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HK00675026 IgE ANTIBODY RESPONSE OF S?•tOKERS, EO\-SPfOKERS AND "SMOKE S?;?:SITIVE" INDIVIDUALS TO TOBACCO LE.yT A.I1D S?IOKE ANTIGENSI

N f{C0675027 S.B. Lehrer, *t.R. Wilson, R.M. Itarr, and J.E. Salvaggio

i Clinical Immunology Section Department of :`:edicine Tulane Med4cal Center 1700 Perdido Sz:reet New Orleans, Louisiana 70112 HIt~~~575028

! h E"JG675029 (1) Supported by Grants from the Council fior Tobacco Research,•USA, Special Project 84 and the National Institute of Nealth AI-1341. Sanuel B. Lebrer is supported by the Young Investigator Award from the NIAID.

•NKC-067503( Sera from cigarette smokers, non-smokers, and individuals reporting "smoke sen- sitivity" were tested for IgE antibodies to tobacco leaf and smoke extracts by the radioallergosorbent test (RAST). Results indicated that none of the serum samples tested contained detectable IgE antibodies to smoke extracts. Occasionally sera from smokers or non-smoker§ demonstrated most significant reaction to leaf antigens was detected in serum reactivity to leaf antigen. The from on4 of the seven "smoke sensitive" individuals tested. These results demonstrate that smoking, non-smoking or clinical "smoke sensitivity" do not correlate wit.h the presence of IgE antibodies to tobacco leaf or smoke antigen.

,_- 11 KE'0675031 There has been a growing concern as to the effects of tobacco smoke on non-smokers (1-7) and it has been proposed by some that atopic individuals may encounter IgE mediated raspiratory difficulties when exposed to cigarette smoke. Since recent studies from our laboratory have demonstrated that immuno- gens are in fact present in tobacco smoke (8) and are capable of stimulating a reaginic antibody regponse tb leaf antigens in mice (9), the following study was undertaken to determine if IgE antibodies to tobacco leaf or smoke antigens are present in human serum. The pr!:paration of all leaf and smoke extracts were reported in detail elsewhere (8,9). Leaf A extiract was obtained from flue-cured tobacco leaf (NC 2326 variety, donated by Wallace J. Dickens, Borderbelt Tobacco Research Station, Whitesville, N.C.), Leaf B from Maryland Leaf (donated by M.K. Ayecock, Jr., Department of Agronomy, University of Maryland, College Park), and Leaf C from air-cured burley tobacco leaf (donated by D.L. Davies, Department of Agronomy University of Kentucky, Lexi.ngton Kentucky). Smoke extracts (SE) were prepared by passing smoke frbm a total of 1500 1R2F cigarettes (Tobacco and Health Institute, Lexington, Kentucky) produced with a 30 port BoNrgwaldt smoking machine (donated by the R.J. Reynolds Company) through a standard gas bubbler. SE-NHS smoke extracts was prepared by pas5ing cigarette smoke through a solution of 1% pooled normal human serum (obtained from non-smokers) as described above. The radioallergosorbent test (RAST) was performed as described by Ceska and co-workers (10). Discs were coated with Leaf A, Leaf B, Leaf C and SE-NHS in the conventional manner. Other discs were prepared by passing smoke from cigarettes

I N1 Nl~C6 75032 through a 50 ml sol.utiord of 0.1*1 haHO03 containing 100 cyanogen bromide activat- ed discs (SE) or through 50 ml PBS containing 100 discs coupled with NHS (SE-NHS- A). These discs were then treated in an identical fashion as those coated with proteins. RAST assay was performed by incubation of antigen coated discs with the test subject's serum, followed by washing and incubation with 1 125 labelled anti- human IgE (Pharmacia). Disc were again washed and total counts per minute (cpm) determined and expressed as an ratio of cpm obtained with discs coated with NHS. Ratios greater thatt 2 were considered to be positive. Sera from 16 smokers and 4 non-snokers were obtained from wmrkers who bad participated in an occupational study at local manufacturing plarlt in Louisiana. Smoking history on each of the individuals tested was available from the study. Any individual who smoked cigarettes presently or in the past was listed as a smoker. Sera from "smoke sensitive" subjects were obtained from selected atopic individuals seen in the Tulane Allergy Clinic who complained of oceular; nasal or lower respiratory tract syir;,toms (tearing of the eyes, thinorrbea, cough or wheezing) when encountering tobacco smoke. RAST values to tobacco leaf and SE obtained for the smoking and non-smoking popu- lation aresrmmarized in'Table I. Results indicate that all smokers and non--smokershad no detectable IgE antibody to smoke extracts, although one nonsmoker, JC, and two smokers, TG and MW had positive yet low responses to leaf extracts. These studies were extended to determine if sera from individuals with a history of clinical smoke sensitivity contained IgE antibodies to leaf or smoke antigens (Table II). Sera from several such individuals were tested for IgE antibodies to leaf extracts and SE-'~'HS, Only one, JD, demonstrated IgE

-3- H,t~G675U35 antibodies and these were only to leaf antigens. Further studies tested sera utilizing discs to which attempts have been made to directly couple smoke extract to activated discs. However these discs were not active in the asszy since IgE antibodies were not detected in any of the individuals tested, even 3D, the "smoke sensitive" subject with IgE antibodies to Leaf B and C allergens. These results arL of interest since they demonstrate that IgE antibody responses cannot be detected to smoke extract in the sera of smokers, non-smokers or reported "smoke sensitive" individuals. Several individuals demonstrated IgE antibodies to leaf attigens, but no correlation between the presence of these antibodies and smoking or clinical smoke sensitivity could be detected. Since our earlier reports (9,10) demonstrated that micE or rabbits can produce antibody to tobacco leaf antigens in response to smoke stimulation, our current demonstra- tion of reaginic antibodies to leaf antigens in nan are still of great interest. Although only 1 of 7 smoke sensitive subjects demonstrated IgE antibodies to leaf antigens, it is possible that "smoke sensitive" in4ividuals are a complex group of subjects, some of whom have IgE antibody to leaf antigen present in cigarette s~noke. These possibilities are being further analyzed in epidemiologcal studies of smoke sensitive and non-sensitive subjects and provocative inhalation challenge s:udies of smoke sensitive and RAST positive individuals with smoke and leaf antigens.

-4- AcknocrlFdo ent HKF,Ob7*503q The authors wish to thank ":iss Julie Mitchell and Mr. Jeffexy Tayloi for their technical assistanceand Ms. Debbie Jackson for her secratarial help in preparation of this iaanuscript.

c_. -. .,. ~. .. :~.._...~_ ~L.. . . . ,, H K00675.035 TABLE I IgE ANTIBODY RESPONSE TO TOBACCO OR SMOKE ANTIGENS IN SMOKERS AND N0\-'_..OI:ERS RAST Ratios Subject Smoking History Tested Leaf A Leaf B Leaf C SE-Iv'HS Smoker Non-Smoker RB 0.71 0.65 0.58 0.75 % RB 0.93 0.74 0.70 0.85 . X AC 0.72 0.61 0.52 0.63 X IiC 0.81 0.62 0.83 0.84 X JC 0.66 0.62 0.60 0.76 X FF 1.31 0.85 0.76 0.78 X TG 2.35 1.82 1.23 0.69 X WG 0.92 0.70 0.60 0.76 X LG 0.64 0.67 0.82 0.88 X JG 0.78 0.60 0.68 0.76 X WH 1.23 0.98 1.08 0.78 X BL 0.71 0.72 0.73 0.81 X , ML 0.89 0.86 0.98 0.88 X SL 0.94 0.94 1.12 1.31 X MW 3.01 2.75 2.06 1.14 X HP 0.71 0.93 0.65 0.65 X DB 1.0 0.93 0.79 0.80 X Jc 3.15 3.40 1.59 0.72 X HG 0.67 0.60 1.03 0.85 X WH 1.06 0.73 0.90 0.85 X The ratios were calculated using normal human serum discs as the control for Leaf A, Leaf B, Leaf C, and smoke extract in NgS.