School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Loughborough, UK.
Department of Veterinary Pathology, College of Veterinary Medicine, China Agricultural University, Beijing, China.
Open Vet J. 2022 Mar-Apr;12(2):221-230. doi: 10.5455/OVJ.2022.v12.i2.10. Epub 2022 Apr 4.
remains a major pathogen of poultry. Most vaccines are inactivated and produced empirically. Although inactivated vaccines have been produced by culture under conditions of Fe deprivation, no vaccines have been produced which are likely to express all the proteins expressed during infection of antigen-presenting cells.
The aim was to produce a more protective inactivated vaccine by culturing the avian in a synthetic medium that resembled the environment of the phagolysosome.
Global gene expression in a pathogenic avian O78:K80 strain of , harvested from infected avian macrophage-like HD11 cells, was compared by microarray with bacteria cultured in a tissue culture medium. A liquid synthetic medium was produced based on the environmental conditions identified to which the bacteria were exposed intracellularly. A bacterin was produced from this strain and its protective ability was assessed in chickens.
The changes in gene expression observed included the use of different electron acceptors and carbon sources such as ethanolamine, β-glucosides, galactonate, dicarboxylic acids, and amino acids, up-regulation of genes associated with Fe and Mn uptake, and up-regulation of type-1 and curli fimbriae, other adhesion genes and down-regulation of sialic acid synthesis genes. The bacterin produced in the synthetic medium was statistically more protective than a bacterin prepared from bacteria cultured in the nutrient broth when tested in vaccinated chickens challenged with a different virulent O78:K80 strain.
The approach of using gene expression to produce synthetic media for the generation of more effective bacterins could be used for a number of intracellular bacteria pathogens including Enteroinvasive , , and the group of organisms.
仍然是家禽的主要病原体。大多数疫苗都是失活的,是凭经验生产的。尽管已经在缺铁条件下通过培养生产了失活疫苗,但尚未生产出可能表达感染抗原呈递细胞期间表达的所有蛋白质的疫苗。
目的是通过在类似于吞噬体环境的合成培养基中培养禽源 ,生产更具保护作用的灭活疫苗。
通过微阵列比较了从感染的禽巨噬样 HD11 细胞中收获的致病性禽 O78:K80 株 和在组织培养基中培养的细菌的全球基因表达。根据细菌在细胞内暴露的环境条件,生产了一种液体合成培养基。从该菌株生产了菌苗,并在鸡中评估了其保护能力。
观察到的 基因表达变化包括使用不同的电子受体和碳源,如乙醇胺、β-糖苷、半乳糖酸盐、二羧酸和氨基酸,与 Fe 和 Mn 摄取相关的基因上调,以及 1 型和卷曲菌毛、其他粘附基因的上调和唾液酸合成基因的下调。在合成培养基中生产的菌苗在接种鸡中受到不同毒力 O78:K80 株挑战时,比在营养肉汤中培养的细菌制备的菌苗更具统计学保护性。
使用基因表达生产用于产生更有效菌苗的合成培养基的方法可用于包括肠侵袭性 、 和 组生物体在内的许多胞内细菌病原体。