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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

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)

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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