Lee Jason W, Parlane Natalie A, Rehm Bernd H A, Buddle Bryce M, Heiser Axel
Institute of Fundamental Sciences, Massey University, Palmerston North, New Zealand.
AgResearch, Hopkirk Research Institute, Palmerston North, New Zealand.
Appl Environ Microbiol. 2017 Feb 15;83(5). doi: 10.1128/AEM.02289-16. Print 2017 Mar 1.
Tuberculosis (TB) is a disease caused by or and still remains one of the world's biggest global health burdens. Recently, engineered polyhydroxyalkanoate (PHA) biobeads that were produced in both and and displayed mycobacterial antigens were found to induce significant cell-mediated immune responses in mice. We observed that such PHA beads contained host cell proteins as impurities, which we hypothesized to have the potential to induce immunity. In this study, we aimed to develop PHA beads produced in mycobacteria (mycobacterial PHA biobeads [MBB]) and test their potential as a TB vaccine in a mouse model. As a model organism, nonpathogenic was engineered to produce MBB or MBB with immobilized mycobacterial antigens Ag85A and ESAT-6 on their surface (A:E-MBB). Three key enzymes involved in the poly(3-hydroxybutyric acid) pathway, namely, β-ketothiolase (PhaA), acetoacetyl-coenzyme A reductase (PhaB), and PHA synthase (PhaC), were engineered into - shuttle plasmids and expressed in Immobilization of specific antigens to the surface of the MBB was achieved by creating a fusion with the PHA synthase which remains covalently attached to the polyester core, resulting in PHA biobeads displaying covalently immobilized antigens.
MBB, A: E-MBB, and an vector control (MVC) were used in a mouse immunology trial, with comparison to phosphate-buffered saline (PBS)-vaccinated and BCG-vaccinated groups. We successfully produced MBB and A:E-MBB and used them as vaccines to induce a cellular immune response to mycobacterial antigens. Tuberculosis (TB) is a disease caused by or and still remains one of the world's biggest global health burdens. In this study, we produced polyhydroxyalkanoate (PHA) biobeads in mycobacteria and used them as vaccines to induce a cellular immune response to mycobacterial antigens.
结核病（TB）是由结核分枝杆菌引起的疾病，至今仍是全球最大的健康负担之一。最近，在大肠杆菌和枯草芽孢杆菌中生产并展示分枝杆菌抗原的工程化聚羟基脂肪酸酯（PHA）生物珠被发现可在小鼠中诱导显著的细胞介导免疫反应。我们观察到这种PHA珠含有宿主细胞蛋白作为杂质，我们推测这些杂质有可能诱导免疫。在本研究中，我们旨在开发在分枝杆菌中生产的PHA珠（分枝杆菌PHA生物珠[MBB]），并在小鼠模型中测试其作为结核病疫苗的潜力。作为模型生物，非致病性耻垢分枝杆菌被工程化以生产MBB或表面固定有分枝杆菌抗原Ag85A和ESAT-6的MBB（A:E-MBB）。参与聚（3-羟基丁酸）途径的三种关键酶，即β-酮硫解酶（PhaA）、乙酰乙酰辅酶A还原酶（PhaB）和PHA合酶（PhaC），被工程化到耻垢分枝杆菌穿梭质粒中并在耻垢分枝杆菌中表达。通过与仍然共价连接到聚酯核心的PHA合酶形成融合，实现将特定抗原固定到MBB表面，从而产生展示共价固定抗原的PHA生物珠。
MBB、A:E-MBB和一个大肠杆菌载体对照（MVC）用于小鼠免疫学试验，并与接种磷酸盐缓冲盐水（PBS）和卡介苗（BCG）的组进行比较。我们成功生产了MBB和A:E-MBB，并将它们用作疫苗来诱导对分枝杆菌抗原的细胞免疫反应。结核病（TB）是由结核分枝杆菌引起的疾病，至今仍是全球最大的健康负担之一。在本研究中，我们在分枝杆菌中生产聚羟基脂肪酸酯（PHA）生物珠，并将它们用作疫苗来诱导对分枝杆菌抗原的细胞免疫反应。
