Department of Medicine, University of Washington, Seattle, Washington, USA.
Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, Washington, USA.
Int J Exp Pathol. 2022 Aug;103(4):140-148. doi: 10.1111/iep.12437. Epub 2022 Mar 4.
Islet autoantibodies, including autoantibodies directed against the 65kDa isoform of glutamate decarboxylase (GAD65Ab), are present in the majority of patients with newly diagnosed type 1 diabetes (T1D). Whereas these autoantibodies are historically viewed as an epiphenomenon of the autoimmune response with no significant pathogenic function, we consider in this study the possibility that they impact the major islet function, namely glucose-stimulated insulin secretion. Two human monoclonal GAD65Ab (GAD65 mAb) (b78 and b96.11) were investigated for uptake by live rat beta cells, subcellular localization and their effect on glucose-stimulated insulin secretion. The GAD65 mAbs were internalized by live pancreatic beta cells, where they localized to subcellular structures in an epitope-specific manner. Importantly, GAD65 mAb b78 inhibited, while GAD65 mAb b96.11 enhanced, glucose-stimulated insulin secretion (GSIS). These opposite effects on GSIS rule out non-specific effects of the antibodies and suggest that internalization of the antibody leads to epitope-specific interaction with intracellular machinery regulating insulin granule release. The most likely explanation for the alteration of GSIS by GAD65 Abs is via changes in GABA release due to inhibition or change in GAD65 enzyme activity. This is the first report indicating an active role of GAD65Ab in the pathogenesis of T1D.
胰岛自身抗体,包括针对谷氨酸脱羧酶 65kDa 同工型的自身抗体(GAD65Ab),存在于大多数新诊断的 1 型糖尿病(T1D)患者中。虽然这些自身抗体在历史上被视为自身免疫反应的伴随现象,没有显著的致病功能,但在本研究中我们考虑了它们是否会影响主要的胰岛功能,即葡萄糖刺激的胰岛素分泌。研究了两种人源单克隆 GAD65Ab(GAD65 mAb)(b78 和 b96.11)对活大鼠β细胞的摄取、亚细胞定位及其对葡萄糖刺激的胰岛素分泌的影响。GAD65 mAb 被活的胰腺β细胞内化,并以表位特异性的方式定位于亚细胞结构中。重要的是,GAD65 mAb b78 抑制,而 GAD65 mAb b96.11 增强葡萄糖刺激的胰岛素分泌(GSIS)。这些对 GSIS 的相反影响排除了抗体的非特异性影响,并表明抗体的内化导致与调节胰岛素颗粒释放的细胞内机制的表位特异性相互作用。GAD65Abs 改变 GSIS 的最可能解释是由于 GABA 释放的改变,这是由于 GAD65 酶活性的抑制或改变。这是第一个表明 GAD65Ab 在 T1D 发病机制中具有积极作用的报告。