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Abstract
We have analysed the expression of interleukin-2 receptor (IL-2R) on a panel of small-cell lung cancer (SCLC) cell lines. None of the 11 SCLC cell lines studied expressed detectable surface IL-2R alpha or beta chains by indirect immunofluorescence. Reverse transcriptase-polymerase chain reaction (RT-PCR) analyses indicated that only one out of 11 cell lines expressed detectable IL-2R beta mRNA while two expressed a weak positivity for IL-2R gamma. Five SCLC cell lines were transfected with the plasmid vector RSV.5 neo containing IL-2 cDNA coding sequence. Stable transfectants secreted biologically active IL-2 (ranging from 25 to 100 U ml-1 in the culture supernatant). IL-2 transfection did not produce significant modifications in the expression of surface molecules such as IL-2R alpha and beta chains, intercellular adhesion molecule-1 (ICAM-1), CD44, HLA class I and II or in IL-2R beta or gamma mRNA. More importantly, IL-2-transfected N592 and NCI H69 cell lines completely lost their tumorigenic potential in nude mice after subcutaneous injection, whereas experimental controls transfected with RSV.5 neo vector only, displayed an in vivo growth pattern identical to that of untransfected cells. In addition, in the N592 model, IL-2-producing N592 inhibited the growth of wild-type N592 injected at the same site, while injection of parental cells on the opposite side did not significantly affect the growth of wild-type tumour cells. Histopathological analysis of the rejection process of IL-2-transfected cells demonstrated the presence of MAC-1+, MAC-3+ macrophages and of RB68C5+ granulocytes, whereas T cells were undetectable and NK cells were scarcely represented. In addition, a reduction of the tumour blood vessels was observed. The possible relevance of these data for the development of vaccination strategies using cytokine-engineered tumour cells in SCLC is discussed.
Highlights
RT PCR analysis of interleukin-2 receptor (IL-2R) f3 and -; mRNA expression indicated that only the NCI H82 cell line displayed a small amount of both IL-2R fl and IL-2R r' amplification products
Table I) In addition, no changes in the IL-2R fl and p mRNA were detected in the N592, NCI H69, NCI H 146, NCI H446 and IST-SLI cell lines after transfection with the IL-2 gene (Figure 2)
The small-cell lung cancer (SCLC) cell lines N592, NCI H69, NCI H 146, NCI H446 and IST-SLI were transfected with RSV.5 neo, L-2 vector by the use of cationic liposomes or by electroporation
Summary
The IST-SLl cell line was derived from a biopsy sample obtained from a supraclavicular lymph node metastasis of a 60-year-old male patient with SCLC by culture in RPMI1640 medium (Hyclone, Cramlington, UK) containing 5% fetal calf serum (FCS) and supplemented with 10-8 M hydrocortisone, S pg ml- insulin, 10-8 M /-oestrodiol and 0.1 jg ml-' bombesin (all from Sigma, St Louis, MO, USA). The other cells were cultured in RPMI-1640 supplemented with 10% FCS, 2 mM glutamine and penicillin-streptomycin. Surface expression of IL-2 receptor a and chains was analysed by indirect immunofluorescence and cytofluorimetric analysis. The MAbs used in this study were: MAR 93, antiIL-2R a (kindly provided by Dr A Moretta, Genoa, Italy), TU27, antil-IL2R # chain (Takeshita et al, 1989); T617, anti-
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