Department of Medicine, Section of Infectious Diseases, School of Medicine, Boston University, Boston, Massachusetts.
SutroVax Inc., Foster City, California.
J Clin Periodontol. 2019 Feb;46(2):197-205. doi: 10.1111/jcpe.13047. Epub 2019 Jan 31.
Periodontal diseases (PD) are complex oral inflammatory diseases initiated by keystone bacteria such as Porphyromonas gingivalis. A vaccine for PD is desirable as clinical treatment involves protracted maintenance strategies aimed to retain dentition. Although prior immunization approaches targeting P. gingivalis have reported variable success in limiting facets of disease such as oral bone loss, it remains that a vaccine for this disease may be attainable.
To investigate cell-free protein synthesis (CFPS) as a platform to produce vaccinable targets suitable for efficacy testing in a P. gingivalis-induced murine oral bone loss model.
Recombinantly generated P. gingivalis minor fimbriae protein (Mfa1), RgpA gingipain hemagglutinin domain 1 (HA1), and RgpA gingipain hemagglutinin domain 2 (HA2) were combined in equivalent doses in adjuvants and injected intramuscularly to immunize mice. Serum levels of protein-specific antibody were measured by ELISA, and oral bone levels were defined by morphometrics.
Recombinantly generated P. gingivalis proteins possessed high fidelity to predicted size and elicited protein-specific IgG following immunization. Importantly, immunization with the vaccine cocktail protected from P. gingivalis elicited oral bone loss.
These data verify the utility of the CFPS technology to synthesize proteins that have the capacity to serve as novel vaccines.
牙周病(PD)是一种复杂的口腔炎症性疾病,由龈卟啉单胞菌等关键细菌引发。由于临床治疗需要采用长期的维护策略来保留牙齿,因此牙周病疫苗的研发非常有必要。尽管先前针对龈卟啉单胞菌的免疫接种方法在限制口腔骨质流失等疾病方面取得了不同程度的成功,但针对这种疾病的疫苗可能仍然是可行的。
研究无细胞蛋白合成(CFPS)作为一种平台,用于生产可接种的目标物,以便在牙龈卟啉单胞菌诱导的小鼠口腔骨质流失模型中进行功效测试。
采用 CFPS 技术生成重组的牙龈卟啉单胞菌小菌毛蛋白(Mfa1)、RgpA 牙龈蛋白酶血凝素结构域 1(HA1)和 RgpA 牙龈蛋白酶血凝素结构域 2(HA2),并将其等量混合于佐剂中,通过肌肉注射对小鼠进行免疫接种。采用 ELISA 法检测血清中蛋白特异性抗体水平,采用形态计量学方法定义口腔骨质水平。
生成的重组牙龈卟啉单胞菌蛋白具有高度的预测大小保真度,免疫接种后可诱导产生蛋白特异性 IgG。重要的是,疫苗混合物的免疫接种可预防牙龈卟啉单胞菌引起的口腔骨质流失。
这些数据验证了 CFPS 技术在合成可作为新型疫苗的蛋白方面的实用性。