330

asbestos among such workers. By contrast, employees with small opacities (= 1K); IL0 classification) experienced a significantly raised risk of lung cancer (nine observed deaths v 2.1 expected), even though their exposures to asbestos were similar to the exposures of long term workers without opacities. In this population, excess risk of lung cancer was restricted to workers with x ray film evidence of asbestosis, a finding consistent with the view that asbestos is a lung carcinogen because of its tibrogenicity.

Mortality from lung cancer among Sardinian patients with silicosis Carta P, Cocco PL, Casula D. Institure of Occupational Medicine, UniversilyofCagliari, CagliariSard nia. Br J IndMed 1991;48:122-9.

The mortality of 724 subjects with silicosis, first diagnosed in 1964- 70 in the Sardinia region of Italy, was followed up through to 31 December 1987. Smoking, occupational history, chest x ray films, and data on lung function were available from clinical records for each member of the cohort. The overall cohort accounted for 10956.5 person-years. The standardised mortality ratios (SMRs) for selected causes of death (International Classification of Diseases (ICD) eighth revision) were based on the age specific regional death rates for each calender year. An excess of deaths for all causes (SMR = 1.40) was found, mainly due to chronic obstructive lung disease, silicosis, and tuberculosis with an upward trend of the SMR with increasing severity of the International Labour Office (ILO) radiological categories. Twenty two subjects died from lung cancer (SMR = 1.29, 95% confidence interval (95% CI) = 0.8-2.0). The risk increased after a 10 and 15 year latency but the SMR never reached statistical significance. No correla- tion was found between lung cancer and severity of the radiological category, the type of silica (coal or metalliferous mines, quarries etc), or the degree of exposure to silica dust. A significant excess of deaths from lung cancer was found among heavy smokers (SMR = 4.11) and subjects with airflow obstruction (SMR = 2.83). A nested case-control study was planned to investigate whether the association between lung cancer and airway obstruction was due to confounding by smoking. No association was found with the IL0 categories of silicosis or the estimated cumulative exposure to silica. The risk estimate for lung cancer by airflow obstruction after adjusting by cigarette consumption was 2.86 for a mild impairment and 7.23 for a severe obstruction. The results do not show any clear association between exposure to silica, severity of silicosis, and mortality from lung cancer. Other environ- mental or individual factors may act as confounders in the association between silicosis and lung cancer. Among them, attention should be given to chronic airways obstruction as an independent risk factor for lung cancer in patients with silicosis.

Lung cancer risk among exsmokers Sobue T. Suzuki T. Fujimoto I. Matsuda M. Doi 0. Mori T et al. Center forAdultDiseases, l-3-3 Nakamichi, Higashimzri-ku, Osaka537. Jpn J CancerRes 1991;82:273-9.

Lung cancer risk among exsmokers according to years since cessa- tion of smoking was assessed by means of a case-control study. The case series consisted of 1,052 lung cancer patients who were newly diag- nosed and admitted to eight hospitals in Osaka in 1986-88. Smoking histories were compared with those of 1,111 controls admitted to the same hospitals during the same period without any diagnosis of smok- ing-related disease. The odds ratio of lung cancer for exsmokers compared tocurrent smokers was estimated tobe 0.90,0.50,0.51,0.59, 0.48 and 0.29, for l-4.5-9, 10-14, 15-19.20-24 and = 25 years after cessation of smoking, respectively. Risk reduction appeared to be greater for those who smoked less than the 1200 cigarette index, compared to those who smoked more. In classification according to histologic type, small cell and large cell carcinoma showed a rapid decrease compared to adenocarcinoma, while squamous cell carcinoma showedan intermediate pattern. Quantitative estimates for reduction of lung cancer risk among exsmokers can be used for projecting lung cancer incidence in the future, by assuming future trends of smoking prevalence, as well as for health education among individual smokers.

Basic biology Association between restriction fragment length polymorphism of the human cytochrome P45OIIEl gene and susceptibility to lung cancer Uematsu F, Kikuchi H, Motomiya M, Abe T, Sagami 1, Ohmachi T et al. Department ofcancerche~therapy andprevention, The Research Instilafe for TuLxrcalosis and Cancer. Tohoku University. 4-1 Seiryoma- chi, Aoba-ku. Sendai 980. Jpn J Cancer Res 1991;82:254-6.

Cytochrome P45OIIEl (P450IIEl) is involved in metabolic activa- tion of carcinogenic nitrosamines, aniline and benzene. We detected a restriction fragment length polymorphism of the human P450IIEl gene with the restriction endonuclease DraI. The population was thus divided into three genotypes, namely, heterozygotes (CD) and two forms of homozygotes (CC and DD). The distribution of these genotypes among lung cancer patients differed from that among controls with statistical significance of Pc0.05(ch&7.01 with 2 degrees of freedom). This result strongly suggests that host susceptibility to lung cancer is associ- ated with the DraI polymorphism of the P4SOIIEl gene.

Oliioclonal T lymphocytes infiltrating human lung cancer tissues YoshinoI,YanoT,Yoshikai Y,MurataM,Sugimachi K, KimuraGet al. Departmenr of Virology, Medical Institute ofBioregulafion, Kyushu Universiry, Maidashi 3-1-I. Higashi-ku, Fukuoka 812. Int J Cancer 1991:47:654-S.

To clarify the nature of tumor-infiltrating lymphocytes (TILs), we investigated the possible clonality of the T cells in TILs freshly isolated from human primary lung cancer tissues by assessing the rearrangement pattern of the T-cell receptor VCR) gene 8 locus using Southern blotting. First, in phenotypic analysis, TILs represented differentpopu- lations among corresponding peripheral blood lymphocytes (PBLs) with an increased proportion of CD20+ (B) cells as well as a decreased proportion of CD16* (natural killer) cells, and a variable CD4/CDS ratio. Considering the central role of T cells in immune responses, we analyzed TCR 8 gene rearrangement patterns in TILs and correspond- ing PBLs from 12 patients. In 10 of the 12 cases, TILs showed one or more TCR gene rearrangement bands with a predominance of the C(O)2 gene, in which 2 types of common rearranged band were observed among the cases with different clinical profiles in terms of histological types and disease stage, with bands at about 9.5 kb in 7 and at 11.5 kb in 8 patients. On the other hand, predominant rearranged bands were hardly detected in corresponding PBLs except in 2 cases. From these results, we conclude that TILs in lung cancer tissues frequently contain oligcclonal T-cell populations, which were probably sensitized by relatively common antigens at the tumor sites.

Characterization and purification of an immunosuppressive factor produced by a small cell lung cancer cell line Ikeda T, Masuno T, Ogura T, Watanabe M, Shirasaka T, Hara H et al. Third Departtnenf of Internal Medicine, Osaka Universiry Medical SchooLI-I-50Fukushima,Fukushima-ku,Osaka553. Jpn JCancerRes 1991:82:332-8.

The present study was undertaken to determine whether small cell lung cancer (SCLC) cell lines produce immunosuppressive factors and, if they do, to characterize the factors. The supematants of SCLC cell lines, H69 and N857, inhibited not only the blastogenic response of human peripheral blood lymphocytes (PBL) to phymhemagglutinin or concanavalinA,butalsothecytotoxicactivityoflymphokine-activated killercells. Neither was inhibited by supematants from non-SCLC cell lines PC9, QG56, and A549. The immunosuppressive activity of H69 suparnatant was stable upon heating to 56°C for 60 min, but labile when heated to 70°C for IO min. The activity was abolished after dialysis at pH 2.0 or pH 11.0, but not at pH 4.5 or pH 9.0. Digestion with trypsin or proteinase eliminated the immunosuppressive activity, whereas treatment with neuraminidase, mixed glycosidase,DNase or RNase had no effect, suggesting that the immunosuppressive activity in H69 supematant is due to a protein factor. This H69-derived immunosup-

331

pressive factor was isolated by ion exchange chromatography using a gradient of 0.04 to 0.08 M NaCl solution, Gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the factor to have molecular weights of 98 kD and 102 kD, respectively. These results suggest that SCLC cells produce a potent immunosuppressive factor which may account for the immunedeficiency in SCLC patients.

Induction of cytokine messenger RNA and secretion in alveolar macrophages and blood monocytes from patients with long cancer receiving granutocyte-macrophage colony-stimulating factor ther- apy Thomassen MJ. Ahmad M, Bama BP, Antal J, Wicdemann HP. Meeker et al. Department ofPulmonary Disease, ClevelandClinic Foundation, 9500 EuclidAvenue, Cleveland, OH4419S-5038. Cancer Res 1991;51:857- 62.

Human granulocyte-macrophage colony-stimulating factor (GM- CSF) promotes the proliferation and differentiation of hematopoietic progenitor cells. Although preliminary data are available from clinical trials, the effect of GM-CSF on gene expression of immunocompetent cells in treated patients has not been studied. We previously demon- strated that in vitro treatment with GM-CSF also enhances maturation- related anti-tumor activities in mononuclear phagccytes. The purpose of the present study was to examine the effects of in viva recombinant GM-CSF therapy on alveolar macrophages and blood monocytes, to determine if these cells demonstrated differential expression of cytok- ine genes, cytokine production, and tumoricidal activity. Alveolar macrophages and blood monocytes were isolated from 13 patienta receiving a range of GM-CSF doses (60-250 pg/m%iay) by continuous infusion over a 2-week period. Both monocytes and macrophages were isolated prior to therapy and at day 10 of the infusion, Monocytes, in addition, were isolatedon day 3 of infusion. Results indicated that GM- CSF therapy enhanced expression of tumor necrosis factor, interleukin 1, and interleukin 6 mRNA in both monocytes and alveolar macroph- ages. Differential responses, however, were observed in cytokine secre- tion; monocytes demonstrated enhanced secretion of all three cytokines by day 3 of treatment, but alveolar macrophages showed only enhanced interleukin 6 secretion at&y IO. Monocyte tumoricidal activity after in vitro lipopolysaccharide stimulation was also significantly elevated by day 3 of treatment. but at day 10 activity was not statistically different from pretreatment values in either monccytes or alveolar macrophages. These data indicate that GM-CSF exerts striking time-dependent modulatory effects on gene expression and functional activities of monocytes and alveolar macrophages in viva, although the responses of the two cell types differ with respect to cytokine secretion.

Retention of activity by selected anthracyclines in a multidrug resistant human brge cell lung carcinoma line without P-glycopro- tein hyperexpression Coley HM, Workman P, Twentyman PR. MRC Clinical Oncology. MRC Centre, Hills Road, Cambridge CB2 ZQH. Br J Cancer 1991:63:351- 7.

A subline (COR-L23/R) of the human large cell lung cancer line COR-L23. derived by in viva exposure to doxontbicin, exhibits an unusual multidrug resistant (MDR) phenotype. This subline shows cross-resistance to daunorubicin, vincristine. colchicine and etoposide but does not express P-glycoprotein. Interestingly, COR-Ll2/R shows little or no resistance to a range of structurally-modified analogues of doxorubicin comprising 9-aIkyl and/or sugar modified anthracyclines. We have previously identified these same compounds as effective agents against P-glycoprotein-positive MDR cell lines. In contrast to typical MDRcelI lines, COR-LI2/R shows only minimal chemosensi- tisation by verapamil and no collateral sensitivity to verapamil. Com- pared to the parental cell line, COR-LlUR displays reduced accumula- tion of doxorubicin and daunorubicin. Accumulation defects were apparent only after 0.5- 1 h of incubation of cells with these agents. The rate of daunorubicin effhtx was shown to be enhanced by COR-Ll2IR and this efflux was demonstrated to be energy-dependent. The use of anthracyclines which retain activity in MDR cells thus appears to be a valid approach for the circumvention of MDR, not only in cells which

express P-glycoprotein, but also where defective drug accumulation is due to other mechanisms possibly involving an alternative multidrug transporter.

Chromosome alterations in 21 non-small cell lung carcinomas Miura I, Siegfried JM, Resau J, Keller SM, Zhou J-Y, Testa JR. Fox Chase Cancer Center, 7701 B&v&e Ave., Philadelphia, PA 19111. Genes Chromosomes Cancer 1990:2:328-38.

Cytogenetic analysis was performed on 16 primary tumors, 2 effu- sions, and 3 cell lines from 21 patients with non-small cell lung cancer (NSCLC). In 20 patients specimens were obtained prior to initiating cytotoxictherapy.Ertensiveclonalchromosomealterationswerefound in all cases. The most frequent numerical changes werepolysomy 7 and polysomy 20 (each seen in 12 specimens). In addition, tumor cells from another six cases exhibited partial trisomy 7, with the shortest region of overlap(SRO)at7pll-pl3.Reat~angementsofchmmosomes I, 3,6,8, 11, 15, 17, and 19 were each observed in nine or more tumors. Breakpoints were clustered at several chromosomal sites, including Ip13,3pl3,15pll-q1l,l7pll,andl9ql3.Recurrentlossrnvolvinglp, 3p,6q, 1 Ip, 15p, 17p,and 19q wereeach seen inat IeasteightcasesThe SRO of 3p losses was at band 3~21. Double minute chromosomes were found in three tumors. Overall, our findings indicate that even though karyotypes in newly diagnosed NSCLC are very complex, recurrent cytogenetic changes can be identified. The high incidence of loss of 17p (14 of21 specimens) appears to be compatible with reports implicating the TP53 gene (at band 17~13) as a frequent site for genetic alteration in lung cancer. Moreover, the recurrence of loss of 3p (12 cases) and I 1 p (10 casea) is also consistent with recent molecular evidence. The existence of other “hot spots” for cytogenetic change, particularly those involving specific regions on chromosomes 7, 15, and 19, war- rants further molecular investigation of these sites in NSCLC.

[Des-Met”]bombesin analogues function as small cell lung cancer bombesin receptor antagonists Staley J, Coy D, Taylor JE, Kim S, Moody TW. GWU Biochemistry Department, 2300 Eye St. N.W., Washington, DC 20037. Peptides 1991:12:145-9.

A series ofbombesin (BN) analogues lacking the C-terminal methion- ine at the 14 position were evaluated as BN receptor antagonists. ID- Phe”]BN(6-13)amide inhibited specific ‘=I-GRP binding to lung cancer cell line NCI-H720 with an IC,, value of 12 nM. In contrast, [D- Phe’]BN(6-13)propylamide. butylamide and methylester were more potent with IC,, values of 3,5 and 5 nM whereas [D-Phe6,Sta”]BN(6- 13)amide was less potent with an IC,, value of 180 nM. [D-Phe6]BN(6- 13)propylamide antagonized the ability of BN to elevate cytosolic Ca”, whereas [D-Phe6]BN(6-13)butyIamide was a partial agonist. In a small cell lung cancer (SCLC) growth assay, [D-Phe6]BN(6-13)propylamide inhibited colony formation. In summary, BN analogues which lack a C- terminal methionine may function as useful SCLC BN receptor antago- nists.

Nearoteosin may function as a regulatory peptide in small cell lung cancer Davis TP, Crowell S, McInturff B, Louis R, Gillespie T. Department of Pharmacology, University ofArizona College ofMedicine. Tucson. AZ 85724. Peptides 1991;12:17-23.

Neurotensin (NT) has been postulated to act as a modulatory agent in the central nervous system. Besides its presence in mammalian brain, NT is produced by small cell carcinoma of the lung (SCLC) and cell lines derived from these tumors. Receptors have also been character- ized in some SCLC cell lines leading to the suggestion that NT could regulate the growth of SCLC in an autocrine fashion similar to bomb- esin/GRP. Previously, we had reported that a 10 nM dose of NT and NT@-13). but not NT(l-8). elevated cytosolic Ca2*, indicating that SCLC NT receptors may use Ca2* as a second messenger. Using intact SCLC cells we report that time-course incubations with NT lead to the formation of the amino-terminal fragment NT(l-8) and small amounts of the C-terminal fragment NT(9- 13). These fragments are formed by