The benefits of component-resolved diagnosis in diagnostics of peanut allergy

Introduction: Why can a peanut allergy cause exceptionally severe reactions, but not to everyone? Is there a way to identify patients who might experience these reactions? What is molecular diagnostics of allergy and what are the benefits of diagnosing patients at risk of anaphylaxis after consuming peanuts?  Purpose: The aim of the study is to show the benefits of molecular diagnostics of allergies based on the example of peanut allergy.Description of the state of knowledge: The diagnosis of peanut allergy is based on the assessment of the clinical presentation. The gold diagnostic standard is to perform a food challenge for suspected allergen. The following tests are helpful in diagnosis: skin tests or determination of allergen-specific E antibodies (asIgE) in blood serum. Until recently, the determination of asIgE concentration was possible only for allergen extracts. A milestone in diagnosing allergies is a serological diagnosis based on allergen molecules. The possibility of testing individual molecules increases the sensitivity and specificity of peanut allergy diagnostics and allows to indicate the risk of anaphylaxis in individual patients.Summary: Molecular diagnostics in individuals with peanut allergy make it possible to determine the individual patient’s allergy profile and indicate the risk of anaphylaxis. The information obtained with the help of this tool allows for precise therapeutic recommendations for people allergic to peanuts. Such diagnostics is a step towards personalized medicine in allergology. Keywords: allergy; molecular diagnostics of allergy; allergen-specific E antibodies; asIg

The general trends of laboratory diagnostic of drug hypersensitivity reactions

BACKGROUND: Drug hypersensitivity reactions (DHRs) are estimated to account for 3% to 6% of all hospital admissions and to occur in 10% to 15% of hospitalized patients resulting in morbidity, prolonged hospitalization, and increased risk of mortality. AIM: This review aims: 1) to cast a look on DHRs drug hypersensitivity reactions as allergic and non-allergic; 2) to establish of immunopathogenesis of various DHRs; 3) to create of new laboratory trends for diagnostic of allergic and non-alergic DHRs. MATERIALS AND METHODS: Clinically patients with DHRs can be classified as a) immediate (urticaria, angioedema, anaphylaxis etc.); b) non-immediate (delayed maculopapular eruptions, SJS/TEN, DRESS, vasculitis and cytopenia). Laboratory diagnostic of DHRs generally starts with in vivo tests (skin prick tests/ skin testing or intradermal testing) and continue with in vitro tests (radio immunoassays, fluor immunoassays, flow cytometry). RESULT: We propose the new trends to differential diagnostic of DHRs. In vitro testing for immediate IgE-dependent allergies 1) tryptase; 2) histamine release test; 3) specific IgE; 4) cellular in vitro tests - BAT, CAST-ELISA; for drug-specific T cell-mediated reactions 1) classical LTT with detecting of stimulation index; b) modern flow cytometry analysis with measuring the expression of activation surface markers on basophils; 2) ELISpot, which determines the number of cells that release relevant cytokines and cytotoxicity markers; 3) ELISA to measure released cytokines. In vitro testing for non-allergic reactions: 1) biochemical investigation of hepatic and renal metabolism; 2) detecting of the level of arachidonic acid metabolites - leukotrienes, prostaglandins; 3) components of complement C3a, C5a, and C5b-9; bradykinin; factor XII of coagulation system; 4) IgG; 5) infections; 6) active products of nitrosative and oxidative stress etc. CONCLUSION: We may conclude, that the various endotypes of DHRs identifying characteristics defined by specific mechanisms with each phenotype, diagnostic in vitro algorithm should be based on the new laboratory technologies. The results of various laboratory tests in DHRs-diagnostic will be taken into consideration to assign modern treatment

Zastosowanie testów aktywacji bazofilów w badaniach kliniczno-laboratoryjnych u pacjentów z alergią na beta-laktamy (badanie pilotażowe)

BACKGROUND: Beta (β)-lactam antibiotics (BLAs) are the first-line therapy for non-nosocomial and nosocomial bacterial infections and are most commonly reported to cause allergic reactions. Approximately 50% of all allergic patients in Europe and the USA suffer from drug allergies and BLA allergies. The AIM of the study was to assess cross-reactivity reactions between 2nd and 3rd generation cephalosporins in patients with a medical history of BLA reactions and the risk of adverse reactions to BLAs based on the results of the basophil activation test. MATERIALS AND METHODS: we examined 48 females and 8 males (in all 56 patients) aged 26 to 61 with primary reactions to BLAs and 24 healthy volunteers (control group). 19 (34%) patients were treated with amoxicillin, 18 (32,1%) patients were receiving amoxicillin+clavulanic acid, 6 (10,7%) patients were treated with cefuroxime, and 13 (23,2%) patients with ceftriaxone. Quantitative determination of the CD63 marker of basophil degranulation upon antigen stimulation in whole blood was performed with the use of Flow CAST (FK-CCR) (Bühlmann Laboratories AG, Switzerland). Based on the obtained ВАТ results, the patients were divided into two subgroups: the first group included 33 patients with positive stimulation index but lower CD63 expression (10 %). THE RESULT: We showed that patients from the second subgroup had the highest level of CD63 expression and stimulation index when amoxicillin, whereas the level of CD63 expression and stimulation index were lower whith ceftriaxone; at the same time, the level of CD63 expression and stimulation index were the lowest with cefuroxime. The patients who treated with and reacted to amoxicillin, as shown by high BAT, also had high CD63 expresiion after ceftriaxone and cefuroxime stimulation. In the first subgroup, urticarial and bronchospasm disappeared within 3 hours of the onset of symptoms in 51.5% of patients, the symptoms persisted for 2-3 days in 42.4% of patients with urticaria and angioedema, whereas maculopapular exanthema persisted for more than a week in 6.1% of the patients. Patients from the first subgroup (with low CD63 expression) had a weak reaction to the culprit antibiotic. Patients from the second subgroup had the strongest reaction to culprit antibiotics: anaphylaxis – 60.0%; Stevens-Johnson syndrome – 6.7%. We established that in patients with hypersensitivity to antibiotics the higher the baseline test scores after in vitro stimulation, the more severe clinical symptoms. CONCLUSION: for patients with clinical manifestations of BLA in case of conflicting anamnesis data, it is recommended to establish true sensitization to antibiotics and to predict the occurrence of cross-reactions between penicillins and cephalosporins not only of the 2nd but also of the 3rd generation. The results of BAT for antibiotics can be used to formulate future antibacterial treatments recommendation.WPROWADZENIE: Antybiotyki beta(β)-laktamowe (BLA) są terapią pierwszego rzutu w pozaszpitalnych i szpitalnych zakażeniach bakteryjnych i są najczęściej zgłaszane jako wywołujące reakcje alergiczne. Około 50% wszystkich alergików w Europie i USA cierpi na alergie na leki, w tym na alergie na BLA. Celem badania była ocena reakcji krzyżowych między cefalosporyną II i III generacji u pacjentów z klinicznym wywiadem reakcji na BLA i ryzyka wystąpienia niepożądanych reakcji BLA na podstawie wyników testu aktywacji bazofilów. MATERIAŁ I METODY: przebadaliśmy 48 kobiet i 8 mężczyzn (razem 56) z pierwotnymi reakcjami BLA w wieku od 26 do 61 lat oraz 24 zdrowych ochotników (grupa kontrolna). Pacjentów 19 (34%) bylo leczonych amoksycyliną, 18 (32,1%) – amoksycylina + kwasem klawulanowym, 6 (10,7%) – cefuroksymem i 13 (23,2%) – ceftriaksonem. W celu oceny markera degranulacji bazofili CD 63 po stymulacji antygenem w pełnej krwi wykorzystano oznaczenie Flow CAST (FK-CCR) (Bühlmann Laboratories AG, Szwajcaria). Na podstawie wyników ВАТ pacjentów podzielono na dwie podgrupy: w pierwszej grupie było 33 pacjentów z dodatnim niższym wynikiem indeksu stymulacji (10%). WYNIK: Nasze wyniki wykazały, że pacjenci z drugiej podgrupy mieli najwyższe wyniki ekspresji CD63 i wskaźnika stymulacji dla amoksycyliny, następnie dla ceftriaksonu, a ostatni dla cefuroksymu. Byli leczeni amoksycyliną i odpowiadali na nią, jak wykazały wysokie wartości ekspresji CD63 w BAT, ci pacjenci mieli również wysoką ekspresję CD63 po stymulacji ceftriaksonem i cefuroksymem. W pierwszej podgrupie u 51,5% pacjentów pokrzywka i skurcz oskrzeli ustąpiły w ciągu 3 godzin od wystąpienia objawów, u 42,4% pacjentów z pokrzywką i obrzękiem naczynioruchowym objawy utrzymywały się przez 2-3 dni, a u 6,1% osutka plamkowo-grudkowa - przez ponad tydzień. Pacjenci z pierwszej podgrupy (z niską ekspresją CD63) wykazywali klinicznie słabe objawy reakcji. Po leczeniu antybiotykami pacjenci z drugiej podgrupy wykazywali silniejsze objawy: u 60,0% - anafilaksja; 6,7% - zespół Stevensa-Johnsona. Wykazaliśmy, że u pacjentów z reakcja nadwrażliwości na leczenie antybiotykami im wyższe wyjściowe wyniki testu po stymulacji in vitro, tym bardziej nasilone są ich objawy kliniczne. WNIOSEK: u pacjentów z klinicznymi objawami BLA w przypadku sprzecznych danych z wywiadu zaleca się ustalenie rzeczywistego uczulenia na antybiotyki i przewidywanie występowania reakcji krzyżowych między penicylinami i cefalosporynami nie tylko drugiej, ale i trzeciej generacji. Wyniki BAT z antybiotykami mogą być wykorzystane do opracowania zaleceń przyszłych terapii przeciwbakteryjnych