Cloning and functional analysis of anti-double strand DNA IgG autoantibodies using the phage-display method

Citation
M. Suzuki et al., Cloning and functional analysis of anti-double strand DNA IgG autoantibodies using the phage-display method, INT J MOL M, 3(4), 1999, pp. 385-390
Citations number
27
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Medical Research General Topics
Journal title
INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE
ISSN journal
1107-3756 → ACNP
Volume
3
Issue
4
Year of publication
1999
Pages
385 - 390
Database
ISI
SICI code
1107-3756(199904)3:4<385:CAFAOA>2.0.ZU;2-#
Abstract
To examine the relationship between pathophysiolo gical effects and molecul ar features of anti-double-stranded (ds)DNA autoantibodies (Abs), we isolat ed anti-dsDNA Ab fragments by using the phage-display method. Fd gamma and light-chain DNA were PCR amplified from peripheral blood lymphocytes of a p atient with systemic lupus erythematosus (SLE), complicated with lupus neph ritis. They were then inserted into a phagemid vector, pComb3-H. We generat ed eight Fab fragments that specifically bound to solid-phase DNA. The Fab clones stained positively using Crithidia luciliae, indicating that they we re anti-dsDNA antibodies. Nucleotide sequences of VH of Fab clones were ver y similar and appeared to be derived from VH26 germline gene. Differences i n the activities of anti-dsDNA Ab of the Fab clones may be ascribed to the diversity of VL. Fab fragments with anti-dsDNA Ab activities exhibited a po sitive charge on isoelectric focusing, consistent with pathogenic features. Our results indicate that anti-dsDNA Ab Fab fragments obtained by the phag e-display method in the present study may possess molecular and functional characteristics of pathogenic anti-dsDNA autoAbs in SLE.