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Detection of the status of tumour-infiltrating CD4+ T-cell subpopulations in squamous cell carcinoma of the head and neck (HNSCC) using real-time RT-PCR detection of CD4 mRNA

Cellular & Molecular Biology Letters volume 12, pages 308–316 (2007)Cite this article

Abstract

Several studies have reported tumour infiltrating CD4+ T cells as a favourable prognostic factor in some types of cancer. We investigated 37 head and neck squamous cell carcinomas (HNSCC) at different stages, using immunohistochemical staining for CD4+ infiltrates and real-time reverse transcription polymerase chain reaction (RT-PCR) detection of CD4 mRNA. The CD4+ infiltrates were evaluated and expressed as a percentage according to the ratio of CD4+ T cells to epithelial cells in the cancer cell nests and to the overall inflammatory cell infiltrate in the tumor stroma. The CD4 mRNA expression level strongly correlated with the CD4+ infiltration score in the cancer epithelium (rs = 0.858, P < 0.001) and in the cancer stroma (rs = 0.797, P < 0.001). These results indicate that the real-time RT-PCR assay is a sensitive and reliable method for the detection of CD4 mRNA, and that it could be used to reassess CD4+ infiltration status in resected specimens from patients with HNSCC.

Abbreviations

ECL:

enhanced chemiluminiscence

EDTA:

ethylendiaminetetraacetic acid

F:

female

HNSCC:

head and neck squamous cell carcinoma

Hpx:

hypopharynx

IHC:

immunohistochemistry

IL:

interleukin

Lrx:

larynx

M:

male

MHC:

major histocompatibility complex

NFDM:

non-fat dry milk

PBS:

phosphate buffer saline

PCR:

polymerase chain reaction

PMSF:

phenylmethylsulfonyl fluoride

pTNM:

pathological Tumour Node Metastasis classification of cancer

RT:

room temperature

RT-PCR:

reverse transcriptase polymerase chain reaction

SD:

standard deviation

SDS:

sodium dodecyl sulfate

Th:

T-helper immune cells

TRI reagent:

TRIZOL reagent

SPSS:

Statistical Package for the Social Sciences

UICC:

International Union Against Cancer (Union Internationale Contre le Cancer)

References

  1. Swain, S.L. T cell subsets and the recognition of MHC class. Immunol. Rev. 74 (1983) 129–142.

    Article  CAS  PubMed  Google Scholar 

  2. Nishimura, M.I., Avichezer, D., Custre, M.C., Lee, C.S., Chen, C., Parkhurst, M.R., Diamond, R.A., Robbins, P.F., Schwartzentruber, D.J. and Rosenberg, S.A. MHC class I-restricted recognition of a melanoma antigen by a human CD4+ tumour infiltrating lymphocyte. Canc. Res. 59 (1999) 6230–6238.

    CAS  Google Scholar 

  3. Veillette, A., Bookman, M.A., Horak, E.M. and Bolen, J.B. The CD4 and CD8 T-cell surface antigens are associated with the internal membrane tyrosine-protein kinase p56lck. Cell 55 (1988) 301–308.

    Article  CAS  PubMed  Google Scholar 

  4. Hung, K., Hayashi, R., Lafond-Walker, A., Lowenstein, C., Pardoll, D. and Levitsky, H. The central role of CD4+ T cells in antitumour immune response. J. Exp. Med. 188 (1998) 2357–2368.

    Article  CAS  PubMed  Google Scholar 

  5. Volpert, O.V., Fong, T., Koch, A.E., Peterson, J.D., Waltenbaugh, C., Tepper, R.I. and Bouck, N.P. Inhibition of angiogenesis by interleukin 4. J. Exp. Med. 188 (1998) 1039–1046.

    Article  CAS  PubMed  Google Scholar 

  6. Mattes, J., Hulett, M., Xie, W., Hogan, S., Rothenberg, M.E., Foster, P. and Parish, C. Immunotherapy of cytotoxic T cell-resistant tumours by T helper 2 cells: an eotaxin and STAT6-dependent process. J. Exp. Med. 197 (2003) 387–293.

    Article  CAS  PubMed  Google Scholar 

  7. Badoual, C., Hans, S., Rodriguez, J., Peyrard, S., Klein, C., Agueznay Nel, H., Mosseri, V., Laccourreye, O., Bruneval, P., Fridman, W.H., Brasnu, D.F. and Tartour, E. Prognostic value of tumour-infiltrating CD4+ T-cell subpopulations in head and neck cancers. Clin. Cancer Res. 15 (2006) 465–472.

    Article  Google Scholar 

  8. Hiraoka, K., Miyamoto, M., Cho, Y., Suzuoki, M., Oshikiri, T., Nakakubo, Y., Itoh, T., Ohbuchi, T., Kondo, S. and Katoh, H. Concurrent infiltration by CD8+ T cells and CD4+ T cells is a favourable prognostic factor in nonsmall-cell lung carcinoma. Br. J. Cancer 94 (2) (2006) 275–280.

    Article  CAS  PubMed  Google Scholar 

  9. Macchetti, A.H., Marana, H.R., Silva, J.S., de Andrade, J.M., Ribeiro-Silva, A. and Bighetti, S. Tumour-infiltrating CD4+ T lymphocytes in early breast cancer reflect lymph node involvement. Clinics 61 (2006) 203–208.

    PubMed  Google Scholar 

  10. Knutson, K.L, Disis, M.L. and Salazar, L.G. CD4 regulatory T cells in human cancer pathogenesis. Cancer Immunol. Immunother. (2006) in press.

  11. Cho, Y., Miyamoto, M., Kato, K., Fukunaga, A., Shichinohe, T., Kawarada, Y., Hida, Y., Oshikiri, T., Kurokawa, T., Suzuoki, M., Nakakubo, Y., Hiraoka, K., Murakami, S., Shinohara, T., Itoh, T., Okushiba, S., Kondo, S. and Katoh, H. CD4+ and CD8+ T cells cooperate to improve prognosis of patients with esophageal squamous cell carcinoma. Cancer Res. 63 (2003) 1555–1559.

    CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Molecular Genetics, Institute of Pathology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia

    Sava Smerkolj, Metka Volavšek & Damjan Glavač

Authors
  1. Sava Smerkolj
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  2. Metka Volavšek
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  3. Damjan Glavač
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Correspondence to Damjan Glavač.

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Smerkolj, S., Volavšek, M. & Glavač, D. Detection of the status of tumour-infiltrating CD4+ T-cell subpopulations in squamous cell carcinoma of the head and neck (HNSCC) using real-time RT-PCR detection of CD4 mRNA. Cell Mol Biol Lett 12, 308–316 (2007). https://doi.org/10.2478/s11658-007-0004-3

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  • DOI: https://doi.org/10.2478/s11658-007-0004-3

Key words

  • Tumour infiltrating CD4+ T cells
  • RT-PCR CD4 mRNA detection
  • HNSCC

Cellular & Molecular Biology Letters

ISSN: 1689-1392

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