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      CCR5-Δ32 gene variant frequency in the Turkish Cypriot population

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          Abstract

          Recent UNAIDS reports (December 2019) indicate that 37.9 million people have been affected by HIV infection around the globe in 2018, of which 1.7 million are cited as new infections. Human immunodeficiency virus-1 (HIV-1) requires both the CD4 receptor, as the primary receptor, and a chemokine co-receptor to gain entry into the cell. In addition to the WT allele for C–C motif chemokine receptor 5 (CCR5-wt), there is another allele with a 32 bp deletion in the protein coding region (CCR5-Δ32). Individuals who are homozygous for the mutant allele are resistant towards M-tropic HIV infections. In the current study, we aimed to determine the CCR5-Δ32 allele frequency in the Turkish Cypriot population with 326 subjects, 141 men (43.1%) and 185 (56.9%) women. The region of the CCR5 gene containing the Δ32 deletion was amplified using flanking primers. The CCR5 gene Δ32 allele frequency was calculated at 3% and only observed in heterozygous individuals. We hope that our current publication could be a point of dialog between the physicians, the government officials and the public set up a more modern and well-structured HIV screening program in an effort to control and hopefully eliminate HIV from the Turkish Cypriot population.

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          The biology of CCR5 and CXCR4.

          We discuss the current knowledge concerning the biology of CXCR4 and CCR5 and their roles in HIV-1 infection. Important research findings reported in the last 2 years have advanced our knowledge in the field of HIV coreceptors and pathogenesis. Novel methods have been used to crystallize two new members of the G-protein coupled receptors. It has been demonstrated that expression and stability of the naturally occurring truncated CCR5 protein is critical for resistance to HIV-1. The first stem cell transplantation of donor cells with the CCR5 mutation provided proof of principle. The Food and Drug Administration approved the first CCR5-based entry inhibitor. New CXCL12 isoforms were discovered, one isoform is a potent X4 inhibitor with weak chemotaxis activity. The coreceptor discoveries revealed new insights into host and viral factors influencing HIV transmission and disease. The HIV/coreceptor interaction has become a major target for the development of novel antiviral strategies to treat and prevent HIV infection. The first CCR5-based entry inhibitor has been recently approved. New drugs that promote CCR5 and CXCR4 internalization, independent of cellular signaling, might provide clinical benefits with minimum side effects.
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            Simultaneous Knockout of CXCR4 and CCR5 Genes in CD4+ T Cells via CRISPR/Cas9 Confers Resistance to Both X4- and R5-Tropic Human Immunodeficiency Virus Type 1 Infection

            Previous research has proven that disruption of either the CCR5 or the CXCR4 gene confers resistance to R5-tropic or X4-tropic human immunodeficiency virus type 1 (HIV-1) infection, respectively. However, the urgent need to ablate both of the co-receptors in individual post-thymic CD4+ T cells for dual protection remains. This study ablated the CCR5 and CXCR4 genes in human CD4+ cell lines and primary CD4+ T cells simultaneously using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9, a well-developed, highly efficient genetic engineering tool. The efficiency of gene modification is as high as 55% for CCR5 and 36% for CXCR4 in CD4+ cell lines through infection of a single lentiviral vector (LV-X4R5), which were markedly protected from both HIV-1NL4-3 (X4-using strain) and HIV-1YU-2 (R5-using strain) infection. Importantly, approximately 9% of the modified GHOST (3) CXCR4+CCR5+ cells harbor four bi-allelic gene disruptions in both the CXCR4 and CCR5 loci. Moreover, co-delivery of two single-guide RNAs loaded with Cas9: ribonucleoprotein (sgX4&R5 Cas9RNP) disrupted >12% of CCR5 and 10% of CXCR4 in primary human CD4+ T cells, which were rendered resistant to HIV-1NL4-3 and HIV-1YU-2 in vitro. Further, the modified cells do not show discernible mutagenesis in top-ranked off-target genes by the Surveyor assay and Sanger sequencing analysis. The results demonstrate the safety and efficacy of CRISPR/Cas9 in multiplex gene modification on peripherally circulating CD4+ T cells, which may promote a functional cure for HIV-1 infection.
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              Is Open Access

              C-C chemokine receptor type five (CCR5): An emerging target for the control of HIV infection☆

              When HIV was initially discovered as the causative agent of AIDS, many expected to find a vaccine within a few years. This has however proven to be elusive; it has been approximately 30 years since HIV was first discovered, and a suitable vaccine is still not in effect. In 2009, a paper published by Hutter et al. reported on a bone marrow transplant performed on an HIV positive individual using stem cells that were derived from a donor who was homozygous for a mutation in the CCR5 gene known as CCR5 delta-32 (Δ32) (Hütter et al., 2009). The HIV positive individual became HIV negative and remained free of viral detection after transplantation despite having halted anti-retroviral (ARV) treatment. This review will focus on CCR5 as a key component in HIV immunity and will discuss the role of CCR5 in the control of HIV infection.
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                Author and article information

                Contributors
                umut.fahrioglu@neu.edu.tr , umutfahrioglu@gmail.com
                mahmutcerkez.ergore@neu.edu.tr
                gamze.mocan@neu.edu.tr
                Journal
                Braz J Microbiol
                Braz. J. Microbiol
                Brazilian Journal of Microbiology
                Springer International Publishing (Cham )
                1517-8382
                1678-4405
                31 July 2020
                : 1-7
                Affiliations
                [1 ]GRID grid.412132.7, ISNI 0000 0004 0596 0713, Department of Medical Biology, Faculty of Medicine, , Near East University, ; 99138 Nicosia, Cyprus
                [2 ]GRID grid.412132.7, ISNI 0000 0004 0596 0713, Near East University, Research Center of Experimental Health Sciences (DESAM), ; Nicosia, Cyprus
                [3 ]GRID grid.412132.7, ISNI 0000 0004 0596 0713, Department of Medical Pathology, Faculty of Medicine, , Near East University, ; Nicosia, Cyprus
                Author notes

                Responsible Editor: Giliane Souza Trindade.

                Author information
                https://orcid.org/0000-0002-7933-2395
                https://orcid.org/0000-0001-9593-9325
                https://orcid.org/0000-0002-7625-4934
                Article
                352
                10.1007/s42770-020-00352-8
                7392619
                9883f305-960e-45ba-83c3-01a134c61e88
                © Sociedade Brasileira de Microbiologia 2020

                This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.

                History
                : 7 May 2020
                : 25 July 2020
                Categories
                Clinical Microbiology - Research Paper

                ccr5-δ32,hiv,allele frequency,turkish cypriots,public health policy

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