von Mensdorff-Pouilly S, Gourevitch M M, Kenemans P, Verstraeten A A, van Kamp G J, Kok A, van Uffelen K, Snijdewint F G, Paul M A, Meijer S, Hilgers J
Department of Obstetrics and Gynaecology, Academic Hospital Vrije Universiteit, Amsterdam, The Netherlands.
Tumour Biol. 1998;19(3):186-95. doi: 10.1159/000030006.
About one-third of breast and ovarian carcinoma patients have circulating antibodies reactive with polymorphic epithelial mucin (MUC1), either free or bound to immune complexes. While the presence of these immune complexes has prognostic significance in breast cancer patients, the significance of free MUC1 antibodies is less clear. The objective of this study was to develop a reliable assay for the accurate determination of circulating free antibodies to MUC1.
We developed an enzyme-linked immunosorbent assay (ELISA) (PEM.CIg) employing a 60 mer peptide (a triple tandem repeat sequence of the MUC1 peptide core) conjugated to bovine serum albumin and peroxidase-labeled antihuman immunoglobulin G or M antibodies. The assay was standardized and its analytical performance evaluated. A total of 492 serum samples were obtained from 40 healthy men, from 201 healthy women (including 55 women without a history of pregnancy and 45 pregnant women), and (before primary treatment) from 62 benign breast tumor patients and 190 breast cancer patients. MUC1 serum levels were determined with commercial CA 15-3 tests.
Circulating antibodies to MUC1 are present both in healthy subjects and in breast cancer patients. The within- and between-assay coefficients of variation were, respectively, 2 and 12% for the IgG determinations and 1.2 and 3% for the IgM determinations. Correlation coefficients for serially diluted serum samples ranged from 0.9998 to 0.9920 for IgG and from 0.9996 to 0.9818 for IgM determinations. The reactivity of serum samples was partially blocked by the addition of various MUC1 peptides and by MUC1 mucin. The inhibiting effect of modified 60 mer peptides suggests the presence of antibodies directed to more than one epitope.
The PEM. CIg assay is a reliable ELISA for measuring free MUC1 antibodies in serum. We are in the process of relating the results obtained in the breast cancer group to disease outcome to evaluate its prognostic significance. In addition, the assay may become a useful tool for vaccine therapy monitoring.
约三分之一的乳腺癌和卵巢癌患者体内存在与多态性上皮粘蛋白（MUC1）反应的循环抗体，这些抗体可游离存在或与免疫复合物结合。虽然这些免疫复合物的存在对乳腺癌患者具有预后意义，但游离MUC1抗体的意义尚不清楚。本研究的目的是开发一种可靠的检测方法，用于准确测定循环中游离的MUC1抗体。
我们开发了一种酶联免疫吸附测定法（ELISA）（PEM.CIg），该方法采用与牛血清白蛋白和过氧化物酶标记的抗人免疫球蛋白G或M抗体偶联的60聚体肽（MUC1肽核心的三重串联重复序列）。对该检测方法进行了标准化，并评估了其分析性能。共采集了492份血清样本，分别来自40名健康男性、201名健康女性（包括55名无妊娠史的女性和45名孕妇），以及62名良性乳腺肿瘤患者和190名乳腺癌患者（初次治疗前）。用商业CA 15-3检测法测定MUC1血清水平。
健康受试者和乳腺癌患者体内均存在循环MUC1抗体。IgG测定的批内和批间变异系数分别为2%和12%，IgM测定的批内和批间变异系数分别为1.2%和3%。系列稀释血清样本的IgG相关系数范围为0.9998至0.9920，IgM相关系数范围为0.9996至（此处原文似乎有误，应为0.9920）0.9818。血清样本的反应性可通过添加各种MUC1肽和MUC1粘蛋白而部分被阻断。修饰的60聚体肽的抑制作用表明存在针对多个表位的抗体。
PEM.CIg检测法是一种可靠的ELISA方法，用于测定血清中游离的MUC1抗体。我们正在将乳腺癌组获得的结果与疾病转归相关联，以评估其预后意义。此外，该检测方法可能成为疫苗治疗监测的有用工具。
