Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences & Yunnan Province, Kunming Institute of Zoology, Jiaochang East Road 32, Kunming, Yunnan Province, 650223 China.
BMC Med Genomics. 2013 May 1;6:15. doi: 10.1186/1755-8794-6-15.
Upon co-stimulation with CD3/CD28 antibodies, activated CD4 + T cells were found to lose their susceptibility to HIV-1 infection, exhibiting an induced resistant phenotype. This rather unexpected phenomenon has been repeatedly confirmed but the underlying cell and molecular mechanisms are still unknown.
We first replicated the reported system using the specified Dynal beads with PHA/IL-2-stimulated and un-stimulated cells as controls. Genome-wide expression and analysis were then performed by using Agilent whole genome microarrays and established bioinformatics tools.
We showed that following CD3/CD28 co-stimulation, a homogeneous population emerged with uniform expression of activation markers CD25 and CD69 as well as a memory marker CD45RO at high levels. These cells differentially expressed 7,824 genes when compared with the controls on microarrays. Series-Cluster analysis identified 6 distinct expression profiles containing 1,345 genes as the representative signatures in the permissive and resistant cells. Of them, 245 (101 potentially permissive and 144 potentially resistant) were significant in gene ontology categories related to immune response, cell adhesion and metabolism. Co-expression networks analysis identified 137 "key regulatory" genes (84 potentially permissive and 53 potentially resistant), holding hub positions in the gene interactions. By mapping these genes on KEGG pathways, the predominance of actin cytoskeleton functions, proteasomes, and cell cycle arrest in induced resistance emerged. We also revealed an entire set of previously unreported novel genes for further mining and functional validation.
This initial microarray study will stimulate renewed interest in exploring this system and open new avenues for research into HIV-1 susceptibility and its reversal in target cells, serving as a foundation for the development of novel therapeutic and clinical treatments.
在与 CD3/CD28 抗体共刺激后，发现激活的 CD4+T 细胞失去了对 HIV-1 感染的易感性，表现出诱导的抗性表型。这种相当出人意料的现象已经被反复证实，但细胞和分子机制仍不清楚。
我们首先使用指定的 Dynal 珠粒复制了报告的系统，并用 PHA/IL-2 刺激和未刺激的细胞作为对照。然后使用安捷伦全基因组微阵列和已建立的生物信息学工具进行全基因组表达和分析。
我们表明，在 CD3/CD28 共刺激后，出现了一个均匀表达激活标志物 CD25 和 CD69 以及记忆标志物 CD45RO 的同质群体，其水平很高。与对照相比，这些细胞在微阵列上表达了 7824 个差异基因。系列聚类分析确定了 6 个不同的表达谱，包含 1345 个基因作为许可和抗性细胞的代表性特征。其中，245 个（101 个潜在许可和 144 个潜在抗性）在与免疫反应、细胞黏附和代谢相关的基因本体类别中具有显著性。共表达网络分析确定了 137 个“关键调控”基因（84 个潜在许可和 53 个潜在抗性），在基因相互作用中处于中心位置。通过将这些基因映射到 KEGG 途径上，诱导抗性中肌动蛋白细胞骨架功能、蛋白酶体和细胞周期停滞的优势显现出来。我们还揭示了一整套以前未报道的新基因，以供进一步挖掘和功能验证。
这项初步的微阵列研究将激发人们重新探索这个系统的兴趣，并为研究 HIV-1 易感性及其在靶细胞中的逆转开辟新的途径，为开发新的治疗和临床治疗方法奠定基础。
