Molecular cloning and the allergenic characterization of tropomyosin from Tyrophagus putrescentiae.
Storage mites have been recognized as a cause of asthma and rhinitis. Studies from several countries have shown that the IgE-mediated allergy to storage mites is of considerable importance, especially in rural populations. This study aimed to identify and characterize new allergens from Tyrophagus putrescentiae. A partial cDNA sequence encoding tropomyosin was isolated from the cDNA library by immunoscreening using anti-mouse IgG1 sera raised against T. putrescentiae whole body extract. The deduced amino acid sequence shares 64-94% identity with previously known allergenic tropomyosins. Its recombinant protein was produced by using a pET 28b expression system and purified by affinity chromatography using Ni-NTA agarose. The IgE reactivities of tropomyosins from T. putrescentiae and Dermatophagoides farinae were compared by enzyme linked immunosorbent assay (ELISA). Recombinant Tyr p 10 showed 12.5% (5/40) IgE-binding reactivity, whereas recombinant Der f 10 showed 25% (10/40) IgE-binding reactivity against the same sera from storage mite-sensitized and house dust mite-sensitized subjects. Both recombinant Tyr p 10 and Der f 10 showed little inhibition of IgE binding to T. putrescentiae crude extract by ELISA. Tropomyosin seems to contribute only a small portion of the cross-reactivity with house dust mites.
Jeong
Kyoung Yong
KY
Department of Parasitology and Institute of Tropical Medicine, Brain Korea 21 Project for Medical Science, Yonsei University, College of Medicine, Seoul, Korea. jeongky@paran.com
Lee
Haeseok
H
Lee
Jae Sik
JS
Lee
Jongweon
J
Lee
In-Yong
IY
Ree
Han-Il
HI
Hong
Chein-Soo
CS
Yong
Tai-Soon
TS
D040041:Acaridae / Q000737:chemistry*; D000293:Adolescent; D000328:Adult; D000368:Aged; D000485:Allergens / Q000096:biosynthesis / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D002648:Child; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism; D051379:Mice; D008875:Middle Aged; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D014335:Tropomyosin / Q000096:biosynthesis / Q000276:immunology*
D000485:Allergens; D011994:Recombinant Proteins; D014335:Tropomyosin; C523282:Tyr p 10 allergen, Tyrophagus putrescentiae; D007073:Immunoglobulin E