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

Este trabalho foi financiado por uma L. Dan Duncan colaborativo bolsa de investigação (CMB e EJS), o Instituto Nacional do Coração, Pulmão e Sangue (US4HL081007), uma Leucemia e Linfoma Sociedade prêmio Acadêmico de Pesquisa Clínica (CMB), e do Instituto Nacional do Câncer (RO1 CA06150816; EJS).

1. Isolamento de células mononucleares (dia 0) <ol><li> Insira espiga de adaptador luer fêmea no orifício de saída da fracção de 20% da unidade de sangue do cordão umbilical, anexar e remover a seringa do sangue. Transferir sangue descongeladas a 20 mL de RPMI aquecido no tubo de centrífuga de 50 mL. Enxaguar saco de sangue do cordão umbilical com 5 mL de meio RPMI e transferir para o tubo de centrífuga mesmo. </li><li> Centrifugar as células durante 10 minutos a 400 x g. Aspirar o sobrenadante. </li><li> R...

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<html> As estratégias actuais no sentido de controlar as infecções virais após TCC pode ser eficaz, mas estão associadas com toxicidade significativa, são caros, e não conferem protecção a longo prazo contra a infecção posterior. Na verdade, o uso de algumas drogas antivirais podem limitar a expansão de células T específicas de vírus que de outra forma seria protectora 14. Outra opção é a infusão de doador derivados de vírus-células T específicas. Nós e outros autores mostraram que essas...

<table border="1"><tbody><tr><td> Nome do reagente </td><td> Companhia </td><td> Número de catálogo </td></tr><tr><td> RPMI 1640 </td><td> Invitrogen </td><td> 21870-076 </td></tr><tr><td> DC mídia </td><td> CellGenix </td><td> 20801-0500 </td></tr><tr><td> EHAA (meio de Click) </td><td> Irvine Scientific </td><td> 9195 </td></tr><tr><td> Soro Humano </td><td> Gémeos Bio Products </td><td> 100-110 </td></tr><tr><td> Gás Permeável Cultureware 18 </td><td> Wilson-Wolf </td><td> 80040S </td></tr><tr><td> IL-2 </td><td> Chiron (TCH Farmácia) </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> CélulaGenix </td><td> 1004-050 </td></tr><tr><td> GM-CSF </td><td> TCH Farmácia </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-alfa </td><td> R &amp; D </td><td> AFL210 </td></tr><tr><td> Ad5f35pp65 </td><td> BCM CAGT Facilidade de Produção Vector </td><td></td></tr><tr><td> Transferência de plasma conjunto com adaptador luer fêmea </td><td> Carta de Medicina </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

Infecções por vírus após o transplante de células-tronco estão entre as causas mais comuns de morte, especialmente depois do sangue de cordão umbilical (CB) transplante (CBT), onde o BC não contém números apreciáveis ​​de células T-vírus experientes que podem proteger o destinatário da infecção 1. - 4 Nós e outros autores mostraram que o vírus-specific CTL gerados a partir de dadores seropositivos e injectada para o receptor são seguras e de protecção. 5-8 No entanto, até há pouco tempo, as células T específicas para o vírus não pode ser gerado a partir de sangue do cordão umbilical, provavelmente devido ao ausência de vírus específicos de células T de memória. Num esforço para melhor imitar a condições de estímulo in vivo de células T naive, foi estabelecido um método que utilizou células CB-derivadas dendríticas (DC) transduzida com um vector adenoviral (Ad5f35pp65) contendo o antigénio imunodominante CMV pp65, portanto dirigir a especificidade de células T para CMV e adenovírus. 9 No início, nós usamos estes maDCs estruturadas, bem como derivados de CB células T na presença das citocinas IL-7, IL-12 e IL-15. 10 Na segunda estimulação foi utilizado EBV-células B transformadas, ou EBV-LCL, que expressam tanto latente e antígenos líticos EBV. Ad5f35pp65-EBV-LCL transduzidas são usados ​​para estimular as células T na presença de IL-15 na segunda estimulação. Estimulações subsequentes usam Ad5f35pp65-EBV-LCL transduzidas e IL-2. De 50x10 6 CB células mononucleares que são capazes de gerar mais de 150 x 10 6 de vírus específicos de células T que lisam alvos pulsados ​​antigénio e libertação de citocinas em resposta a estimulação antigénica. 11 Estas células foram fabricadas de um modo compatível com GMP utilizando somente a fracção de 20% de uma unidade de fraccionamento de sangue do cordão e foram traduzidos para o uso clínico.

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

Aqui descrevemos a prática de fabricação primeiro (BPF) compatível com método de produção de vírus específicos linfócitos T citotóxicos (CTL) de sangue de cordão umbilical, fonte de células T predominantemente ingênua.

Expandindo linfócitos T citotóxicos do sangue do cordão umbilical que citomegalovírus Target, vírus de Epstein-Barr, e Adenovirus

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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