Balestrieri G, Tincani A, Spatola L, Allegri F, Prati E, Cattaneo R, Valesini G, Del Papa N, Meroni P
Servizio di Immunologia Clinica, Spedali Civili, Brescia, Italy.
Lupus. 1995 Apr;4(2):122-30. doi: 10.1177/096120339500400208.
Anticardiolipin (aCL) and anti-beta 2-glycoprotein I(anti beta 2GPI) antibodies have been shown in animal models as not cross-reacting antibody populations. This observation prompted us to prove if anti-beta 2GPI exist in human sera by using a reliable method and then to investigate if these are independent from aCl antibodies. We have developed a new ELISA for the detection of anti-beta 2GPI antibodies employing the coating of the protein in carbonate buffer to irradiated microtitre plates and the filtration of serum samples, that makes irrelevant the binding to the uncoated wells. IgG F(ab)2 fragments from IgG positive sera were shown bind beta 2GPI, providing that the binding was a specific antibody binding, mediated by the antigen binding site of the antibody molecule: moreover the antibodies were not able to differentiate native and delipidated beta 2GPI coated plates, making a possible role of a phospholipid contaminant unlikely. On the other hand, the phosphorus content of native as well as delipitated beta 2GPI was undetectable. IgG, but not IgM, anti-beta 2GPI antibodies were classically inhibited by the addition of soluble beta 2GPI, while cardiolipin liposomes appear to modify the reaction in a completely different way, possibly by the described interaction between cardiolipin and beta 2GPI.(ABSTRACT TRUNCATED AT 250 WORDS)
抗心磷脂(aCL)抗体和抗β2糖蛋白I(抗β2GPI)抗体在动物模型中已显示为不交叉反应的抗体群体。这一观察结果促使我们通过可靠的方法来证明人血清中是否存在抗β2GPI抗体,然后研究这些抗体是否独立于aCL抗体。我们开发了一种新的酶联免疫吸附测定法(ELISA)来检测抗β2GPI抗体,该方法采用在碳酸盐缓冲液中将蛋白质包被到经辐射的微量滴定板上,并对血清样本进行过滤,这使得血清与未包被孔的结合无关紧要。来自IgG阳性血清的IgG F(ab)2片段显示能结合β2GPI,这表明该结合是由抗体分子的抗原结合位点介导的特异性抗体结合:此外,这些抗体无法区分天然和脱脂的β2GPI包被板,这使得磷脂污染物发挥作用的可能性不大。另一方面,天然和脱脂的β2GPI的磷含量均无法检测到。经典的情况是,加入可溶性β2GPI可抑制IgG而非IgM抗β2GPI抗体,而心磷脂脂质体似乎以完全不同的方式改变反应,可能是通过心磷脂与β2GPI之间描述的相互作用。(摘要截选至250字)