J ALLERGY CLIN IMMUNOL

FEBRUARY 2014

AB114 Abstracts

SUNDAY

398 Utility Of Probability Curves Using 3gAllergy For Diagnosis OfWheat Allergy

Sakura Sato1, Kiyotake Ogura1, Yasunori Sato2, Motohiro Ebisawa, MD,

PhD, FAAAAI1; 1Clinical Research Center for Allergy and Rheuma-

tology, Sagamihara National Hospital, Kanagawa, Japan, 2Department

of Biostatistics, Clinical Research Center Chiba University.

RATIONALE: Previously we presented the utility of allergen specific IgE

(sIgE) measurements by 3gAllergy using probability curves for diagnosis

of hen’s egg and cow’s milk allergy. In this study, we assessed the utility of

probability curves in the diagnosis of wheat allergy.

METHODS: Serum samples were collected from 330 children (mean age

10.5 months) suspected of having wheat allergy. Allergen sIgE testing for

wheat (W) and Gluten (G) was performed using the IMMULITE� 2000

3gAllergyTM and ImmunoCAP� sIgE assays. Final diagnosis of food al-

lergy (FA) was confirmed by oral food challenge (OFC) and convincing

allergic reactions to wheat products; acquisition of tolerance was defined

as patients who had ingested 100 g of Japanese wheat noodle without

any symptoms. Serum samples were drawn 6 months before the FA diag-

nosis and then stored frozen.

RESULTS: Of the 330 patients, 49 children were diagnosed with W

allergy. W and G sIgE measurements by 3g were correlated with CAP W

(W: r50.91, G: r50.93, p<0.0001); In comparison to CAP, 3g W values

were likely to be lower than Cap W. Probability curves using the sIgE

values from the FA positive negative groups were established using logistic

regression analysis, and the 95%predictive decision point were determined

by 3g (W: 234 IUA/mL, G: 92 IUA/mL.) The 95% decision point for W by

CAP was not determined because sIgE results exceeded the range of the

assay.

CONCLUSIONS: Measurement of sIgE to Wand G using 3g and the PC

are beneficial for diagnosis of W allergy.

399 Skin Prick Test and Specific IgE To Purified Peanut AllergensAre Related To The Age Of Onset Of Sympstons

Dr. Maria Salas, MD, PhD1, Dr. Francisca Gomez, MD, PhD2, Dr. Ana

Aranda, PhD3, Dr. Carmen Rondon, MD, PhD1, Dr. Natalia Blanca-

L�opez, MD, PhD4, Dr. Gabriela Canto, MD, PhD4, Dr. Maria J

Torres, MD, PhD1, Dr. Cristobalina Mayorga, PhD3, Dr. Miguel

Blanca, MD, PhD1, Maria Isabel S�anchez Rivas5; 1Allergy Service, Carlos

Haya Hospital, M�alaga, Spain, 2Allergy Service, Carlos Haya Hospital,

Spain, 3Research Laboratory, Carlos Haya Hospital-FIMABIS, M�alaga,

Spain, 4Allergy Service, Infanta Leonor Hospital, Madrid, Spain, 5Allergy

Service Carlos Haya Hospital, Malaga, Spain.

RATIONALE: We aim to analyse changes in the skin prick test (SPT) to

peanut and IgE levels to Ara h1; Ara h2; Arah3; Ara h9 and Pru p3 in

relationship with the age of patients and the onsent of symptons.

METHODS: We studied 167 peanut allergic patients confirmed by SPTor

prick by prick to fresh extract, in vitro specific IgE and/or a double blind

placebo control challenge with peanut. Patients were divided into 3 ages

groups: 15-30, 31-45 and 46-80. IgE levels were determined by ELISA to

Ara h1, Ara h2, Ara h3, Ara h9 and Pru p3.

RESULTS: Thirty-three patients evaluated in the first group (54%) were

positive to peanut in SPT, 34 (42,50%) in second group and 4 (15%) in the

last group. ELISA showed 50%positivity toAra h1, Ara h2; Ara h3, 60% to

Pru p3 and 80% to Ara h9 in the first group. 40% to Ara h1, Ara h3, 50% to

Ara h9 and Pru p3 and less than 30% to Arah2 in the second group. In the

third group IgE levels fall below 20% to Ara h2, Ara h3 and Ara h9. Ara h1

and Pru p3 were not detected.

CONCLUSIONS: We observed a decrease in the reactivity response of

SPT to peanut and IgE levels in the third age group with and onset of

symptoms over tan ten years. Levels to Ara h 2 were lower onmedian ages,

although Ara h9 and Pru p3 were similar in the the first groups.

400 The Relationship Between Mitochondrial HaplogroupsVariant on Children with Cow Milk Allergy Expressed AsAtopic Dermatitis and Gastrointestinal Disease

Dr. Juan Carlos Muino, MD, PhD, FAAAAI1, Dr. Raul Boudet1,

Dr. Maria Chaig1, Dr. Roberto Chaig1, Prof. Nelida Gerez2, Prof. Juan

Carlos Copioli1; 1FAC CS MED UNC, Cordoba, Argentina, 2FAC CS

MED UNC, C�ordoba, Argentina.

RATIONALE: Genotypes associated to cow’s milk allergy are unknown.

They have not been replicated in independent population, and could be

responsible for themarked variability in individual clinical response to cow

milk proteins.The objectivewas to characterize haplogroups of the D-Loop

region of mitochondrial DNA in a group of children allergic to cow’s milk

in order to arrive to a better knowledge of biological and genetic

heritability in the etiology of the disease

METHODS: .We studied 41 children of both sex aged 0 – 2 years, 11

allergic to cow’s milk demonstrated by challenge and 30 healthy subjects

(controls), from the urban area of Rio Cuarto City, C�ordoba, Argentina.We

performed Analysis of variants of D-loop region of the mitochondrial

genome. The D-Loop region HVI, II and II of the mitochondrial genome

was amplified by PCR, for which we used specific primers. Phylogenetic

analysis was calculated using the program CLUSTAL OMEGA, the

Neighbor–Joining, BLOSUM62 with data studied and recorded by Jukes-

Cantor and then with Kimura-2

RESULTS: The cow milk allergic patients were divided in: (a) Atopic

Dermatitis (AD) + Gastrointestinal Disease (GID) (n: 6) and (b) Rhinitis

and Asthma (n: 5). We found the non described variant in transition of

haplogroups, T16519C associated with (a) in 6/6 cases when compared

with (b) negative in 5/5 cases and the control group (6/30), p5 0, 0312, RR:

2,900

CONCLUSIONS: These features suggest that this variant probably

increases the possibility of suffering cow milk allergy associated with

AD +GID.

401 Detection Of Peanut Allergens In Breast Milk and SalivaDr. Kelli M. Rose, MD1, Christian Plaisance2, Casey C.

Grimm, PhD2, Hsiaopo Cheng, MS2, Tysheena Charles, MS2, Saeed A.

Jortani3, Soheila J. Maleki, PhD2; 1Tulane University, New Orleans,

LA, 2USDA-ARS-SRRC, New Orleans, LA, 3University of Louisville,

Louisville, KY.

RATIONALE: Infants have been reported to react to peanut upon their

first known exposure and peanut proteins have been detected in breast milk.

Identification of the allergens or fragments thereof in breast milk may

allow us to determine if they are sensitizing or tolerizing.

METHODS: Various immunoassays were optimized and utilized to

analyze for the presence of peanut proteins in breast milk. Breast milk

samples from healthy lactating mothers were collected following a 48 hour

peanut fast and spiked with known amounts of peanuts and subjected to

immunoprecipitation, SDS-PAGE, western blot and ELISA with anti-

peanut, anti-Ara h 1, 2 and 3 antibodies.Mass spectrometry was performed

to identify peanut peptides in breast milk.

RESULTS: We found that we were able to detect peanut allergen, Ara h 1,

Ara h 2 and Ara h3, in peanut-spiked breast milk and saliva at nM levels.

We were able to identify specific peanut peptides of Ara h 1, Ara h 2 and

Ara h 3/4 using mass spectrometry.

CONCLUSIONS: The fact that allergic proteins or peptides survive in

digestive enzymes, and are likely to be secreted in biological fluids

indicates they are most likely the sensitizing or tolerizing agents within an

allergic food. Developing methods to detect these allergens in breast milk

is a preliminary step in identification of allergens or fragments thereof that

are secreted and may contribute to the original development of allergic

disease.