Kok Marc R, Yamano Seichii, Lodde Beatrijs M, Wang Jianghua, Couwenhoven Ross I, Yakar Shoshana, Voutetakis Antony, Leroith Derek, Schmidt Michael, Afione Sandra, Pillemer Stanley R, Tsutsui Marjorie T, Tak Paul P, Chiorini John A, Baum Bruce J
Gene Therapy and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
Hum Gene Ther. 2003 Nov 20;14(17):1605-18. doi: 10.1089/104303403322542257.
Female nonobese diabetic (NOD) mice develop spontaneous autoimmune sialadenitis and loss of salivary flow, and are a widely used model of Sjögren's syndrome. We examined the feasibility of local salivary gland immunomodulatory gene delivery to alter these sequelae in NOD mice. We constructed recombinant adeno-associated virus (rAAV) vectors encoding either human interleukin 10 (rAAVhIL-10) or beta-galactosidase (rAAVLacZ, control vector). Mice received rAAVhIL-10 or rAAVLacZ by retrograde submandibular ductal instillation either at age 8 weeks (early, before onset of sialadenitis), or at 16 weeks (late, after onset of sialadenitis). As a systemic treatment control, separate mice received intramuscular delivery of rAAVhIL-10 at each time point. Both submandibular and intramuscular delivery of vector led to low circulating levels of hIL-10. After submandibular administration of rAAVhIL-10, salivary flow rates at 20 weeks for both the early and late treatment groups were significantly higher than for both rAAVLacZ-administered and untreated mice. Systemic delivery of rAAVhIL-10 led to improved salivary flow in the late treatment group. Inflammatory infiltrates in submandibular glands, however, were significantly reduced only in mice receiving rAAVhIL-10 locally in the salivary gland compared with mice receiving this vector intramuscularly, or rAAVLacZ or no treatment. In addition, after submandibular rAAVhIL-10 delivery, NOD mice exhibited significantly lower blood glucose, and higher serum insulin, levels than all other groups, indicating some systemic benefit of this treatment. These studies show that expression of hIL-10 by rAAV vectors can have disease-modifying effects in the salivary glands of NOD mice, and suggest that local immunomodulatory gene transfer may be useful for managing the salivary gland pathology in Sjögren's syndrome.
雌性非肥胖型糖尿病（NOD）小鼠会自发发生自身免疫性涎腺炎并出现唾液分泌减少，是一种广泛应用的干燥综合征模型。我们研究了局部唾液腺免疫调节基因递送以改变NOD小鼠这些后遗症的可行性。我们构建了编码人白细胞介素10（rAAVhIL-10）或β-半乳糖苷酶（rAAVLacZ，对照载体）的重组腺相关病毒（rAAV）载体。小鼠在8周龄（早期，涎腺炎发作前）或16周龄（晚期，涎腺炎发作后）通过逆行下颌下导管滴注接受rAAVhIL-10或rAAVLacZ。作为全身治疗对照，在每个时间点分别有小鼠通过肌肉注射接受rAAVhIL-10。载体的下颌下和肌肉注射均导致循环中hIL-10水平较低。在通过下颌下给予rAAVhIL-10后，早期和晚期治疗组在20周时的唾液流速均显著高于给予rAAVLacZ和未治疗的小鼠。全身给予rAAVhIL-10使晚期治疗组的唾液分泌得到改善。然而，与通过肌肉注射接受该载体、接受rAAVLacZ或未接受治疗的小鼠相比，仅在唾液腺局部接受rAAVhIL-10的小鼠下颌下腺中的炎性浸润显著减少。此外，在通过下颌下给予rAAVhIL-10后，NOD小鼠的血糖水平显著低于所有其他组，血清胰岛素水平则高于所有其他组，表明这种治疗具有一定的全身益处。这些研究表明，rAAV载体表达hIL-10可对NOD小鼠的唾液腺产生疾病改善作用，并提示局部免疫调节基因转移可能有助于治疗干燥综合征的唾液腺病变。
