Optimised combination therapies in order to reverse the diabetic state in type 1 diabetes including the LADA type based on the pathomechanisms: studies in the LEW.1AR1-iddm rat with a comparative view to the human situation

糖尿病表现后胰岛免疫细胞的细胞组成和细胞因子模式与人类1型糖尿病动物模型LEW.1AR1.iddm大鼠的细胞组成和细胞因子模式最相似，与患者的胰腺相比。因此，该模型特别适合于成功的免疫调节预防疗法的验证。本项目的目的是利用生化和分子形态学方法，与成功建立的抗TCR和抗肿瘤坏死因子-α联合治疗相比，评估新的合理抗体组合的有效性，作为胰腺和血液中的参考。重点将放在T细胞抗体抗TCR与抗促炎细胞因子IL-1β、干扰素-γ、IL-17和趋化因子MCP-1的抗体的组合上，以缓解糖尿病的代谢状态。此外，联合使用这两种抗体对抗主要的促炎细胞因子--肿瘤坏死因子-α和白介素1-β的疗效将被确定；然而，在没有抗-TCR的情况下。此外，将在最小化两种抗体的不良相互作用的风险这一特殊方面下，分析顺序应用抗TCR和抗肿瘤坏死因子-α的有效性。此外，将对抗TCR联合抗肿瘤坏死因子-α的参考组合进行为期一年的长期监测，以了解糖尿病高血糖日后复发的风险。缓慢进行性的自身免疫性糖尿病被称为LADA(成人潜伏性自身免疫性糖尿病)，其特点使其特别适合于将免疫调节联合疗法移植到人类。在LEW.1AR1-IDDM大鼠模型中，首次有可能在动物模型中验证这种LADA形式。到目前为止，只有少数LADA患者描述了胰腺的变化。因此，目前还不可能有明确的病理生理描述。LADA患者的胰腺和缓慢进展的自身免疫性糖尿病大鼠模型的胰腺将可用于使用分子形态学方法进行基因和蛋白质水平的分析，重点是胰岛浸润和β细胞变化。同时，贝塔细胞的再生过程将以新生和增殖分析为特征。这些比较形态分析的结果将提供区分自身免疫性糖尿病形式的标准。为了区分早发性1型糖尿病患者和潜伏型1型糖尿病患者的胰腺病变，两者均有免疫细胞浸润的征象，我们将对2型糖尿病患者的胰腺进行分析。这些研究将为将来转移治疗具有不同疾病进展速度的1型糖尿病患者的最佳抗体组合的治疗方法奠定基础。

Cellular composition and cytokine pattern of the immune cell infiltrate in pancreatic islets after diabetes manifestation show the greatest similarities in an animal model of human type 1 diabetes mellitus, the LEW.1AR1.iddm rat, when compared with the pancreas of patients. Thus, this model is particularly suited for validation of successful immunomodulatory prevention therapies.Using biochemical and molecular morphological methods it is the aim of this project to evaluate the effectiveness of new rational antibody combinations in comparison to the successfully established combination therapy of anti-TCR and anti-TNF-alpha as a reference in pancreas and in blood. The particular focus will be on combinations of the T-cell antibody anti-TCR with antibodies against the proinflammatory cytokines IL-1beta, IFN-gamma, IL-17 and the chemokine MCP-1 with respect to the remission of the diabetic metabolic state. Furthermore the efficacy of a combined administration of the two antibodies against the main proinflammatory cytokines, TNF-alpha and IL-1beta, will be determined; however, in the absence of anti-TCR. Additionally the effectiveness of a sequential application of anti-TCR and anti-TNF-alpha will be analysed under the special aspect of a minimisation of the risk of undesirable interactions of the two antibodies. Furthermore, a one-year long-term monitoring of the reference combination anti-TCR plus anti-TNF-alpha will be assessed with respect to the risk of a later reccurrence of diabetic hyperglycemia.The slowly progressive form of autoimmune diabetes called LADA (Latent Autoimmune Diabetes in Adults) has features that make it particularly attractive for possible translation of immunomodulatory combination therapies to humans. In the LEW.1AR1-iddm rat model it was possible for the first time to verify this LADA form in an animal model. So far pancreatic changes have only been described in a few patients with LADA. Therefore a clear pathophysiological delineation is not yet possible. Pancreases from LADA patients and in parallel pancreases from the rat model with a slowly progressing form of autoimmune diabetes will be available for analyses on the gene and protein level using molecular morphology methods with special emphasis on islet infiltration and beta cell changes. In parallel beta cell regeneration processes will be characterised by neogenesis and proliferation analyses. The results of these comparative morphological analyses will provide criteria for distinction between the autoimmune diabetes forms. For the purpose of distinction of the pathological changes in pancreases of patients with early onset type 1 diabetes and the latent form, both with signs of immune cell infiltration, pancreases of patients with type 2 diabetes will be analysed. These studies will be the basis for a future transfer of curative therapy approaches with optimal antibody combinations for type 1 diabetes patients with different speed of disease progression.