Blank Miri, Anafi Liat, Zandman-Goddard Gisele, Krause Ilan, Goldman Shlomit, Shalev Eliezer, Cervera Ricard, Font Josep, Fridkin Mati, Thiesen Hans-Jurgen, Shoenfeld Yehuda
The Autoimmune Disease Center, Sheba Medical Center, Tel-Aviv University, Israel.
Int Immunol. 2007 Jul;19(7):857-65. doi: 10.1093/intimm/dxm052. Epub 2007 Jun 18.
Administration of intravenous Ig (IVIG) is a recognized, safe and efficient mode of immunomodulatory therapy for many autoimmune diseases. Anti-idiotypic antibody binding to pathogenic autoantibodies and hence inhibition of binding to the corresponding antigen is one postulated mechanism of the beneficial effect of IVIG. The aim of this study was to fractionate the anti-beta-2-glycoprotein-I (beta(2)GPI) anti-idiotypic antibodies from a commercial IVIG preparation and use it as a treatment in an experimental antiphospholipid syndrome (APS) mouse model.
Anti-beta(2)GPI polyclonal antibodies were purified on a beta(2)GPI column. The purified antibodies were bound to CN-Br-activated sepharose and employed for purification of IVIG-anti-anti-beta(2)GPI (anti-idiotypic antibodies), defined as specific intravenous Ig (sIVIG). The idiotype specificities were confirmed by competition assays. The effect of sIVIG in vitro was tested in a trophoblast and choriocarcinoma matrigel/invasion assay (i.e. proliferation and metalloproteinase (MMP)2/MMP9 expression) and in vivo in a fetal loss model of APS.
Anti-beta(2)GPI antibodies inhibited human trophoblast cell invasion in vitro. The function was attributed to the Fab portion of the anti-beta(2)GPI Igs, since beta(2)GPI-related synthetic peptides specific for the Fab part of the anti-beta(2)GPI antibodies neutralized its activity. APS sIVIG fraction reduce human trophoblast invasion in vitro by 560 times more than the whole IVIG compound and improved the MMP2 and MMP9 production by trophoblast cells. sIVIG improved significantly (200 times more) the pregnancy outcome in BALB/c mice passively infused with anti-beta(2)GPI antibodies, in comparison to treatment with IVIG (P < 0.02).
Based on the current results, we propose that APS sIVIG may be considered as potential specific therapeutic safe compound for developing a treatment for APS patient's early fetal loss.
静脉注射免疫球蛋白（IVIG）的应用是治疗多种自身免疫性疾病公认的、安全有效的免疫调节治疗方式。抗独特型抗体与致病性自身抗体结合，从而抑制其与相应抗原的结合，这是IVIG发挥有益作用的一种推测机制。本研究的目的是从一种市售IVIG制剂中分离抗β2糖蛋白I（β2GPI）抗独特型抗体，并将其用于实验性抗磷脂综合征（APS）小鼠模型的治疗。
抗β2GPI多克隆抗体在β2GPI柱上进行纯化。纯化后的抗体与溴化氰活化的琼脂糖结合，用于纯化IVIG抗抗β2GPI（抗独特型抗体），定义为特异性静脉注射免疫球蛋白（sIVIG）。通过竞争试验确认独特型特异性。sIVIG的体外作用在滋养层和绒毛膜癌基质胶/侵袭试验（即增殖和金属蛋白酶（MMP）2/MMP9表达）中进行测试，并在APS胎儿丢失模型中进行体内测试。
抗β2GPI抗体在体外抑制人滋养层细胞侵袭。该功能归因于抗β2GPI免疫球蛋白的Fab部分，因为针对抗β2GPI抗体Fab部分的β2GPI相关合成肽可中和其活性。APS的sIVIG组分在体外使人类滋养层侵袭减少的程度比整个IVIG化合物高560倍，并改善了滋养层细胞的MMP2和MMP9产生。与IVIG治疗相比，sIVIG显著改善了（提高了200倍）被动注入抗β2GPI抗体的BALB/c小鼠的妊娠结局（P<0.02）。
基于目前的结果，我们提出APS的sIVIG可被视为开发治疗APS患者早期胎儿丢失的潜在特异性治疗安全化合物。
