289 Early T-cell changes following local allergen challenge of asthmatic bronchial mucosa

212 Abstracts 289

291

DISTINCT PATTERNS OF APPEARANCEOF NEUROPEPTlDES IN ACUTE IN-TORY REACTIONS OF THE HUMAN LUNG. P.F. Esoiritu. J. Seltzer. JA Weiner-Kronish. D. C. Adelsan. M.A. Matthav. H.A. Boushev. and E.J.Goetzl. San Francisco, California The adult respiratory distress :~;ndron~i* :IKDS; and O,-induced bronchial hyperreactivi:y (OIBH) both are characterized by nrutrophil;: inflammation and increased lesiona; fluid concentrations of arachidonic acid.derived capable of releasing neurupepLidcs mediators, from pulmonary neurons. The cDllcentrations of substance P (SP), calcitonin gene-related peptide peptide (GRP) WBI-P (CGW, and gastrin-releasing determined by specific radioimmunoass,iys iti cell-free edema fluid from patients with ARliS (n=7) and control patients with congestive heart failure (CHF, n=6), nnd in bronchoalvrolai I avege fluids from normal subjects after 0. ;xposurr and after breathing Filtered air during idonticai exercise (control) (n-7). The mean (iS.D concentrations of SP and GRP were higher :n .ARD:; than in CHF, with 73_+28 vs. 33~17 pg/nl of SP (pO.l). In contrast, the hi@: meail (2S.D.) CGRP concentration of 19+17 pg/ml ir! OIBH, as compared to E&8 pg/ml in cnnrroli was not accompanied by ;~ny differrncc (p-0.07), between the respective levels of 1156 and l?+lb pattern of pg/ml of SP. A distinctive steady-state concentrations of individual neuropeptides in human pulmonary aircxag fluids may be one characteristjc of differe::( inflammatory reactions

292

OUTWARDAND~ARD

Southampton,U.K. To investigate cellular events involved in allergen induced bronchoconstriction we have performed bronchoalveolarlavage (BAL) 1Ominafter local allergen challengeof bronchial mucosa.Inboth 10 mild asthmatics (PC20>0.5mg/ml) and 10 normal subjects allergen was instilled in RML through a fine nylon catheter during a fiberoptic bronchoscopy followed IOmin later by BAL (with 6x20ml prewarmed saline). At the same bronchoscopy a control challenge with 0.9% sodium chloride followed by lavage was undertakenin RUL. In all of the asthmatics but in none of the normals a visible bronchoconstriction was seen following allergen challenge.The volume of BAL was measured,the differential cell count estimatedand the cells subjectedto flow cytometric (FACS) analysis. No differences were found in the volume of BAL recovered or the differential cell count between the two challenged areas in normals and asthmatics. The FACS analysis showedno differencesin level of T-cells (CD3+) or their CD4+ and CD&t subsetsbetweenthe two groups in the non allergen challenged area.However,there was a significant change in T-cell subsets recovered from asthmaticsafter allergen chal1enge.Arise of CD8+ cells (~~0.05) and a fall in CD4+ cells(P
was no such changes seen between the

two areasin normal individualsIn addition no differences were found in T-cell activation markers11-2and HLA-DR in normals and asthmaticsor the control and challenged sites. Thesefindings are further evidence to support the role of T-cells as an initiator cell in the inflammatory processof allergeninducedbronchoconsrriction.

290

EPITHELIAL CELLS FUNCTION OF OBTAINED BY BRONCHIAL BRUSHING IN ASTHMATIC PATIENTS. A.M.-. P.Chana . .Michel. J,Bow P.Godiud, Montpellier, I. Couret . FB France. Airway epithelial cells may LX involved in the pathogenesis of asthma but their role remains to he determined. In normal subjects, these cells release eicosanoids(IS-I-IETE). Epithelial cells obtained by brushing during fiberoptic bronchoscopy were studied in 6 asthmatics (mean age: 449 yr) and 6 normal subjects (mean age: 5M7 yr). None of the subjects had received any anti-inflammatov treatments.The percentage of epithelial cells was examined after cytocentrifugation by immunocytcchemistry with anti-cytokeratin antibody (Dako). The viability of the cells was studied by trypan blue exclusion. The release of 15HETE was studied in resting cells and after stimulation with 5pM of A23 187. The mediator was measured by HPLC and RIA using a monoclonai antibody (Arnersham, UK). The percentage of epithelial cells was always over 86% of cells recovered. Viability of epithelial cells was significantly (pcO.05, Mann-Whitney U test) greater in normal subjects (m rt sd: 64z!z446) than in asthmatics (15rlr3 %). 15-HE’IE RELEASE BY F.PITHFLIAL CELLS cm?/106cells) baseline A23187 Q value viable cells: asthma controls total cells: asthma controls

7213-l 4k 1
lof 2A 2+ 1

94k81 ;g; co.05 8f2l

NS
(statistical analysis by non-parametric tests) This study suggests that epithelial cells are activated and less viable in asthma. supported by the Fond Sptfcial des Maladies Respiratoires

This study tests the mels that mucosal &udation of plasma solutes may occur without affecting airway absorption permeabiilty (Int Arch Allergy Appl lmmunol 1990, 92, 14Q-53). Guinea-pig tracheobronohkl mucosa was superfused in vivo with 40 ILI saline, with or without Mstamine 5 nmol, and subsequently lavaged in vitro by validated non-traumatic techniques (pulm Pharmacol 1Q8Q,2,Q31~2). Mucosal exudation of intravenously I-atbumin was expressed as ml piasma. administer c-DTPA (MW 492) was co-adm@stered in Separately, % the superfusate and its tracheobronchiai half-life determined as previously descni (Thorax 1990 in press). Histamine produced mucosal exudation of 1.571rO.41 ml I. pO.O5). A Nasal Fool technique was employed for simultaneous provocation and lailage of human nasal rnuqqa (Clin Exp Allergy 1990, 20, 253-9). Cr-EDTA (MW 372) with -or without Saline containing histamine (2 mg/ml) was instilled in the nasal cavity for 10 min and subsequently recovered. Absorption of 5’CrEDTA was determined as urinary radio-activity and expressed ae mt in&ate. Histamine increased the lavage fluid levei of albumin from 2223 p~,rnl (con@&) to 377&O fig/ml (~~0.01) but absorption of Cr-EDTA was unaffected (control: 0.10~0.03 ml; h&amine: O.OQ1tO.02 ml. p >0.05). These data support the view that mucosal exudation of plasma occurs in airway defence and in inflammatory airway diseases (Lancet tQQ6,2,1126-9) without increasing absofption of luminai solutes.