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Diagnostic Procedures Of Delayed-type Hypersensitivity (DTH) Reactions To Low Molecular Weight Heparins (LMWHs)
Abstracts AB191
J ALLERGY CLIN IMMUNOL VOLUME 127, NUMBER 2
Immediate Hypersensitivity Reactions To Quinolones N. Blanca-Lopez1, M. Torres2, M. Seoane1, F. Ruano1, I. Do~ na3, M. Garcimartin1, M. Vazquez1, R. Barranco1, M. Blanca3, G. Canto1; 1Hospital Infanta Leonor, Madrid, SPAIN, 2Hopital Carlos Haya, Malaga, SPAIN, 3Hospital Carlos Haya, Malaga, SPAIN. RATIONALE: Hypersensitivity reactions (HSR) to quinolones (Q) seems to be increasing in the last decade being most of the cases immediate reactions. Our objective was to describe a group of patients diagnosed of immediate HSR to Q and verify cross-reactivity between them. METHODS: We studied a group of patients with confirmed immediate HSR to Q. Those with anaphylaxis were considered allergic by clinical history once other causes were ruled out and those with urticaria by control drug provocation. Tolerance to other different Q was assessed. RESULTS: From a total of 90 patients with confirmed diagnosis of HSR to antibiotics 24 cases (26,6%) were induced by Q. Eighteen were females and six males, with a mean age of 60.41 years, being the culprit Q levofloxacin (N5 12), moxifloxacin (N5 6), ciprofloxacin (N5 4) and norfloxacin (N52). Ten cases developed anaphylaxis (6 to moxiflocin and 4 to levofloxacin) and fourteen urticaria (8 with levofloxacin, 4 with ciprofloxacin and 2 with norfloxacin). Considering anaphylaxis this appeared in 100% of reactions induced by Moxifloxacin, 17% induced by levofloxacin and none with ciprofloxacin and norfloxacin. Regarding response to other Q 42,8% responded to more than one quinolone and 57,1% just to the culprit. CONCLUSIONS: Quinolones have become the second group of antibiotics responsible of immediate HSR in our population, being levofloxacin the most frequently involved. However moxifloxacin induced most severe reactions with 100% of cases developing anaphylaxis. The pattern of response was variable with nearly half of the patients responding to more than one Q suggesting cross-reactivity among them.:
732
Nano-technological Improvement in the Design of Radioimmunoassay to Detect IgE to Betalactams by Using Oligo(ethylenglycol)-Spacer to anchor Dendrimeric Conjugates to a Solid Phase M. I. Monta~ nez1, E. Perez-Inestrosa2, R. Suau2, C. Mayorga1, J. L. Rodri1 guez-Bada , M. J. Torres3, M. Blanca3; 1IMABIS Foundation-Carlos Haya Hospital, Malaga, SPAIN, 2Organic Chemistry, Faculty of Science, Universidad de Malaga, Malaga, SPAIN, 3Allergy Service, Carlos Haya Hospital, Malaga, SPAIN. RATIONALE: IgE specific to benzylpenicillin can be determined using radioimmunoassay (RAST) with benzylpenicilloyl-dendrimer conjugate coupled to cyanogen bromide activated cellulose surfaces. Herein we study how the activation of solid phases with reagents that act as a spacer, providing a well-defined distance between the nanoconjugate and the surface, can improve the IgE detection. METHODS: Hydroxyl groups of cellulose solid phase were activated with: cyanogen bromide (A), a difunctionalized pentaethylenglycol spacer (B) and (3-aminopropyl)trimethoxysilane reagent followed by pentaethylenglycol spacer (C), and reacted with 2nd and 4th generation (G) PAMAM dendrimers. The dendrimerized surfaces were penicilloylated. The dendrimer bound to the discs and the penicilloylation were assessed with ninhydrin method. The materials were immunologically tested by RAST using 20 sera from betalactam allergic patients. RESULTS: The results showed different amount of haptens bound to solid phases expressed as benzylpenicilloyl nmols/mg of cellulose, being for G2 and G4, 10 and 4 respectively, method A; 12 and 15, method B; and 40 and 38, method C. Results achieved positivity in 55%, 80% and 70% for materials A, B and C respectively. High/medium RAST level sera showed positive results with all the evaluated materials. However, IgE detected in low RAST level sera increased significantly with B and C compared to A materials. CONCLUSIONS: The different activated cellulose showed differences in the content of hapten-dendrimer conjugates. RAST results suggest that these nanoconjugates spaced from surfaces incorporate the appropriate structures for specific IgE recognition and improve the sensitivity for clinical diagnostic tests for allergic reactions to betalactams.
733
Prostaglandins Receptors Polymorphisms In Patients With NSAIDs-induced Urticaria J. A. Cornejo-Garcia1, L. R. Jagemann1, I. Do~na2, N. Blanca-Lopez3, J. J. Laguna4, J. Fernandez5, A. Rosado6, S. Terrados7, M. Gandolfo8, M. Blanca2, G. Canto3; 1Fundacion IMABIS, Malaga, SPAIN, 2Servicio de Alergologia, Hospital Carlos Haya, Malaga, SPAIN, 3Servicio de Alergologia, Hospital Infanta Leonor, Madrid, SPAIN, 4Servicio de Alergologia, Hospital de la Cruz Roja, Madrid, SPAIN, 5Servicio de Alergologia, Hospital de Elche, Alicante, SPAIN, 6Unidad de Alergia, Hospital de Alcorcon, Madrid, SPAIN, 7Servicio de Alergologia, Hospital Ramon y Cajal, Madrid, SPAIN, 8Unidad de Alergia, Hospital de Fuenlabrada, Madrid, SPAIN. RATIONALE: NSAIDs are actually the compounds most frequently involved in hypersensitivity drug reactions, and there are at least three different mechanisms involved: IgE- mediated, and T cell-mediated (immunological specific mechanisms) and cross-intolerance (CI) (immunological non-specific). In the latter group skin is the main affected organ. Associations between CI and different SNPs in arachidonic acid pathway or prostaglandins receptors (PGsR)related genes had been described in patients with respiratory symptoms or chronic urticaria, but up today the number of cases is rather limited and selected populations are heterogeneous. In this study, we analysed the association between CI and several SNPs in PGsR in a large group of patients with acute urticaria/angioedema, defined as skin episode in absence of airways involvement or coexistent chronic urticaria. METHODS: The population was obtained from several Allergy Services integrated into the Spanish network RIRAAF. Cases included had to have at least two episodes of CI with cutaneous symptoms after the intake of two or more NSAIDs from different chemical groups. We studied several SNPs in PTGER1, PTGER2, PTGER3, PTGER4, PTGDR, and PTGFR genes with the TaqManÒOpenArrayä System. RESULTS: We included 249 subjects with urticaria/angioedema, and 247 age- and sex-matched healthy controls. We found statistically significant differences in genotypic frequencies in PTGDR rs8004654 (p50.01). CONCLUSIONS: Genotypic and allelic frequencies in the SNPs analyzed were similar to others studied populations. Although an association was found between acute urticaria/angioedema and the rs8004654 polymorphism, further studies are required to analyze the role of genetic variants in PGsR and other related genes in this clinical entity.
734
Diagnostic Procedures Of Delayed-type Hypersensitivity (DTH) Reactions To Low Molecular Weight Heparins (LMWHs) C. Vila, A. Rosado, W. L. Almanzar, M. M. Moro, M. D. Alonso, M. Acosta; Hospital Universitario Fundacion Alcorcon, Alcorcon (Madrid), SPAIN. RATIONALE: Cross-reactivity is a widespread phenomenon in patients who develop cutaneous delayed-type hypersensitivity (DTH) reactions to low molecular weight heparins (LMWHs). Applying adequate diagnostic tools and with a correct interpretation we can identified a safe alternative treatment. Skin tests with immediate and late reading are the most reliable diagnostic tools for heparin-induced DTH reactions. METHODS: We reviewed 3 patients with DTH reactions caused by enoxaparin. Using patch-test (PT) and delayed-reading intradermal skin test (IST) we evaluated the responsability of enoxaparin in the reaction and also cross-reactivities among different LMWHs and heparin sodium as unfractioned heparin. The list of LMWHs tested including: enoxaparin, dalteparin, nadroparin, bemiparin. PTs were performed with undiluted compounds and the readings were realized at 48 hours and 96 hours. ISTs were done with undiluted and 10 folds diluted heparins in normal saline. Readings were performed at 20 minutes, 48 hours and 96 hours. RESULTS: 2 of 3 patients were sensitized to all tested heparins when PTand IST results were read up to 96 hours. In addition, the tested substances caused local eczematous reactions in these patients. PTs were safer than ISTs, showing milder local reactions; however, PTs were less sensitive than ISTs. CONCLUSIONS: The most sensitive test for diagnosis of DTH reactions to heparins was the IST; higher sensitivity was showed when the results were reading at 96 hours. Therefore, we recommend studies of tolerance to alternative anticoagulants after a minimum interval of 5 days from the realization of skin tests.
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731
J ALLERGY CLIN IMMUNOL VOLUME 127, NUMBER 2
Immediate Hypersensitivity Reactions To Quinolones N. Blanca-Lopez1, M. Torres2, M. Seoane1, F. Ruano1, I. Do~ na3, M. Garcimartin1, M. Vazquez1, R. Barranco1, M. Blanca3, G. Canto1; 1Hospital Infanta Leonor, Madrid, SPAIN, 2Hopital Carlos Haya, Malaga, SPAIN, 3Hospital Carlos Haya, Malaga, SPAIN. RATIONALE: Hypersensitivity reactions (HSR) to quinolones (Q) seems to be increasing in the last decade being most of the cases immediate reactions. Our objective was to describe a group of patients diagnosed of immediate HSR to Q and verify cross-reactivity between them. METHODS: We studied a group of patients with confirmed immediate HSR to Q. Those with anaphylaxis were considered allergic by clinical history once other causes were ruled out and those with urticaria by control drug provocation. Tolerance to other different Q was assessed. RESULTS: From a total of 90 patients with confirmed diagnosis of HSR to antibiotics 24 cases (26,6%) were induced by Q. Eighteen were females and six males, with a mean age of 60.41 years, being the culprit Q levofloxacin (N5 12), moxifloxacin (N5 6), ciprofloxacin (N5 4) and norfloxacin (N52). Ten cases developed anaphylaxis (6 to moxiflocin and 4 to levofloxacin) and fourteen urticaria (8 with levofloxacin, 4 with ciprofloxacin and 2 with norfloxacin). Considering anaphylaxis this appeared in 100% of reactions induced by Moxifloxacin, 17% induced by levofloxacin and none with ciprofloxacin and norfloxacin. Regarding response to other Q 42,8% responded to more than one quinolone and 57,1% just to the culprit. CONCLUSIONS: Quinolones have become the second group of antibiotics responsible of immediate HSR in our population, being levofloxacin the most frequently involved. However moxifloxacin induced most severe reactions with 100% of cases developing anaphylaxis. The pattern of response was variable with nearly half of the patients responding to more than one Q suggesting cross-reactivity among them.:
732
Nano-technological Improvement in the Design of Radioimmunoassay to Detect IgE to Betalactams by Using Oligo(ethylenglycol)-Spacer to anchor Dendrimeric Conjugates to a Solid Phase M. I. Monta~ nez1, E. Perez-Inestrosa2, R. Suau2, C. Mayorga1, J. L. Rodri1 guez-Bada , M. J. Torres3, M. Blanca3; 1IMABIS Foundation-Carlos Haya Hospital, Malaga, SPAIN, 2Organic Chemistry, Faculty of Science, Universidad de Malaga, Malaga, SPAIN, 3Allergy Service, Carlos Haya Hospital, Malaga, SPAIN. RATIONALE: IgE specific to benzylpenicillin can be determined using radioimmunoassay (RAST) with benzylpenicilloyl-dendrimer conjugate coupled to cyanogen bromide activated cellulose surfaces. Herein we study how the activation of solid phases with reagents that act as a spacer, providing a well-defined distance between the nanoconjugate and the surface, can improve the IgE detection. METHODS: Hydroxyl groups of cellulose solid phase were activated with: cyanogen bromide (A), a difunctionalized pentaethylenglycol spacer (B) and (3-aminopropyl)trimethoxysilane reagent followed by pentaethylenglycol spacer (C), and reacted with 2nd and 4th generation (G) PAMAM dendrimers. The dendrimerized surfaces were penicilloylated. The dendrimer bound to the discs and the penicilloylation were assessed with ninhydrin method. The materials were immunologically tested by RAST using 20 sera from betalactam allergic patients. RESULTS: The results showed different amount of haptens bound to solid phases expressed as benzylpenicilloyl nmols/mg of cellulose, being for G2 and G4, 10 and 4 respectively, method A; 12 and 15, method B; and 40 and 38, method C. Results achieved positivity in 55%, 80% and 70% for materials A, B and C respectively. High/medium RAST level sera showed positive results with all the evaluated materials. However, IgE detected in low RAST level sera increased significantly with B and C compared to A materials. CONCLUSIONS: The different activated cellulose showed differences in the content of hapten-dendrimer conjugates. RAST results suggest that these nanoconjugates spaced from surfaces incorporate the appropriate structures for specific IgE recognition and improve the sensitivity for clinical diagnostic tests for allergic reactions to betalactams.
733
Prostaglandins Receptors Polymorphisms In Patients With NSAIDs-induced Urticaria J. A. Cornejo-Garcia1, L. R. Jagemann1, I. Do~na2, N. Blanca-Lopez3, J. J. Laguna4, J. Fernandez5, A. Rosado6, S. Terrados7, M. Gandolfo8, M. Blanca2, G. Canto3; 1Fundacion IMABIS, Malaga, SPAIN, 2Servicio de Alergologia, Hospital Carlos Haya, Malaga, SPAIN, 3Servicio de Alergologia, Hospital Infanta Leonor, Madrid, SPAIN, 4Servicio de Alergologia, Hospital de la Cruz Roja, Madrid, SPAIN, 5Servicio de Alergologia, Hospital de Elche, Alicante, SPAIN, 6Unidad de Alergia, Hospital de Alcorcon, Madrid, SPAIN, 7Servicio de Alergologia, Hospital Ramon y Cajal, Madrid, SPAIN, 8Unidad de Alergia, Hospital de Fuenlabrada, Madrid, SPAIN. RATIONALE: NSAIDs are actually the compounds most frequently involved in hypersensitivity drug reactions, and there are at least three different mechanisms involved: IgE- mediated, and T cell-mediated (immunological specific mechanisms) and cross-intolerance (CI) (immunological non-specific). In the latter group skin is the main affected organ. Associations between CI and different SNPs in arachidonic acid pathway or prostaglandins receptors (PGsR)related genes had been described in patients with respiratory symptoms or chronic urticaria, but up today the number of cases is rather limited and selected populations are heterogeneous. In this study, we analysed the association between CI and several SNPs in PGsR in a large group of patients with acute urticaria/angioedema, defined as skin episode in absence of airways involvement or coexistent chronic urticaria. METHODS: The population was obtained from several Allergy Services integrated into the Spanish network RIRAAF. Cases included had to have at least two episodes of CI with cutaneous symptoms after the intake of two or more NSAIDs from different chemical groups. We studied several SNPs in PTGER1, PTGER2, PTGER3, PTGER4, PTGDR, and PTGFR genes with the TaqManÒOpenArrayä System. RESULTS: We included 249 subjects with urticaria/angioedema, and 247 age- and sex-matched healthy controls. We found statistically significant differences in genotypic frequencies in PTGDR rs8004654 (p50.01). CONCLUSIONS: Genotypic and allelic frequencies in the SNPs analyzed were similar to others studied populations. Although an association was found between acute urticaria/angioedema and the rs8004654 polymorphism, further studies are required to analyze the role of genetic variants in PGsR and other related genes in this clinical entity.
734
Diagnostic Procedures Of Delayed-type Hypersensitivity (DTH) Reactions To Low Molecular Weight Heparins (LMWHs) C. Vila, A. Rosado, W. L. Almanzar, M. M. Moro, M. D. Alonso, M. Acosta; Hospital Universitario Fundacion Alcorcon, Alcorcon (Madrid), SPAIN. RATIONALE: Cross-reactivity is a widespread phenomenon in patients who develop cutaneous delayed-type hypersensitivity (DTH) reactions to low molecular weight heparins (LMWHs). Applying adequate diagnostic tools and with a correct interpretation we can identified a safe alternative treatment. Skin tests with immediate and late reading are the most reliable diagnostic tools for heparin-induced DTH reactions. METHODS: We reviewed 3 patients with DTH reactions caused by enoxaparin. Using patch-test (PT) and delayed-reading intradermal skin test (IST) we evaluated the responsability of enoxaparin in the reaction and also cross-reactivities among different LMWHs and heparin sodium as unfractioned heparin. The list of LMWHs tested including: enoxaparin, dalteparin, nadroparin, bemiparin. PTs were performed with undiluted compounds and the readings were realized at 48 hours and 96 hours. ISTs were done with undiluted and 10 folds diluted heparins in normal saline. Readings were performed at 20 minutes, 48 hours and 96 hours. RESULTS: 2 of 3 patients were sensitized to all tested heparins when PTand IST results were read up to 96 hours. In addition, the tested substances caused local eczematous reactions in these patients. PTs were safer than ISTs, showing milder local reactions; however, PTs were less sensitive than ISTs. CONCLUSIONS: The most sensitive test for diagnosis of DTH reactions to heparins was the IST; higher sensitivity was showed when the results were reading at 96 hours. Therefore, we recommend studies of tolerance to alternative anticoagulants after a minimum interval of 5 days from the realization of skin tests.
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731