402 Abstracts

6.

GALACTOSYLTRANSFERASE IN MALIGNANT EFFUSIONS Eric G. Ber er'

--+, - E.F. Holdener', K.L. w' and

A..,. Gehler . 'Medizinisch-chemisches Institut, UniversitBt Bern, Postfach, CH-3000 Bern 9, Switzerland. 'Medizinische Klinik C, Kantonsspital, St. Gallen, Switzerland.

Galactosyltransferase activity (GT.E.C.2.4.1.22) and immunoactivity were determined in 69 malignant, cytology positive effusions. Enzyme activity was measured using free N-acetylglucosamine as acceptor substrate: immunoreactivity was determined by ELISA using a polyclonal rabbit antiserum against the so- luble human milk enzyme.

The tumor types analyzed included ovarian (9). breast (15). lung (12), gastrointestinal (12) and miscellaneous cancers (21). The median value of specific GT-activitv was hishest for ovarian cancer (b.57 U/L) as well is the &an value (0.80 U/L) with a range from 0.11 to 2.25 U/L. For comparison mean serum reference value for GT is 0.29 U/L. These values suggest that effusion GT is generated by cancer cells and that the determination of its level in effusion may be helpful in the diagnosis of ovarian cancer.

Supported by grants FOR 195AK81(2) of the Swiss Cancer League, and 3.873.0.79 of the Swiss National Science Foundation. X.L.H. was a recipient of a fellowship of the Roche Studienstiftung.

7.

THE MEMBRANE-ASSOCIATED I-GLUTAMYL TRANSPEPTIOASE (I-GT) IN THE 01AGtdos1s OF ACUTE LEIJKEMIAS (AL). 0. Heumann, V. van Fliedner. A. Morel]. G. Loss. Ludulg Institute for Cancer Research Lausanne; Instltut fur Kllnisch-ExperlmentelIe'Tumorforschung Bern and Istltuto Cantonale dl PatoIogia.Laboratory of Cellular Pathology, Locarno, Switzerland.

Flcoll-enriched blast suspensions from 92 patients with AL were classified using surface markers. mor- phology and cytochemistry. The cells were biochemically as- sayed for the plasma membrane associated I-GT and terminal transferax (TdT). In myeloid leukemias (n-37)we observed an increase of I-GT activity in immature AML (FAB-Ml, median: m = 18.3 u;u = nmole/hr/l06 cells). This activity gradually decreased in more mature AML (FAB-t42-~3, m = 10.211). The level of I-GT in granulocytes was 11.5~. Myelomonocytic and monocytlc AL showed the highest I-GT values (m = 29.7u in M4 and 30.2~ in M5). Extremely high values (150-250~1 were only recorded in M4 and MS AL. The value for monocytes was 2.5~. In lymphoblastic Ieukemias (n=41), however. the i-GT was very low (m=l.Su for C-ALL and 2.1~ for T-ALL). The median was 9.Ou and 14.0~ I" normal T and B lymphocytes. Interestingly, I-GT values had a bimodal distribution in oatients with acute undlfferentlated leukemlas (AUL. ~14): b patients had values between 0 and 2.8~ and 5 between 7.6 and 21.0~. The correlation with TdT allowed to define 3 types of AUL: a) lymphold-like (TdT+. low I-GT); b) myelo- monocytic-like (TdT-, high I-GT); C) undifferentiated type (TdT-, low I-GT). 13 of 14 AUL cases fitted in these 3 categories. As l-GT activity is nearly absent in lymphoid AL and elevated in myeloid and monocytic AL, this enzyme may be a helpful marker for the distlnction between myeloid and lymphold leukemias.

8. ENZYME CHARAXTERISTICS OF PLAW MEMWANE IN CELLS FROM NON-HODGKIN LYMPHOMAS (NHL). G.A.Losa. Laboratory of Cellular Pathology,Istituto cantonale diPatologia.6604 Locarno , CH.

The activity profiles of marker enzymes associated with the plasma membrane were investigated on cells isolated from lymph nodes of patients with malignant NHL.The composition of the population was assessed by testing cells for the presence of surface irmwnoglobulins(SIg) and T antigens. and of non specific esterase.Activity levels of plasma membrane enzymes involved in purine metabolism,as 5'-nucleotidase(S'-APase) and nucleotide phosphodiesterase.in ion transport as adenosi- ne triphosphatase and alkaline phosphcmonoesterase.and in a- minoacid transport as&glutamyltranspeptidase(GLUPAase) were assayed at saturating-concentration of substrate.Cases of NHL with a majority of cells bearino SIo denoted a sionificant lower activity-level of 5'-AMP&e in comparison oi cells of non malignant lymph nodes.In contrast.NHL with abundant cyto- plasmic inrnunoglobrlins(CIg) displayed a significant higher activity of this enzyme.A third group included NHL cases with a varlable proportion of lymphoid cells bearing either B or T markers but without detectable CIg.Correlations be-

tween the various enzyme activities were absent with the ex- ception for 5'-AMPase and GLUPAase.These findinas indicate .- ---- that the cellular heterogeneity of NHL is reflected by the enzymatic heterogeneity of the plasma membrane which in turn may relate to biological peculiarities of malignant cells.

IS THIS MtjlW2D ? G.K. y. Schulthe& Ch. L& U. H& R. An&es F. &wzheggerw J.P. Mach' W. WelheO R. Am+wd2 and A. Bekler~, te of ~uclearne1 end In&it rlewrtement Of surqery Y Kaotonesoital CH-9~7 St. Gallen EIR SWISS wct0jj in&t~te3 CH 5303 wpilngen, ~loche-' mistry Institute and Ludwig Institute , c&la66 Epalrges and Central Rnlmal Laboratory6 Univ. Hospital c&&91 &inch, Switzerland.

With the developrent of rronoclonal antibodies and new Nuclear Medicine tecbruws. there 1st areat hone that ~m)ro-

antibody may be better detected by sclntlgraphy than by mar- phologlcal abnormalities. We report here on the lnltlal cllnical expnence with this mthcd at our hospital as well a.5 on experucental results obtained in nude mre bearing

bdy the coUnt rates are so low that exennnatlon by emission co-ted tomography ~111 present some problems. Rlrthermore, the tomorauhlc localization of the radloactlve uDtake I" tumxs‘ofte~ requn-es the use of an atitional sc&grapby with a bone or liver seeklng Technetium labslled agent. Clearly. this t-r detection method let et111 at a stage of cl~nlcal research. The lcqstxs needet are formidable, it requres expert producers of ant1bcclles. expenenced radio- chemists, anstant quality controls, experimental msdels of human tlUB5r grafted in nude race. and an optimum collabora- tnn between surgeons and nuclear pbyaxw,s.

10.

RCINOMA BY EMISSION COMPUTERIZED LABELED F(ab')2 FRAGMENTS FROM

MONOCLONAL ANTI-CEA ANTIBODIES. J.-Ph. Grab, B. Delalo e, A: IW;f-""'a;;~;;,;. :""::~~:~;,'.V'ld~~~el.

.-. ac. u L dwig Institute for Cancer Research, CH-1006 Epalinges, Div. of Nuclear Medicine and Dept. of Surgery CHUV, Institute of Biochemistry and Policlinique Medicale Universitaire, CH-ID11 Lausanne.

Eleven patients with known colorectal carcinomas were injected each with 1.5 mg of F(ab')2 of awnoclonal ant{&:" antibodies No 35 and 202 labeled with 3-4 mCi of P-5n ECT was performed in all patients 6, 24 and 48 h after injection using a double head Rotacamera. In all 5 patients with localized carcinoma (4 primary tumors from the caecum, right colon, sigmoid and rectum, respectively, and 1 signmid recurrence), the tumors were clearly detected by ECT. In 2 patients, turrar to normal tissue radioactivity ratios were measured on surgically resected material at day 5 post injection and found to be 6, 8, 15 and 9 for one patient and 3, 5, 7 and 2 for the other, in comparison with normal mucosa, serosa, fat and blood, respectively. Two out of 2 bone metastases, in the sacroiliac area and in the scapula were also detected, one of which was previously unknown. In 1 patient under chemotherapy 2 small lung metastases were not detected. Out of 6 patients with liver metastases, 2 remained negative, !&ile in the 4 others, the metastases presented as cold area at the 6 h ECT. These defects progressively filled with radioactivity at later ECT in 2 cases and gave doubtful uptakes in the 2 others. Briefly, 9 of the 14 turrnr sites were localized by ECT, 2 were doubtful and 3 negative. The excellent definition of the tumor in the positive cases represents an improvement over previously reported results.

II.

IMMUNOSCANNING OF GI TRACT ADENOCARCINOMAS USnJG TUMOR SPECI-

FIC “ONOCLONAL ANTIBODIES (MoAb): A PROSPECTIVE STUDY. JY.Douillard. PA.Le SW* and .JF.Chata*. CRLC- INSERM U.211- -Y ??Hopltal se -- Jacques, CGI (Pr "Isset) 44035 NANTES CEDEX F-

Abstracts 403

Anti-CEA mou*e NoAb, produced by JP Mach (Ludwig Inst.. Lau-

sanne, Switzerland) were injected IV as 500 yg of intact im- munoglobulins, 1311 labeled according m rbe iodoge” technic to a specific activity of 5 to IO ucilup. Mouse M”Ab 19-9 and 17-1-A provided by H. Koprovski (Wistar Inst., Philadelphia, USA) labeled with 1311 LO a specific activity af 2 pCi/ug were injected IV as I mg of F(ab’)2 fragments. A mixture of 2 out of the three antibadies menfianned above was infused 22 out of 27 Limes, and scanning was perfamed up to 7 days af- ter injection. Final disgnasis was obtained by pathology (141 27) or 0” clinical evolueio” (13127). Overall specificity af the method was loozl (10110) with no false positive results, sensitivity yielded to 59% (10117) with 7 false negative re- sults and accuracy happened to be 74% (20127). According to our limited experience, three major indications could be se- lected Drosoectivelv:,)Isalated elevation of serum eumar mar- kers, 2jNo”‘co”vi”ci”g evidence a” co”ve”tio”“a1 methods, 3) Staging of GI carcinoma prior 10 surgery. I” mmt cases. fai- lure of the method ~85 observed in peritoneal carcinoaatosis (4/7) or when recurrence accured behind the bladder, making imaae analysis difficult due to urinarv clearance of 1311. _ Improvement of the technic could be expected fram mre appro- priated radiolabel than Iodine and possibly from single pho- to” mission tomography along with ehe assaciacio” of several MoAb sharing similar tumcx cell specificity but reacting with distinct epitopes.

12.

ESTIMTION OF CIRCUATING IWINE CWPLEXES IN CANCER PATIENTS: SENSE OR MMSERSE? II. Bcrtschann, J.P. Splth. U.E. Nydq9w and E.F. LUSChW, Theoddh Kocher Institute, University of Bwme and Blood Transfusion Service, Swiss Red Cross, Central laboratory, W-3000 Bern 9. Switzerland. Since the detection of blocking factors in serum of cancer patients and the identification of circulating tumor-antigen- antibody complexes (CIC) as the active entities a large body of data has accumulated from experimental animal models and cancer patients in the hope to use levels of CIC as para- meters for diagnosis and/or prognosis of malignancies. In the present experiments CIC were sequentially determined in serum samples from DBA/Z mice bearing histoco.patible P-815 masto- cytomas. 1251~Clq binding assays and Raji cell binding assays were used according to standard procedures. The tumor model is characterized by a transient phase of spontaneous regres- sion correlated with proliferation and differentiation of cytotoxic T cells. However, regression normally changes to progression and not only T- but also B cell proliferation is observed. Since no functional correlate of B cell stimulation in the form of crrculating or tumor cell bound antibody could be detected so far, tests were performed for detecting antibody in the form of CIC and for correlating CIC levels wth tumor development. Despite the wide variation in tumor development elevated levels of CIC were not detected, neither in the 1251-Clq binding nor in the Raji cell binding assay. Reference complexes formed between soluble tumor cell membrane proteins and allo-antibody, however, were easily detected. These negative results are in accordance with recent negative results in mannary cancer patients (Krieger et al., Int. J. Cancer 31, 207, 19831. They support a more crltlcal evaluation of the usefulness of CIC determination for diagnosis and/or prognosis of malignancies.

13. MONOCLONAL ANTIBODIES AGAINST HUMAN LUNG CARCINCMAS. R. Stahel and S. Bernal. Dana-Farber Cancer Institute and Harvard Medical Schaol, Bosta”, MA 02115, U.S.A.

House monoclonal aneibodies were generated against cell lines of human lung adenocarcinma (SLC40) and small cell carcinoma toHI). SK‘0 growr as cellular aggregates in sus- pension culture and forms elandular structures in nude mice. The antibodies LAn2 and L&3 are strongly reactive vith the surface memb,-a”e of SLC40, as demonstraced by indirect immu- nofluorescence and radioimmunoassay. I&,3 is also reactive with several lung adenocarcinmas SrowinS as adherent or suspension cells in culture, but ““t with other adenocareino- mas, including ml”” and pancreas. LAM3 may be helpful in distinguishing adenacarcinmas of Lhz lung fram other carci- nomas. I” mntrast, LAN2 is reactive with several carcinomas chat grow as suspension cells in culture, including small cell carcinomas, lung adenocarcinmnas, mesothelima and semi- “mm. Little or “O reactivirv was observed wirh 18 carcinma cell lines that grow adhere”; t~1 the culture dish. Thus, LAM2 recognizes a surface membrane antigen that is present

marrow cells and leukemic cells. These antibodies may be use- ful in studies of differentiario” and biologic behavior of lung cancer cells; they could also be important in the diag- nosis, staging and treatment af lung cancers.

Evaluable patients 24 16 COnplete remission (CR) 6 (25%) o (0.2) Partial retiesion (PR) 15 (62%) 9 (56%) CF. + PR 21 (87X) 9 (56X)

evide”ce of pro ressive diaearre ; rhe corresponding figure f”r PR’B i* 141 4. s

Although final c~nclusims ca” only be drew” after

16

TOT&L BODY IRRADII\TION (TBIi AND CYCLOPHOSPHAMIDE tcy, AS A CONOITIONNING REGIIIEN OF ALLOGENIC MARROW TRANSPL.*NTATION (ALL0 M.T.I : INVESTIGATION OF SINGLE DOSE VERSUS FRACTIONNPI- TED REGIHEN OF TBI. D. Maraninchi, D. BPUR~, J.A. ~a~taut, B. Mascret, J.P. Guillet, R. Amalric and Y. Carcasaonne, narrow Transplant Unit. I”XZXTZli-Calmettes (IPCl. Marseille 13273 - WB”C.2 -

we investigated I” 33 patients (pt8) with bematologlc mall- ~“ancies different TBI regimen as a preparatlo” of allo M.T.. 5 pts received 1000 rads %I (lungshielding 8001. 4 pt6 recei- ved Zoo rads per day for 5 days. 11 pt~ received 220 rads ,,er day for 5 days. 7 pts received 240 per day for S days. Irra- diation source was linear accelerator seturne. Lungehielding