109 INDUCTION OF DIFFERENTIATION OF A549 HUMAN ALVEOLAR TUMOR CELLS BY BASEMENT MEMBRANE

" 107 I N V ITRO PROLIFERATION OF ACTIVATED LYMPHOCYTES INFILTPATING SMALL BOWEL IN COELIAC DISZASE. " M.StegagnO,M.C~biaggi,M.B~namic~,P.Mariani,E.Carapella Departments of Child Health and Paediatrics, University "La Sapienza" Rome - ITALY. 110 THE AMIBODY (antlCD 2, antiCD 3) INDUCED LYMPHOCYTE MITOGENESIS IS DEFECTIVE IN INFANCY AND EARLY CHILDHOOD N. Remy.S.Krusche.T. Grass. M. Oberreit. U. Wahn Univ. Childrens Hospital. FU Beriln,Germany The ability of interleukin 2 (IL-2) to stimulate cellular division of actrvated T lymphocytes allows to propagate3'invitro" the T cells ~nfiltratinginflammatory tlssues. we developed a system that allows to propagate relevant amounts of activated T lyxphocytes from minute fragments of peroral small bowel bropsies performed for diagnostic purposes. In preliminary experrments on three different patients with gluten enteropathy on a gluten containing diet, the culture of bioptic tissue in the presenceof IL-2 allowed us to recover as many as lo8 lymphocytes. The evaluation of surface markers with monoclonal antibodies showed that these cells were virtually all T Lymphocytes (CD3+): the two different T cells subsets, CD4+ (helper / inducer) and CD8c (suppressor / cytotox~c)were almost equally represented in the two patients challenged with gluten while in the third one, who was at the first diagnostic biopsy, there was a marked predominance of CD8+ cells. This difference could be related to the different phases of the disease in the patients examined. The relevant numbers of cells obtained with ourexperimentalprocedure can be used in functional assays to further investigate the pathogenesis of coeliac disease. Recent findingsIndicatea defectivemitogenesis ofcord Mood lymphocytes as demonstrated by stimulation with the monoclonal ant bodies antiCD 212a (aitemativepthwayfand antiCD 3 (Tcell-rezeptorcomplex)(Gelli 89). In orderto investigatetheage dependencyofthisantibody Inducedtransformationand a possible assocbtion withchangesof lymphocyte su&ts. Mood was obtained from healthy subjects: cord blood samples.5 day dd neonates,4 month dd infants.2-4 year dd children as well as aduits (n= 10 In each group). Lymphocytes were incubated for 96 h with optimal concentrations d anti CD-2/2a and antlGD 3. In addition lymphocyte subsets C D 2 and C D 3 were determined by flow cytometry. Only cord Mood samples were found to have a slgnificanliy (p<O,M)l) reduced percentage d CD 2 and CD 3 positive lymphocytes, wheras from day 5 on percentages were within the range d adults.. In contk~st,proliferative responses of lymphocytes induced by antlCD 2/2a and antlCD 3 were signiilcantly reduced in all pediatric groups (p<O,WI)compareto adub. Wihin the &iatric groupsthere were no differencesin lymphocyte responses. From our data w e condude that In infancy and early childhood there is a functional defectof lymphocytes detectable by stimulationwkh monoclonal antibodies,which Is not related to differences in lymphocyte subsets. COGNITIVE EFFECT AT 5 YEARS OF AGE IN INFANTS WHO WERE ANEMIC AT 12 MONTHS: A LONGITUDINAL STUDY. T. Walter, I.de Andraca, M.Castillo, F.Rivera, C.Cobo. Hematology and Psychoneurology Units. INTA, U.of Chile, Santiago. Anemic infants from a longitudinal study of iron nutrition who were evaluated with the Bayley Scales of Infant Development showed significantly lower scores than their iron sufficient controls at 12 mo and after therapy for complete correction of iron status. Anemia was determined by random assignment to non fortified diets (Peds.84:l-17,1989).At 5f years of age they were reevaluated with a general intelligence test (I), fine and gross motor abilities (2), psycho-linguistic capabilities (3), visual-motor integration (4) and an educational preschool scale (5). Maternal and environmental variables have been controlled for. Results are: Former iron status(at 12 mo) Anemic n=43 p< control n=29 Current age(months) 66 _+ 2.4 NS 66.5 + 4.8 (1)Stanford-Binet(1Q) 87 2 8.1 .05 92.4 A 8.0 (2)Bruininks-Oseretsky(Composite) 39 2 9 -04 44 + 8 (3)Illinois(composite quotient) 83 _+ 11 .02 89.6 5 8 (1)Visual Motor(percenti1e) 31 _+ 24 .02 45 + 24 (5)Woodcock(Score) 86 3 19 .02 96 11 Unfavourable effect of iron deficiency anemia on development at 12 mo persists in cognition at 5 1 despite timely and adequate iron therapy.Although causality cannot be proven,anemia in infancy may be a marker for disadvantaged cognition in childhood. LUNG E D E M DECREASES THE RESPONSIVENESS TO EXOGENOUS SURFACTANT. Mikko Hallman, Veeraiah Chundu, William Cvetnic, Stefan Slivka, University of California, Irvine, Medical Ctr., Department of Pediatrics, Orange, CA. There is a large variability in the effect of exogenous surfactant in RDS. We have tested the hypothesis that hydrostatic lung edema decreases the responsiveness to surfactant. Respiratory failure was induced to rabbits, wt. 1-1.3 kg,by alveolar lavage (BAL, 10 mllkg x 4). Thereafter, natural surfactant (NS, 100 mglkg in 4 ml saline, n=10), saline carrier (n=8), or air (n=8) was administered.The animals were paralyzed and ventilated at constant tidal volume. They received either 1 (low) or 20 mllkglh (high) saline i.v. during 155 min. The lung function was measured and BALs were analyzed for phosphatidylcholine (PC), concentration of PC in epithelial lining fluid (PCelf), protein, and surface activity. Saline carrier to airways decreased the gas exchange and compliance (p<0.01). In animals on low i.v. fluids, NS improved the respiratory function as compared to air-treated controls. However, in animals on high i.v. fluids, NS neither improved the gas exchange nor the compliance, despite the increase in PC (p< 0.05) and the decrease in soluble protein (p<0.05) in BAL. The lack of surfactant responsiveness was due to low PCelf and to surfactant inhibitors. Thus, variability in surfactant responsiveness may be caused by factors affecting lung liquid. 108 INDUCTION OF DIFFERENTIATION OF A549 HUMAN ALVEOLAR TUMOR CELLS BY BASEMENT MEMBRANE. Paul Stevens1, Ursula Brands1, Bernd Rustow2, Michael Obladenl. 1 Univ. Children's Hospital, Berlin ,FRG; 2 Inst. Pathol. Clin. Biochemistry. Charit6 Hospital, Berlin , GDR. A 549 tumor cells may be of type II cell origin, but do not synthesize surfactant under normal conditions. We cultured this cell line either on tissue culture-treated plastic or on top of an extracellular matrix, obtained from the Englebreth-Holm-Swarm(EHS) tumor. We found that cells cultured on EHS matrix synthesized surfactant protein A (SP-A) as shown by Western blot and by slot blot hybridization with an SP-A cDNA probe (n=5). Compared to cells cultured on plastic, cells cultured on EHS for 3-5 days also contained a higher percentage of 16:O-16:O phosphatidylcholine(10 vs. 2 O/o. n=2) and incorporated more 3H-glycerol into this lipid species (10 vs. 2.8 O h , n=2). On EHS matrix, the cells became cuboidal, and formed alveoli-like spherical structures surrounding a closed lumen. Also (...truncated)