Detection of allergen-induced genes in peripheral blood mononuclear cells of patients with allergic asthma using subtractive hybridization.
When stimulated with mite antigens, peripheral blood mononuclear cells obtained from patients with mite allergy release eosinophil chemotactic factor, a type of protein. Complementary DNA libraries were formulated from both peripheral blood mononuclear cells stimulated with mite antigens and those not stimulated with mite antigens and underwent subtraction and differential screening. The authors obtained 13 genes for which expression had increased because of mite antigens. Of these, nine were mitochondrial genes; two, h-satellite DNA III; one, factor XIII; and one, ferritin heavy subunit. As a result of polymerase chain reaction on both cDNA libraries, IL-5 genes were amplified only from the cDNA library that had been stimulated with mite antigens.
Department of Internal Medicine and Clinical Immunology, Dokkyo University School of Medicine, Tochigi-ken, Japan.
T
T
Fukuda
S
S
Mochida
Y
Y
Fukushima
S
S
Makino
D000485:Allergens / Q000494:pharmacology*; D000818:Animals; D000941:Antigens / Q000494:pharmacology*; D001249:Asthma / Q000097:blood / Q000235:genetics*; D001483:Base Sequence; D002631:Chemotactic Factors, Eosinophil / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000097:blood / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D015848:Interleukin-5 / Q000096:biosynthesis / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D008925:Mites / Q000276:immunology*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013268:Stimulation, Chemical
D000485:Allergens; D000941:Antigens; D002631:Chemotactic Factors, Eosinophil; D018076:DNA, Complementary; D015848:Interleukin-5; D012333:RNA, Messenger