Laboratory Medicine, Immunology, University Hospitals Leuven, Leuven, Belgium.
Clin Chim Acta. 2012 Jan 18;413(1-2):105-8. doi: 10.1016/j.cca.2011.09.001. Epub 2011 Sep 22.
Anti-Golgi antibodies are rare autoantibodies that have been described in systemic autoimmune diseases. Not all Golgi auto-antigens are known. The objective of this study was to identify a novel auto-antigen associated with anti-Golgi immune reactivity. Sera from a patient with Golgi immune reactivity and from a control individual were used for Western blotting after 2-dimensional gel separation of a rat Golgi-enriched extract. Betaine homocysteine S-methyltransferase 1 (BHMT1) was identified as an auto-antigen by MALDI-TOF/TOF mass spectrometry. Using human recombinant BHMT1, a strong positive blotting signal was obtained with serum from the patient but not from a control. Pre-absorption of the serum sample with reactivity to BHMT1 with recombinant human BHMT1 resulted in decreased reactivity on Western blotting and in disappearance of the Golgi-like pattern on indirect immunofluorescence. Using immunocytochemistry, we confirmed the subcellular localization of BHMT1 to the Golgi apparatus. Antibodies to BHMT1 were found in four of 80 samples with a Golgi-pattern on indirect immunofluorescence. The antibodies were not associated with a specific clinical condition. We identified BHMT1 as a novel auto-antigen associated with anti-Golgi immune reactivity.
抗高尔基抗体是一种罕见的自身抗体，已在系统性自身免疫性疾病中描述过。并非所有高尔基自身抗原都已知。本研究的目的是鉴定与抗高尔基免疫反应相关的新型自身抗原。用 2- 维凝胶分离大鼠高尔基体富集提取物后，用来自抗高尔基免疫反应患者和对照个体的血清进行 Western 印迹。通过 MALDI-TOF/TOF 质谱鉴定甜菜碱同型半胱氨酸 S- 甲基转移酶 1（BHMT1）为自身抗原。用人重组 BHMT1，与患者血清而非对照血清进行强阳性印迹信号。用重组人 BHMT1 对血清样本进行预吸收以与 BHMT1 反应，导致 Western 印迹反应性降低，并在间接免疫荧光中消失高尔基样模式。通过免疫细胞化学，我们证实了 BHMT1 在高尔基器中的亚细胞定位。在 80 个具有间接免疫荧光高尔基模式的样本中发现了四种 BHMT1 抗体。这些抗体与特定的临床状况无关。我们鉴定出 BHMT1 是与抗高尔基免疫反应相关的新型自身抗原。
