Efficacy of Tuberculosis Vaccine Candidate pcDNA3.1-rpfB in Inhibiting the Growth of Mycobacterium tuberculosis In Vitro with Mycobacterial Growth Inhibition Assay

IF 0.8 Q3 MULTIDISCIPLINARY SCIENCES
Ratih Pujilestari, Andriansjah Rukmana, A. Karuniawati
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引用次数: 0

Abstract

Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb). Bacille Calmette-Guérin (BCG) is the only licensed vaccine against TB, and it is effective in children but not in adults. The Vaccine Research Team, Department of Microbiology FKUI has developed a DNA-based TB vaccine candidate pcDNA3.1-rpfB. This candidate induces immune responses in mice, but its potency is unknown. The gold standard for potency testing of TB vaccine is the challenge method. The BSL3 animal laboratory for the challenge method is currently unavailable at FKUI. Therefore, mycobacterial growth inhibition assay (MGIA) was used as a preliminary test before the in vivo challenge test was conducted. The principle of MGIA is to reculture Mtb in a Mycobacteria Growth Indicator Tube (MGIT) from co-cultured Mtb with mammalian cells that have been previously treated with pcDNA3.1-rpfB, pcDNA3.1 (negative control), and BCG (positive control). MGIT shows the time to positivity, which is the time that has lapsed until a positive growth of Mtb is detected. In addition, measurements of interferon (IFN)γ levels by enzyme-linked immunosorbent assay were carried out. This study concluded that pcDNA3.1-rpfB can inhibit the growth of Mtb in vitro and showed no statistical difference from BCG. The IFNγ levels from co-culturing did not correlate with the level of inhibition of the growth of Mtb in vitro.
结核分枝杆菌生长抑制试验研究结核候选疫苗pcDNA3.1-rpfB体外抑制结核分枝杆菌生长的效果
结核病是由结核分枝杆菌(Mtb)引起的。卡介苗(Bacille calmette - gusamrin, BCG)是唯一获得许可的结核病疫苗,它对儿童有效,但对成人无效。FKUI微生物系疫苗研究小组开发了一种基于dna的结核病候选疫苗pcDNA3.1-rpfB。该候选物在小鼠中诱导免疫反应,但其效力尚不清楚。结核病疫苗效价检测的金标准是挑战法。FKUI目前没有用于挑战方法的BSL3动物实验室。因此,在进行体内攻毒试验之前,采用分枝杆菌生长抑制试验(MGIA)作为初步试验。MGIA的原理是将结核分枝杆菌生长指示管(mycobacterium Growth Indicator Tube, MGIT)中的结核分枝杆菌与哺乳动物细胞共培养,这些细胞先前已接受pcDNA3.1- rpfb、pcDNA3.1(阴性对照)和BCG(阳性对照)的处理。MGIT显示到阳性的时间,这是到检测到结核分枝杆菌阳性生长的时间。此外,采用酶联免疫吸附法测定干扰素(IFN)γ水平。本研究得出pcDNA3.1-rpfB对Mtb体外生长有抑制作用,与BCG无统计学差异。共培养的IFNγ水平与体外抑制结核分枝杆菌生长的水平无关。
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来源期刊
Makara Journal of Science
Makara Journal of Science MULTIDISCIPLINARY SCIENCES-
CiteScore
1.30
自引率
20.00%
发文量
24
审稿时长
24 weeks
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