Brzozowska Ewa, Świętnicki Wiesław, Sycz Jordan, Kołodziejczak Monika, Stachowicz Łukasz, Wzorek Anna, Korzeniowska-Kowal Agnieszka, Skowicki Michał, Lipiński Tomasz
Department of Immunology of Infectious Diseases, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland.
Department of Chemistry, Wroclaw University of Environmental and Life Sciences, Poland.
Adv Clin Exp Med. 2025 Jul 25. doi: 10.17219/acem/200882.
Porphyromonas gingivalis is a major human oral opportunistic pathogen and a key etiological agent of periodontal disease, contributing to inflammation and bone loss in the oral cavity. Periodontitis is not limited to oral health complications; it has also been associated with a range of systemic conditions, including coronary heart disease (CAD), respiratory disease, rheumatoid arthritis, chronic kidney disease (CKD), and certain types of cancer.
Immunization-based prevention of periodontitis appears to be a promising strategy; however, no vaccine is currently available for commercial use. In the present study, a novel vaccine candidate against P. gingivalis was proposed, consisting of a P. gingivalis protein, gingipain, glycosylated with the carbohydrate moiety of P. gingivalis lipopolysaccharide (LPS).
Glycosylation of gingipain was achieved in Escherichia coli by introducing the Campylobacter jejuni N-glycosylation system, the P. gingivalis LPS biosynthetic pathway and the gingipain gene.
The neoglycoprotein was purified using column chromatography to a purity exceeding 99%, yielding a soluble antigen. The modified protein was recognized by commercial antibodies targeting the protein backbone, the carbohydrate moiety, and a custom monoclonal antibody specific to the purified LPS of P. gingivalis American Type Culture Collection (ATCC) 33277. The glycoprotein was used to immunize mice, and the resulting sera were analyzed for their ability to opsonize bacterial cells. The absence of detectable opsonization suggests that the elicited antibodies are more likely directed against the protein component of the vaccine rather than the glycan surface antigen.
The final product was most likely assembled correctly, as it was recognized by LPS-specific antibodies. Further evaluation in an animal model of induced periodontitis is necessary to determine whether the elicited antibodies can effectively inhibit gingipain released by the pathogen. If this vaccine candidate demonstrates protective efficacy, the approach could accelerate and enhance the safety of vaccine design against a wide range of other pathogens.
牙龈卟啉单胞菌是主要的人类口腔机会性病原菌，也是牙周疾病的关键病因，可导致口腔炎症和骨质流失。牙周炎不仅局限于口腔健康并发症；它还与一系列全身性疾病有关，包括冠心病（CAD）、呼吸系统疾病、类风湿性关节炎、慢性肾脏病（CKD）以及某些类型的癌症。
基于免疫的牙周炎预防似乎是一种有前景的策略；然而，目前尚无用于商业用途的疫苗。在本研究中，提出了一种新型抗牙龈卟啉单胞菌疫苗候选物，其由牙龈卟啉单胞菌蛋白牙龈蛋白酶组成，该蛋白经牙龈卟啉单胞菌脂多糖（LPS）的碳水化合物部分糖基化。
通过引入空肠弯曲菌N-糖基化系统、牙龈卟啉单胞菌LPS生物合成途径和牙龈蛋白酶基因，在大肠杆菌中实现牙龈蛋白酶的糖基化。
使用柱色谱法将新糖蛋白纯化至纯度超过99%，得到可溶性抗原。修饰后的蛋白可被靶向蛋白骨架、碳水化合物部分的商业抗体以及针对牙龈卟啉单胞菌美国模式培养物保藏中心（ATCC）33277纯化LPS的定制单克隆抗体识别。用该糖蛋白免疫小鼠，并分析所得血清调理细菌细胞的能力。未检测到调理作用表明所诱导的抗体更可能针对疫苗的蛋白质成分而非聚糖表面抗原。
最终产物很可能正确组装，因为它可被LPS特异性抗体识别。有必要在诱导性牙周炎动物模型中进行进一步评估，以确定所诱导的抗体是否能有效抑制病原体释放的牙龈蛋白酶。如果该疫苗候选物显示出保护效力，该方法可加速并提高针对多种其他病原体的疫苗设计的安全性。
