Moulds J M, Rowe K E
Department of Internal Medicine, University of Texas-Houston Medical School, USA.
Transfusion. 1996 Jun;36(6):517-20. doi: 10.1046/j.1537-2995.1996.36696269510.x.
Antibodies of the Knops system have been referred to as nonneutralizable because they cannot be inhibited with serum, saliva, or urine. Because the Knops system antigens have been located on complement receptor 1 (CR1), the question of whether the antibodies could be neutralized with soluble CR1 (sCR1) produced by recombinant DNA techniques was studied.
First, radiolabeled immunoprecipitation techniques were used to test sCR1 for the expression of the high-incidence Knops system antigens. Then, a total of 45 antibodies were neutralized with sCR1, including the following: one each of anti-Cr(a), -Dr(a), -Do(b), -Hy, -Ge, -Jr(a), -Sc1, -Jk(a), -Cs(a), and -Kp(b); two each of anti-Lu(b), -Yt(a), and -JMH; three each of anti-McC(a), -Rg, and -Sl(a); and four each of anti-Ch, -Kn(a), -Yk(a), -Kn/McC. In addition, two examples of anti-Kn(a) + K, one example of anti-Sl(a) + K + Fy(a), and one example of anti-Yk(a) + E were tested. The sCR1 was added to each test serum and 6-percent albumin was added to the control; this was followed by neutralization incubation for 5 minutes at 25 degrees C. The antibody samples were then tested by a low-ionic-strength solution, anti-human globulin technique.
The sCR1 expressed Kn(a), McC(a), Sl,a and Yk(a). All Knops system antibodies (n = 22) were neutralized by the sCR1, but none of the other 23 alloantibodies decreased in reactivity. The samples containing antibodies of two specificities showed inhibition of the Knops system antibody but not of the second antibody.
This neutralization method, in which recombinant protein is used, provides an expedient and definitive method of identifying Knops system antibodies.
诺普斯系统的抗体被认为是不可中和的,因为它们不能被血清、唾液或尿液抑制。由于诺普斯系统抗原定位于补体受体1(CR1)上,因此研究了这些抗体是否能用重组DNA技术产生的可溶性CR1(sCR1)中和。
首先,使用放射性标记免疫沉淀技术检测sCR1中高频率诺普斯系统抗原的表达。然后,用sCR1中和总共45种抗体,包括以下抗体:抗Cr(a)、抗Dr(a)、抗Do(b)、抗Hy、抗Ge、抗Jr(a)、抗Sc1、抗Jk(a)、抗Cs(a)和抗Kp(b)各1种;抗Lu(b)、抗Yt(a)和抗JMH各2种;抗McC(a)、抗Rg和抗Sl(a)各3种;抗Ch、抗Kn(a)、抗Yk(a)、抗Kn/McC各4种。此外,还检测了2例抗Kn(a)+K、1例抗Sl(a)+K+Fy(a)和1例抗Yk(a)+E。将sCR1加入到每份测试血清中,向对照中加入6%的白蛋白;随后在25℃下进行中和孵育5分钟。然后通过低离子强度溶液抗人球蛋白技术检测抗体样本。
sCR1表达Kn(a)、McC(a)、Sl(a)和Yk(a)。所有诺普斯系统抗体(n = 22)均被sCR1中和,但其他23种同种抗体的反应性均未降低。含有两种特异性抗体的样本显示诺普斯系统抗体受到抑制,但第二种抗体未受抑制。
这种使用重组蛋白的中和方法为鉴定诺普斯系统抗体提供了一种便捷且确定的方法。