Devendra D, Galloway T S, Horton S J, Wilkin T J
Department of Endocrinology & Metabolism, Peninsula Medical School, Plymouth campus, Plymouth, UK.
Diabet Med. 2004 Dec;21(12):1316-24. doi: 10.1111/j.1464-5491.2004.01344.x.
Complete or partial remission can occur in newly diagnosed Type 1 diabetes patients. We created idiotype-specific reagents to explore the idiotypes of insulin antibodies (IA) in a patient in remission, and to compare with a patient who was not.
Phage display was used to create a library of phagotopes specific to insulin binding in four sera. Sera from a Type 1 diabetes subject deemed to have undergone remission were taken at diagnosis and again during remission. Sera from a non-remitter were taken at diagnosis and after 3 months on insulin. Phagotopes from the four sera were randomly selected and tested for insulin specificity in a radiobinding assay by using sera from remitters and non-remitters.
IA-binding phagotope selected from serum during remission displaced insulin binding in all nine IA(+) remitters and all 10 IA(+) non-remitters. IA-binding phagotope selected from the non-remission patient (3 months after insulin therapy) displaced insulin binding in 8/9 IA(+) remitters and 8/10 IA(+) non-remitters. The consensus peptide sequences adduced from the phages were identical for both these phagotopes. Phagotopes derived from insulin autoantibody-positive individuals at diagnosis were unable to displace insulin binding in the IA(+) sera 3 months later, whether in remission or not.
We have established the principle of using phage display in the investigation of insulin antibodies during remission in Type 1 diabetes. The immunological characteristics of IA 3 months after the introduction of insulin treatment were different from those at diagnosis of Type 1 diabetes (IAA). Using phage display technology, it was not possible to distinguish insulin antibodies according to remission status.
新诊断的1型糖尿病患者可能会出现完全或部分缓解。我们制备了独特型特异性试剂，以探究一名缓解期患者体内胰岛素抗体（IA）的独特型，并与未缓解的患者进行比较。
利用噬菌体展示技术构建针对四种血清中胰岛素结合的噬菌体位点文库。选取一名被认为已缓解的1型糖尿病患者在诊断时和缓解期的血清。选取一名未缓解患者在诊断时和胰岛素治疗3个月后的血清。从这四种血清中随机选择噬菌体位点，并使用缓解者和未缓解者的血清通过放射结合试验检测其对胰岛素的特异性。
从缓解期血清中选出的IA结合噬菌体位点在所有9名IA（+）缓解者和所有10名IA（+）未缓解者中均能取代胰岛素结合。从未缓解患者（胰岛素治疗3个月后）血清中选出的IA结合噬菌体位点在8/9名IA（+）缓解者和8/10名IA（+）未缓解者中能取代胰岛素结合。从这些噬菌体中推导的共有肽序列在这两种噬菌体位点中是相同的。诊断时来自胰岛素自身抗体阳性个体的噬菌体位点在3个月后无论是在缓解期还是未缓解期都无法在IA（+）血清中取代胰岛素结合。
我们确立了利用噬菌体展示技术研究1型糖尿病缓解期胰岛素抗体的原则。胰岛素治疗3个月后IA的免疫特性与1型糖尿病诊断时（IAA）不同。利用噬菌体展示技术无法根据缓解状态区分胰岛素抗体。
