Assessment of recombinant dog allergens Can f 1 and Can f 2 for the diagnosis of dog allergy

Saarelainen, Soili; Taivainen, Antti; Rytkönen‐Nissinen, Marja; Auriola, Seppo; Immonen, Anu K.; Mäntyjärvi, Rauno; Rautiainen, J. Mikko; Kinnunen, Tuure; Virtanen, Tuomas

doi:10.1111/j.1365-2222.2004.02071.x

Cited by 50 publications

(70 citation statements)

References 26 publications

(34 reference statements)

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“…The study included 25 allergic patients whose allergy to dog was confirmed at the Pulmonary Clinic of Kuopio University Hospital, as described in detail elsewhere (12). For a person to be classified as dog-allergic, the following criteria were adopted: specific IgE by the dog UniCAP fluoroenzyme-immunometric assay (Pharmacia) of Ͼ0.7 kU/l and a skin prick test (SPT) 3 with dog allergen (epithelial preparation from ALK Abelló) of Ն3 mm.…”

Section: Subjectsmentioning

confidence: 99%

“…One of the subjects who had clinical symptoms upon exposure to dog and a positive SPT with dog allergen had no measurable IgE Ab to dog. Thirteen of the dog-allergic subjects were found to be sensitized to Can f 1 by SPTs with rCan f 1, as described previously (12). Twelve randomly selected healthy nonatopic dog owners served as control subjects.…”

Section: Subjectsmentioning

confidence: 99%

“…From day 5 onward, the cultures were refed with fresh medium and rIL-2 (25 U/ml) at 2-3 day intervals and on day 7 with feeder cells (2-3 ϫ 10 5 cells/well) and rIL-2. When the growth of the clones became visible (days [12][13][14][15][16][17][18][19][20], the cells were tested for specificity in the proliferation assay.…”

Section: Generation Of T Cell Lines and Clonesmentioning

confidence: 99%

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T Cell Epitope-Containing Peptides of the Major Dog Allergen Can f 1 as Candidates for Allergen Immunotherapy

Immonen

Farci

Taivainen

et al. 2005

The Journal of Immunology

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One prerequisite for developing peptide-based allergen immunotherapy is knowing the T cell epitopes of an allergen. In this study, human T cell reactivity against the major dog allergen Can f 1 was investigated to determine peptides suitable for immunotherapy. Seven T cell epitope regions (A–G) were found in Can f 1 with specific T cell lines and clones. The localization of the epitope regions shows similarities with those of the epitopes found in Bos d 2 and Rat n 1. On average, individuals recognized three epitopes in Can f 1. Our results suggest that seven 16-mer peptides (p15–30, p33–48, p49–64, p73–88, p107–122, p123–138, and p141–156), each from one of the epitope regions, show widespread T cell reactivity in the population studied, and they bind efficiently to seven HLA-DRB1 molecules (DRB1*0101, DRB1*0301, DRB1*0401, DRB1*0701, DRB1*1101, DRB1*1301, and DRB1*1501) predominant in Caucasian populations. Therefore, these peptides are potential candidates for immunotherapy of dog allergy.

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Section: Subjectsmentioning

confidence: 99%

Section: Subjectsmentioning

confidence: 99%

Section: Generation Of T Cell Lines and Clonesmentioning

confidence: 99%

See 1 more Smart Citation

T Cell Epitope-Containing Peptides of the Major Dog Allergen Can f 1 as Candidates for Allergen Immunotherapy

Immonen

Farci

Taivainen

et al. 2005

The Journal of Immunology

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“…The yeast P. pastoris has been used to express many allergens (Barral et al 2004;Saarelainen et al 2004;Barderas et al 2005). Because P. pastoris secretes few of its own proteins, recombinant proteins extracted from the medium are less contaminated with endogenous e-mail: Marcie_Moehnke@baylor.edu.…”

Section: Introductionmentioning

confidence: 99%

The expression of a mountain cedar allergen comparing plant-viral apoplastic and yeast expression systems

et al. 2008

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Jun a 3, a major allergenic protein in mountain cedar pollen, causes seasonal allergic rhinitis in hypersensitive individuals. Recombinant Jun a 3 was expressed in Nicotiana benthamiana interstitial fluid (300 μg/g leaf material) and Pichia pastoris (100 μg/ml media). Polyclonal anti-Jun a 3 and IgE antibodies from the sera of allergic patients both reacted with the recombinant protein. Of the two systems, recombinant protein from the plant apoplast contained fewer contaminating proteins. This method allows for a more convenient and inexpensive expression of the recombinant allergen, which will allow for further structural studies and may prove useful in diagnostic and/or immunotherapeutic strategies for cedar allergy.

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“…Can f 1 is more allergenic than Can f 2: 70% of the patients showed positive allergenic signals to Can f 1, but only 20% of the patients reacted to Can f 2 [6]. Recent research in Finland showed that the allergenic dominancy of Can f 1 was not found [8]. …”

Section: Introductionmentioning

confidence: 99%

Characterization of Dog Allergens Can f 1 and Can f 2. 1. Preparation of Their Recombinant Proteins and Antibodies

Kamata

Miyanomae

Nakayama

et al. 2006

Int Arch Allergy Immunol

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Background: Recombinant dog allergens, rCan f 1 and rCan f 2, and their antibodies are good tools for the characterization of dog allergens in order to develop modern therapeutic and preventive methods for dog allergy. Methods: In this study, cDNA was synthesized from the mRNA of dog salivary glands and cloned into the pGEX4T vector. rCan f 1 and rCan f 2 containing glutathione S-transferase were prepared by an Escherichia coli expression system. The antibodies against the recombinant allergens were prepared in rabbit. The serum of patients with dog allergy was evaluated by ELISA and immunoblot, using the recombinant allergens, goat anti-human immunoglobulin (Ig) E (Ε) labeled with biotin, and enzyme-labeled streptavidin. The binding of IgE in the serum of patients with dog allergy to dog saliva as a natural antigen was determined in the presence or absence of dog saliva, rCan f 1 and rCan f 2 as competitors. The anaphylactic potential of rCan f 1 and rCan f 2 was evaluated. The body temperature of the mice sensitized with rCan f 1 and rCan f 2 was monitored after intravenous injection of the allergens. The passive cutaneous anaphylaxis reaction was examined for rCan f 1 and rCan f 2. Dog salivary glands, dog saliva and dog hair/dander extracts were analyzed with antibodies by means of an immunoblot assay. The expression of the mRNA of Can f 1 and Can f 2 was verified in various dog tissues by reverse transcription polymerase chain reaction. Results: The E. coli expression system revealed the yield of rCan f 1 and rCan f 2 in 36 and 30 mg/l of culture. The molecular weights of rCan f 1 and rCan f 2 were 18 and 20 kDa in SDS-PAGE, respectively. rCan f 1 and rCan f 2 were found to bind to specific IgE in the serum of dog allergy patients. The binding of IgE in the patient serum for dog saliva was partially inhibited in the presence of rCan f 1 and rCan f 2. These recombinant allergens showed positive signals in passive cutaneous anaphylaxis reaction and induced anaphylactic shock in the mouse model, resulting in a decrease in body temperature. The polyclonal rabbit antibody for rCan f 1 bound to a protein of 20 kDa in the salivary gland, saliva and hair/dander extracts of dogs. The rabbit antibody for rCan f 2 bound to proteins in the saliva and the hair/dander extracts. The proteins possessed a molecular weight of 22/ 23 kDa. Reverse transcription polymerase chain reaction showed the presence of mRNA expression of Can f 1 and Can f 2 not only in the salivary gland but also in dog skin. A clear expression of Can f 2 mRNA was observed in dog skin. Conclusions: The recombinant allergens and antibodies for Can f 1 and Can f 2 are available for immunological and biochemical characterization of dog allergens. The molecular weight of the natural Can f 1 and Can f 2 in dog saliva and hair/dander extracts showed a higher molecular weight than that of rCan f 1 and rCan f 2. The significance of dog skin as the tissue producing dog allergens, especially Can f 2, should be considered in further studies.

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Assessment of recombinant dog allergens Can f 1 and Can f 2 for the diagnosis of dog allergy

Abstract: The recombinant allergens can be used reliably to identify Can f 1 and Can f 2-sensitized individuals. However, the two allergens are insufficient as reagents for diagnosing dog allergy.

Cited by 50 publications

References 26 publications

T Cell Epitope-Containing Peptides of the Major Dog Allergen Can f 1 as Candidates for Allergen Immunotherapy

T Cell Epitope-Containing Peptides of the Major Dog Allergen Can f 1 as Candidates for Allergen Immunotherapy

The expression of a mountain cedar allergen comparing plant-viral apoplastic and yeast expression systems

Characterization of Dog Allergens Can f 1 and Can f 2. 1. Preparation of Their Recombinant Proteins and Antibodies

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