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COMBINED USE OF IMMUNOEISTOCREMISTRY AND M SITU IiYBRlDIZATIONFORCBROMOGRANlNATO DETECTSMALL CELL LUNGCANCERWITB NEUROENDOCRINE FEATURES G.v. Scsgliotti, E. Lemardo*,S. Cappia’, P. De Giuli*, P. Masiem, L. Gubetta*, F. Gozzelino, E. Pozzi. University of Torino, Department of Clinical & Biological Sciences and *Laboratory of Pathology, S. Luigi Gonzaga Hospital, Orbassano (Torino). ITALY. r

One hundred and twentyseven Small Cell Lung Cancers (SCLC) were pro-

spectively evaluated for the expression of Immunohistochemical (IHC) markers of squamous-, adenomatous- and neuroendocrine (NE) differentiation. Neuron Specific Enolase (NSE) was the IHC marker of NE differentiation more commonly positive (57%) while the positivity for Chromogranin A (ChrA) and synaptophysin was confined to 22% and 14% of the turnours, respectively. MC results allow to distiguish two subsets of SCLC : the first (1~72) characterized by mild/strong positivity for NSE and/or ChrA and/or synaptophysin. negativity for High Molecular Weight (HMW) cytokeratins (CKs) and positivity for Low MW CKs. the second subgroup (n=55) with negativity for NE markers, controversial results for HMW CKs (positive or negative) and constant positivity for LMW CKs. To characterize farther the latter subgroup a 24-base oligonncleotide probe complementary to human mP.NA ChrA coding for aminoacids 277-284 was synthesized. purified by gel electrophoresis on 20% polyacrilamide and the In Situ Hybridizatron (ISH) evaluation performed on 5 pm paraffin-embedded sections. 20 out of 55 resulted positive for mRNA ChrA probe and the evaluation of the distribution of the response rate to induction therapies according to the IHC and ISH results revealed a superior. nearly significant. percentage of respon- ses in the group with NE features (p=O.O59). After performing ISH in the re- maining 35 negative turnours were gathered all the 12 SCLCs positive for epi- thelial membrane antigen and strong pxitivie for HMW CKs indicating a ten- dency toward sqnamons differentiation while the other 23 tomours all displayed mild/strong reactivity for carcinoembryonic antigen as nsnally detected in adecarcinoma cells. In conclusion: IHC and ISH combined results seem to contribute to the screening of a wider group of SCLCs with NE features and the clinical relevance of these information should be further evaluated in prospective randomized trials.

087

MODIFICATIONS OF PLASMA GLYCOSYLATED RESIDUES IN LUNG CANCER DETECTED BY PROTON MRS : EFFECT OF TUMOR EXTENSION. Kleisbauer J.P.**, P.Thomas **, J. Vion-Dury *, M. Sciaky*, S. Confort-Gotmy* and P.J.Cozzone.* (*) Centre de R&onance Magnc?tique Biologique et Medicaie, (URA CNRS 1186). FacultC de Medecine, Bd J.Moulin, (**), Service d’Oncologie Respiratoire, Haprtal Ste Marguerite, 13000 Marseille (Fmnce)..

We have documented the variations of the concentration of glycosylated residues (mmnly N-acetylglucosamine (NAG) and N-acetyl-neumminic acid (sialic acid, NANA)) residues detected by MRS of plasma as a function of the extension of lung cancer described usmg the TNM staging. Plasma samples were collected from 110 patients sufferinp from SCLC and NSCLC. proton MR spectroscopy of plasma samples was performed at 400 MHz on a Bruker AM 400 spectrometer, using a 5 mm probe as previously described (1). The results are : 1) All lung cancers induce a strong increase tn plasma total glycosylatcd residues (GRt) and Nag and Nana resonances but different lung primary cancers or metastases cannot be differentmtcd using thcsc mctabolitcs. ?-) GRt 1s increasal in the same manner whatever the nature and importance of cancer invasion, 3) NANA concentration progressively increases when tumor is growing 4) NAG and NANA concentrations conslitute sensitive pammeters to describe the involvement of the nodes in the tumor 5) NAG and NANA constitute metabolic markers able to differentiate small cancer without mediastinal nodes to 3a, 3b. This result suggests that the propagation of the tumor into the nodes generates a complex immunological/ inflammatory reaction, with production of large quantities of glycosylated compounds related to the production of large amounts of TNF, interleukins and interferon by immuno competent and phagocytic cells lying in the nodes.

086

EXPRESSION OF MAGE GENES BY NON-SMALL CELL LUNG CARCINOMAS. P. Wemants*‘. B. Lethe*. F. Brasseur+, M. MM*, f. Boo”*. fLadwigIastiMefaCstlcerResercb,l3NssslsBnoch,&ussets.Belgirnn

Uniwsit6 Catbdiqw de Low& H&&l de Mont-Gcdime, Swice de Pnmmologie, Yvoir, Ek+um

We have re@ed t4e idmtification of RI human gme “MAGEl” which directstbeexpressiooofaaantigenMZ2-Emw&edmahunsomelaaomseetl line by autolegoas cytolic T Lympbwytes @XL) (1). This antigen is expessed on 4o%dmel~asmdis~byHLA-Al.Ibc~~~ byMAGElhasbeaidmt&dNoexPrwimdtb&geaetusb%mobwvedso farinwrmsltissueswimmeexoeptiandtestis(l).~enmdedbyMAGE- 1maytbereforeemstiMeas&dtwgetsforspecifiefic.Thegoalof thisshdywastoaoplyse~~mof~MAGE1,2Qd3(twoclosaly related genes) in LUNG caminmm RNAw8sisdatedfmmmplesdlm~ hrmorsmolhdbeea~quicklyafterresedion.Tbs~dMAOEl,2.3 mRNAW%SevaluDtedbylW~hagoiptmnsodpd~dU&l~a*ian (PCRjmn@i&musingoliganrieo(id~pirners~foreSmd meseg~.Wetested46svrgicnlsrrmplesdwrcsarllcellLuagcrciwm~ @XXLC)mdobsewdMAGEl ~in16oftbsm05%~.GmesMAoE2 wd3were~sedbysimilar~mdthesetumars35%md30% rrspectively. In a&litim we exemiwd the erpesdon of hiAGE-1 in a qamtitative PCRasaymdweobsemdawyvui&eleveld erprersioarl&llghorn1tO 1~dthepmolmt~by~acellIheMzZE.lo(hisshdy,5art of46(1O.846)prtieMswereHLA-Alrd~~MAGElae~armOr. lheprcptimdtumcm ex~mss&MAGElsuggesrrtbatNSCLCp&entsmay cm&ate the lrgest gmup of @eats suscepiie to be eligible fn Pilot stadies involving hiAGE inrmunization (2). 1. VanderBn~gga, P.,Trswmari C.. Chmw P. and all. A gme encoding m mtigea reco@tized by eyiolytic T @n@ecytes m a buman melaaane Science 1991; 254 : 1643-1647. 2. Boon T. Tumor aotigens racogaieed by cytolytic T IpnPbocytes : mt PeRpectives for specific immunothempy. bt. J. Cancer 1993; 54 : 177-160.

088

CYTOKINES AND ADHESION MOLECULES IN LUNG CaNCEA . D. Vitolo, M.B. Palmieri, G. Bonsignore*, C.D. Baroni. Department of Experimental Medicine, University La Sapienza, Rome and *Institute of Respiratory Pathophysiology, CNR, Palermo, Italy.

Cytokines regulate recruitment of circulating leukocytes in all tissues through the up-regulation of ICAM-1 and VCAM-1 on endothelial cells (EC) and of their ligands LFA-1 and VLA-4 on leukocytes. We studied in cryostat sections of lung cancer the distribution of these molecules on EC and of cytokines' mRNA in tumor infiltrating lymphomononuclear cells (TIL). ICAM- I+ EC were present in neoplastic and tumor-free tissues. V-CAM-l+ EC were observed only when VLA- 4+ TIL were present. In all cases of squamous cell carcinoma (SCC), neoplastic nests consisted of ICAM-l+ cells infiltrated by LFA-lt TIL. ICAW 1+ neoplastic cells were observed in only 2 of 5 cases of adenocarcinoma (AC). Few TNFc(+ and IL- lp+ TIL were present in neoplastic as well as in normal lung. In XC, IFNy+, IL-4+, and IL-5+ TIL were more numerous than in AC. These results suggest that in lung cancer the differentiation of Thl (IFNy+) and Th2 (IL-4+, IL-5+) helper lymphocytes is more efficient in SCC than in AC. Furthermore IFNr might determine ICAM- up- regulation on SCC, so far facilitating the intratumoral migration of LFA-1+ lymphocytes.

Supported by CNR (92.02250.30). AIRC (1993) and CNR PFO (ACRO) UA Responsabile: D D'Amico.