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Characterization of a monoclonal antibody to cis-urocanic acid: detection of cis-urocanic acid in the serum of irradiated mice by immunoassay.

机译:顺式尿烷酸单克隆抗体的表征:通过免疫测定法检测辐照小鼠血清中的顺式尿烷酸。

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摘要

Cis-urocanic acid (cis-UCA), which is formed from the naturally occurring trans-isomer on ultraviolet (UV) irradiation, has been suggested as a photoreceptor for and mediator of the suppressive effects of UV irradiation on systemic immune responses. Trans-UCA is located predominantly in the stratum corneum, and the extent of isomerization to cis-UCA may be analysed by high-performance liquid chromatography (HPLC) of skin extracts. Such an analysis is not suitable for other tissues. In this study a murine monoclonal antibody to cis-UCA was prepared and tested by ELISA using UCA isomers conjugated to protein as antigens. The interaction of the antibody with structural analogues of UCA was assessed by competitive inhibition ELISA which indicated that the antibody had a high specificity for cis-UCA. Screening of sera at various times after UVB irradiation of mice by competitive inhibition ELISA using the monoclonal antibody showed that cis-UCA was present, probably in an unbound form, for at least 2 days after the exposure. Thus, cis-UCA produced in the epidermis following UVB irradiation reaches the serum a few hours later. The implications of this finding for the generation of suppressed immune responses are discussed.
机译:顺式尿烷酸(cis-UCA)是由紫外线(UV)照射下的天然反式异构体形成的,已被建议用作紫外线照射对全身免疫应答的抑制作用的光感受器和调节剂。反式-UCA主要位于角质层,可以通过皮肤提取物的高效液相色谱(HPLC)分析异构化成顺式-UCA的程度。这样的分析不适用于其他组织。在这项研究中,制备了针对顺式-UCA的鼠类单克隆抗体,并使用与蛋白质结合的UCA异构体作为抗原,通过ELISA进行了测试。通过竞争抑制ELISA评估了抗体与UCA的结构类似物的相互作用,这表明该抗体对顺式-UCA具有高度的特异性。使用单克隆抗体通过竞争性抑制ELISA对小鼠进行UVB照射后,在不同时间进行血清筛选,结果显示,暴露后至少2天,顺式-UCA可能以未结合的形式存在。因此,UVB照射后在表皮中产生的顺式-UCA在数小时后到达血清。讨论了这一发现对于产生抑制的免疫反应的意义。

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