14 Une autre option est l&#39;infusion de cellules virales spécifiques des ba...

<table border="1"><tbody><tr><td> Nom du réactif </td><td> Entreprise </td><td> Numéro de catalogue </td></tr><tr><td> RPMI 1640 </td><td> Invitrogen </td><td> 21870-076 </td></tr><tr><td> DC médias </td><td> CellGenix </td><td> 20801-0500 </td></tr><tr><td> EHAA (Moyen cliquable) </td><td> Irvine Scientific </td><td> 9195 </td></tr><tr><td> Sérum humain </td><td> Gémeaux Produits Bio </td><td> 100-110 </td></tr><tr><td> Perméables aux gaz Cultureware 18 </td><td> Wilson-Loup </td><td> 80040S </td></tr><tr><td> IL-2 </td><td> Chiron (TCH pharmacie) </td><td></td></tr><tr><td> IL-12 </td><td> NCI / CTEP </td><td></td></tr><tr><td> IL-15 </td><td> CellGenix </td><td> 1013-050 </td></tr><tr><td> IL-7 </td><td> R &amp; D </td><td> AFL207 </td></tr><tr><td> IL-1beta </td><td> R &amp; D </td><td> AFL201 </td></tr><tr><td> IL-6 </td><td> CelluleGenix </td><td> 1004-050 </td></tr><tr><td> GM-CSF </td><td> TCH pharmacie </td><td></td></tr><tr><td> IL-4 </td><td> R &amp; D </td><td> <a href="http://www.rndsystems.com/pdf/AFL204.pdf" target="_blank">AFL204</a> </td></tr><tr><td> TNF-alpha </td><td> R &amp; D </td><td> AFL210 </td></tr><tr><td> Ad5f35pp65 </td><td> BCM CAGT installation de production de vecteur </td><td></td></tr><tr><td> Transfert au plasma avec adaptateur Luer femelle </td><td> Charte médical </td><td> 89-550-66j </td></tr><tr><td> Lymphoprep </td><td> Nycomed </td><td> 1114550 </td></tr></tbody></table>

expanding cytotoxic t lymphocytes from umbilical cord blood that target cytomegalovirus, epstein-barr virus, and adenovirus

Les infections virales après transplantation de cellules souches sont parmi les causes les plus fréquentes de décès, en particulier après le sang du cordon (CB) transplantation (TCC) où le CB ne contient pas un nombre appréciable de cellules T du virus expérimentés qui peuvent protéger le destinataire de l&#39;infection 1. - 4 Nous et d&#39;autres ont montré que le virus de CTL spécifique généré à partir de donneurs séropositifs et infusé au bénéficiaire sont en sécurité et de protection. 8.5 Cependant, jusqu&#39;à récemment, le virus de cellules T spécifiques ne pouvaient pas être générée à partir de sang de cordon, probablement en raison de la absence de virus-cellules T spécifiques de la mémoire. Dans un effort pour mieux imiter les conditions d&#39;amorçage in vivo des cellules T naïves, nous avons établi une méthode qui a utilisé des cellules dendritiques dérivées de CB (DC) transduites avec un vecteur adénoviral (Ad5f35pp65) contenant l&#39;antigène pp65 CMV immunodominant, par conséquent, de conduire la spécificité des lymphocytes T vers le CMV et l&#39;adénovirus. 9 Au début, nous utilisons ces machinesPED structurés, ainsi que CB dérivées des cellules T en présence des cytokines IL-7, IL-12 et IL-15. Au 10 secondes, nous avons utilisé la stimulation transformées par EBV des cellules B, ou EBV-LCL, qui expriment à la fois latente et lytiques antigènes d&#39;EBV. Ad5f35pp65-transduites EBV-LCL sont utilisés pour stimuler les cellules T en présence d&#39;IL-15 à la seconde stimulation. Stimulations ultérieures utilisent Ad5f35pp65 transduites par EBV-LCL et de l&#39;IL-2. De 6 CB 50x10 cellules mononucléaires nous sommes en mesure de générer plus de 150 x 10 6 virus de cellules T spécifiques de l&#39;antigène qui lysent des cibles pulsées et des cytokines libération en réponse à une stimulation antigénique. 11 Ces cellules ont été fabriqués d&#39;une manière conforme aux BPF en utilisant seulement la fraction 20% d&#39;une unité de fractionnement de sang de cordon et qui ont été convertis pour un usage clinique.

Watch this Scientific Journal Video about Expanding Cytotoxic T Lymphocytes from Umbilical Cord Blood that Target Cytomegalovirus, Epstein-Barr Virus, and Adenovirus at JoVE.com

Expanding Cytotoxic T Lymphocytes from Umbilical Cord Blood that Target Cytomegalovirus, Epstein-Barr Virus, and Adenovirus

Nous décrivons ici la première bonne pratique de fabrication (BPF) conforme à la méthode de production spécifiques du virus de lymphocytes T cytotoxiques (CTL) à partir de sang de cordon ombilical, source de cellules T naïves principalement.

L&#39;expansion des lymphocytes T cytotoxiques à partir de sang de cordon ombilical qui cytomégalovirus cible, le virus Epstein-Barr Virus et Adénovirus

Virus infections after stem cell transplantation are among the most common causes of death, especially after cord blood (CB) transplantation (CBT) where ...

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