Kadir Nur-Ayuni, Acosta Armando, Sarmiento Maria E, Norazmi Mohd-Nor
School of Biomedicine, Faculty of Health Sciences, Universiti Sultan Zainal Abidin, Kuala Nerus 21300, Terengganu, Malaysia.
School of Health Sciences, Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia.
Pathogens. 2020 Nov 29;9(12):1000. doi: 10.3390/pathogens9121000.
Tuberculosis (TB) causes more than 1.5 million deaths each year, remaining a significant global health problem. () and () share features, which support the use of the former use in new generation TB vaccine development. In a previous study, the specific humoral and cellular immunogenicity of a recombinant strain expressing epitopes from Ag85B protein (rMs064), was demonstrated in mice. In the current study, the immunomodulatory capacity of rMs064 was determined in a J774A.1 murine macrophage cell line. To determine the immunomodulatory effect of rMs064 in J774A.1 macrophages, the expression of inducible nitric oxide synthase (iNOS) and production of nitric oxide (NO) was evaluated. The expression of activation surface markers (MHC-II, CD40, CD80 and CD86) and the production of cytokines (IL-1β, TNF-α, IL-12p70 and IL-6) was also determined in rMs064 infected J774A.1 macrophages. Our findings showed the ability of rMs064 to induce substantial increases in macrophage activation markers expression; MHC class II and CD40, compared with ransformed with the empty vector (rMs012) and uninfected cells. rMs064 induced significant increases in IL-12p70 compared to uninfected cells. The expression of iNOS and CD86, and the production of IL-1β, and TNF-α were increased in rMs064 and rMs012, compared to uninfected cells. rMs064 demonstrated its immunomodulatory ability by stimulating the innate immune response, which supports its further evaluation as a TB vaccine candidate.
结核病（TB）每年导致超过150万人死亡，仍然是一个重大的全球健康问题。（）和（）具有共同特征，这支持在新一代结核病疫苗研发中使用前者。在先前的一项研究中，在小鼠体内证明了表达来自Ag85B蛋白表位的重组菌株（rMs064）的特异性体液免疫原性和细胞免疫原性。在当前研究中，在J774A.1小鼠巨噬细胞系中测定了rMs064的免疫调节能力。为了确定rMs064对J774A.1巨噬细胞的免疫调节作用，评估了诱导型一氧化氮合酶（iNOS）的表达和一氧化氮（NO）的产生。还测定了感染rMs064的J774A.1巨噬细胞中活化表面标志物（MHC-II、CD40、CD80和CD86）的表达以及细胞因子（IL-1β、TNF-α、IL-12p70和IL-6）的产生。我们的研究结果表明，与用空载体转化的（rMs012）和未感染细胞相比，rMs064能够显著增加巨噬细胞活化标志物的表达；MHC II类分子和CD40。与未感染细胞相比，rMs064诱导IL-12p70显著增加。与未感染细胞相比，rMs064和rMs012中iNOS和CD86的表达以及IL-1β和TNF-α的产生均增加。rMs064通过刺激先天免疫反应展示了其免疫调节能力，这支持将其作为结核病疫苗候选物进行进一步评估。
