The pattern of cytokine gene expression in freshly excised human metastatic melanoma suggests a state of reversible anergy of tumor‐infiltrating lymphocytes

Abstract

Expression of an extended panel of cytokine genes was investigated by reverse polymerase chain reaction (PCR) in 10 freshly excised melanoma metastases infiltrated by lymphocytes (TIL). cDNA encoding for CD3-delta and tyrosinase could be amplified in all samples, confirming the presence of T lymphocytes and melanoma cells. Cytokine genes possibly transcribed by both cell types, such as GM-CSF, IL-6 and IL-10 could be amplified from 5, 2 and 2 samples respectively. In contrast, IL-1 beta and TNF-alpha mRNA were never detectable, IL-1 alpha, IL-3 and IL-7 mRNA could be observed only in one case each. Transcripts encoding for TGF-beta 1 were observed in 8 samples, while TGF-beta 2 and 3 mRNA were detectable in only 2 specimens. mRNA encoding for cytokine genes typically transcribed by antigen-stimulated T lymphocytes, such as IL-2, IL-4 and IFN-gamma were rarely or never detectable (none, none and 1 of the samples respectively). In one case, where no cytokine gene transcription was detectable at the time of surgery, we addressed the question of the antigenicity of the tumor and of the functional competence of TIL. A primary tumor cell line was generated and cultured TIL were induced to transcribe IL-2 and IFN-gamma genes by incubation with the autologous irradiated tumor cell line, but not with autologous EBV-transformed cells. In these conditions, tumor-specific cytotoxic T lymphocytes (CTL) could be generated only after 3 weekly re-stimulations. In contrast, if autologous irradiated EBV-transformed cells were added to the cultures, specific CTL could be detected after one single tumor stimulation. Thus, signs of active responsiveness in terms of lymphokine gene mRNA are seldom detectable in melanoma metastases. Tumor-specific responses, however, including IL-2 and IFN-gamma gene expression and generation of CTL can be produced in vitro from specimens in which no cytokine gene mRNA is detectable ex vivo.

Predominant Th2/Tc2 Polarity of Tumor-Infiltrating Lymphocytes in Human Cervical Cancer

“…Our results confirm that cancer cells may attain immune escape by changing the effector composition of the host immune cells. We further explore the issues of proposed cancer-host immune interactions based on previous in vivo murine models and in vitro cell line results (27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40).…”

“…TGF-␤ and IL-10 are secreted spontaneously by a variety of human malignant tumors, including breast, colon, ovary, cutaneous basal/squamous cancers, metastatic melanoma, fibrosarcoma, and pancreatic carcinoma (27)(28)(29)(30)(31)(32)(33)(34)(35). The mechanisms responsible for the increased expression by tumor cells are not known.…”

High density oligonucleotide array analysis of interferon-α2a sensitivity and transcriptional response in melanoma cells

“…ME15, ME51, ME59 and ME67 cell lines were generated in our laboratory upon culture of cell suspensions derived from surgically excised melanoma metastases (Lüscher et al, 1994). A375 cell line was a gift from Dr Eberle (Basel, Switzerland) where as D10 cell line was provided by Dr Rimoldi (Lausanne, Switzerland).…”

Elevated plasma levels of transforming growth factor (TGF)-β1 and TGF-β2 in patients with disseminated malignant melanoma

“…Overexpression of TGF-P and loss of growth inhibition have been described in melanoma cell lines in vitro (MacDougall et al, 1993;Krasagakis et al, 1994;Rodeck et al, 1994). In vivo, TGF-f31 mRNA has been detected in metastatic nodules, and increased TGF-12 mRNA expression has been reported in deep invasive primary tumours (Luscher et al, 1994;Reed et al, 1994).…”