Mouse salivary glands and human beta-defensin-2 as a study model for antimicrobial gene therapy: technical considerations.
Mouse salivary glands and human beta-defensin-2 as a study model for antimicrobial gene therapy: technical considerations.
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小鼠唾液腺和人 β-防御素-2 作为抗菌基因治疗的研究模型：技术考虑。
DOI：
10.1016/j.ijantimicag.2006.08.003
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发表时间：
2006
影响因子：
10.8
通讯作者：
Huang,GeorgeT-J
中科院分区：
文献类型：
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作者：
Yin,Chunyi;Dang,HoaN;Gazor,Farzad;Huang,GeorgeT-J
唾液腺转抗菌肽基因在口腔感染控制中具有巨大的潜力。我们的最终目标是将抗菌肽基因引入唾液腺，唾液腺将这些肽分泌到唾液中以控制口腔中的细菌/真菌感染。然而，尚未建立测试这种潜力的动物研究模型。因此，我们决定测试（i）有效的抗微生物肽人β-防御素-2（hBD-2）是否可以在唾液腺转导后在唾液中过表达和（ii）口腔真菌感染是否可以在NOD/SCID鼠模型中发展。将携带hBD-2 cDNA的慢病毒载体SIN 18 cPPTRhMLV通过插管导入SCID小鼠下颌下腺。采用逆转录聚合酶链反应（RT-PCR）、免疫组化和酶联免疫吸附试验（ELISA）检测唾液和腺体中hBD-2的表达。将白色念珠菌613 p经口接种到SCID小鼠中以建立口腔念珠菌病。虽然通过RT-PCR和免疫组织化学在慢病毒递送后1天或1周在小鼠唾液腺中检测到hBD-2的表达，但在唾液中未检测到hBD-2。有可恢复的C。感染后4天至4周，SCID小鼠口腔和胃肠道均出现白色念珠菌感染，但体视显微镜下未观察到SCID小鼠口腔念珠菌感染。我们的数据表明，慢病毒载体抑制小鼠唾液腺，但没有达到足够的水平，使唾液中的hBD-2检测。为了利用小鼠唾液腺测试抗微生物基因疗法，需要用于基因转导的其他载体和对SCID小鼠建立口腔念珠菌病的额外治疗。
Transduction of salivary glands with antimicrobial peptide genes has great potential for oral infection control. Our ultimate goal is to introduce antimicrobial peptide genes into salivary glands that secrete these peptides into saliva to control bacterial/fungal infection in the oral cavity. However, an animal study model to test this potential has not been established. Therefore, we determined to test (i) whether the potent antimicrobial peptide human β-defensin-2 (hBD-2) can be overexpressed in saliva after transduction of salivary glands and (ii) whether oral fungal infection can be developed in a NOD/SCID murine model. Lentiviral vector SIN18cPPTRhMLV bearing hBD-2 cDNA was introduced into SCID mouse submandibular glands via cannulation. Reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry or enzyme-linked immunosorbent assay (ELISA) were performed to detect hBD-2 expression in glands or in saliva. Candida albicans 613p was inoculated orally into SCID mice to establish oral candidiasis. Whilst expression of hBD-2 was detected in mouse salivary glands by RT-PCR and immunohistochemistry 1 day or 1 week following delivery of lentivirus, hBD-2 was not detected in saliva. There was recoverable C. albicans from the oral cavity and gastrointestinal tract 4 days to 4 weeks after infection, but there was no establishment of observable oral candidiasis in SCID mice under a stereomicroscope. Our data indicate that lentiviral vectors transduce mouse salivary glands, but not at a sufficient level to allow hBD-2 detection in saliva. Other vectors for gene transduction and additional treatment of SCID mice to establish oral candidiasis are needed in order to utilise mouse salivary glands to test antimicrobial gene therapy.