Ignacio Arrieta

Molecular cloning of major allergen from Cupressus arizonica pollen: Cup a 1

The family Cupressaceae is a relevant source of allergens that causes winter respiratory allergies. Cloning and sequencing the major antigen of Cupressus arizonica is important for a better diagnosis and treatment of sensitized patients.

A double-blind, placebo-controlled oral challenge study with lyophilized larvae and antigen of the fish parasite, Anisakis simplex

Background: The third‐stage larvae of Anisakis simplex may be a hidden source of allergens in fish. The objective was to determine whether the ingestion of lyophilized A. simplex larvae, or antigen, induces clinical symptoms in a group of A. simplex‐sensitized patients.

Immunopathogenesis of human gastrointestinal infection by Anisakis simplex

BACKGROUND Anisakis simplex is a parasite of fish, and in the case of human infestation, it should be considered as a possible cause of gastrointestinal disease, especially in countries where raw or undercooked fish is a frequent food. Clinical features of anisakiasis may simulate acute abdominal pain, such as that found in patients with gastric ulcers, appendicitis, and Crohns disease. Furthermore, many cases of anisakiasis are diagnosed as eosinophilic gastroenteritis, which is a broad term for a specific disease. OBJECTIVE The purpose of this study was to investigate the immunopathogenesis of human gastrointestinal infestation by A simplex. METHODS Thirteen intestinal biopsy specimens from patients with anisakiasis were analyzed for the presence of messenger (m)RNA for different cytokines and inflammatory mediators by RT-PCR. Specific IgE, eosinophil cationic protein, eosinophil protein X, and tryptase levels were measured in each patients serum. Also, cell cultures were set up with lymphocytes from some patients and stimulated in vitro with Anisakis and Ascaris antigens. RESULTS We performed immunologic phenotyping in 13 patients. All patients underwent biopsy after emergency surgery caused by episodes of acute abdominal pain. In all cases inflammatory infiltrate composed of eosinophils and lymphocytes was found in the intestinal wall. We demonstrated that after infestation, a T(H2)-type immune response occurred. Also, major basic protein, nitric oxide, and eotaxin were found in the tissue, and eosinophil cationic protein and eosinophil protein X levels were elevated in sera. CONCLUSION These data and in vitro lymphocyte cultures indicate that a T(H2) mechanism plays an important role in the inflammatory infiltrate produced by the anchorage of parasites in the gastrointestinal wall.

Beer-induced anaphylaxis : identification of allergens

Background: We report on a 21‐year‐old atopic woman who developed urticaria, angioedema of the face, and wheezy dyspnea shortly after drinking beer and after eating a corn‐made snack.

Molecular cloning and characterization of an IgE-reactive protein from Anisakis simplex: Ani s 1.

Ingestion of the parasitic nematode Anisakis simplex in undercooked fish can cause severe allergic reactions in some individuals. Using pooled human sera from sensitized patients we have probed an expression library for A. simplex antigens. One positive clone was found to encode a full length 21 kDa protein with strong homology to nematode troponins. The recombinant protein was expressed as a GST-fusion protein and found by immunoblot analysis to react with sera from 20% of allergic patients. The presence of functional EF-hand Ca(2+) binding motifs was demonstrated by gel-shift analysis.

Genetic restrictions in olive pollen allergy

BACKGROUND The major antigen of olive tree pollen, Ole e 1, produces an IgE response restricted by DQ2. OBJECTIVE Our purpose was to further analyze the genetic restrictions associated with IgE and IgG antibodies against Ole e 1 and IgE against the recently described antigen Ole e 3. METHODS Twenty-two nuclear olive pollen-allergic families (n = 88) were selected. DRB1 and DQB1, TCR-Valpha 8.1, the high-affinity receptor of IgE (FcepsilonRI-beta) Rsa I exon 7 and intron 2 and TNF-beta (LTalpha-Nco I) polymorphisms were determined by PCR and analyzed for association with allergic traits by the multiallelic transmission disequilibrium test. RESULTS Significant associations were found among HLA-DQB1*0201 (n = 29) and high levels of IgG (P =.023) and IgE (P =.0136) antibodies to Ole e 1 and with IgE specific to Ole e 3 (P =.0368). DRB1*0701 was associated with high levels of total serum IgE (P =.04) and IgG against Ole e 1 (P =.025). The FcepsilonRI-beta Rsa I exon 7, allele 1 (n = 39), was associated with high levels of total serum IgE (P =. 01), IgE antibodies against Olea europaea extract (P =.004), and specific antibodies to Ole e 1, IgG (P =.04), and IgE (P =.006). The FcepsilonRI-beta Rsa I intron 2, allele 2 (n = 33), was associated with IgE antibodies to O europaea extract (P =.003) and specific antibodies to Ole e 1, IgG (P =.025), and IgE (P =.05). CONCLUSIONS We found a new association between IgE antibody response to Ole e 3 and DQB1*0201 and verified the previously reported association between Ole e 1-specific response and DQB1*0201. Also, the association between FcepsilonRI-beta and IgE antibodies against Ole e 1 was demonstrated.

Allergy to dermatophagoides in a group of Spanish gypsies: genetic restrictions.

Background: Spanish gypsies have traditionally lived as nomads, a reason why few epidemiological studies were done in this ethnic group. However, the high prevalence of asthmatic diseases demonstrated in a population residing in the North of Spain induces us to analyse whether it was due to the influence of genetic loci previously implicated in other population studies as causing the disorders. Methods: DRB1* and DQB1* HLA class II, TCR-Vα8.1, FcΕRI-β Rsa I exon 7 and intron 2, TNF-β (LTα-Nco I) and CD14, were tested for association with asthma and atopy by multiple regression analysis, in 5 families comprising 87 individuals. Results: Significant associations were found with DQB1*02 (p = 0.02) and DQB1*0301 (p = 0.008) and elevated levels of total serum IgE. A negative association (p = 0.02) was found between total serum IgE and DRB1*14. FcΕRI-β Rsa I-In2 allele 1 was associated with high levels of total serum IgE (p = 0.04). Levels of Der p 1 IgE antibodies were negatively associated with DRB1*11-DQB1*0301 (p = 0.007), and positively with TCR Vα-8 allele 1 (p = 0.04) and with FcΕRI-β Rsa I-In2 allele 1 (p = 0.009). Conclusions: Our results do not show any association between asthma and the genetic loci studied although they do suggest the existence of multiple genetic influences on the allergic response in these families.

Immunological Aspects of the Toxic Oil Syndrome

The Toxic Oil Syndrome (TOS), which occurred in Spain in 1981, was caused by the ingestion of rapeseed oil denatured with 2% aniline, sold illegally as edible oil. The number of unaffected individuals exposed to the toxic oil is unknown, but more than 20,000 people suffered from the disease.

Eosinophils transcribe and translate messenger RNA for inducible nitric oxide

Because of the involvement of nitric oxide (NO) in inflammatory states such as parasitic and hypersensitivity disorders and the fact that eosinophils are one of the cell types implicated, we asked whether eosinophils were able to express mRNA specific to inducible NO synthase (iNOS) and iNOS protein and to secrete nitric oxide. iNOS protein was detected on eosinophil preparations by immunocytochemistry using iNOS mAb. Expression of iNOS protein was also detected by immunoblotting in human purified eosinophils and an eosinophilic leukemia cell line, Eol-3. Nitrite production was detected in the supernatant of human eosinophils and Eol-3 cells cultured for 24 h, and was completely inhibited in the presence of the NOS inhibitor N-methylester-L-arginine. iNOS cDNA was obtained by reverse transcription-PCR. After subcloning, sequencing of the 259-bp fragment from three different human eosinophils cDNAs revealed 97% identity with macrophage/monocyte iNOS. Our studies describe for the first time the presence of iNOS on eosinophil and a putative new role for this cell in inflammatory states such as asthma and parasitic disease.