title: Potent and Broad Inhibition of HIV-1 by a Peptide from the gp41 Heptad Repeat-2 Domain Conjugated to the CXCR4 Amino Terminus.
abstract: HIV-1 entry can be inhibited by soluble peptides from the gp41 heptad repeat-2 (HR2) domain that interfere with formation of the 6-helix bundle during fusion. Inhibition has also been seen when these peptides are conjugated to anchoring molecules and over-expressed on the cell surface. We hypothesized that potent anti-HIV activity could be achieved if a 34 amino acid peptide from HR2 (C34) were brought to the site of virus-cell interactions by conjugation to the amino termini of HIV-1 coreceptors CCR5 or CXCR4. C34-conjugated coreceptors were expressed on the surface of T cell lines and primary CD4 T cells, retained the ability to mediate chemotaxis in response to cognate chemokines, and were highly resistant to HIV-1 utilization for entry. Notably, C34-conjugated CCR5 and CXCR4 each exhibited potent and broad inhibition of HIV-1 isolates from diverse clades irrespective of tropism (i.e., each could inhibit R5, X4 and dual-tropic isolates). This inhibition was highly specific and dependent on positioning of the peptide, as HIV-1 infection was poorly inhibited when C34 was conjugated to the amino terminus of CD4. C34-conjugated coreceptors could also inhibit HIV-1 isolates that were resistant to the soluble HR2 peptide inhibitor, enfuvirtide. When introduced into primary cells, CD4 T cells expressing C34-conjugated coreceptors exhibited physiologic responses to T cell activation while inhibiting diverse HIV-1 isolates, and cells containing C34-conjugated CXCR4 expanded during HIV-1 infection in vitro and in a humanized mouse model. Notably, the C34-conjugated peptide exerted greater HIV-1 inhibition when conjugated to CXCR4 than to CCR5. Thus, antiviral effects of HR2 peptides can be specifically directed to the site of viral entry where they provide potent and broad inhibition of HIV-1. This approach to engineer HIV-1 resistance in functional CD4 T cells may provide a novel cell-based therapeutic for controlling HIV infection in humans.
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: George J
authors.initials: GJ
authors.lastname: Leslie
authors.affiliation: Sangamo BioSciences Inc., Richmond, CA, United States of America.
authors.forename: Jianbin
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: Max W
authors.initials: MW
authors.lastname: Richardson
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: Beth S
authors.initials: BS
authors.lastname: Haggarty
authors.affiliation: Sangamo BioSciences Inc., Richmond, CA, United States of America.
authors.forename: Kevin L
authors.identifier: http://orcid.org/0000-0003-2186-6674
authors.initials: KL
authors.lastname: Hua
authors.affiliation: Sangamo BioSciences Inc., Richmond, CA, United States of America.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Duong
authors.affiliation: Department of Pathology & Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: Anthony J
authors.identifier: http://orcid.org/0000-0001-6646-368X
authors.initials: AJ
authors.lastname: Secreto
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: Andrea P O
authors.initials: AP
authors.lastname: Jordon
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: Josephine
authors.initials: J
authors.lastname: Romano
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: Kritika E
authors.identifier: http://orcid.org/0000-0002-6066-9174
authors.initials: KE
authors.lastname: Kumar
authors.affiliation: Sangamo BioSciences Inc., Richmond, CA, United States of America.
authors.forename: Joshua J
authors.initials: JJ
authors.lastname: DeClercq
authors.affiliation: Sangamo BioSciences Inc., Richmond, CA, United States of America.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Gregory
authors.affiliation: Department of Pathology & Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: Carl H
authors.identifier: http://orcid.org/0000-0003-0241-3557
authors.initials: CH
authors.lastname: June
authors.affiliation: Department of Biochemistry and Molecular Biology, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, United States of America.
authors.forename: Michael J
authors.identifier: http://orcid.org/0000-0002-6844-7170
authors.initials: MJ
authors.lastname: Root
authors.affiliation: Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: James L
authors.initials: JL
authors.lastname: Riley
authors.affiliation: Sangamo BioSciences Inc., Richmond, CA, United States of America.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Holmes
authors.affiliation: Department of Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States of America.
authors.forename: James A
authors.identifier: http://orcid.org/0000-0002-6257-4902
authors.initials: JA
authors.lastname: Hoxie
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism / Q000821:virology*; D005434:Flow Cytometry; D057809:HEK293 Cells; D015700:HIV Envelope Protein gp41 / Q000378:metabolism*; D015658:HIV Infections / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D010446:Peptide Fragments / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: D015700:HIV Envelope Protein gp41; D010446:Peptide Fragments; D019718:Receptors, CXCR4; C407149:peptide C34