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通过免疫印迹法检测人乳腺癌血清中针对与乳腺癌细胞相关抗原的(自身)抗体。

(Auto)antibodies in human breast cancer sera against antigens associated with breast cancer cells, detected by immunoblotting.

作者信息

Ronai Z, Sulitzeanu D

出版信息

J Natl Cancer Inst. 1986 Dec;77(6):1203-9.

PMID:3540417
Abstract

Sera of patients with breast cancer (as well as control normal sera and sera of patients with ovarian cancer or melanoma) were screened for the presence of antibodies against antigens expressed by the MDA breast cancer cell line. The techniques employed were radioimmunoassay with radioiodinated protein A and immunodotting with peroxidase-conjugated anti-human immunoglobulin antibodies. Sera reacting strongly by immunodotting were subsequently tested against antigens of the MDA and T47D cell lines in immunoblotting experiments. Both the breast cancer and the control sera yielded highly complex band patterns, which varied from serum to serum. The cancer sera differed from the normal sera, however, as they produced in most cases one or several bands that were distinctly stronger than the others. One of the strong bands, in fact a doublet of approximately 50 kilodaltons (kd), was produced preferentially (although not exclusively) when breast cancer sera were reacted with T47D cell membrane antigens. Absorption of selected sera with normal tissue or MDA antigens abolished or greatly reduced the intensity of some of the bands. It is concluded that, with the possible exception of the 50-kd band, most (probably all) of the bands seen in immunoblots resulted from the binding of autoantibodies to normal antigens expressed by the breast cancer cell lines. The main difference between cancer and normal sera would seem to be an increased content of autoantibodies in cancer, the specificity of these autoantibodies varying, however, from serum to serum.

摘要

对乳腺癌患者的血清(以及对照正常血清、卵巢癌或黑色素瘤患者的血清)进行筛查,以检测是否存在针对MDA乳腺癌细胞系所表达抗原的抗体。所采用的技术包括用放射性碘化蛋白A进行放射免疫测定,以及用过氧化物酶偶联的抗人免疫球蛋白抗体进行免疫斑点法。通过免疫斑点法反应强烈的血清随后在免疫印迹实验中针对MDA和T47D细胞系的抗原进行检测。乳腺癌血清和对照血清均产生高度复杂的条带模式,且不同血清之间存在差异。然而,癌症血清与正常血清不同,因为在大多数情况下,它们会产生一条或几条明显强于其他条带的条带。事实上,其中一条强带是一条约50千道尔顿(kd)的双峰带,当乳腺癌血清与T47D细胞膜抗原反应时,这条带会优先产生(尽管不是唯一产生)。用正常组织或MDA抗原吸收选定的血清可消除或大大降低某些条带的强度。得出的结论是,除了50-kd条带可能例外,免疫印迹中看到的大多数(可能是所有)条带是由自身抗体与乳腺癌细胞系所表达的正常抗原结合产生的。癌症血清与正常血清之间的主要差异似乎在于癌症血清中自身抗体含量增加,然而这些自身抗体的特异性因血清而异。

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