Malaria is a tropical human disease, caused by protozoan parasites, wherein a significant number of the world's population is at risk. Annually, more than 219 million new cases are reported. Although there are prevention treatments, there are no highly and widely effective licensed anti-malarial vaccines available for use. Opportunities for utilization of plant-based vaccines as novel platforms for developing safe, reliable, and affordable treatments offer promise for developing such a vaccine against malaria. In this study, a Malchloroplast candidate vaccine was designed, composed of segments of AMA1 and MSP1 proteins, two epitopes of Plasmodium falciparum, along with a GK1 peptide from Taenia solium as adjuvant, and this was expressed in tobacco chloroplasts. Transplastomic tobacco lines were generated using biolistic transformation, and these were confirmed to carry the synthetic gene construct. Expression of the synthetic GK1 peptide was confirmed using RT-PCR and Western blots. Furthermore, the GK1 peptide was detected by HPLC at levels of up to 6 µg g⁻¹ dry weight of tobacco leaf tissue. The plant-derived Malchloroplast candidate vaccine was subsequently tested in BALB/c female mice following subcutaneous administration, and was found to elicit specific humoral responses. Furthermore, components of this candidate vaccine were recognized by antibodies in Plasmodium falciparum malaria patients and were immunogenic in test mice. Thus, this study provided a ‘proof of concept’ for a promising plant-based candidate subunit vaccine against malaria.

疟疾是一种由原生动物寄生虫引起的热带人类疾病，其中世界上大量人口处于危险之中。每年，报告的新病例超过2.19亿。尽管有预防措施，但没有高度有效且广泛有效的许可抗疟疾疫苗可供使用。利用植物疫苗作为开发安全，可靠和负担得起的治疗方法的新平台的机会为开发这种抗疟疾疫苗提供了希望。在这项研究中，一个Malchloroplast候选疫苗的设计，AMA1和MSP1蛋白，的两个表位的段组成的恶性疟原虫，与来自GK1肽沿着猪带绦虫作为佐剂，这在烟草叶绿体中表达。使用基因枪转化产生了转质体烟草系，并证实这些携带了合成的基因构建体。使用RT-PCR和Western印迹证实了合成的GK1肽的表达。此外，通过HPLC检测到GK1肽的水平高达6μgg ^-1烟叶组织干重。皮下施用后，随后在BALB / c雌性小鼠中测试了植物来源的Malchloroplast候选疫苗，发现该疫苗引起特定的体液反应。此外，该候选疫苗的成分被恶性疟原虫中的抗体识别疟疾患者，并且在测试小鼠中具有免疫原性。因此，这项研究为有前途的基于植物的疟疾候选亚单位疫苗提供了“概念证明”。