161 What Specific IgE Levels to Milk Protein Fractions May Contribute to Cow’s Milk (CM) Allergy Diagnosis?

A. P. M. Castro, A. K. F. Gushken, A. C. Pastorino, E. Ciccone, A. C.

Brandao, C. M. A. Jacob; Department of Pediatrics, University of Sao

Paulo, Sao Paulo, BRAZIL.

RATIONALE: Specific IgE levels to milk protein fractions have been

evaluated in association with specific IgE to.whole milk protein, but their

importance is not well established for CM allergy diagnosis.

METHODS: We evaluated 23 patients (14 male, mean 4yrs 2mo, range

11mo - 14y) with CM IgE mediated allergy confirmed by anaphylaxis or

positive DBPCFC. All of them performed Pharmacia ImmunoCAP® sys-

tem to whole milk (WM), casein, �-lactoalbumin and �-lactoglobulin . A

group of 23 equivalent healthy children was considered as control. Sensi-

tivity (Se), specificity (Sp), PPV, NPV for all levels from 0,35 to 15 kU/L

were determined. As the objective of this analysis was to consider the

diagnosis and not only a screening test, PPV and NPV were the main val-

orized measures.

RESULTS: For �-lactoalbumin, the best result considering PPV = 100%,

was ImmunoCap level of 3 kU/L (Se =65,2%, Sp= 100%, PPV = 100%,

NPV= 74,1%). For �-lactoglobulin the best level was 2 kU/L (Se =56,5%,

Sp= 100%, PPV = 100%, NPV= 69,7%) and for casein the best level was

4 kU/L (Se =78,2%, Sp= 100%, PPV = 100%, NPV= 82,1%). Consider-

ing specific IgE to WM =11kU/L as a decision point for CM allergy diag-

nosis, five patients with IgE < 11kU/L presented positive protein fraction

IgE.

CONCLUSIONS: This study showed that the decisions points for pro-

tein fractions were lower than to WM protein and they can improve the

diagnosis in patients whose specific IgE levels to whole milk are lower

than the established decision point. More studies are necessary to confirm

this hypothesis.

Funding: University of Sao Paulo

162 Cow’s Milk Allergy: A Pilot Trial of Sublingual Immuno-therapy

d. de boissieu, C. Dupont; St Vincent de Paul Hospital, Paris, FRANCE.

RATIONALE: To evaluate the safety and efficacy of sublingual

immunotherapy in children with protracted mediated cow’s milk allergy.

METHODS: 3 children (7, 12 and 17 years) with protracted IgE mediat-

ed milk allergy previously manifested by anaphylactic shock or edema

were submitted to a food challenge. The cumulated dose at which the

child reacted was: case 1: 8ml with asthma; case 2: 44ml with urticaria,

rhinitis and conjonctivitis; case 3: 37ml with edema and urticaria. Chil-

dren were then asked to put under the tongue everyday, during 2 minutes,

an increasing amount of milk from 0.1ml to 1ml, below the triggering

level, during 6 months. A second food challenge analysed the modifica-

tion of the volume tolerated. Casein specific IgE were measured before

each challenge (CAP system, Pharmacia®).

RESULTS: No adverse reaction was reported at home. In case 1, a bron-

chospasm was observed for a cumulated dose of 93ml. In case 2 and 3, no

reaction was observed until a cumulated dose of 200ml of milk and the

diet was progressively normalized at home without any manifestation. In

the mean time, a decrease of casein specific IgE was observed, from 33 to

15KUI/L in case 1, from 7.5 to 5.5KUI/L in case 2, from 2.5 to 1.6KUI/L

in case 3.

CONCLUSIONS: In this preliminary pilot trial, a 6 months sublingual

immunotherapy is a safe procedure, easy to perform at home. It allows the

child to get free of any anaphylactic reaction and in some cases to rein-

troduce dairy products into the diet.

163 Seafood Allergic Reactions in Restaurants

T. J. Furlong1, J. M. Maloney2, S. H. Sicherer2; 1Food Allergy Network,

Fairfax, VA, 2Allergy/Immunology, Mount Sinai Medical Center, NY, NY.

RATIONALE: Seafood allergy is estimated to affect 2.3% of the US

population; restaurant reactions are common, but little is known about the

circumstances.

METHODS: A structured questionnaire was administered to a random

sample selected from 415 registrants reporting restaurant reactions in the

National Seafood Allergy Registry (n=1000).

RESULTS: Participants (125 registrants describing 182 reactions [42%

first lifetime reactions]) included 58% allergic adults and 42% surrogates

for children. Allergies included: shellfish (60%), fish (17%), or both

(22%). The most frequent restaurant types were seafood (23%) and Asian

(18%). Reactions were attributed to oral ingestion (90%), skin contact

(4%) or inhalation (6%). Symptom distribution was: gastrointestinal

(67%), cutaneous (61%), respiratory (47%), and cardiovascular (9.4%).

Treatment was given for 70% of reactions (epinephrine-17%, antihista-

mines-81%). Only 21% with a known allergy disclosed their allergy to the

restaurant. Mistakes were attributed to cross-contact (same oil, grill or

pan) in 34% of reactions. It was common (61%) for persons with a prior

reaction to re-order seafood. A subsample who did so indicated their rea-

sons: allergy not clearly diagnosed (64%), attributed to a different cate-

gory of seafood (23%), or curiosity about persistence (13%). Restaurants

were rarely (19%) notified about reactions. Participants did not reduce

their frequency of restaurant dining, but 95% indicate they ask more ques-

tions or avoid particular types of foods/restaurants.

CONCLUSIONS: Seafood restaurant reactions can be attributed to poor

communication, misunderstanding of the allergy by those affected, and

cross-contact in preparation. These problems could be rectified through

education of patients and the restaurant industry.

Funding: Food Allergy and Anaphylaxis Network

164 Detection of Parvalbumin in Various Fish Species by 1D and2D Electrophoresis and Western Blots Using a CommerciallyAvailable Anti-frog Parvalbumin MAb

L. Chen, S. L. Hefle, R. E. Goodman; Food Allergy Research and

Resource Program, University of Nebraska-Lincoln, Lincoln, NE.

RATIONALE: Parvalbumin is the major cross-reactive food allergen

found in muscle tissue of several fish and frog species. We tested the bind-

ing of a commercially available anti-frog parvalbumin MAb to evaluate

its potential utility in detecting parvalbumin in commercially important

fish and fish-derived ingredients.

METHODS: Proteins from twelve commonly consumed fish species

were extracted and separated by one dimensional (1D) SDS polyacry-

lamide gel electrophoresis (SDS-PAGE). Proteins were stained with col-

loidal blue, or blotted onto nylon membranes for detection using an anti-

frog parvalbumin MAb, followed by goat anti-mouse IgG-horseradish

peroxidase and electrochemiluminescence (ECL) detection. Extracts with

detectable putative parvalbumin (10-12 kDa) were then separated by two

dimensional (2D) electrophoresis, with staining and western blotting to

investigate parvalbumin isoforms.

RESULTS: Most fish species, including catfish, carp, cod, flounder, hal-

ibut, mahi mahi, orange roughy, salmon, sole and tilapia, exhibited one or

two protein bands in the range of 10-12kDa in 1D SDS-PAGE and the

corresponding western blot. However, no putative parvalbumin was

detected in tuna or swordfish. The 2D electrophoresis and immunoblots

revealed from one to five potential isoforms of parvalbumins in different

species. Based on differences in relative intensities of protein staining

compared to ECL band intensities, parvalbumin epitopes of various iso-

forms probably differ, as might be expected from published sequences of

characterized parvalbumins.

CONCLUSIONS: These results suggest that the frog anti-parvalbumin

antibody may be a useful reagent for detecting parvalbumin from many of

the fish tested in this study, but further work is needed to evaluate appar-

ent differences in isoform recognition.

Funding: Food Allergy Research and Resource Program, University of

Nebraska

J ALLERGY CLIN IMMUNOL Abstracts S41VOLUME 117, NUMBER 2

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