Cytokine biosynthesis by tumor-infiltrating T lymphocytes from human non-small-cell lung carcinoma

Cancer Immunol Immunother. 2000 Feb;48(11):627-34. doi: 10.1007/s002620050011.


Purpose: The purpose of this work was to assess the capacity of tumor-infiltrating leukocytes (TIL) from human non-small-cell lung carcinoma (NSCLC) specimens to synthesize type-1 and type-2 cytokines.

Methods: TIL were isolated from tumors following digestion with collagenase/DNase and further enriched by ficoll-hypaque gradient centrifugation. Membrane phenotypes and intracellular cytokine protein expression of TIL were assessed by flow cytometry.

Results: The majority of TIL expressed the CD3 antigen with a CD4:CD8 ratio of approximately 2:1. Other leukocytes such as macrophages (CD14), B lymphocytes (CD20), and natural killer (NK) cells (CD56) were also found to infiltrate the tumors, but in significantly lower numbers. Owing to the limited recovery of non-CD3(+) leukocytes, our analysis of cytokine biosynthesis has focused on T lymphocytes. In the absence of activation, a small percentage of CD3(+) TIL synthesized cytokines ( <4%). Following activation with anti-CD3+interleukin-2 (IL-2), CD3(+) TIL synthesized predominantly a type-1 cytokine profile; however, the type-2 cytokines, IL-6 and IL-10, were also detected in a small percentage of infiltrating cells. Following activation with phorbol 12-myristate 13-acetate + ionomycin, CD3(+) TIL also expressed more type-1 than type-2 cytokines and in significantly greater numbers of cells. The CD3(+)CD8(+) component of the TIL synthesized only type-1 cytokines, whereas the CD3(+)CD4(+) component synthesized both type-1 and type-2 cytokines.

Conclusion: These results show that the majority of the TIL isolated from NSCLC specimens are T lymphocytes with the capacity to synthesize type-1 cytokines.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenocarcinoma / metabolism
  • Adenocarcinoma / pathology
  • CD3 Complex / immunology
  • Carcinoma, Non-Small-Cell Lung / metabolism*
  • Carcinoma, Non-Small-Cell Lung / pathology
  • Carcinoma, Squamous Cell / metabolism
  • Carcinoma, Squamous Cell / pathology
  • Cytokines / biosynthesis*
  • Cytokines / genetics
  • Gene Expression Regulation
  • Humans
  • Interleukin-2 / pharmacology
  • Ionomycin / pharmacology
  • Lung Neoplasms / metabolism*
  • Lung Neoplasms / pathology
  • Lymphocyte Activation
  • Lymphocytes, Tumor-Infiltrating / metabolism*
  • Muromonab-CD3 / pharmacology
  • T-Lymphocytes / metabolism*
  • Tetradecanoylphorbol Acetate / pharmacology


  • CD3 Complex
  • Cytokines
  • Interleukin-2
  • Muromonab-CD3
  • Ionomycin
  • Tetradecanoylphorbol Acetate