title: Diagnosis of eight groups of xeroderma pigmentosum by genetic complementation using recombinant adenovirus vectors.
abstract: Because patients with xeroderma pigmentosum (XP) must avoid ultraviolet (UV) light from an early age, an early diagnosis of this disorder is essential. XP is composed of seven genetic complementation groups, XP-A to -G, and a variant type (XP-V). To establish an easy and accurate diagnosis of the eight disease groups, we constructed recombinant adenoviruses that expressed one of the XP cDNA. When fibroblasts derived from patients with XP-A, -B, -C, -D, -F or -G were infected with the adenovirus expressing XPA, XPB, XPC, XPD, XPF or XPG, respectively, and UV-C at 5-20 J/m2was irradiated, cell viability was clearly recovered by the corresponding recombinant adenoviruses. In contrast, XP-E and XP-V cells were not significantly sensitive to UV irradiation and were barely complemented by the matched recombinant adenoviruses. However, co-infection of Ad-XPA with Ad-XPE increased survival rate of XP-E cells after UV-C exposure. When XP-V cell strains, including one derived from a Japanese patient, were infected with Ad-XPV, exposed to UV-B and cultured with 1 mmol/L of caffeine, flow cytometry detected a characteristic decrease in the S phase in all the XP-V cell strains. From these results, the eight groups of XP could be differentiated by utilizing a set of recombinant adenoviruses, indicating that our procedure provides a convenient and correct diagnostic method for all the XP groups including XP-E and XP-V.
authors.affiliation: Department of Dermatology, Sapporo Medical University School of Medicine, Sapporo, Japan. yamasita@sapmed.ac.jp.
authors.forename: Toshiharu
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: Department of Dermatology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Masae
authors.initials: M
authors.lastname: Okura
authors.affiliation: Department of Dermatology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Yasue
authors.initials: Y
authors.lastname: Ishii-Osai
authors.affiliation: Department of Dermatology, Sapporo Medical University School of Medicine, Sapporo, Japan.
authors.forename: Tokimasa
authors.initials: T
authors.lastname: Hida
mesh_terms: D000256:Adenoviridae; D002460:Cell Line; D002470:Cell Survival; D018076:DNA, Complementary / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D005347:Fibroblasts / Q000502:physiology / Q000528:radiation effects; D005434:Flow Cytometry; D015870:Gene Expression; D005816:Genetic Complementation Test / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D014466:Ultraviolet Rays / Q000009:adverse effects; D014983:Xeroderma Pigmentosum / Q000175:diagnosis* / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D004274:DNA, Recombinant
keywords: DNA polymerase eta; complementation; recombinant adenovirus; ultraviolet light; xeroderma pigmentosum