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Comparative biochemical and immunological studies of commercial alternaria tenuis batches
265
URINARY AND EPITHELIAL RAST TESTING IN SYMPTOMATIC LABORATORY WORKERS. J.Fink~M.D., J.Kidd,M.D., I. Lutsky~V.M.D., and J.Yunginger~ M.D., Milwaukee, WI., Jerusalem, Israel and Rochester, MN. We have previously reported that urinary extracts of rats are more sensitive than epithelium in demonstrating skin test reactivity in symptomatic laboratory workers. Eight of 16 workers skin tested with both urinary and epithelial antigens had positive skin tests only to the urinary antigen (p.<.025). Eight were positive to both antigens. This study was done to compare rat urine RAST with epithelial RAST in these 16 workers with rhinitis and/or asthma on exposure to rats. Of these, 3 had negative RAST to both antigens, 4 had positive RAST to both antigens and none were positive only to epithelial antigen. However, 9 had positive RAST only to urinary antigen (p.<.01). RAST inhibition studies in 2 workers with positive RAST to both antigens showed different degrees of cross inhibition between the two antigens. Inhibition with rat epithelium was greater than with urinary antigens. These results show that RAST to urinary antigens are more sensitive than epithelial RAST in showing in vitro sensitivity to rats. The urine may contain the major antigen causing symptoms in some workers. Urinary BAST appear to be more specific but less sensitive than skin tests with urinary antigen. In 2 workers, inhibition studies showed that the antigens responsible for the immunologic response are present in both urine and epithelium. In doing RAST testing to assist in the diagnosis of allergy to rats, we suggest including both urinary and epithelial EAST.
266 COMPARATIVE BIOCHEMICAL AND IMX~UNOLOGICAL
267 EXACERBATION OF RESPIRATORYALLERGIES RELATED TO AUTOMOBILE AIR CONDITIONERS: AN EPIDEMIOLOGIC STUDY. P. Kumar, M.D.~ R. Marier, M.D. and S.Ho Leech, M.D., Ph.D., New Orleans, Louisiana. The present study of 207 patients with a proven diagnosis of respiratory a l l e r g i e s (bronchial asthma, a l l e r g i c r h i n i t i s ) was conducted to determine: a) i f exposure to car a i r conditioners (a/c) was associated with exacerbations of t h e i r symptoms and changes in pulmonary functions and b) whether such exacerbations were associated with the presence of molds in t h e i r car a / c ' s . One hundred and f i f t e e n of 207 patients were evaluable. Twenty of these 115 (17.7%) evaluable patients reported exacerbation of symptoms while using t h e i r car a/c's. Seven of these patients were studied f u r t h e r . A v a r i ety of molds including P e n i c i l l i u m , A l t e r n a r i a , Aspergillus and others were isolated from the outflows of t h e i r a/c's. Peak expiratory flow rates (PEFR) were measured before, during and up to 6 hours a f t e r exposure to t h e i r own and a control (culture negative) car a/c. Results were as follows: Baseline A f t e r turning a/c on PEFR (MeaniSEM) 15 mins 89hr 1 hr Patients' 300.00• 216.7• 235.33• 298.0• cars 12.47 9.24 9.24 17.10 Control 272.50• 267.50• 285.0• 290.0• car 23.17 33.12 26.87 27.89 A s i g n i f i c a n t decline in PEFR between baseline and 15 mins (P
268 SPECIFICITY OF SERUM IgE ANTIBODY REACTIVE WITH
STUDIES OF COMMERCIAL ALTERNARIA TENUIS BATCHES. H.M. Vijay, Ph.D., H. Huan$, B.A., N.M. Young, Ph.D., Ottawa, Canada and I.L. Bernstein, M.D., Cincinnati, Ohio. Samples of A. tenuis from two commercial sources were extracted with buffer and separated by Sephadex G-100 gel-filtration. There was considerable variation in the UV absorbance spectra and carbohydrate profiles, even within samples from the same company. The extracts contained a significant amount of high mol. wt. polysaccharides devoid of allergenic activity. GLC analysis showed approx, equal amounts of galactose and mannose, but widely varying amounts of glucose. It appears that A. tenuis contains two polysaccharides in varying amounts; galactomannan and glucan. In contrast, extensive antigenic cross-reactivity between the batches, even from different companies, was found by immunodiffusion studies. In direct RAST assays, they also had similar reactivities but there was 8 - fold variation in PAST inhibition activities between the batches of the same company and i0 - fold between inter-company batches. In rat-lgE PCA tests, extensive cross-reactivity was observed among the batches from the same company but this was less marked among the inter-company batches. The lack of correlation between allergenic activity and biochemical properties of different batches suggest that simple biochemical analysis cannot be substituted for more sophisticated tests of allergenic potency.
TOBACCO LEAF ANTIGENS. P. W. Welsh, B.A., and G. J. Gleich, M.D., Rochester, MN. Because IgE antibodies (Ab) to tobacco leaf antigens may be associated with hypersensitivity to tobacco, we determined the frequency and specificity of these Ab's. IgE Ab to green tobacco leaf extract (GTE) and cured tobacco leaf extract (CTE) were measured by the radioallergosorbent test. IgG Ab was detected by radiolabeled Protein A and IgA Ab by radiolabeled purified anti-IgA. Sera from 145 of 1200 patients (12%) had significant levels of IgE Ab to GTE, while 86 of 1029 (8%) had IgE Ab to CTE. Serum IgG Ab to GTE was present in 54 of 1081 (5%). Elevated serum IgA to GTE was not found. Only 19 of 268 patients with a clinical history of tobacco allergy actually had elevated levels of IgE Ab to GTE. Of these 19, six also had elevated IgE Ab to both tomato and potato. Further study of these 19 sera showed two patterns of IgE reactivity. In one, IgE binding to GTE was inhibited by GTE and crossreacting antigens from the botanical family Solanaceae. In the other, IgE binding to GTE was not only inhibited by GTE and botanically related antigens, but also by unrelated antiens, such as ragweed (RW), banana, green pea, green coffee and cherries, but not by cow's milk. IgE Ab in these sera also bound to RW, but this reaction was inhibited only by RW. These results suggest that IgE Ab to tobacco leaf antigen is present in many sera regardless of whether or not the patients are clinically sensitive and that certain sera contain IgE Ab reactive with antigenic determinants common to many plant sources.
159
URINARY AND EPITHELIAL RAST TESTING IN SYMPTOMATIC LABORATORY WORKERS. J.Fink~M.D., J.Kidd,M.D., I. Lutsky~V.M.D., and J.Yunginger~ M.D., Milwaukee, WI., Jerusalem, Israel and Rochester, MN. We have previously reported that urinary extracts of rats are more sensitive than epithelium in demonstrating skin test reactivity in symptomatic laboratory workers. Eight of 16 workers skin tested with both urinary and epithelial antigens had positive skin tests only to the urinary antigen (p.<.025). Eight were positive to both antigens. This study was done to compare rat urine RAST with epithelial RAST in these 16 workers with rhinitis and/or asthma on exposure to rats. Of these, 3 had negative RAST to both antigens, 4 had positive RAST to both antigens and none were positive only to epithelial antigen. However, 9 had positive RAST only to urinary antigen (p.<.01). RAST inhibition studies in 2 workers with positive RAST to both antigens showed different degrees of cross inhibition between the two antigens. Inhibition with rat epithelium was greater than with urinary antigens. These results show that RAST to urinary antigens are more sensitive than epithelial RAST in showing in vitro sensitivity to rats. The urine may contain the major antigen causing symptoms in some workers. Urinary BAST appear to be more specific but less sensitive than skin tests with urinary antigen. In 2 workers, inhibition studies showed that the antigens responsible for the immunologic response are present in both urine and epithelium. In doing RAST testing to assist in the diagnosis of allergy to rats, we suggest including both urinary and epithelial EAST.
266 COMPARATIVE BIOCHEMICAL AND IMX~UNOLOGICAL
267 EXACERBATION OF RESPIRATORYALLERGIES RELATED TO AUTOMOBILE AIR CONDITIONERS: AN EPIDEMIOLOGIC STUDY. P. Kumar, M.D.~ R. Marier, M.D. and S.Ho Leech, M.D., Ph.D., New Orleans, Louisiana. The present study of 207 patients with a proven diagnosis of respiratory a l l e r g i e s (bronchial asthma, a l l e r g i c r h i n i t i s ) was conducted to determine: a) i f exposure to car a i r conditioners (a/c) was associated with exacerbations of t h e i r symptoms and changes in pulmonary functions and b) whether such exacerbations were associated with the presence of molds in t h e i r car a / c ' s . One hundred and f i f t e e n of 207 patients were evaluable. Twenty of these 115 (17.7%) evaluable patients reported exacerbation of symptoms while using t h e i r car a/c's. Seven of these patients were studied f u r t h e r . A v a r i ety of molds including P e n i c i l l i u m , A l t e r n a r i a , Aspergillus and others were isolated from the outflows of t h e i r a/c's. Peak expiratory flow rates (PEFR) were measured before, during and up to 6 hours a f t e r exposure to t h e i r own and a control (culture negative) car a/c. Results were as follows: Baseline A f t e r turning a/c on PEFR (MeaniSEM) 15 mins 89hr 1 hr Patients' 300.00• 216.7• 235.33• 298.0• cars 12.47 9.24 9.24 17.10 Control 272.50• 267.50• 285.0• 290.0• car 23.17 33.12 26.87 27.89 A s i g n i f i c a n t decline in PEFR between baseline and 15 mins (P
268 SPECIFICITY OF SERUM IgE ANTIBODY REACTIVE WITH
STUDIES OF COMMERCIAL ALTERNARIA TENUIS BATCHES. H.M. Vijay, Ph.D., H. Huan$, B.A., N.M. Young, Ph.D., Ottawa, Canada and I.L. Bernstein, M.D., Cincinnati, Ohio. Samples of A. tenuis from two commercial sources were extracted with buffer and separated by Sephadex G-100 gel-filtration. There was considerable variation in the UV absorbance spectra and carbohydrate profiles, even within samples from the same company. The extracts contained a significant amount of high mol. wt. polysaccharides devoid of allergenic activity. GLC analysis showed approx, equal amounts of galactose and mannose, but widely varying amounts of glucose. It appears that A. tenuis contains two polysaccharides in varying amounts; galactomannan and glucan. In contrast, extensive antigenic cross-reactivity between the batches, even from different companies, was found by immunodiffusion studies. In direct RAST assays, they also had similar reactivities but there was 8 - fold variation in PAST inhibition activities between the batches of the same company and i0 - fold between inter-company batches. In rat-lgE PCA tests, extensive cross-reactivity was observed among the batches from the same company but this was less marked among the inter-company batches. The lack of correlation between allergenic activity and biochemical properties of different batches suggest that simple biochemical analysis cannot be substituted for more sophisticated tests of allergenic potency.
TOBACCO LEAF ANTIGENS. P. W. Welsh, B.A., and G. J. Gleich, M.D., Rochester, MN. Because IgE antibodies (Ab) to tobacco leaf antigens may be associated with hypersensitivity to tobacco, we determined the frequency and specificity of these Ab's. IgE Ab to green tobacco leaf extract (GTE) and cured tobacco leaf extract (CTE) were measured by the radioallergosorbent test. IgG Ab was detected by radiolabeled Protein A and IgA Ab by radiolabeled purified anti-IgA. Sera from 145 of 1200 patients (12%) had significant levels of IgE Ab to GTE, while 86 of 1029 (8%) had IgE Ab to CTE. Serum IgG Ab to GTE was present in 54 of 1081 (5%). Elevated serum IgA to GTE was not found. Only 19 of 268 patients with a clinical history of tobacco allergy actually had elevated levels of IgE Ab to GTE. Of these 19, six also had elevated IgE Ab to both tomato and potato. Further study of these 19 sera showed two patterns of IgE reactivity. In one, IgE binding to GTE was inhibited by GTE and crossreacting antigens from the botanical family Solanaceae. In the other, IgE binding to GTE was not only inhibited by GTE and botanically related antigens, but also by unrelated antiens, such as ragweed (RW), banana, green pea, green coffee and cherries, but not by cow's milk. IgE Ab in these sera also bound to RW, but this reaction was inhibited only by RW. These results suggest that IgE Ab to tobacco leaf antigen is present in many sera regardless of whether or not the patients are clinically sensitive and that certain sera contain IgE Ab reactive with antigenic determinants common to many plant sources.
159