Decker P, Briand J P, de Murcia G, Pero R W, Isenberg D A, Muller S
Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
Arthritis Rheum. 1998 May;41(5):918-26. doi: 10.1002/1529-0131(199805)41:5<918::AID-ART20>3.0.CO;2-W.
To characterize autoantibody response to poly(ADP-ribose) polymerase (PARP) and to assess the significance of autoantibodies to the 2 zinc fingers of this enzyme in patients with autoimmune rheumatic and bowel diseases.
The specificity of antienzyme autoantibodies was established by dot immunoassay with recombinant human PARP and by enzyme-linked immunosorbent assay using the recombinant N-terminal fragment containing the DNA binding domain of PARP, the recombinant C-terminal catalytic domain (40-kd fragment), a peptide containing the nuclear localization signal (NLS) of PARP, 2 synthetic peptides (and mutated peptides) corresponding to zinc-finger motifs F1 and F2 that are present in the DNA binding domain, zinc fingers from other self antigens (e.g., peptides from Ro60, Ro52, and U1C proteins), and poly(ADP-ribose). Sera from patients with autoimmune rheumatic and bowel diseases were tested, as were affinity-purified antibodies. Histocompatibility typing of systemic lupus erythematosus (SLE) patients was performed by serology.
Antibodies from the patient sera reacted only weakly with the recombinant N- and C-terminal domains and with the NLS peptide. In contrast, the 2 synthetic peptides corresponding to zinc-finger motifs F1 and F2 represented immunodominant targets for IgG antibodies from patients with SLE, mixed connective tissue disease (MCTD), Crohn's disease, and ulcerative colitis. The sera from patients with SLE and MCTD showed much weaker reactivity with mutant peptides F1 and F2, which contain mutations at the cysteine residues involved in zinc coordination. F1/F2 antibodies did not cross-react with zinc fingers from other self proteins. No correlation was found between the presence of F1/F2 autoantibodies in SLE sera and the presence of other autoantibodies typical of this disease (e.g., anti-double-stranded DNA and poly[ADP-ribose] antibodies). The presence of F2 antibodies in the serum of SLE patients was negatively associated with HLA-DR6.
An autoimmune response to PARP is potentially important because this enzyme is involved in DNA repair and is rapidly cleaved during the "execution phase" of apoptosis. The high prevalence in certain autoimmune rheumatic and bowel diseases of antibodies to F1 and F2, which are directly involved in this process, is further evidence implicating involvement of the DNA repair system in chronic inflammatory diseases.
鉴定针对聚（ADP - 核糖）聚合酶（PARP）的自身抗体反应，并评估自身抗体对该酶的2个锌指结构在自身免疫性风湿性疾病和肠道疾病患者中的意义。
通过用重组人PARP进行斑点免疫测定，以及使用包含PARP DNA结合域的重组N端片段、重组C端催化域（40-kd片段）、含PARP核定位信号（NLS）的肽、对应于DNA结合域中存在的锌指基序F1和F2的2种合成肽（及突变肽）、来自其他自身抗原的锌指（如Ro60、Ro52和U1C蛋白的肽）以及聚（ADP - 核糖）进行酶联免疫吸附测定，确定抗酶自身抗体的特异性。检测自身免疫性风湿性疾病和肠道疾病患者的血清以及亲和纯化抗体。通过血清学对系统性红斑狼疮（SLE）患者进行组织相容性分型。
患者血清中的抗体与重组N端和C端结构域以及NLS肽反应较弱。相反，对应于锌指基序F1和F2的2种合成肽是SLE、混合性结缔组织病（MCTD）、克罗恩病和溃疡性结肠炎患者IgG抗体的免疫显性靶点。SLE和MCTD患者的血清与在锌配位中涉及的半胱氨酸残基处含有突变的突变肽F1和F2的反应性弱得多。F1/F2抗体不与来自其他自身蛋白的锌指交叉反应。在SLE血清中F1/F2自身抗体的存在与该疾病的其他典型自身抗体（如抗双链DNA和聚[ADP - 核糖]抗体）的存在之间未发现相关性。SLE患者血清中F2抗体的存在与HLA - DR6呈负相关。
对PARP的自身免疫反应可能很重要，因为该酶参与DNA修复并在凋亡的“执行阶段”迅速裂解。在某些自身免疫性风湿性疾病和肠道疾病中，直接参与此过程的F1和F2抗体的高患病率进一步证明了DNA修复系统参与慢性炎症性疾病。
