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Cancer Immunology, Immunotherapy : CII logoLink to Cancer Immunology, Immunotherapy : CII
. 1995 Jan;41(1):1–9. doi: 10.1007/BF01788953

High expression of adhesion molecules/activation markers with little interleukin-2, interferon γ, and tumor necrosis factor β gene activation in fresh tumor-infiltrating lymphocytes from lung adenocarcinoma

Eugène Roussel 1,2, Marie-Claude Gingras 3,2, Elizabeth A Grimm 4, Jack A Roth 1,4,✉
PMCID: PMC11037739  PMID: 7641214

Abstract

Little is known about the activation level of tumor-infiltrating lymphocytes (TIL) in human lung adenocarcinoma. We investigated the activation of fresh TIL at cellular and molecular levels and compared it with autologous and healthy normal peripheral blood lymphocytes (PBL) for baseline level. TIL were extracted from 12 primary lung adenocarcinomas by mechanical disruption without enzyme use and isolated by double-density Ficoll gradients. Flow-cytometry analysis of TIL subset distribution revealed that the majority was composed of T lymphocytes and double labeling with α-CD3 and adhesion/activation markers revealed T cell subsets expressing CD49a, CD49b, CD54, and CD15, each of which was almost absent in autologous T peripheral blood lymphocytes (T-PBL). Moreover, the proportions of T-TIL expressing CD58, CD65, or CD25 were increased severalfold compared to T-PBL. Lymphokine gene activation in TIL was assessed by mRNA reverse transcriptase/polymerase chain reaction (RT-PCR) and primers for interleukin(IL)-2, IL-4, interferon (IFN) γ, granulocyte/macrophage-colonystimulating factor (GM-CSF), and tumor necrosis factor (TNF) β. Semiquantitative comparisons between patients' TIL and PBL and healthy normal and activated PBL were performed by computerized image analysis. RT-PCR gel band products were quantified in relative units as a function of their size and intensity. TIL expressed detectable lymphokine mRNA but seemed poorly activated with respect to the total number of lymphokine genes and the amount of mRNA compared with α-CD3-activated healthy PBL. IL-2, IFNγ, and TNFβ did not appear to be expressed at higher levels in TIL than in autologous or healthy normal PBL. However, two-thirds of the patients had TIL distinguishable from autologous PBL by specific expression of GM-CSF and from healthy normal PBL by expression of IL-4. These results show that lung adenocarcinoma TIL populations had little lymphokine gene activation despite the presence of several T cell subsets expressing different adhesion/activation markers. The lack or deficient combination of lymphokine production may be a factor that prevented efficient activation of TIL in these tumors.

Key words: Cell adhesion molecules, Lung adenocarcinoma, Lymphokines, Polymerase chain reaction, Tumor-infiltrating lymphocytes

Footnotes

Partially supported by grants from the National Cancer Institute (RO1 CA45187) (J.A.R.), by gifts to the Division of Surgery from Tenneco and Exxon for the Core Lab Facility, by the M. D. Anderson Cancer Center Core Grant (NCI CA16672), by a generous gift from the Mathers Foundation (J.A.R.), and by a grant from the M. D. Anderson Cancer Center Physicians Referral Service (E.R., M.C.G.). E.R. is supported by NCI SPORE grant 1P50-CA58204-03PP5

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