Targeted cloning of cytotoxic T cells specific for minor histocompatibility antigens restricted by HLA class I molecules of interest

Yoshiki Akatsuka, Eisei Kondo, Hirohumi Taji, Yasuo Morishima, Makoto Yazaki, Yuichi Obata, Yoshihisa Kodera, Stanley R. Riddell, Toshitada Takahashi

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background. T cells specific for minor histocompatibility antigens (mHAgs) play a major role in both graft-versus-host disease and the graft-versus-tumor effect after HLA-identical bone marrow transplantation (BMT). However, characterization of individual T-cell responses to mHAgs is difficult and has generally involved extensive screening of T-cell clones isolated from bulk T-cell cultures generated from BMT recipients. In this report, we describe a new approach that permits both direct visualization of CD8+ T-cell responses to mHAgs and cloning of T cells reacting with mHAgs presented by individual HLA alleles of interest. Methods and Results. Panels of Epstein-Barr virus-transformed B-cell lines (B-LCL) expressing retrovirally transduced HLA cDNA were used as stimulator cells in an enzyme-linked immunospot (ELISPOT) assay to identify CD8+ T cells reacting with mHAgs in cultures generated from postBMT recipient peripheral blood. T cells specific for mHAgs presented by selected HLA alleles could then be captured and cloned using an interferon-y secretion assay and magnetic bead selection. A majority of T-cell clones thus isolated exhibited cytolytic activity against the same HLA-transfected B-cell lines used for the ELISPOT assay. Conclusion. The ELISPOT assay was useful for identification of the HLA alleles presenting mHAgs recognized by individual T-cell lines. This approach for isolating mHAgs-specific CD8+ T-cell clones should assist in characterizing responses restricted by HLA alleles of interest, which are common in a certain ethnic group.

Original languageEnglish
Pages (from-to)1773-1780
Number of pages8
JournalTransplantation
Volume74
Issue number12
DOIs
Publication statusPublished - 27-12-2002
Externally publishedYes

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Minor Histocompatibility Antigens
Organism Cloning
T-Lymphocytes
Enzyme-Linked Immunospot Assay
Alleles
Clone Cells
Bone Marrow Transplantation
B-Lymphocytes
Cell Line
Transformed Cell Line
HLA-B Antigens
Graft vs Host Disease
Human Herpesvirus 4
Ethnic Groups
Interferons

All Science Journal Classification (ASJC) codes

  • Transplantation

Cite this

Akatsuka, Yoshiki ; Kondo, Eisei ; Taji, Hirohumi ; Morishima, Yasuo ; Yazaki, Makoto ; Obata, Yuichi ; Kodera, Yoshihisa ; Riddell, Stanley R. ; Takahashi, Toshitada. / Targeted cloning of cytotoxic T cells specific for minor histocompatibility antigens restricted by HLA class I molecules of interest. In: Transplantation. 2002 ; Vol. 74, No. 12. pp. 1773-1780.
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abstract = "Background. T cells specific for minor histocompatibility antigens (mHAgs) play a major role in both graft-versus-host disease and the graft-versus-tumor effect after HLA-identical bone marrow transplantation (BMT). However, characterization of individual T-cell responses to mHAgs is difficult and has generally involved extensive screening of T-cell clones isolated from bulk T-cell cultures generated from BMT recipients. In this report, we describe a new approach that permits both direct visualization of CD8+ T-cell responses to mHAgs and cloning of T cells reacting with mHAgs presented by individual HLA alleles of interest. Methods and Results. Panels of Epstein-Barr virus-transformed B-cell lines (B-LCL) expressing retrovirally transduced HLA cDNA were used as stimulator cells in an enzyme-linked immunospot (ELISPOT) assay to identify CD8+ T cells reacting with mHAgs in cultures generated from postBMT recipient peripheral blood. T cells specific for mHAgs presented by selected HLA alleles could then be captured and cloned using an interferon-y secretion assay and magnetic bead selection. A majority of T-cell clones thus isolated exhibited cytolytic activity against the same HLA-transfected B-cell lines used for the ELISPOT assay. Conclusion. The ELISPOT assay was useful for identification of the HLA alleles presenting mHAgs recognized by individual T-cell lines. This approach for isolating mHAgs-specific CD8+ T-cell clones should assist in characterizing responses restricted by HLA alleles of interest, which are common in a certain ethnic group.",
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Akatsuka, Y, Kondo, E, Taji, H, Morishima, Y, Yazaki, M, Obata, Y, Kodera, Y, Riddell, SR & Takahashi, T 2002, 'Targeted cloning of cytotoxic T cells specific for minor histocompatibility antigens restricted by HLA class I molecules of interest', Transplantation, vol. 74, no. 12, pp. 1773-1780. https://doi.org/10.1097/00007890-200212270-00023

Targeted cloning of cytotoxic T cells specific for minor histocompatibility antigens restricted by HLA class I molecules of interest. / Akatsuka, Yoshiki; Kondo, Eisei; Taji, Hirohumi; Morishima, Yasuo; Yazaki, Makoto; Obata, Yuichi; Kodera, Yoshihisa; Riddell, Stanley R.; Takahashi, Toshitada.

In: Transplantation, Vol. 74, No. 12, 27.12.2002, p. 1773-1780.

Research output: Contribution to journalArticle

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T1 - Targeted cloning of cytotoxic T cells specific for minor histocompatibility antigens restricted by HLA class I molecules of interest

AU - Akatsuka, Yoshiki

AU - Kondo, Eisei

AU - Taji, Hirohumi

AU - Morishima, Yasuo

AU - Yazaki, Makoto

AU - Obata, Yuichi

AU - Kodera, Yoshihisa

AU - Riddell, Stanley R.

AU - Takahashi, Toshitada

PY - 2002/12/27

Y1 - 2002/12/27

N2 - Background. T cells specific for minor histocompatibility antigens (mHAgs) play a major role in both graft-versus-host disease and the graft-versus-tumor effect after HLA-identical bone marrow transplantation (BMT). However, characterization of individual T-cell responses to mHAgs is difficult and has generally involved extensive screening of T-cell clones isolated from bulk T-cell cultures generated from BMT recipients. In this report, we describe a new approach that permits both direct visualization of CD8+ T-cell responses to mHAgs and cloning of T cells reacting with mHAgs presented by individual HLA alleles of interest. Methods and Results. Panels of Epstein-Barr virus-transformed B-cell lines (B-LCL) expressing retrovirally transduced HLA cDNA were used as stimulator cells in an enzyme-linked immunospot (ELISPOT) assay to identify CD8+ T cells reacting with mHAgs in cultures generated from postBMT recipient peripheral blood. T cells specific for mHAgs presented by selected HLA alleles could then be captured and cloned using an interferon-y secretion assay and magnetic bead selection. A majority of T-cell clones thus isolated exhibited cytolytic activity against the same HLA-transfected B-cell lines used for the ELISPOT assay. Conclusion. The ELISPOT assay was useful for identification of the HLA alleles presenting mHAgs recognized by individual T-cell lines. This approach for isolating mHAgs-specific CD8+ T-cell clones should assist in characterizing responses restricted by HLA alleles of interest, which are common in a certain ethnic group.

AB - Background. T cells specific for minor histocompatibility antigens (mHAgs) play a major role in both graft-versus-host disease and the graft-versus-tumor effect after HLA-identical bone marrow transplantation (BMT). However, characterization of individual T-cell responses to mHAgs is difficult and has generally involved extensive screening of T-cell clones isolated from bulk T-cell cultures generated from BMT recipients. In this report, we describe a new approach that permits both direct visualization of CD8+ T-cell responses to mHAgs and cloning of T cells reacting with mHAgs presented by individual HLA alleles of interest. Methods and Results. Panels of Epstein-Barr virus-transformed B-cell lines (B-LCL) expressing retrovirally transduced HLA cDNA were used as stimulator cells in an enzyme-linked immunospot (ELISPOT) assay to identify CD8+ T cells reacting with mHAgs in cultures generated from postBMT recipient peripheral blood. T cells specific for mHAgs presented by selected HLA alleles could then be captured and cloned using an interferon-y secretion assay and magnetic bead selection. A majority of T-cell clones thus isolated exhibited cytolytic activity against the same HLA-transfected B-cell lines used for the ELISPOT assay. Conclusion. The ELISPOT assay was useful for identification of the HLA alleles presenting mHAgs recognized by individual T-cell lines. This approach for isolating mHAgs-specific CD8+ T-cell clones should assist in characterizing responses restricted by HLA alleles of interest, which are common in a certain ethnic group.

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