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首页> 外文期刊>ACS Omega >Comparative Study of 5′- and 3′-Linked CpG–Antigen Conjugates for the Induction of Cellular Immune Responses
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Comparative Study of 5′- and 3′-Linked CpG–Antigen Conjugates for the Induction of Cellular Immune Responses

机译：诱导细胞免疫应答诱导的5'-和3'-连接的CpG-antigen缀合物的比较研究

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Conjugation of CpG to an antigen induces a stronger immune response compared to that of the mixture. This study compares the in vitro immunostimulatory activity of CpG conjugated via either its 5′ or 3′ end to the model antigen ovalbumin (OVA). CpG modified with an amine at either the 5′ or 3′ end was conjugated to OVA via a stable bis-aryl hydrazone bond. Similar levels of CpG conjugation to OVA were observed for both conjugates on the basis of the absorbance at 360 nm for the formation of the bis-aryl hydrazone bond, which determined 2.8 ± 0.3 CpGs linked per OVA. Both the 5′ and 3′ CpG–OVA conjugates had similar size-exclusion chromatography elution profiles. The immunostimulatory properties of the conjugates were determined by dendritic cells (DCs) and T-cells isolated from mice. The activation of DCs was determined by the upregulation of activation markers CD86 and CD40. T-cells were co-cultured with stimulated DCs, and the immunogenicity was determined by measuring T-cell proliferation and interferon γ production. Both the CpG 5′- and 3′-linked conjugates induced the same level (p > 0.5) of DC activation markers, which were significantly higher than those of the untreated control. Similarly, T-cell assays showed no significant difference (p > 0.5) between the 5′ and 3′ conjugates with respect to T-cell proliferation and interferon γ production. The 5′ and 3′ conjugates induced T-cell activation significantly higher than the mixture of CpG and OVA. This study showed that the end at which CpG is conjugated to an antigen has no influence on the generation of a T-cell-based immune response in vitro.
机译：与混合物相比，CpG与抗原的缀合诱导更强烈的免疫应答。该研究将CPG通过其5'或3'末端缀合的体外免疫刺激活性与模型抗原卵蛋白（OVA）进行比较。通过稳定的双芳基腙键将用5'或3'末端用胺改性的Cpg与OVA缀合。在360nm处的吸光度下，在形成双芳基腙键的吸光度的基础上，观察到与胚轴相似的CPG缀合水平的CPG缀合物，所述缀合物为每种OVA连接的2.8±0.3cpg。 5'和3'CPG-OVA缀合物都具有相似的尺寸排阻色谱炼金曲线。缀合物的免疫刺激性质由树突细胞（DC）和来自小鼠分离的T细胞测定。通过激活标志物CD86和CD40的上调来确定DC的激活。用刺激的DC共同培养T细胞，通过测量T细胞增殖和干扰素γ产生来确定免疫原性。 CPG 5'-和3'-连接的缀合物均诱导DC活化标志物的相同水平（ p> 0.5），其显着高于未处理的对照。类似地，T细胞测定在与T细胞增殖和干扰素γ产生方面显示在5'和3'缀合物之间没有显着差异（ P> 0.5）。 5'和3'缀合物诱导T细胞活化明显高于CpG和OVA的混合物。该研究表明，CPG与抗原缀合的结束对体外的T细胞免疫反应产生没有影响。

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