132 natural history of large local reactions (llr) to stinging insects

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CROSS-REACTIVITY OF PURIFIED VENOM ANTI- GENS OF HORNETS, YELLOWJACKETS, AND WASPS. T.P. King, New York, N.Y.

Venom from each vespid contains three known biochemically homologous proteins which are allergens in man: hyaluroni- dase (HYA), phospholipase (PLA)and anti- gen 5(Ag5) of 23K to 45 K daltons. PLA from yellowjacket (YJ)or wasp (W)venom can be isolated by affinity chromato- graphyonadsorbent containing substrate analog of Pm (Mol. Immunol. 1983, in press), but that from hornets canbeiso- lated only by chromatography on cellu- lose cation exchangers. HYA can be iso- lated by affinity chromatography on heparin-Sepharose.

Immunodiffusion studies with venom- specific rabbit sera showed extensive cross-reaction of HYAs from white-faced H (WFH), yellow H (YH), and different species of YJ. Also, they showed partial cross-reaction of PlAs from WFH and YH but no cross-reaction of PLAs from H,YJ or W. Studies of Ag5s with Ag5-specific mouse sera showed results similar to those of PLAs. The findings suggest that the known cross-reactivity of vespid venoms resides mainly in the HYA com- ponent. Furthermore, they suggest that purified PLA or Ag5 c:n be useful for diagnosis of patients true sensitivity to individual vespid venom.

PROTEIN MAPS OF VENOM AND VENOM PROTEIN EXTRACTS Catherine L. Wood, M.D.. PhAD&m..L. Timnons IV, and Donald R. Hoffman. Ph.D.,Greenville, NC.

Protein maps of vespid venoms were prepared by non-equilibrium gel electrophoresis, a pl dependent stacking technique, in the first di- mension and SDS-polyacrylamide gel electropho- resis in the second dimension. A polychromatic silver stain, Gelcode I was used to increase sensitivity over 30 fold and to eliminate the non-reproducible dye uptake of venom proteins. The map of honeybee venom was used as a refer- ence; bee venom was found to be more complex than was previously supposed. Fresh pure venom was obtained from insects of a single species by stinging into micro test tubes. Venom from lo- 20 insects was pooled and used to prepare maps. Venom protein extracts from single species were obtained from Vespa Laboratories. Comparison of the venoms and protein extracts from Vespula maculifrons, Polistes exclamans, P. fuscatus and P. metricus showed that venom protein extracts have many more components than venoms. Most of these components were more acidic and of higher molecular weight than the venom proteins. Venom maps for the closely related species, P. fuscatus and P. metricus were almost identi- cal. It was possible to renature and extract allergens with intact IgE binding activity from Coomassie blue stained maps of bee and yellow jacket venoms. Protein mapping provides a detailed comparison of components present in biological fluids and can be used for isolation of individual components as well. Venom protein extracts used for diagnosis and therapy contain a significant amount of proteins not found in pure fresh venoms.

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USEFUINESS OF ~PTE&+SPEUF'IC I&MEASURE- ?4fZNTS. M.J. Genthik, M.D., R.G. I&iiltm, Ph.D., N.F. Fdkinson, Jr., M.D. ai-d H.C. Mans- mmn, Jr., M.D. Philadelphia, Pennsylvania

Venrn specific 1s antibody (Ab) response was evaluated in 32 patients using m solid phase radio ircmmassay (SPRIA) n-ethcds, before and after one year ofv

95 imnunotherapy. An

agarose-based test using I-Staph. Protein A (SPRIA) was used to determine specific Ic$3 for five Hymenoptera species: yellow jacket (YJ), honeybee (HE), yellow-faced hornet (YX), white-

~~~~~~,~hbt~~i~'~~~~~~~~ ~*gG RAST) was available only for YJ and HB vencmm.

Bothmethods yieldedacceptable HEAblevels Por YJ, 25% of positive Protein A-SPFUA were negati~byIg+RAST,yetconversenotseen. 3i-1 31 treated with 90 venans, 90/90 Ab levels were increasedormaintained attbebighpre- treatment levels. L!evelsofAbtiuntreated ~specieswere undetectable or declined in 50160. Yet, YJ stimulated a WH and/or YH response in four. In one, YJ caused a rise in J%l. Treatment resulted in)5 ug/ml of Ab to 61 vencrns, yet in 25 the level was < 5 ug/'ml. One had no Ab response.

SPRIA can provide specific and quantitative assessment to vencm imnunctberapy. These results suggest that scme or all of the less responsive 26 casesmy representsub-optil therapybasedon data fromGolden etal.

NATURAL HISTORY OF LARGE LOCAL REACTIONS (LLR) TO STINGING INSECTS. Paul M. Mauriello, M.D., Susan H. Barde, M.D., John W. Georgitis, M.D. and Robert E. Reisman, M.D., Buffalo, New York

In ongoing studies of stinging insect allergy, 133 ots with LLR have been f0110WedDrOSpeCtiVelY for up to 6 yr. Pts were divided, based upon - RAST analysis with honeybee and vespid venoms.

Sixty-six pts were RAST negative, with venom skin tests being positive in 58%. Seventy-five re-stings in this group led to no systemic and 74 local reactions. Follow-up RASTs and skin tests were unchanged.

Sixty-seven pts had detectable serum venom- specific IgE. There was a wide range in anti- body titers, indistinguishable from pts with systemic reactions. Skin tests were positive in almost all. Twenty-four of 67 received venom immunotherapy (VIT). RASTs decreased similarly in VIT and untreated groups. At follow-up, half the pts were RAST negative. There were 101 re- stings, 46 before and 55 after evaluation, 80% resulting in recurrent local reactions in both treated and untreated groups. One systemic re- action occurred in an untreated pt.

In reviewina 118 ots with stina anaphvlaxis, a prior LLR occurred'in 5.

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Following repeat stings, pts with LLR tend to have subsequent LLR regardless of the presence of venom-specific IgE or immunotherapy. There is a small risk of anaphylaxis.

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