- Email: [email protected]
287 Neutrophil chemotactic activity in laboratory animal-induced asthma
--IN VIVO RELEASE OF HISTAMINE BY HYPEROSMOLAR M.D., P. G. Silber, M.D., R. Naclerio, STIMULI. Eggleston, M.D., D. Proud, Ph.D., A. Togias, M.D., L. Lichtenstein, M.D., P. Norman, M.D., Baltimore, MD. Hyperosmolar stimuli induce hista--in vitro mine release from mast cells and basophils. To assess whether the same stimuli induce mediator nine healthy human volunteers release in vivo, underwent nasal lavage with mannitol solutions Washes before and of different osmolarities. after isosmolar (ISO) and hyperosmolar (HYPER) (800 m0sm) challenges were assayed for histamine and TAME-esterase(s) (TAMe). -IS0 -HYPER Histamine (rig/ml) n=22 2.1iO.7 8.1t1.7 pc.002 1.7kO.4 4.2AO.8 pc.01 TAMe (CPMx10z3) n=17 4.420.2 6.2tO.3 pc.001 Volume Recovered (ml) A greater than 50% increase in histamine was obtained in 86% of the challenges; associated with this histamine increase was a significant Three increase in the lavage fluid recovered. of four subjects tested demonstrated increasing release of histamine with incremental increases When two HYPER challenges in osmotic stimuli. were given sequentially, the histamine level of the second challenge was also increased over baseline (~~0.02) but was less than the first challenge (x1=10.7 vs. X2=6.9). In twelve experiments matched for A histamine, the ratio of histamine to TAMe was reversed when comparing stimuli (1:0.6 vs. 1:4.4). osmotic vs. antigenic We conclude that mast cells and/or basophils can be stimulated stim--in vivo by hyperosmolar The mechanism of this uli to release histamine. release appears to be different than that induced by antiger
286
287
NEDTROPHIL CHEMOTACTIC ACTIVITY IN LABORATORY ANIMAL-INDUCED ASTHMA. A.J. Newman Taylor, T.H. Lee, M.D., S.R. Durham, M.R.C.P. M.R.C.P., and A.B. Kay, M.D., Ph.D., London, U.K. Allergic reactions to laboratory animals develop in approximately 15-20% of exposed workers of whom less than half develop asthma. Those who develop asthma, unlike those with urticaria and rhinitis, are very likely to have positive skin prick test reactions and specific IgE antibodies to animal extracts, particularly urine (A.J. Newman Taylor et al,Thorax 1981, 36: 229). We have undertaken bronchial provocation studies and measurements of serum neutrophil chemotactic activity (NCA) in 4 laboratory animal handlers. The subjects were atopic with positive skin prick tests and elevated allergen specific IgE to the appropriate animal urine extracts. Three rat handlers (immediate asthmatic reactions) demonstrated increases in NCA {maximal 175 t 38 (mean + SEM) per cent at 10 min). The NCA
ENDOGENCUS HEPARIN-LIKE MA’IERIAL (EM) ELEVATIONS FOLLCWINGANI’IGEN CHALLENGE Elliott C Lasser, M.D., Ronald A. Simon, M.D. ; Sandra G. Lyon, B.S., A. Elizabeth Hamblin, B.S., and Rosalyn Stein, B.S., San Diego, California 7 have reported that the average baseline EHMplasma level in 19 atopic asthmatics was elevated. In the current study, five patients with atopic asthma were successfully challenged with appropriate inhalational antigens to determine anti-Xa EIM plasma levels before and after the onset of an induced reaction (>25% +FEVl). E.HMassay was carried out by determining the effect of this substance together with AT-III on Factor Xa, utilizing a chromogenic peptide as substrate. Three of the five patients displayed increased EHMwithin lo-15 minutes following the onset of the induced reaction. IGo elevations were detected in the other two patients, and none in two additional non-reacting patients. A technique was developed to utilize low molecular weight (8,000) dextran sulfate to displace EHM from plasma protein (ET+! neutralizing) binding sites in vitro. Maximum displacement values occurred,at the zenith of EMi concentrations. The heparin-neutralizing potential of these plasmas was assayed by adding a known amount of heparin and determining the resultant heparin concentrations. An expanded heparin-neutralizing potential occurred at the time of maximal plasma EHM levels. The physiological significance of the increase in both anti-ERM proteins and EW, follcn&rg antigen-induced bronchospasm, has not yet been determined.
EFFECT OF SEX STEROIDS ON BASOPHIL HISTAMINE RELEASE. Jay E. Slater, M.D., and Michael A. Maryland. Kaliner, M.D., Bethesda, Alleroic disease worsens in some females at menstruaeion, and progesterone (P) sensitivity has been reported in a woman with recurrent idiopathic anaphylaxis (RIA). Therefore, the effects of P 40 rig/ml, B-estradiol 10 rig/ml (E), and dexamethasone 4 rig/ml (D) on basophil histamine release (BHR) were studied. The protocol involved challenge of basophils with antigen E or anti-IgE on day 1 to establish reactivity, and overnight incubation of remaining cells in buffer, P, E, or D followed by antigen E or anti-IgE challenge on day 2. Results, expressed as % release (mean⦥ *p<.O5) on day 2, are listed below. All 5 subjects demonstrated BHR after overnight culture, although there was some reduction in responses. Pretreatment with P had no effect; E resulted in a small stimulation of BHR in 1 male control; 0 suppressed BHR by approximately 50% in a patient with premenstrual RIA. Sex Control P Subject ! E 4BZ4 MTTZ5G.R.(normal) 45*0 45*3 P.R.(normal) F 46*3 41+3 51k2” 40*1 K.G.(normal) M 21*0 33*0 B.K.(normal) F 22*0 29*0 11*0x F 22*2 2Ok2 21&l C.W.(RIA) Thus far, sex steroids have had little effect on in vitro BHR in normals as well as in a patient with RIA. D inhibition of BHR, previously reported by Schleimer et al., suggests the possibility of in vivo glucocorticoid control of histamine release which may be blocked by sex steroids. These experiments are being conducted.
176
286
287
NEDTROPHIL CHEMOTACTIC ACTIVITY IN LABORATORY ANIMAL-INDUCED ASTHMA. A.J. Newman Taylor, T.H. Lee, M.D., S.R. Durham, M.R.C.P. M.R.C.P., and A.B. Kay, M.D., Ph.D., London, U.K. Allergic reactions to laboratory animals develop in approximately 15-20% of exposed workers of whom less than half develop asthma. Those who develop asthma, unlike those with urticaria and rhinitis, are very likely to have positive skin prick test reactions and specific IgE antibodies to animal extracts, particularly urine (A.J. Newman Taylor et al,Thorax 1981, 36: 229). We have undertaken bronchial provocation studies and measurements of serum neutrophil chemotactic activity (NCA) in 4 laboratory animal handlers. The subjects were atopic with positive skin prick tests and elevated allergen specific IgE to the appropriate animal urine extracts. Three rat handlers (immediate asthmatic reactions) demonstrated increases in NCA {maximal 175 t 38 (mean + SEM) per cent at 10 min). The NCA
ENDOGENCUS HEPARIN-LIKE MA’IERIAL (EM) ELEVATIONS FOLLCWINGANI’IGEN CHALLENGE Elliott C Lasser, M.D., Ronald A. Simon, M.D. ; Sandra G. Lyon, B.S., A. Elizabeth Hamblin, B.S., and Rosalyn Stein, B.S., San Diego, California 7 have reported that the average baseline EHMplasma level in 19 atopic asthmatics was elevated. In the current study, five patients with atopic asthma were successfully challenged with appropriate inhalational antigens to determine anti-Xa EIM plasma levels before and after the onset of an induced reaction (>25% +FEVl). E.HMassay was carried out by determining the effect of this substance together with AT-III on Factor Xa, utilizing a chromogenic peptide as substrate. Three of the five patients displayed increased EHMwithin lo-15 minutes following the onset of the induced reaction. IGo elevations were detected in the other two patients, and none in two additional non-reacting patients. A technique was developed to utilize low molecular weight (8,000) dextran sulfate to displace EHM from plasma protein (ET+! neutralizing) binding sites in vitro. Maximum displacement values occurred,at the zenith of EMi concentrations. The heparin-neutralizing potential of these plasmas was assayed by adding a known amount of heparin and determining the resultant heparin concentrations. An expanded heparin-neutralizing potential occurred at the time of maximal plasma EHM levels. The physiological significance of the increase in both anti-ERM proteins and EW, follcn&rg antigen-induced bronchospasm, has not yet been determined.
EFFECT OF SEX STEROIDS ON BASOPHIL HISTAMINE RELEASE. Jay E. Slater, M.D., and Michael A. Maryland. Kaliner, M.D., Bethesda, Alleroic disease worsens in some females at menstruaeion, and progesterone (P) sensitivity has been reported in a woman with recurrent idiopathic anaphylaxis (RIA). Therefore, the effects of P 40 rig/ml, B-estradiol 10 rig/ml (E), and dexamethasone 4 rig/ml (D) on basophil histamine release (BHR) were studied. The protocol involved challenge of basophils with antigen E or anti-IgE on day 1 to establish reactivity, and overnight incubation of remaining cells in buffer, P, E, or D followed by antigen E or anti-IgE challenge on day 2. Results, expressed as % release (mean⦥ *p<.O5) on day 2, are listed below. All 5 subjects demonstrated BHR after overnight culture, although there was some reduction in responses. Pretreatment with P had no effect; E resulted in a small stimulation of BHR in 1 male control; 0 suppressed BHR by approximately 50% in a patient with premenstrual RIA. Sex Control P Subject ! E 4BZ4 MTTZ5G.R.(normal) 45*0 45*3 P.R.(normal) F 46*3 41+3 51k2” 40*1 K.G.(normal) M 21*0 33*0 B.K.(normal) F 22*0 29*0 11*0x F 22*2 2Ok2 21&l C.W.(RIA) Thus far, sex steroids have had little effect on in vitro BHR in normals as well as in a patient with RIA. D inhibition of BHR, previously reported by Schleimer et al., suggests the possibility of in vivo glucocorticoid control of histamine release which may be blocked by sex steroids. These experiments are being conducted.
176