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285 Bronchial mucosal intercellular adhesion molecule-1 [ICAM-1] and endothelial leukocyte adhesion molecule-1 [ELAM-1] expression in normals and asthmatics
VOLUME NUMBER
285
87 1, PART 2
BRONCHIAL mDOTIJ&IAL
Abstracts
MUmAL INTERCEJJJLAR MOLECI,&& 1 IICAM-11 LEUKOCYTE AD&%
287
ENHANCEDBASOPHIL HISTAMINR RELEASE (HR) FOLLOWING IN VITRO INCUBATION WITH LIVE INFLUENZA A VIRUS IS ATTENUATED BY T-CELL DEPLETION. M.A. Huftel, M.D., C.A. Swenson, B.S., W.R. Borcherding, B.S., E.C. Dick, Ph.D., R. Hong, M.D., W.W. Busse, M.D., Madison, WI A number of mechanisms participate in virusPreviously, we described eninduced asthma. hanced basophil HR during an experimentally induced rhinovirus infection and following & .vitro incubation of peripheral blood mononuclear cells (PBMC) with Influenza virus. Furthermore, incubation with interferon produced similar results, suggesting cytokines may play a role in this process. To further define the role of Tenhanced basophil HR, PBMC (basocells in virus monocytes, phils, and lymphocytes) were isolated from ragweed allergic subjects and divided into 2 aliquots. In one aliquot, T-cells were labeled with mouse anti-CD3 IgG, followed by goat anti-mouse IgG bound to magnetic beads. T-cells bound to the magnetic beads were removed by placing the cell suspension in a magnetic field; depletion was verified by flow cytometry. Tcell depleted and nontreated PBMC suspensions were incubated with either Influenza A or an appropriate control medium for 2 hr at 37'C, collected and resuspended in fresh culture medium for an additional 22 hr incubation. Following incubation, the cells were re-collected and challenged with ragweed antigen E. Basophil HR was enhanced in the virus treated group of PBMC which had not undergone T-cell depletion. In contrast, virus incubation did not enhance HR in the T-cell depleted fraction. This suggests that T-cells play an integral role in the process by which viruses enhance basophil HR.
288
THE EFFECT OF RHINOVIRUS (RV) UPPER RESPIRATORY INFECTION (URI) ON AIRWAY REACTIVITY IN NONALLERGIC SUBJECTS. C. Swenson, E. Olck. W. Calhoun, W. Busse, Madlson, WI Viral URIS provoke wheezing in some patients wlth asthma and enhance bronchlal responslveIn previous studies, we have found ext-ass. perlmental RV URIS enhance alrway responsiveness and promote the appearance of late asthmatlc reactlons. All these observations were made In patients wlth allergic alrway disease. Consequently, the followlng study was undsrtaken to determlne whether allergic alrway disease may be a predlsposlng factor In the development of bronchlal hyperresponslveness with viral non-allergic subJects. free of URIS. Thirteen, RV neutrallzlng antlbody, were experlmentally Infected wlth RV type 16. Bronchlal reactlvlty, as deteralned by the PC90 value to Inhaled hlstamlne. prlor to RV lnfectlon was 110 breath unlts (geometric means) and dld not change durlng the acute RV URI, 102 breath unlts. When changes In lndlvldual responses were analyzed, 403 had enhanced bronchlal reactlvlty to hlstaml’ne durlng the RV URI; thls compared a frequency of enhanced alrway responsiveness In patlents wlth allergic alrway disease of 18/18 (p-0.003). These data suggest that the exlstence of allergic alrways disease Is a predlspOSlnQ factor for the development of bronchlal reactlvlty durlng the viral URI. Furthermore, these observations support our hypothesis that viral URIS promote bronchlal reactlvlty by enhanclng exlstlng allergic alrway Inflatwnatlon.
Immunopharmacology Group, University of Southampton, U.K.*U.M.D.S., London, U.K. Bronchial lavage and biopsy studies have provided evidence for the specific involvement of eosinophils and T-lymphocytes in allergic inflammation of the airways in asthma.There is accumulating evidence suggestingthat the expression of adhesion molecules on endothelial cells and their receptors on leukocytes are involved in this process. To investigate these mechanismswe obtained bronchial biopsies from 10 normal subjects [5m, Sf, mean age 26yrs, Pc28Mch>l6mg/ml] and from 10 asthmatics [6m, 4f, mean age 28yrs, Pc2$4ch 0.2-4.2mg/ml] Six of the asthmatics were rebiopsied after 6wks of inhaled beclamethasonedipropionate[BDP] [2mgsJday for 2wks and lmg/day for 4wks]. Frozen sections of the biopsies were stained by the immunoperoxidase method using MoAb 6SB5 for ICAM- and MoAb 1.2B6 for ELAM-1. Two independent observers graded the slides semiquantitatively for intensity [O-3+] and extent of stainiig for both epithelium and endothelium. No major differences were noted between normals and asthmatics,and between the pm and post-steroid asthmatic biopsies, both in the epithelium and endothelium. We conclude that there is basal expression of ICAM- 1 and ELAM-1 in normal human bronchial mucosa and that there is no detectable difference in the expression of these 2 adhesion molecules on frozen sections of bronchial bi;gzs in the groups of subjects studied using this
286
MECHANISMS OF NOCTURNALASTHMA: THE EFFECT OF DIURNAL CONCENTRATIONSOF CIRCULATING EPINEPHRINE. K. Geiger, M.E. Bates, L. Sehrader. T. Schultz, W.J. Calhoun, W. Busse, Madison, WI Patients wlth asthma frequently have nocturnal wheezing. Although the mechanisms of nocturnal asthma are not fully establlshed. dlurnal cycles In catecholamlnes have been considered a maJor factor. In the followlng study, 10 asthma patlents were hospltallred for 3 consecutive days; every 6 hours the following measurements were made on each sublect over the 3 days: FEVT, plasma CortlSOl and eplnephrlne, and white blood cell count and eoslnophll count, as well as the eoslnophll density profile. Nocturnal asthma was defined as a 15% difference In AM to PM FEVT. Plasma cortlsol had dlurnal varlatlons which had no relatlonshlp to the appearance of nocturnal asthma. Plasma eplnephrlne also had AM to PM varlatlons; however, 10 PM circulating eplnephrlne concentrations were slgnlflcantly lower wlth the appearance of nocturnal asthma ClS.O+l.S vs. 31.026.8 pg/ml, p-0.05). Furthermore, circulating eoslnophlls were proportlonately greater 4 AM to 4 PM only when the subjects experienced nocturnal asthma. No slgnlflcant varlatlon In eoslnophll density proflles were noted In relatlonshlp to nocturnal asthma. These data Indicate that plasma concentrations of eplnephrlne may be an Important factor In the development of nocturnal asthma. Furthermore, we postulate that the degree to which eplnephrlne falls at nlghtmay not only lead to bronchospasm but also promote alrway obstruction and Inflammation as reflected In elevated concentrations of eoslnophlls.
211
285
87 1, PART 2
BRONCHIAL mDOTIJ&IAL
Abstracts
MUmAL INTERCEJJJLAR MOLECI,&& 1 IICAM-11 LEUKOCYTE AD&%
287
ENHANCEDBASOPHIL HISTAMINR RELEASE (HR) FOLLOWING IN VITRO INCUBATION WITH LIVE INFLUENZA A VIRUS IS ATTENUATED BY T-CELL DEPLETION. M.A. Huftel, M.D., C.A. Swenson, B.S., W.R. Borcherding, B.S., E.C. Dick, Ph.D., R. Hong, M.D., W.W. Busse, M.D., Madison, WI A number of mechanisms participate in virusPreviously, we described eninduced asthma. hanced basophil HR during an experimentally induced rhinovirus infection and following & .vitro incubation of peripheral blood mononuclear cells (PBMC) with Influenza virus. Furthermore, incubation with interferon produced similar results, suggesting cytokines may play a role in this process. To further define the role of Tenhanced basophil HR, PBMC (basocells in virus monocytes, phils, and lymphocytes) were isolated from ragweed allergic subjects and divided into 2 aliquots. In one aliquot, T-cells were labeled with mouse anti-CD3 IgG, followed by goat anti-mouse IgG bound to magnetic beads. T-cells bound to the magnetic beads were removed by placing the cell suspension in a magnetic field; depletion was verified by flow cytometry. Tcell depleted and nontreated PBMC suspensions were incubated with either Influenza A or an appropriate control medium for 2 hr at 37'C, collected and resuspended in fresh culture medium for an additional 22 hr incubation. Following incubation, the cells were re-collected and challenged with ragweed antigen E. Basophil HR was enhanced in the virus treated group of PBMC which had not undergone T-cell depletion. In contrast, virus incubation did not enhance HR in the T-cell depleted fraction. This suggests that T-cells play an integral role in the process by which viruses enhance basophil HR.
288
THE EFFECT OF RHINOVIRUS (RV) UPPER RESPIRATORY INFECTION (URI) ON AIRWAY REACTIVITY IN NONALLERGIC SUBJECTS. C. Swenson, E. Olck. W. Calhoun, W. Busse, Madlson, WI Viral URIS provoke wheezing in some patients wlth asthma and enhance bronchlal responslveIn previous studies, we have found ext-ass. perlmental RV URIS enhance alrway responsiveness and promote the appearance of late asthmatlc reactlons. All these observations were made In patients wlth allergic alrway disease. Consequently, the followlng study was undsrtaken to determlne whether allergic alrway disease may be a predlsposlng factor In the development of bronchlal hyperresponslveness with viral non-allergic subJects. free of URIS. Thirteen, RV neutrallzlng antlbody, were experlmentally Infected wlth RV type 16. Bronchlal reactlvlty, as deteralned by the PC90 value to Inhaled hlstamlne. prlor to RV lnfectlon was 110 breath unlts (geometric means) and dld not change durlng the acute RV URI, 102 breath unlts. When changes In lndlvldual responses were analyzed, 403 had enhanced bronchlal reactlvlty to hlstaml’ne durlng the RV URI; thls compared a frequency of enhanced alrway responsiveness In patlents wlth allergic alrway disease of 18/18 (p-0.003). These data suggest that the exlstence of allergic alrways disease Is a predlspOSlnQ factor for the development of bronchlal reactlvlty durlng the viral URI. Furthermore, these observations support our hypothesis that viral URIS promote bronchlal reactlvlty by enhanclng exlstlng allergic alrway Inflatwnatlon.
Immunopharmacology Group, University of Southampton, U.K.*U.M.D.S., London, U.K. Bronchial lavage and biopsy studies have provided evidence for the specific involvement of eosinophils and T-lymphocytes in allergic inflammation of the airways in asthma.There is accumulating evidence suggestingthat the expression of adhesion molecules on endothelial cells and their receptors on leukocytes are involved in this process. To investigate these mechanismswe obtained bronchial biopsies from 10 normal subjects [5m, Sf, mean age 26yrs, Pc28Mch>l6mg/ml] and from 10 asthmatics [6m, 4f, mean age 28yrs, Pc2$4ch 0.2-4.2mg/ml] Six of the asthmatics were rebiopsied after 6wks of inhaled beclamethasonedipropionate[BDP] [2mgsJday for 2wks and lmg/day for 4wks]. Frozen sections of the biopsies were stained by the immunoperoxidase method using MoAb 6SB5 for ICAM- and MoAb 1.2B6 for ELAM-1. Two independent observers graded the slides semiquantitatively for intensity [O-3+] and extent of stainiig for both epithelium and endothelium. No major differences were noted between normals and asthmatics,and between the pm and post-steroid asthmatic biopsies, both in the epithelium and endothelium. We conclude that there is basal expression of ICAM- 1 and ELAM-1 in normal human bronchial mucosa and that there is no detectable difference in the expression of these 2 adhesion molecules on frozen sections of bronchial bi;gzs in the groups of subjects studied using this
286
MECHANISMS OF NOCTURNALASTHMA: THE EFFECT OF DIURNAL CONCENTRATIONSOF CIRCULATING EPINEPHRINE. K. Geiger, M.E. Bates, L. Sehrader. T. Schultz, W.J. Calhoun, W. Busse, Madison, WI Patients wlth asthma frequently have nocturnal wheezing. Although the mechanisms of nocturnal asthma are not fully establlshed. dlurnal cycles In catecholamlnes have been considered a maJor factor. In the followlng study, 10 asthma patlents were hospltallred for 3 consecutive days; every 6 hours the following measurements were made on each sublect over the 3 days: FEVT, plasma CortlSOl and eplnephrlne, and white blood cell count and eoslnophll count, as well as the eoslnophll density profile. Nocturnal asthma was defined as a 15% difference In AM to PM FEVT. Plasma cortlsol had dlurnal varlatlons which had no relatlonshlp to the appearance of nocturnal asthma. Plasma eplnephrlne also had AM to PM varlatlons; however, 10 PM circulating eplnephrlne concentrations were slgnlflcantly lower wlth the appearance of nocturnal asthma ClS.O+l.S vs. 31.026.8 pg/ml, p-0.05). Furthermore, circulating eoslnophlls were proportlonately greater 4 AM to 4 PM only when the subjects experienced nocturnal asthma. No slgnlflcant varlatlon In eoslnophll density proflles were noted In relatlonshlp to nocturnal asthma. These data Indicate that plasma concentrations of eplnephrlne may be an Important factor In the development of nocturnal asthma. Furthermore, we postulate that the degree to which eplnephrlne falls at nlghtmay not only lead to bronchospasm but also promote alrway obstruction and Inflammation as reflected In elevated concentrations of eoslnophlls.
211