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T-cell receptor BV gene usage in colorectal carcinoma patients immunised with recombinant Ep-CAM protein or anti-idiotypic antibody

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Abstract

The tumour-associated antigen, Ep-CAM, is over-expressed in colorectal carcinoma (CRC). In the present study, a recombinant Ep-CAM protein or a human anti-idiotypic antibody (anti-Id) mimicking Ep-CAM, either alone or in combination, was used for vaccination of CRC patients (n=9). GM-CSF was given as an adjuvant cytokine. A cellular immune response was assessed by measuring anti-Ep-CAM lymphoproliferation, IFN-γ production (ELISPOT) and by analysing the TCR BV gene usage within the CD4+ and CD8+ T-cell subsets followed by CDR3 fragment analysis. A proliferative and/or IFN-γ T-cell response was induced against the Ep-CAM protein in eight out of nine patients, and against Ep-CAM-derived peptides in nine out of nine patients. Analysis of the TCR BV gene usage showed a significantly higher usage of BV12 family in CD4+ T cells of patients both before and after immunisation than in those of healthy control donors (p<0.05). In the CD8+ T-cell subset, a significant (p<0.05) increase in the BV19 usage was noted in patients after immunisation. In individual patients, a number of TCR BV gene families in both CD4+ and CD8+ T cells were over-expressed mainly in post-immunisation samples. Analysis of the CDR3 length polymorphism revealed a higher degree of clonality in post-immunisation samples than in pre-immunisation samples. In vitro stimulation with Ep-CAM protein confirmed the expansion of anti-Ep-CAM T-cell clones. The results indicate that immunisation with the Ep-CAM protein and/or anti-Id entails the induction of an anti-Ep-CAM T-cell response in CRC patients, and suggest that BV19+ CD8+ T cells might be involved in a vaccine-induced immune response.

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Abbreviations

AJCC:

American Joint Committee on Cancer

Anti-Id:

Anti-idiotypic antibody

BCP:

Baculovirus control protein

CEA:

Carcinoembryonic antigen

CRC:

Colorectal carcinoma

CTL:

Cytotoxic T lymphocyte

ELISA:

Enzyme-linked immunosorbent assay

ELISPOT:

Enzyme-linked immunospot

Ep-CAM:

Epithelial-cell adhesion molecule

GM-CSF:

Granulocyte-macrophage colony-stimulating factor

IFN-γ:

Gamma interferon

mAb:

Monoclonal antibody

MHC:

Major histocompatibility complex

PBMC:

Peripheral blood mononuclear cells

PHA:

Phytohemagglutinin

PPD:

Purified protein derivative of tuberculin

SDS:

Sodium dodecyl sulfate

SFU:

Spot-forming unit

SI:

Stimulation index

TAA:

Tumour-associated antigen

TCR:

T-cell receptor

TIL:

Tumour-infiltrating lymphocytes

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Acknowledgements

This study was supported by grants from the Swedish Research Council, Swedish Cancer Society, Cancer Society in Stockholm, King Gustav V Jubilee Fund, Cancer and Allergy Foundation, Torsten and Ragnar Söderberg Foundation, Gunnar Nilsson Cancer Foundation and Karolinska Institute Foundations. We thank Dr H. Wigzell for the scientific discussion.

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Authors and Affiliations

  1. Immune and Gene Therapy Laboratory, Department of Oncology (Radiumhemmet), Karolinska Institute, Stockholm, Sweden

    Szilvia Mosolits, Katja Markovic, Jan Fagerberg, Jan-Erik Frödin, Mohammad-Reza Rezvany, Shahryar Kiaii, Håkan Mellstedt & Mahmood Jeddi-Tehrani

  2. Department of Immunology, Monoclonal Antibody Research Center, Avesina Research Center, Tehran, Iran

    Mahmood Jeddi-Tehrani

  3. Immune and Gene Therapy Laboratory, Cancer Centre Karolinska, Karolinska Hospital, 171 76, Stockholm, Sweden

    Håkan Mellstedt

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  1. Szilvia Mosolits
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  2. Katja Markovic
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  4. Jan-Erik Frödin
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Correspondence to Håkan Mellstedt.

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Mosolits, S., Markovic, K., Fagerberg, J. et al. T-cell receptor BV gene usage in colorectal carcinoma patients immunised with recombinant Ep-CAM protein or anti-idiotypic antibody. Cancer Immunol Immunother 54, 557–570 (2005). https://doi.org/10.1007/s00262-004-0620-y

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