Yin Chunyi, Dang Hoa N, Gazor Farzad, Huang George T-J
Division of Associated Clinical Specialties, Section of Endodontics, UCLA School of Dentistry, Los Angeles, CA, USA.
Int J Antimicrob Agents. 2006 Oct;28(4):352-60. doi: 10.1016/j.ijantimicag.2006.08.003. Epub 2006 Sep 11.
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 beta-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.
用抗菌肽基因转导唾液腺在口腔感染控制方面具有巨大潜力。我们的最终目标是将抗菌肽基因导入唾液腺，使其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，尚未建立用于测试这种潜力的动物研究模型。因此，我们决定测试：（i）在唾液腺转导后，强效抗菌肽人β-防御素-2（hBD-2）是否能在唾液中过表达；（ii）在NOD/SCID小鼠模型中是否能引发口腔真菌感染。携带hBD-2 cDNA的慢病毒载体SIN18cPPTRhMLV通过插管被导入SCID小鼠的下颌下腺。进行逆转录聚合酶链反应（RT-PCR）、免疫组织化学或酶联免疫吸附测定（ELISA）以检测腺体或唾液中hBD-2的表达。将白色念珠菌613p经口接种到SCID小鼠中以建立口腔念珠菌病。虽然在慢病毒递送后1天或1周通过RT-PCR和免疫组织化学在小鼠唾液腺中检测到了hBD-2的表达，但在唾液中未检测到hBD-2。感染后4天至4周，口腔和胃肠道中可检测到白色念珠菌，但在体视显微镜下SCID小鼠中未形成可观察到的口腔念珠菌病。我们的数据表明，慢病毒载体可转导小鼠唾液腺，但转导水平不足以在唾液中检测到hBD-2。为了利用小鼠唾液腺来测试抗菌基因疗法，需要其他用于基因转导的载体以及对SCID小鼠进行额外处理以建立口腔念珠菌病。
