Recombinant allergens for diagnosis and therapy of allergic disease

Chapman, Martin D.; Smith, Anderson D.; Vailes, Lisa D.; Arruda, L. Karla; Dhanaraj, V.; Pomés, Anna

doi:10.1067/mai.2000.109832

Cited by 205 publications

(148 citation statements)

References 87 publications

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“…Recently a major allergen from C. lunata was cloned, expressed, and characterized as subtilisin-like serine protease (14). Recombinant DNA technology has provided the opportunity to study the specific allergenic proteins, which may be modified to reduce allergenicity for safer treatments (15,16). Proteolytically inactive Per a 10 regulated the inflammatory parameters in mice (11).…”

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confidence: 99%

Characterization of Recombinant Per a 10 from Periplaneta americana

Govindaraj

Gaur

Arora

2013

Clin Vaccine Immunol

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bCockroach allergen is a major risk factor for IgE-mediated allergic response and asthma in sensitized individuals. Serine proteases have been identified from various sources and characterized as major allergens. The present study was aimed to express and characterize recombinant allergen Per a 10 (rPer a 10) from Periplaneta americana. rPer a 10 was expressed in Escherichia coli and purified in soluble form, yielding 0.75 mg/liter of culture. Homology of the Per a 10 protein sequence exhibited 27 to 38% similarity to the mite serine protease and 41 to 52% similarity to other insect trypsins. The purified rPer a 10 protein resolved at 28 kDa on SDS-PAGE and was recognized by cockroach-hypersensitive patients' sera by immunoblotting and enzymelinked immunosorbent assay (ELISA). In competitive ELISA, rPer a 10 required 96 ng of purified protein for 50% inhibition of IgE binding, whereas 34 ng of native protein (nPer a 10) was required for the same inhibition. rPer a 10 and nPer a 10 induced basophil histamine release in the range of 47 to 64% and 60 to 85%, respectively, when sensitized with cockroach-hypersensitive patients' sera. In conclusion, Per a 10 was subcloned, and the protein was purified to homogeneity. rPer a 10 showed reduced IgE binding and histamine release and showed no proteolytic activity. These data suggest that rPer a 10 has potential for immunotherapy.

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confidence: 99%

Characterization of Recombinant Per a 10 from Periplaneta americana

Govindaraj

Gaur

Arora

2013

Clin Vaccine Immunol

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“…This is mainly due to the complexity of many allergenic sources [5] and to the ability of the immune system of atopic individuals to mount IgE immune responses to a wide variety of molecular structures. Various allergens have been cloned and characterized using either direct PCR amplification or screening of bacteriophage λ cDNA [6]. However, these approaches suffer from serious methodological limitations.…”

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confidence: 99%

Rapid Identification of Allergen-Encoding cDNA Clones by Phage Display and High-Density Arrays

Kodzius¹,

Rhyner²,

Konthur

et al. 2003

CCHTS

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Abstract:We describe a high-throughput, quantitative technology for fast identification of all different clones present in selectively enriched phage surface-displayed cDNA libraries. The strategy is based on a combination of phage display and high-density arrays. To demonstrate the utility of the method cDNAs of Aspergillus fumigatus cloned into phagemid pJuFo were expressed on the tip of filamentous M13 phage and affinityselected on solid phase-immobilized serum IgE from allergic patients. Enriched phagemid libraries were amplified in bacteria, plated and arrayed into 384-well microtitre plates by robotic colony picking. cDNA inserts were amplified by high-throughput PCR and gridded onto high-density filter membranes. Filters were iteratively probed with randomly-sequenced inserts until all clones were identified. Eighty-one different sequences encoding IgE-binding proteins likely to cover a large part of the allergen repertoire of the mould were found. This approach represents a widely applicable method for rapid high-throughput identification of all individual cDNAs present in selectively enriched libraries.

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“…These methods should combine the analysis of individual proteins responsible for cross-reactions (both those contained in eHF and those forming de novo, as well as those found in various allergen extracts) with the simultaneous assessment of the antibodies contained in the investigated sera. In the future the diagnosis of allergy to eHF should be confirmed by means of tests utilizing recombinant allergens and new diagnostic techniques such as immunoblotting or ImmunoCAP ISAC (Immuno Solid-Phase AllergoChip) [52][53][54][55][56][57][58]. When deciding on which additional tests should be ordered it should therefore be kept in mind that hypersensitivity reactions to eHF have an unknown IgE-mediated, non-IgE-mediated or mixed underlying mechanism.…”

Section: Discussionmentioning

confidence: 99%

Patient characteristics of children with cow’s milk allergy presenting with an allergy to extensively hydrolyzed whey and/or casein formulae used as part of an elimination diet

Nowacki¹

2012

pdia

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A Ad dd dr re es ss s f fo or r c co or rr re es sp po on nd de en nc ce e: : Zygmunt Nowacki MD, PhD, Allergological-Pediatric Practice, Original paper A b s t r a c t I In nt tr ro od du uc ct ti io on n: : In the recent years the number of children with cow's milk protein allergy (CMPA) presenting with allergies to extensively hydrolyzed whey (eHF-W) and casein formulae (eHF-C) respectively has increased. A Ai im m: : To present the patient characteristics of children with CMPA who presented with allergic reactions to eHF-W and/or eHF-C used as part of an elimination diet. M Ma at te er ri ia al l a an nd d m me et th ho od ds s: : A retrospective study was carried out for a group of 65 selected from of 359 children (3 to 12 months) affected with CMPA and allergy to eHF. R Re es su ul lt ts s: : 18.1% (65/359) and 7.8% (28/359) from the group presented with allergic reactions to eHF-W and eHF-C, respectively. An allergic reaction to eHF-C occurred in 43% of the patients from the group (28/65). IgE-mediated CMPA presented with one of the following clinical manifestations: atopic dermatitis (AD) 94%, gastrointestinal symptoms (GI) 47%, respiratory symptoms (RS) 44%, anaphylactic reactions (AR) 26%. Non-IgE-mediated CMPA, on the other hand, manifested with: AD 97%, GI 22%, RS 10% .The observations showed that all 31 from the non-IgE mediated CMPA group with GI and AD manifestations presented with symptoms of eHF-C allergy (p = 0.001). In the IgE-mediated CMPA (n = 34) the following factors were statistically significant with relation to eHF-C allergy: CMPA manifesting with GI (p = 0.014) and AD. The occurrence of AR in response to cow's milk protein (CMP) decreased the probability of an eHF-C allergy (p = 0.028) in a statistically significant way. C Co on nc cl lu us si io on ns s: : Children with CMPA who presented with allergies to eHF-W and eHF-C had a higher frequency of clinical manifestation with GI and AD. K Ke ey y w wo or rd ds s: : cow's milk allergy, elimination diet.

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Recombinant allergens for diagnosis and therapy of allergic disease

Cited by 205 publications

References 87 publications

Characterization of Recombinant Per a 10 from Periplaneta americana

Characterization of Recombinant Per a 10 from Periplaneta americana

Rapid Identification of Allergen-Encoding cDNA Clones by Phage Display and High-Density Arrays

Patient characteristics of children with cow’s milk allergy presenting with an allergy to extensively hydrolyzed whey and/or casein formulae used as part of an elimination diet

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