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Diagnosis of eight groups of xeroderma pigmentosum by genetic complementation using recombinant adenovirus vectors
Author(s) -
Yamashita Toshiharu,
Okura Masae,
IshiiOsai Yasue,
Hida Tokimasa
Publication year - 2016
Publication title -
the journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.9
H-Index - 65
eISSN - 1346-8138
pISSN - 0385-2407
DOI - 10.1111/1346-8138.13333
Subject(s) - xeroderma pigmentosum , recombinant dna , complementation , microbiology and biotechnology , biology , virology , dna , dna repair , genetics , gene , phenotype
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/m 2 was 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.