Method of inhibiting enveloped virus binding to target cells by incorporating P-selectin glycoprotein ligand-1 (PSGL-1) into virions
Inventors
Wu, Yuntao • DABBAGH, Deemah • He, Sijia
Assignees
Publication Number
US-12194087-B2
Publication Date
2025-01-14
Expiration Date
2040-04-02
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Embodiments relate to a method comprising (a) expressing a vector comprising a PSGL-1 (P-selectin glycoprotein ligand-1) or a mutant thereof in a VPC (virus producing cell); and blocking a virus infection by inactivating an infectivity of a released virions from the VPC; or (b) expressing a glycoprotein or a mutant thereof in the VPC; blocking the virus infection by preventing binding of the released virions to a target cell; inactivating infectivity of the released virions; and targeting a viral infection. Other embodiments relate to (1) a broad-spectrum anti-viral product comprising: a vector expressing a glycoprotein or a mutant thereof in a VPC; and blocking a virus infection by inactivating infectivity of a released virion from the VPC; and (2) a vaccine comprising a viral particle is configured to a live attenuated or an inactivated or a non-infectious, wherein the viral particle are produced in a VPC.
Core Innovation
The invention provides a method for inhibiting viral infection, specifically targeting enveloped viruses, by expressing P-selectin glycoprotein ligand-1 (PSGL-1) or a mutant thereof in virus producing cells (VPCs). When PSGL-1 is incorporated into assembling virions, it is displayed on the surface of the released viral particles, resulting in the inactivation of their infectivity. This approach is demonstrated to be effective against retroviruses such as human immunodeficiency virus type 1 (HIV-1) and murine leukemia virus (MLV), as well as influenza virus, representing both retroviral and non-retroviral enveloped viruses.
The background problem addressed by the invention is the lack of effective broad-spectrum viral vaccines and the absence of prior literature detailing an anti-viral effect of PSGL-1 expressed within virus producing cells. Prior to this invention, it was unclear how HIV-1 could overcome PSGL-1 restriction at early stages of infection, and the mechanism of PSGL-1-mediated inactivation of virion infectivity had not been elucidated. There remains a need for strategies that can inactivate a broad range of enveloped viruses for vaccine development and antiviral applications.
The core of the method involves co-expressing a PSGL-1 expression vector and a viral vector in a human-derived virus producing cell, leading to the incorporation of PSGL-1 into the envelope of progeny virions. The presence of PSGL-1 on the virion surface blocks binding of the virions to their cognate receptors on target cells, thereby neutralizing their infectivity. Experimental evidence showed that the extracellular N-terminal domain of PSGL-1, particularly containing decameric repeats, is necessary for this antiviral effect, and the mechanism of action includes steric hindrance of viral attachment.
Claims Coverage
The patent contains several inventive features as reflected in its independent claims, mainly focusing on the use of PSGL-1 or its mutants for inactivating viral infectivity by incorporation into virions.
Incorporation of PSGL-1 with decameric repeat region into virions to block viral binding
The inventive method comprises expressing PSGL-1 or a mutant thereof containing an extracellular N-terminal domain with at least one decameric repeat (DR) region and a transmembrane region. The DR region specifically contains at least one proline and more than one O-glycosylated threonine, and the mutant exhibits antiviral activity. A retroviral vector and a PSGL-1 expression vector are co-expressed in a virus-producing cell (VPC) derived from a human cell line. This results in PSGL-1 (or mutant) incorporation into assembling virions, displaying the glycoprotein on the virion surface during assembly. The presence of PSGL-1 or its mutant on the virion surface blocks the binding of the virions to their cognate receptor, inactivating viral infection of the retrovirus.
Dose-dependent inactivation of viral infectivity by PSGL-1 expression in virus-producing cells
The method describes that the expression vector for PSGL-1 or its mutant can be used at specific dose ranges (about 0.5 ng to 500 ng), and blocking viral infection occurs in a dose-dependent manner. Specific quantities are cited, such as 1 μg of PSGL-1 leading to a 100-fold reduction in virion infectivity, and 3 μg causing about an 8000-fold decrease.
PSGL-1-mediated inactivation of virion infectivity is independent of viral envelope glycoprotein
The inventive feature covers that the inactivation of released virion infectivity by PSGL-1 (or mutant) is independent of the envelope glycoprotein (Env-1) of the virus-producing cell. Virions lacking envelope proteins can still be inactivated by PSGL-1 incorporation.
Preparation of non-infectious virions for broad-spectrum antiviral and vaccine use
The method includes harvesting virions produced from virus-producing cells that have been co-transfected with vectors expressing PSGL-1 or a mutant. The resultant virions, with incorporated PSGL-1, are non-infectious due to blocked binding to target cells. This method applies to both retroviral (such as HIV-1, MLV) and non-retroviral (such as influenza virus) enveloped viruses.
The inventive features claimed cover the method of incorporating PSGL-1 or its mutants into virions during assembly in virus-producing human cell lines, leading to steric inhibition of virion binding and thus inactivating viral infectivity in a dose-dependent and envelope-independent manner. This supports the use of the approach for broad-spectrum antiviral or vaccine production.
Stated Advantages
PSGL-1 incorporation in virus producing cells inactivates virion infectivity without chemically destroying the virus-producing cells.
The method allows for production of live attenuated, inactivated, or non-infectious viral particles for vaccines or antiviral products.
PSGL-1-mediated inactivation of viral infectivity is effective against multiple viruses, including HIV-1, murine leukemia virus, and influenza virus.
The approach provides a broad-spectrum antiviral method that does not rely on modifications to viral envelope glycoproteins.
The technique enables dose-dependent and highly potent inhibition of viral infectivity.
Documented Applications
Use in the preparation of live attenuated, inactivated, or non-infectious viral vaccines by expressing PSGL-1 or its mutant in virus producing cells.
Therapeutic production of non-infectious virion particles for treatment or prevention of diseases related to HIV, murine leukemia virus, and influenza virus infections.
Application for inactivation of a broad spectrum of enveloped viruses during virus particle production for anti-viral products.
Interested in licensing this patent?