Induction of human cytotoxic T cells by peptide-pulsed universal antigen presenting cells

• Project/Area Number: 16390294
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Tohoku University
• Principal Investigator: TSUCHIYA Shigeru Tohoku University, Graduate School of Medicine, Professor, 大学院医学系研究科, 教授 (30124605)
• Co-Investigator(Kenkyū-buntansha): KUMAKI Satoru Tohoku University, Graduate School of Medicine, Associate Professor, 大学院医学系研究科, 助教授 (20311566) ; MINEGISHI Masayoshi Tohoku University, Hospital, Associate Professor, 病院・助教授 (20211592) ; SASAHARA Yoji Tohoku University, Hospital, Research Associate, 病院・助手 (60372314)
• Project Period (FY): 2004 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥14,400,000 (Direct Cost: ¥14,400,000); Fiscal Year 2006: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2005: ¥3,500,000 (Direct Cost: ¥3,500,000); Fiscal Year 2004: ¥7,400,000 (Direct Cost: ¥7,400,000)
• Keywords: peptide antigens / B-LCL / cytotoxic T cells / tetoramer / EBV / 抗原提示細胞 / 人工抗原提示 / 汎用性人工抗原提示細胞 / HLA-A24 / CD28抗体 / フローサイトメーター / 細胞障害活性定量

Research Abstract

We examined the presence of EB virus (EBV) specific cytotoxic T cells, induction of EBV specific CTLs and the cytotoxic activities of induced CTLs in the peripheral CD8+ cells from the patient with chronic active EBV infection (CAEBV) and the results were as follows,
1.Demonstration of EBV specific CTLs by HLAA24-specific and EBV-specific tetramer analysis
BRLF1 peptides specific CTLs were found in the peripheral lymphocytes of healthy volunteers and those of patients with infectious mononucleosis. Interestingly, we were also able to demonstrate BRLF1 peptides specific CTLs in the peripheral blood of the patients with CAEBV.
2.Induction of antigen specific CTLs :
We used HLAA24 positive autologous B lymphobastoid cells (B-LCLs) as antigen presenting cells and peripheral mononuclear cells were stimulated with B-LCLs, pulsed with or without BRLF1 peptide. We were successfully able to induce BRLF1 tetramer positive CTLs even in the absence of BRLF1 specific peptides. This suggested that enoug h BRLF1 specific stimulation were given from the EBV infected B-LCLs to specific CTLs without further peptide pulse. These CD8 positive BRLF1 tetramer specific induction and proliferation of CTLs were observed in the mononuclear cells from both healthy volunteers and the CAEBV patient.
3.Cytotoxic activity of induced CTLs using interferon gamma producing ability of CTLs :
Then we examined the cytotoxic activities of induced CTLs. First we confirmed that BRLF1 tetramer specific CD8+ CTLs from healthy volunteers could produce interferon gamma only after BRLF1 specific peptides. We found that the mononuclear cells from a CAEBV patient also produce BRLF1 tetramer specific CTLs after stimulation with specific peptides. However, BRLF1 specific CTLs from the patient produced interferon gamma only approximately 30%, as compared with those from the healthy volunteers approximately 50%.
These results indicated the possibility that mononuclear cells from both healthy volunteers and the CAEBV patients are able to induce and proliferate specific CTLs, but cytotoxic activities of specific CTLs from the patient is reduced as compared with those of healthy volunteers. To elucidate the mechanism of the reduced CTL-activity of the CAEBV patient will have great implications to understand the etiology of CAEBV and the CTL therapy.

Research Products

• [Journal Article] Quality of Umbilical Cord Blood CD34+ Cells in a Double-Compartment Freezing Bag Cryopreserved without a Rate-Controlled Programmed Freezer. (2007)
  • Author(s): Minegishi M et al.
  • Journal Title: Int J Hematol 652 Pages: 78-84
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Cell cycle-dependent oscillation of GATA2 expression in hematopoietic cells. (2007)
  • Author(s): Koga S et al.
  • Journal Title: Blood Jan 25
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Quality of Umbilical Cord Blood CD34+ Cells in a Double-Compartment Freezing Bag Cryopreserved without a Rate-Controlled Programmed Freezer. (2007)
  • Author(s): Minegishi M, Itoh T, Fukawa N, Kitaura T, Miura J, Takahashi H, Suzuki A, Kudo Y, Narita A, Sato Y, Suzuki M, Watanabe T, Wada Y, Takeyama Y, Tsuchiya S.
  • Journal Title: Int J Hematol. 652 Pages: 78-84
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Cell cycle-dependent oscillation of GATA2 expression in hematopoietic cells. (2007)
  • Author(s): Koga S, Yamaguchi N, Abe T, Minegishi M, Tsuchiya S, Yamamoto M, Minegishi N.
  • Journal Title: Blood [Epub ahead of print]
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Quality of Umbilical Cord Blood CD34+ Cells in a Double-Compartment Freezing Bag Cryopreserved without a Rate-Controlled Programmed Freezer. (2007)
  • Author(s): Minegishi M, et al.
  • Journal Title: Int J Hematol 652 Pages: 78-84
• [Journal Article] Cell cycle-dependent oscillation of GATA2 expression in hematopoietic cells. (2007)
  • Author(s): Koga S, et al.
  • Journal Title: Blood Jan 25(E pub)
• [Journal Article] Human tyrosine kinase 2 deficiency reveals its requisite roles in multiple cytokine signals involved in innate and acquired immunity. (2006)
  • Author(s): Minegishi Y et al.
  • Journal Title: Immunity 25 Pages: 745-755
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] A second-site mutation in the initiation codon of WAS (WASP) gene results in expansion of subsets of lymphocytes in an Wiskott-Aldrich Syndrome patient. (2006)
  • Author(s): Du W et al.
  • Journal Title: Hum Mutat 272 Pages: 370-375
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Phage φC31 integrase-mediated genomic integration of the common cytokine receptor gamma chain n human T-cells. (2006)
  • Author(s): Ishikawa Y et al.
  • Journal Title: J Gene Med 8 Pages: 646-653
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Application of HSVtk suicide gene to X-SCID gene therapy : Ganciclovir treatment offsets gene corrected X-SCID B cells. (2006)
  • Author(s): Uchiyama T et al.
  • Journal Title: Biochem Biophys Res Commun 341 Pages: 391-398
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Human tyrosine kinase 2 deficiency reveals its requisite roles in multiple cytokine signals involved in innate and acquired immunity. (2006)
  • Author(s): Minegishi Y, Saito M, Morio T, Watanabe K, Agematsu K, Tsuchiya S, Takada H, Hara T, Kawamura N, Ariga T, Kaneko H, Kondo N, Tsuge I, Yachie A, Sakiyama Y, Iwata T, Bessho F, Ohishi T, Joh K, Imai K, Kogawa K, Shinohara M, Fujieda M, Wakiguchi H, Pasic S, Abinun M, Ochs HD, Renner ED, Jansson A, Belohradsky BH, Metin A, Shimizu N, Mizutani S, Miyawaki T, Nonoyama S, Karasuyama H.
  • Journal Title: Immunity. 25 Pages: 745-55
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A second-site mutation in the initiation codon of WAS (WASP) gene results in expansion of subsets of lymphocytes in an Wiskott-Aldrich Syndrome patient. (2006)
  • Author(s): Du W, Kumaki S, Uchiyama S, Yachi A, Chung Yeng L, Kawai S, Minegishi M, Ramesh N, Geha RS, Sasahara Y, Tsuchiya S.
  • Journal Title: Hum Mutat. 27 Pages: 370-5
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Phage ψC31 integrase-mediated genomic integration of the common cytokine receptor gamma chain n human T-cells. (2006)
  • Author(s): Ishikawa Y, Tanaka N, Murakami K, Uchiyama T, Kumaki S, Tsuchiya 5, Kugoh H, Oshima M, Calos MP, Sugamura K.
  • Journal Title: J Gene Med 8 Pages: 646-53
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Application of HSVtk suicide gene to X-SCID gene therapy : Ganciclovir treatment offsets gene corrected X-SCID B cells. (2006)
  • Author(s): Uchiyama T, Kumaki S, Ishikawa Y, Onodera M, Sato M, Du W, Sasahara Y, Tanaka N, Sugamura K, Tsuchiya S.
  • Journal Title: Biochem Biophys Res Commun. 341 Pages: 391-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Human tyrosine kinase 2 deficiency reveals its requisite roles in multiple cytokine signals involved in innate and acquired immunity. (2006)
  • Author(s): Minegishi Y, et al.
  • Journal Title: Immunity 25 Pages: 745-755
• [Journal Article] A second-site Hum Mutat. 27 : 370-5, 2006 mutation in the initiation codon of WAS (WASP) gene results in expansion of subsets of lymphocytes in an Wiskott-Aldrich Syndrome patient. (2006)
  • Author(s): Du W, et al.
  • Journal Title: Hum Mutat 272 Pages: 370-375
• [Journal Article] Phage φC31 integrase-mediated genomic integration of the common cytokine receptor gamma chain n human T-cells. (2006)
  • Author(s): Ishikawa Y, et al.
  • Journal Title: J Gene Med 8 Pages: 646-653
• [Journal Article] Application of HSVtk suicide gene to X-SCID gene therapy : Ganciclovir treatment offsets gene corrected X-SCID B cells. (2006)
  • Author(s): Uchiyama T, et al.
  • Journal Title: Biochem Biophys Res Commun 341 Pages: 391-398
• [Journal Article] Application of HSVtk suicide gene to X-SCID gene therapy : Ganciclovir treatment offsets gene corrected X-SCID B cells. (2006)
  • Author(s): Uchiyama T, Kumaki S, Ishikawa Y, Onodera M, Sato M, Du W, Sasahara Y, Tanaka N, Sugamura K, Tsuchiya S
  • Journal Title: BBRC 341 Pages: 391-398
• [Journal Article] Phage ΦC31 integrase-mediated genomic integration of the common cytokine receptor gamma chain n human T-cells. (2006)
  • Author(s): Ishikawa Y, Tanaka N, Murakami K, Uchiyama T, Kumaki S, Tsuchiya S, Kugoh H, Oshima M, Calos MP, Sugamura K
  • Journal Title: J Gene Med (In Press)
• [Journal Article] A second-site mutation in the initiation codon of WAS (WASP) gene results in expansion of subsets of lymphocytes in an Wiskott-Aldrich Syndrome patient. (2006)
  • Author(s): Du W, Kumaki S, Uchiyama S, Yachi A, Chung Yeng L, Kawai S, Minegishi M, Ramesh N, Geha RS, Sasahara Y, Tsuchiya S.
  • Journal Title: Human Genet (In Press)
• [Journal Article] A novel JAK3 mutation in a Japanese patient with severe combined immunodeficiency. (2005)
  • Author(s): Uchiyama T, Kumaki S, Fujiwara M, Nishida Y, Hakozaki I, Imai K, Du W, Yoshinari M, Sasahara Y, Tsuchiya S.
  • Journal Title: Pediatr Int. 47 Pages: 575-578
  • NAID: 10016939145
• [Journal Article] Functional and physical interactions between ERCC1 and MSH2 complexes for resistance to cis-diamminedichloroplatinum(II) in mammalian cells (2004)
  • Author(s): Lan L
  • Journal Title: DNA Repair 3 Pages: 135-143
• [Journal Article] A unique immunofluorescence method promotes accurate diagnosis in MYH9 disorders : a case report. (2004)
  • Author(s): Yoshinari M
  • Journal Title: J Pediatr Hematol Oncol 26 Pages: 579-583
• [Journal Article] Characterization of a novel nonsense mutation in the interleukin-7 receptor α gene in a Korean Patient with severe combined immunodeficiency. (2004)
  • Author(s): Jo EK
  • Journal Title: Int J Hematol 80 Pages: 332-335
• [Journal Article] Lentiviral vector-mediated gene transfer in T cells from Wiskott-Aldrich syndrome patients leads to functional correction (2004)
  • Author(s): Dupre L
  • Journal Title: Mol Ther 10 Pages: 903-915
• [Journal Article] Fludarabine- and cyclophosphamide-based nonmyeloablative conditioning regimen for transplantation of chronic granulomatous disease : possible correlation with prolonged pure red cell aplasia (2004)
  • Author(s): Fujiwara T
  • Journal Title: Int J Hematol 79 Pages: 293-307
• [Journal Article] X-linked severe combined immunodeficiency syndrome : the first Korean case with gamma c chain gene mutation and subsequent genetic counseling (2004)
  • Author(s): Jo EK
  • Journal Title: J Korean Med Sci 19 Pages: 123-126