Efficient identification of HLA-A*2402–restricted cytomegalovirus-specific CD8+ T-cell epitopes by a computer algorithm and an enzyme-linked immunospot assay

Kuzushima, Kiyotaka; Hayashi, Nobuyuki; Kimura, Hiroshi; Tsurumi, Tatsuya

doi:10.1182/blood.v98.6.1872

Cited by 146 publications

(152 citation statements)

References 46 publications

(51 reference statements)

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“…One day after each restimulation, IL-2 (Shionogi, Osaka, Japan) was added to a final concentration of 20 U/mL. To establish T cell clones, limiting dilution of polyclonal CTL was performed using round-bottomed 96-well plates as previously described [55]. After 2 wk of culture, growing wells were split into three replicates and used as effectors in the CTL assay against either DLMP1 mRNA-or EGFP mRNA-transduced autologous CD40-B cells.…”

Section: Ctl Inductionmentioning

confidence: 99%

“…Wells were scored as positive when the counts per minute from DLMP1 mRNA-transduced CD40-B cells exceeded the mean counts per minute from EGFP mRNA-transduced CD40-B cells by three standard deviations. Positive wells were transferred into flasks and expanded as previously described [55].…”

Section: Ctl Inductionmentioning

confidence: 99%

“…ELISPOT assays were performed as described earlier [48,50,55]. Briefly, CD8 + T cells were co-cultured with various stimulators in wells of Multiscreen-HA plates (MAHA S4510; Millipore, Billerica, MA) coated with anti human IFN-c mAb (M700A; Pierce Biotechnology, Philadelphia, PA).…”

Section: Elispot Assaymentioning

confidence: 99%

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Epstein‐Barr virus (EBV) latent membrane protein‐1‐specific cytotoxic T lymphocytes targeting EBV‐carrying natural killer cell malignancies

et al. 2006

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Epstein‐Barr virus (EBV)‐encoded latent membrane protein (LMP) 1 is a potential target for immunotherapy of some proportion of Hodgkin's disease cases, nasopharyngeal carcinomas, EBV‐associated natural killer (NK)/T lymphomas, and chronic active EBV infection (CAEBV). Since it is unknown whether EBV‐infected NK/T cells are susceptible to lysis by LMP1‐specific cytotoxic T lymphohcytes (CTL), we here tested the ability of mRNA‐transduced antigen‐presenting cells (APC) to stimulate rare LMP1‐specific CTL. A 43‐amino acid N‐terminal deletion mutant LMP1 (ΔLMP1) could be efficiently expressed in dendritic cells and CD40‐activated B cells upon mRNA electroporation. ΔLMP1‐expressing APC were found to stimulate LMP1‐specific CTL from a healthy donor and a CTL clone recognized a peptide, IIIILIIFI, presented by HLA‐A*0206 molecules. Processing and presentation of the antigenic peptide proved dependent on expression of an immunoproteasome subunit, low‐molecular‐weight protein‐7, as confirmed by RNA interference gene silencing. Furthermore, an EBV‐infected NK cell line derived from a patient with CAEBV, and another from an NK lymphoma with enforced HLA‐A*0206 expression, were specifically lysed by the CTL. Overall, these data suggest that immunotherapy targeting LMP1 in EBV‐associated NK lymphomas and CAEBV might serve as an alternative treatment modality.

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Section: Ctl Inductionmentioning

confidence: 99%

Section: Ctl Inductionmentioning

confidence: 99%

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Epstein‐Barr virus (EBV) latent membrane protein‐1‐specific cytotoxic T lymphocytes targeting EBV‐carrying natural killer cell malignancies

et al. 2006

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“…Isotypematched mouse IgG1 mAbs (BD Biosciences) were used as a control. Cells were also stained with PE-labeled HLA-A*2402 tetramers complexed with the peptide, QYDPVAALF, derived from CMV-pp65 (A24/pp65 328 -336 ), or PE-labeled HLA-A*0201 tetramers complexed with the peptide, NLVP-MVATV, from CMV-pp65 (A2/pp65 495-503 ), both of which were prepared as described previously (16). Flow cytometric analysis of the cells was performed using a FACScan (BD Biosciences), and EGFP emission was detected in the FL1 channel.…”

Section: Flow Cytometric Immunofluorescence Analysismentioning

confidence: 99%

Efficient Generation of Antigen-Specific Cytotoxic T Cells Using Retrovirally Transduced CD40-Activated B Cells

Kondo

Topp²,

Kiem

et al. 2002

The Journal of Immunology

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107

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The development of rapid, efficient, and safe methods for generating Ag-specific T cells is necessary for the clinical application of adoptive immunotherapy. We show that B cells stimulated with CD40 ligand and IL-4 (CD40-B cells) can be efficiently transduced with retroviral vectors encoding a model Ag, CMV tegument protein pp65 gene, and maintain high levels of costimulatory molecules after gene transfer. CTL lines specific for pp65 were readily generated in all four healthy CMV-seropositive donors by stimulating autologous CD8+ T cells with these transduced CD40-B cells, both of which were derived from 10 ml peripheral blood. ELISPOT assays revealed that the CTL lines used multiple HLA alleles as restricting elements. Thus, CD40-B cells transduced retrovirally with Ag-encoding cDNA can be potent APC and facilitate to generate Ag-specific CTL in vitro.

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“…Donor-derived mature DC were generated from frozen CD14 1 cells, pulsed with one of the indicated antigens, and used as a stimulator. As a positive control, HLA-A*2402-restricted CMV pp65 peptide (QYDPVAALF) [10] was used. PBMC of the patient were plated at 2 3 10 5 cells/well with stimulator cells (PBMC: stimulator 5 10:1) in 96-well, capture antibody-precoated PVDF plates (Millipore).…”

Section: Case Reportmentioning

confidence: 99%

Potential of dendritic cell immunotherapy for relapse after allogeneic hematopoietic stem cell transplantation, shown by WT1 peptide‐ and keyhole limpet hemocyanin‐pulsed, donor‐derived dendritic cell vaccine for acute myeloid leukemia

et al. 2007

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Induction of leukemia-specific immune responses is a promising treatment for acute myeloid leukemia. A 58-year-old woman received Wilms' tumor 1 (WT1) peptide-and keyhole limpet hemocyanin (KLH)-pulsed, donor-derived dendritic cell (DC) vaccination for AML relapse after allogeneic stem cell transplantation. The vaccination induced immune responses to the naïve antigen KLH, whereas definitive immune responses to WT1 were not detected. Leukemia gradually progressed despite of vaccination. This study indicates that DC vaccination can induce an antigen-specific immune response in a patient after allogeneic stem cell transplantation, thus representing a viable strategy to induce antigen-specific immune responses in such patients. Am. J. Hematol. 83:315-317, 2008. V

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Efficient identification of HLA-A*2402–restricted cytomegalovirus-specific CD8+ T-cell epitopes by a computer algorithm and an enzyme-linked immunospot assay

Cited by 146 publications

References 46 publications

Epstein‐Barr virus (EBV) latent membrane protein‐1‐specific cytotoxic T lymphocytes targeting EBV‐carrying natural killer cell malignancies

Epstein‐Barr virus (EBV) latent membrane protein‐1‐specific cytotoxic T lymphocytes targeting EBV‐carrying natural killer cell malignancies

Efficient Generation of Antigen-Specific Cytotoxic T Cells Using Retrovirally Transduced CD40-Activated B Cells

Potential of dendritic cell immunotherapy for relapse after allogeneic hematopoietic stem cell transplantation, shown by WT1 peptide‐ and keyhole limpet hemocyanin‐pulsed, donor‐derived dendritic cell vaccine for acute myeloid leukemia

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