Orderly pattern of development of the autoantibody response in (New Zealand White x BXSB)F1 lupus mice: characterization of target antigens and antigen spreading by two-dimensional gel electrophoresis and mass spectrometry.
Immunoblots of a two-dimensional PAGE-separated HL-60 cell proteomic map and mass spectrometry were combined to characterize proteins targeted by autoantibodies produced by male (New Zealand White x BXSB)F(1) (WB) mice that develop lupus and anti-phospholipid syndrome. Analysis of sera sequentially obtained from seven individual mice at different ages showed that six proteins, vimentin, heat shock protein 60, UV excision-repair protein RAD23, alpha-enolase, heterogeneous nuclear ribonucleoprotein L, and nucleophosmin, were the targets of the B cell autoimmune response, and that autoantibodies to them were synthesized sequentially in an orderly pattern that recurred in all the male WB mice analyzed: anti-vimentin first and anti-nucleophosmin last, with anti-RAD23 and anti-heat shock protein 60, then anti-alpha-enolase and anti-heterogeneous nuclear ribonucleoprotein L Abs occuring concomitantly. Anti-vimentin reactivity always appeared before anti-cardiolipin and anti-DNA Abs, suggesting that vimentin is the immunogen initiating the autoimmune process. The pattern of HL-60 proteins recognized by female WB sera differed from that of male sera, indicating that the Y chromosome-linked autoimmune acceleration gene is not an accelerator but a strong modifier of the autoimmune response. Thus, 1) combining two-dimensional PAGE and mass spectrometry constitutes a powerful tool to identify the set of Ags bound by autoantibodies present in a single serum and the whole autoantibody pattern of an autoimmune disease; 2) the diversification of the autoimmune response in male WB mice occurs in a predetermined pattern consistent with Ag spreading, and thus provides a useful model to further our understanding of the development of the autoantibody response in lupus.
Thébault
Sandrine
S
Institut de la Santé et de la Recherche Médicale Unité 519, Faculté de Médecine et de Pharmacie, Hôpital Charles-Nicolle, Rouen, France.
Gilbert
Danièle
D
Hubert
Marie
M
Drouot
Laurent
L
Machour
Nadine
N
Lange
Catherine
C
Charlionet
Roland
R
Tron
François
F
D000818:Animals; D000937:Antigen-Antibody Reactions; D001323:Autoantibodies / Q000096:biosynthesis* / Q000097:blood / Q000737:chemistry; D001324:Autoantigens / Q000097:blood / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D001666:Binding Sites, Antibody; D003433:Crosses, Genetic; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D018922:HL-60 Cells / Q000276:immunology; D006801:Humans; D007106:Immune Sera / Q000737:chemistry; D008180:Lupus Erythematosus, Systemic / Q000235:genetics* / Q000276:immunology*; D008297:Male; D051379:Mice; D008814:Mice, Inbred NZB; D009363:Neoplasm Proteins / Q000276:immunology; D012727:Sex Characteristics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
D001323:Autoantibodies; D001324:Autoantigens; D007106:Immune Sera; D009363:Neoplasm Proteins