Anti-CCR5 monoclonal antibody, clone U32/9 [APC]

Background The chemokine receptor CCR5 is one of the co-receptor of HIV-1 infection. People with homozygousCCR5 Delta 32deletion resist HIV-1 infection, which makes theCCR5an important target for HIV-1 gene therapy. Although the CRISPR/Cas9 has ever been used for HIV-1 study, the newly developed CRISPR/AsCpf1 has never been utilized in HIV-1 co-receptor disruption. The CRISPR/Cpf1 system shows many advantages over CRISPR/Cas9, such as lower off-target, small size of nuclease, easy sgRNA design for multiplex gene editing, etc. Therefore, the CRISPR/Cpf1 mediated gene editing will confer a more specific and safe strategy in HIV-1 co-receptor disruption. Results Here, we demonstrated that CRISPR/AsCpf1 could ablate the main co-receptor of HIV-1 infection-CCR5efficiently with two screened sgRNAs via different delivery strategies (lentivirus, adenovirus). The edited cells resisted R5-tropic HIV-1 infection but not X4-tropic HIV-1 infection compared with the control group in different cell types of HIV-1 study (TZM.bl, SupT1-R5, Primary CD4(+)T cells). Meanwhile, the edited cells exhibited selective advantage over unedited cells while under the pressure of R5-tropic HIV-1. Furthermore, we clarified that the predicted off-target sites of selected sgRNAs were very limited, which is much less than regular using sgRNAs for CRISPR/Cas9, and no evident off-target was observed. We also showed that the disruption ofCCR5by CRISPR/AsCpf1 took no effects on cell proliferation and apoptosis. Conclusions Our study provides a basis for a possible application ofCCR5-targeting gene editing by CRISPR/AsCpf1 with high specific sgRNAs against HIV-1 infection.

Background: We aimed to explore potential gene biomarkers of renal interstitial fibrosis (RIF) due to a lack of effective and non-invasive methods for diagnosis. Methods: Three data sets (GSE22459, GSE76882 and GSE57731) including 350 samples were acquired from Gene Expression Omnibus (GEO) database. We used bioconductor limma package to perform background adjustment. Cluster analysis was conducted by 'edgeR' package to identify the differentially expressed genes (DEGs). We generated heat maps with using heatmap package in R software. Function annotation of genes was performed by Gene Ontology (GO) enrichment analysis. STRING (Search Tool for the Retrieval of Interacting Genes) database was employed to construct the protein-protein interaction (PPI) network and the results were visualized by Cytoscape 3.6.1. At last, we applied Graphpad Prism 7.0. to explore the correlation between three hub genes and pathological degrees of RIF. Results: By applying the "edgeR" package in R, we detected 116 DEGs with three data sets. These genes were enriched in 19 GO biological process categories. Three main hub genes (CD2, CCL5 and CCR5) were identified after construction of PPI network. In Pearson correlation coefficient, CD2, CCL5 and CCR5 was found to hold higher expression patterns in RIF samples based on independent data set GSE57731. Besides, their gene expression levels were found significantly positive correlation with the degree of RIF (CD2: P<0.05, r=0.29; CCL5: P<0.05, r=0.31; CCR5: P<0.05, r=0.38). Conclusions: CD2, CCL5 and CCR5 might serve as potential early biomarkers of RIF. The mechanism between these genes and RIF remains to be further studied.