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

この作品は、ダン·L·ダンカン共同研究助成金（CMBとEJS）、国立心肺血液研究所（US4HL081007）、白血病とリンパ腫協会臨床研究奨学生賞（CMB）、および国立癌研究所によってサポートされていました（RO1 CA06150816; EJS）。

1。単核細胞分離（0日目） <ol><li>臍帯血ユニットの20％分の出口ポートにメスルアーアダプタのスパイクを挿入し、シリンジを接続して、血液を除去する。 50 mLの遠心分離管中で温めたRPMI 20mLに解凍した血液を転送します。 RPMI 5mLで臍帯血袋を洗浄し、同じ遠心チューブに移す。 </li><li> 400 x gで10分間細胞を遠心分離します。上清を吸引します。 </li><li>暖かいRPMI 20mlに細胞を再懸濁する。 50...

<ol>
	<li>Kennedy-Nasser, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparable+outcome+of+alternative+donor+and+matched+sibling+donor+hematopoietic+stem+cell+transplant+for+children+with+acute+lymphoblastic+leukemia+in+first+or+second+remission+using+alemtuzumab+in+a+myeloablative+conditioning+regimen.">Comparable outcome of alternative donor and matched sibling donor hematopoietic stem cell transplant for children with acute lymphoblastic leukemia in first or second remission using alemtuzumab in a myeloablative conditioning regimen.</a> Biol. Blood Marrow Transplant. 14, 1245-1245 (2008).</li><li>Hanley, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+clinical+outcomes+using+adoptively+transferred+immune+cells+from+umbilical+cord+blood.">Improving clinical outcomes using adoptively transferred immune cells from umbilical cord blood.</a> Cytotherapy. 12, 713 (2010).</li><li>Szabolcs, P., Cairo, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unrelated+umbilical+cord+blood+transplantation+and+immune+reconstitution.">Unrelated umbilical cord blood transplantation and immune reconstitution.</a> Semin. Hematol. 47, 22 (2010).</li><li>Canto, E., Rodriguez-Sanchez, J. L., Vidal, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinctive+response+of+naive+lymphocytes+from+cord+blood+to+primary+activation+via+TCR.">Distinctive response of naive lymphocytes from cord blood to primary activation via TCR.</a> J. Leukoc. Biol. 74, 998-998 (2003).</li><li>Leen, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoculture-derived+T+lymphocytes+specific+for+multiple+viruses+expand+and+produce+clinically+relevant+effects+in+immunocompromised+individuals.">Monoculture-derived T lymphocytes specific for multiple viruses expand and produce clinically relevant effects in immunocompromised individuals.</a> Nat. Med. 12, 1160-1160 (2006).</li><li>Riddell, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restoration+of+viral+immunity+in+immunodeficient+humans+by+the+adoptive+transfer+of+T+cell+clones.">Restoration of viral immunity in immunodeficient humans by the adoptive transfer of T cell clones.</a> Science. 257, 238 (1992).</li><li>O'Reilly, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adoptive+transfer+of+antigen-specific+T-cells+of+donor+type+for+immunotherapy+of+viral+infections+following+allogeneic+hematopoietic+cell+transplants.">Adoptive transfer of antigen-specific T-cells of donor type for immunotherapy of viral infections following allogeneic hematopoietic cell transplants.</a> Immunol. Res. 38, 237-237 (2007).</li><li>Peggs, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adoptive+cellular+therapy+for+early+cytomegalovirus+infection+after+allogeneic+stem-cell+transplantation+with+virus-specific+T-cell+lines.">Adoptive cellular therapy for early cytomegalovirus infection after allogeneic stem-cell transplantation with virus-specific T-cell lines.</a> Lancet. 362, 1375-1375 (2003).</li><li>Sili, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+expansion+of+dendritic+cell-primed+polyclonal+human+cytotoxic+T-lymphocyte+lines+using+lymphoblastoid+cell+lines+for+adoptive+immunotherapy.">Large-scale expansion of dendritic cell-primed polyclonal human cytotoxic T-lymphocyte lines using lymphoblastoid cell lines for adoptive immunotherapy.</a> J. Immunother. 26, 241 (2003).</li><li>Bollard, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Good+manufacturing+practice-grade+cytotoxic+T+lymphocytes+specific+for+latent+membrane+proteins+(LMP)-1+and+LMP2+for+patients+with+Epstein-Barr+virus-associated+lymphoma.">Good manufacturing practice-grade cytotoxic T lymphocytes specific for latent membrane proteins (LMP)-1 and LMP2 for patients with Epstein-Barr virus-associated lymphoma.</a> Cytotherapy. 13, 518 (2011).</li><li>Hanley, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functionally+active+virus-specific+T+cells+that+target+CMV,+adenovirus,+and+EBV+can+be+expanded+from+naive+T-cell+populations+in+cord+blood+and+will+target+a+range+of+viral+epitopes.">Functionally active virus-specific T cells that target CMV, adenovirus, and EBV can be expanded from naive T-cell populations in cord blood and will target a range of viral epitopes.</a> Blood. 114, 1958 (1958).</li><li>Hanley, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expansion+of+T+cells+targeting+multiple+antigens+of+cytomegalovirus,+Epstein-Barr+virus+and+adenovirus+to+provide+broad+antiviral+specificity+after+stem+cell+transplantation.">Expansion of T cells targeting multiple antigens of cytomegalovirus, Epstein-Barr virus and adenovirus to provide broad antiviral specificity after stem cell transplantation.</a> Cytotherapy. , (2011).</li><li>Gerdemann, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+Multivirus-specific+T+Cells+to+Prevent/treat+Viral+Infections+after+Allogeneic+Hematopoietic+Stem+Cell+Transplant.">Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant.</a> J. Vis. Exp. (51), e2736 (2011).</li><li>Mori, T., Kato, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytomegalovirus+infection/disease+after+hematopoietic+stem+cell+transplantation.">Cytomegalovirus infection/disease after hematopoietic stem cell transplantation.</a> Int. J. Hematol. 91, 588 (2010).</li><li>Einsele, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infusion+of+cytomegalovirus+(CMV)-specific+T+cells+for+the+treatment+of+CMV+infection+not+responding+to+antiviral+chemotherapy.">Infusion of cytomegalovirus (CMV)-specific T cells for the treatment of CMV infection not responding to antiviral chemotherapy.</a> Blood. 99, 3916 (2002).</li><li>Bao, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expansion+of+cytomegalovirus+pp65+and+IE-1+specific+cytotoxic+T+lymphocytes+for+cytomegalovirus-specific+immunotherapy+following+allogeneic+stem+cell+transplantation.">Expansion of cytomegalovirus pp65 and IE-1 specific cytotoxic T lymphocytes for cytomegalovirus-specific immunotherapy following allogeneic stem cell transplantation.</a> Biol. Blood Marrow Transplant. 14, 1156 (2008).</li><li>Shpall, E. J., Bollard, C. M., Brunstein, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+cord+blood+transplant+therapies.">Novel cord blood transplant therapies.</a> Biol. Blood Marrow Transplant. 17, S39-S45 (2011).</li><li>Vera, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accelerated+production+of+antigen-specific+T+cells+for+preclinical+and+clinical+applications+using+gas-permeable+rapid+expansion+cultureware+(G-Rex.">Accelerated production of antigen-specific T cells for preclinical and clinical applications using gas-permeable rapid expansion cultureware (G-Rex.</a> J. Immunother. 33, 305 (2010).</li></ol>

<html> CBTの後にウイルス感染を制御することを目的と現在の戦略が効果的ですが、彼らはかなりの毒性に関連付けられている、高価なもので、その後の感染症に対する長期の保護を与えない。実際には、いくつかの抗ウイルス薬の使用は、他の方法で保護されるであろうウイルス特異的T細胞の増殖を制限することがあります。14もう一つのオプションは、ドナー由来のウイルス特異的T...

<table border="1"><tbody><tr><td> 試薬の名称 </td><td> 会社 </td><td> カタログ番号 </td></tr><tr><td> RPMI 1640 </td><td>インビトロジェン</td><td> 21870-076 </td></tr><tr><td> DCのメディア</td><td> CellGenix </td><td> 20801-0500 </td></tr><tr><td> EHAA（クリックのミディアム） </td><td>アーバイン·サイエンティフィック</td><td> 9195 </td></tr><tr><td>ヒト血清</td><td>ジェミニバイオ製品</td><td> 100から110 </td></tr><tr><td>ガス透過性Cultureware 18 </td><td>ウィルソン - ウルフ</td><td> 80040S </td></tr><tr><td> IL-2の</td><td>カイロン（TCH薬局） </td><td></td></tr><tr><td> IL-12の</td><td> NCI / CTEP </td><td></td></tr><tr><td> IL-15の</td><td> CellGenix </td><td> 1013-050 </td></tr><tr><td> IL-7の</td><td>研究開発</td><td> AFL207 </td></tr><tr><td> IL-1βの</td><td>研究開発</td><td> AFL201 </td></tr><tr><td> IL-6の</td><td>セルGenix </td><td> 1004-050 </td></tr><tr><td> GM-CSF </td><td> TCH薬局</td><td></td></tr><tr><td> IL-4 </td><td>研究開発</td><td> <a href="http://www.rndsystems.com/pdf/AFL204.pdf" target="_blank">AFL204</a> </td></tr><tr><td> TNF-α </td><td>研究開発</td><td> AFL210 </td></tr><tr><td> Ad5f35pp65 </td><td> BCM CAGTベクトル製造施設</td><td></td></tr><tr><td>プラズマ転送が雌型ルアーアダプターを使用して設定</td><td>チャーターメディカル</td><td> 89から550-66j </td></tr><tr><td> Lymphoprep </td><td>ナイコメッド</td><td> 1114550 </td></tr></tbody></table>

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

幹細胞移植後のウイルス感染は、特にCBは感染から受信者を保護することができ、ウイルス経験豊富なT細胞のかなりの数字を含まない臍帯血（CB）移植（CBT）の後に、死の最も一般的な原因の一つである1 -我々や他のウイルス特異的CTL抗体陽性ドナーから生成され、受信者に注入が安全かつ保護的であることが示されている。5-8しかし、最近まで、ウイルス特異的T細胞が臍帯血から生成することができませんでした、おそらくのために4ウイルス特異的メモリーT細胞が存在しない。 良いナイーブT細胞の in vivoでのプライミング条件で模倣するための努力で、我々は、したがってT細胞特異性を駆動する、CB由来樹状細胞（DC）は免疫CMV抗原pp65抗原を含むアデノウイルスベクター（Ad5f35pp65）で形質導入に使用する方法を確立し開始時9。CMV、およびアデノウイルスに向かって、我々はこれらのmaを使うtured樹と同様に、CB由来T細胞サイトカインの存在下で、IL-7、IL-12およびIL-15 10我々は両方を発現するEBV形質転換B細胞、またはEBV-LCLを、使用される第二の刺激で潜伏と溶解EBV抗原。 Ad5f35pp65形質のEBV-LCLは第2の刺激でのIL-15の存在下でT細胞を刺激するために使用されます。後続の刺激は、EBV-LCLおよびIL-2 Ad5f35pp65形質を使用しています。 50x10 6、CB単核細胞から、我々は抗原刺激に応答して、抗原をパルス目標とリリースサイトカインを溶解さ150×10 6のウイルス特異的T細胞の上方に生成することができます。11これらの細胞は、使用して、GMPに準拠した方法で製造されただけ画し臍帯血ユニットの20％画分および臨床使用のために翻訳されています。

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

ここでは、最初の良い製造基準（GMP）に準拠した臍帯血、主にナイーブT細胞のソースからウイルス特異的細胞傷害性Tリンパ球（CTL）を製造する方法を説明します。

臍帯血から細胞傷害性Tリンパ球の拡大をそのターゲットサイトメガロウイルス、エプスタイン - バーウイルス、およびアデノウイルス

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

Research

JoVE Journal

Immunology and Infection

Preparation and Use of HIV-1 Infected Primary CD4+ T-Cells as Target Cells in Natural Killer Cell Cytotoxic Assays

ナチュラルキラー細胞傷害性アッセイにおけるHIV - 1感染プライマリCD4 + T細胞を標的細胞としての準備と使用

 Natural killer (NK) cells are a vital component of the innate immune response to virus-infected cells. It is important to understand the ability of NK ...

Methods to Assess Beta Cell Death Mediated by Cytotoxic T Lymphocytes

傷害性Tリンパ球によって媒介ベータ細胞死を評価するための方法

Type 1 diabetes (T1D) is a T cell mediated autoimmune disease.  During the pathogenesis, patients become progressively more insulinopenic as 
insulin ...

Introducing Shear Stress in the Study of Bacterial Adhesion

細菌付着の研究ではせん断応力を導入する

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

Establishment of Epstein-Barr Virus Growth-transformed Lymphoblastoid Cell Lines

Epstein - Barrウイルスの成長形質転換リンパ芽球様細胞株の樹立

Infection of B cells with Epstein-Barr virus (EBV) leads to proliferation and subsequent immortalization, resulting in establishment of lymphoblastoid ...

Clinical Application of Sleeping Beauty and Artificial Antigen Presenting Cells to Genetically Modify T Cells from Peripheral and Umbilical Cord Blood

Clinical Application of Sleeping Beauty and Artificial Antigen Presenting Cells to Genetically Modify T Cells from Peripheral and Umbilical Cord Blood

の臨床応用<em&gt;眠れる森の美女</em遺伝的に末梢及び臍帯血由来のT細胞を変更する&gt;と人工抗原提示細胞

The potency of clinical-grade T cells can be improved by combining gene therapy with immunotherapy to engineer  a biologic product with the potential for ...

Isolation of Precursor B-cell Subsets from Umbilical Cord Blood

臍帯血から前駆B細胞サブセットの分離

Umbilical cord blood is highly enriched for hematopoietic progenitor cells at different lineage commitment stages. We have developed a protocol for ...

Isolation and Th17 Differentiation of Na&iuml;ve CD4 T Lymphocytes

Isolation and Th17 Differentiation of NaÃƒÆ’Ã‚Â¯ve CD4 T Lymphocytes

離とナイーブCD4 Tリンパ球のTh17分化

Th17 cells are a distinct subset of T cells that have been found to produce interleukin 17 (IL-17), and differ in function from the other T cell subsets ...

Development of an IFN-&#947; ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

Development of an IFN-ÃƒÅ½Ã‚Â³ ELISpot Assay to Assess Varicella-Zoster Virus-specific Cell-mediated Immunity Following Umbilical Cord Blood Transplantation

臍帯血移植後の水痘帯状疱疹ウイルス特異的細胞性免疫を評価するために、IFN-γELISPOTアッセイの開発

Varicella zoster virus (VZV) is a significant cause of morbidity and mortality following umbilical cord blood transplantation (UCBT). For this reason, ...

Generation of Human Alloantigen-specific T Cells from Peripheral Blood

末梢血からのヒトアロ抗原特異的T細胞の生成

The study of human T lymphocyte biology often involves examination of responses to activating ligands. T cells recognize and respond to processed peptide ...

Radial Mobility and Cytotoxic Function of Retroviral Replicating Vector Transduced, Non-adherent Alloresponsive T Lymphocytes

ラジアルモビリティおよびレトロウイルス複製ベクター形質導入、非付着Alloresponsive Tリンパ球の細胞傷害機能

We report a novel adaptation of the Radial Monolayer Cell Migration assay, first reported to measure the radial migration of adherent tumor cells on ...