Virion Proteomics of Wildtype HCMV Reveals a Novel Regulator of Envelope Glycoprotein Composition that Protects Against Humoral Immunity
Authors: Bentley Kirsten , Statkute Evelina , Murrell Isa , Fielding Ceri A. , Antrobus Robin , Preston Hannah , Kerr-Jones Lauren , Cochrane Daniel , Brizic Ilija , Lehner Paul J. , Wilkinson Gavin W.G. , Wang Eddie C.Y. , Graham Stephen C. , Weekes Michael P. , Stanton Richard J. 
Published: December 2024

Human cytomegalovirus (HCMV) is a clinically important herpesvirus that has co-evolved for millions of years with its human host, and establishes lifelong persistent infection. A substantial proportion of its 235kb genome is dedicated to manipulating host immunity through targeting antiviral host proteins for degradation or relocalisation. Quantitative proteomics of the infected cell has extensively characterised these processes, but the cell-free virion has been less well studied. We therefore carried out proteomic analysis of a clinical HCMV strain virion. This revealed 18 novel components, including the viral protein gpUL141, which is recognised as an NK immune-evasin that targets several host proteins (CD155, CD112, and TRAILR) when expressed in the cell. Co-Immunoprecipitation of gpUL141 from virions identified interactions with viral entry glycoproteins from the trimer (gH/gL/gO), pentamer (gH/gL/UL128/UL130/UL131A), and gH/gpUL116 complexes, as well as gB. These interactions occurred via direct interactions between gpUL141 and either gH or gB. Analysis supported a model in which gpUL141 homodimers independently interacted with separate gB/gH-containing complexes. gpUL141 encodes an ER retention domain that restricts trafficking through the ER/golgi, and limited the transport of glycoprotein complexes bound by gpUL141. As a result, gpUL141 reduced levels of multiple glycoprotein complexes on the infected cell surface as well as in the virion. This reduced syncytia formation, inhibited antibody-dependent cellular cytotoxicity (ADCC), and reduced susceptibility to neutralising antibodies. Thus, gpUL141 represents an immune-evasin that not only targets host proteins to limit NK-cell attack, but also alters the trafficking of multiple viral glycoprotein complexes in order to evade humoral immunity.

Significance

The large number of immune-modulators encoded by HCMV has led to it becoming a paradigm for pathogen-mediated immune-modulation. These mechanisms have informed on virus pathogenesis, the evolution and function of host defences, and identified therapeutic targets. Previously discovered immune-evasins functioned by modifying host proteins. gpUL141 represents a novel strategy in which multiple immunological functions are targeted through manipulating eight different viral entry glycoproteins, via interactions with two proteins common to multiple complexes. Entry glycoproteins define virus tropism, mechanisms of infection and spread, and susceptibility to neutralising and non-neutralising antibody activities, yet UL141 is frequently mutated in passaged viruses. This demonstrates the need to work with virus strains encoding the complete repertoire of viral accessory proteins during preclinical therapeutic development.