Inflammatory mediators in otits media with effusions

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INF-TORY MEDIATORS IN OTITS MEDIA WITH _. . EFFTJSIONS. Y. Faculty of Health Science, Ben-Gurion Univ., Beer-Sheva. Israel. VOotkis media wit% effusion can be a silent condition following subclinical or prolonged inflammation of the middle ear which might result in chronic otitis media with effusion (OME) and chronic suppurative otitis media without cholestatoma (CSOMWC). The factors that contribute to OME have not been established definitively. In an attempt to further identify the pathophysiology of these diseases we examined the ear effusion from 23 patients with OME and 28 patients with CSOMWC for the presence of three potential inflammatory mediators that have been shown to participate in tissue damage

PGEQ in the ear effisions ofCSOMWC patients might. contribute to the irreversible stuctural damage in the middle ear (mucosal orbony)observedinthis disease.

EPIDERMAL CYTOKINE? PRODUXION DURING ONlWZ;ENy AND AGING. S.St&i&GrujiEid and M.L.Lukid. Inst.Biol.Res. and Inst. Microbiol.Immunol.,Sch.Med.,ll060 Belgrade, Yugoslavia. The capacity of epidermal cells (ECs) to produce cytokines that play a role in immune and inflammatory responses is well established. As the ability to elicit these responses in the skin vary with the age, we set up a model to analyze, at different. age, the in vitro capacity of ECs to produce IL-l, IL-~ and IL-2. ECs wereobtained from A0 and DA rats after separation of epidermis from day 16 old fetuses to 20 months old adults. After 24 hr ECs culture supernatants were harvested and tested for IL-l(DlOS assay), IL-2(CTLL) and IL-6(B9) bioactivities. Cytokines able to induce proliferation of CTLL cells were not present. IL-l and IL-6 were detected after day 16 of fetal development and showed sharp increase thereafter. The constitutive production of relevant. cytokines peaked one day prior to the birth. In young adults IL-l and IL-~ expression declined gradually and was very low in 20 months old rats. We then tested the ability of ECs to produce IL-l and IL-6 receptor antagonists (t-a), produced as previously shown by us after exposing cells to various glucocorticoids. To this end DIOS and B9 assays were performed in the presence of exogenous hrIL-1 and hrIL-6, respectively. It. appeared that the production of ra was also age related: fetal ECs and cells from younger animals being higher producers that cells from aged animals. The homeostatic balance of ILs and ILs-ra during ontogenesis, aging and pathologic events remains to be established. (Supported by RSF, Belgrade).

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IN VIVO BIO-DISTRIBUTION AND METABOLISM OF RADIOLABELLED IL2 IN NORMAL AND DIABETES PRONE MICE. A. Sianore Servizio Speciale Medicina Nucleare, II Clinica Medica, University of Rome “La Sapienza”, Italy. Little is known about IL2 kinetics and metabolism in normal and pathological conditions such as autoimmune diabetes. The aim of this study was to investigate the in vivo bio-kinetics of radiolabelled IL2 in Balb/c and diabetes prone NOD mice with particular regard to its binding to lymphocytes homing into lymphoid organs (such as thymus and spleen) and pancreas. Interleukin-2 was labelled with l*slodine and injected i.v. in 32 NOD female mice of 15 weeks of age and in 20 normal Balb/c female mice. Animals were sacrificed after 2, 5, 10, 20, 40, 60 and 90 minutes and liver, kidneys, spleen, pancreas and thymus were removed, weighted and counted for radioactivity before freezing in liquid nitrogen for subsequent histological examination. Results of this study show that IL2 has a bi-esponential plasma disappearance curve with a first rapid half life of 8.03?0.8 mins and a second of 203k6.2 mins in both groups of animals but NOD mice show a higher accumulation in the pancreas, spleen and thymus from the 20th to the 90th minute after injection, as compared with Balb/c mice (at 40 min: Pa NOD=5.31+1.71 vs Pa Balb/c=3.61+0.83, ~~0.04; Sp NOD=15.39*4.46 vs Sp Balb/c=6.38*1.77, pcO.004; Th NOD=6.16+3.89 vs Th Balb/c=2.24?0.73, ~~0.04; data are % of injected dose/gr of tissue *SD). The high accumulation of IL2 in the thymus indicates a high expression of IL2R on NOD thymocytes and this may be of great relevance for the pathogenesis of diabetes.

THE CONTRIBUTION OF HOST INFLAMMATORY CYTOKINESINEXPERIMENTALCANCERCACHEXIA. G. Strassmann, M. Fong, and R. Evans. Otsuka America Pharmaceutical, Rockville, MD 20850 and TheJackson Laboratory, BarHarbor,ME 04609. The goal of this study was to evaluate the contribution of host

and remodeling: tumor necrosis factor (TNF), interleukin-1 (IL1) and prostaglaudin E2 (PGEP). The mean values of IL-1 and PGE2 concentrations in the CSOMWC group were significantly higher than those in the OME group (50f71fmoVml and 1172k766

pgfml

as opposed

to 9k39

fmol/ml

and

315ti22

pg/ml

respectively). In this group IL-1 could be detected in 100% ofthe cases while only 9% were positive in the OME group.The TNF values in the two study groups were not statistically different. (61f132 pg/ml as opposed to lOf17 pg/ml) nor was the number of positive

cases.

The

elevated

concentrations

of IL-l,

TNF,

and

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THE SIGNIFICANCE OF INFECTIONS IN MICE.

inflammatory cytoldnes to the development of experimental cancer Few cell lines were derived from colon-26 cachexia. adenocarcinoma tumor. In viva, these lines developed small tumors which caused severe wasting (up to 40% of body weight), complete depletion of epididymal fat, and hypoglycemia. In contrast, B-16 melanoma produced large tumors and did not cause cachexia.

Concentrated and dialyzed serum-free conditioned medium from

C-26 and B-16 lines did not contain IL-l, TNF, or GM-CSF as measured by radiorecepter assay and bioassays. The C-26 conditioned medium contained significant levels of IL-6 (which was completely neutralized by anti-IL-6 antibody) while B-16 produced

30-fold less IL-6 on a per cell basis. Next, we used peritoneal macrophages as indicator cells to evaluate potential host response to tumor products present in the conditioned medium. Northern blot analyses revealed that both C-26 and B-16 conditioned medium induced a high level of IL-la, IL-U, TNF and IL-6 gene transcription, while culture medium (control) induced only low level (S-fold less). Thus, the induction of host inflammatory genes (i.e. IL-l, TNF and IL-6) occurs in response to both cache&c and noncachetic tumor products and therefore do not appear to be specific events in cancer related cachexia.

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S. Steinshamn and A. Waaqe Institute of Cancer Research, University of Trondheim. N-7006 Trondheim. Norwav. We ha& studied the prod&ion and significance of TNF and IL-6 during C. albicans infections in mice rendered granulocytopenic with cyclophosphamide and in normal mice. TNF was systemically produced in a dose-dependent manner, being detectable between 6 and 60 hours after infecting the mice with peak levels at 24 hours. IL-6 was also systemically produced during C. albicans infections. However. kinetics of IL-6 oroduction did not show any reprbducable regularicy. The levels of both TNF and IL-6 were higher in cyclophosphamidetreated than in normal mice. Heat-inactivated C. albicans induced a TNF-response different from that of viable C. albicans with peak-levels occuring at 3-4 hours declining rapidly to non-detectable levels at lo-12 hours. Furthermore, peak-levels after heat-inactivated C. albicans were lower than after viable C. albicans. Growth of C. albicans from the kidnevs was auamented in normal, but not in granulocytopenic, mice treated with a mbnoclonal antibody to TNF compared with control mice. This study thus shows that TtiF and IL-6 are produced during C. albicans infections in mice and that TNF to the granulocytes' anti-fungal is essential activity against C. albicans in vivo.

INCREASED ACTlVlPl OF THE PITUITARY-ADRENAL AXIS FOLLOWING CHRONIC TREATMENT OF RATS WITH INTERLEUKIN 1 -BETA. C.G.J. Sweep, A.R.M.M. Hermus, M.J.M. van der Meer, A.G.H. Smals, J.W.M. van der Meer and P.W.C. Kloppenborg. Department of Medicine, University Hospital, 6500 HB Nijmegen, The Netherlands. It has been shown that acute administration of IL10 in rats elicits an increase in plasma ACTH and cotiicosterone (B) levels. We investigated the effects of chronic administration of ILlI on the activity of the pituitaryadrenal axis in rats. Rats were implanted with jugular cannulas and minipumps loaded with either ILli. (delivery rate 2 or 4 pg/24h, i.p.) or saline. Blood was collected daily for 7 days for ACTH, 6, prolactin (PRL), epinephrine (E) and norepinephrine (NE) determinations. After decapitation on day 8 the adrenals were weighed and the in vitro release of O-endorphin (BE) by the anterior pituitaries and of B by the adrenal glands was studied. Results: administration of 2 and 4 rg IL1 0/24h induced significant increases in plasma ACTH and B values, and in adrenal weights. These treatments also increased the basal in vitro release of BE and B, whereas in vitro pituitary/adrenal responses to CRF (1 O-‘M) respectively ACTH (250 pg/ml) were significantly decreased. Significant elevations of rectal temperatures were observed 1, 2, 4 and 6 days following start of the 4 &g IL1 0 treatment. At the dose of 2 #g/24h, IL1 I3 temperatures were significantly elevated only the first two days. Plasma levels of NE and E were elevated the first day following treatment with 2 and 4 ,,g IL1 I3 and had returned to control levels two days after implantation. IL1 13treatment did not affect plasma PRL levels. Conclusions: our data show the potential of IL10 to induce a specific and long-term activation of pituitary-adrenal activity upon chronic administration.