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Principal Investigator: Jeremy Phillip Kamil
Organization: LOUISIANA STATE UNIV HSC SHREVEPORT
Fiscal Year: 2024
Award: $142,711
Funding agency: National Institute of Allergy and Infectious Diseases
This project focuses on the role of glycobiology in human cytomegalovirus (HCMV) immune evasion strategies.
Our long-term goal is to understand how the virus exploits host protein glycosylation pathways to shield its
virions and decorate the surfaces of infected cells with oligosaccharide patterns that prevent antibody binding
and/or suppress immune responses. The Specific Aims are (1) to evaluate how the HCMV ER resident protein
UL148 impacts the glycocalyx, or “sugar cloak,” of virions and infected cells, (2) to carry out a structure
function analysis of UL148 to determine the mechanisms by which UL148 activates sculpts the host cell to
enhance glycosylation of virion envelope proteins, and (3) to evaluate how UL148 contributes to evasion of
antibody responses. Throughout the three Specific Aims, wild-type (WT) and UL148-null mutant viruses are
evaluated in comparison to each other. In Specific Aim 1, mass spectrometry based ‘glycomics’ is applied to
profile N-linked oligosaccharide structures conjugated to virion envelope proteins and surface proteins of
infected cells in the presence versus absence of UL148. A second aspect of Aim 1 makes use of mass
spectrometry-based proteomics together with genetic and pharmacological perturbations targeting the unfolded
protein response (UPR) and cellular genes whose expression is positively regulated by UL148 to determine
how the viral protein harnesses the host cell’s stress response circuitry toward the generation of virions that
resist neutralization by antibodies. In Specific Aim 2, computational predictions of UL148’s three-dimensional
protein structure are used to guide mutagenesis studies aimed at understanding the molecular basis for its
biological functions, focusing in large part on the roles of (i) a consensus metal binding motif and (ii) two non-
conserved cysteine residues. Aim 2 also leverages structural models non-human primate cytomegaloviruses
homologs to elucidate the determinants of HCMV UL148’s unique biological functions. These studies will make
use of recombinant HCMVs to test the impact of selected mutations and chimeric UL148 proteins in the context
of infected cells. In Specific Aim 3, we will (i) characterize UL148’s impact on neutralization resistance of
virions while targeting glycan modification enzymes hypothesized to be important for biogenesis of
neutralization resistant virions, (ii) map specific antigenic domains on virion glycoproteins whose reactivity to
neutralizing antibodies is impacted by UL148, (iii) compare the humoral immunogenicity of UL148 competent
versus UL148-null HCMV virions in a mouse model, testing for neutralizing antibody titers, and overall
polyclonal antibody titers against whole virions, viral envelope proteins and internal virion antigens.
Collectively, these studies will address a key knowledge gap concerning how enveloped large DNA viruses
exploit glycobiology to evade adaptive immunity and establish persistent infection.
Terms: <0-11 years old><3-D><3-Dimensional><3D><Acylneuraminyl hydrolase><Address><Amino Acids><Antibodies><Antibody Response><Antibody titer measurement><Antigens><Binding><Binding Proteins><Biogenesis><Biological><Biological Function><Biological Process><Birth Defects><Body Tissues><CD140a Antigens><CMV><CMV infection><CMV vaccine><Cancers><Carbohydrates><Cell Body><Cell Coat><Cells><Cellular Stress><Cellular Stress Response><Charge><Child><Child Youth><Children (0-21)><Complex><Congenital Abnormality><Congenital Anatomical Abnormality><Congenital Defects><Congenital Deformity><Congenital Malformation><Consensus><Cysteine><Cytomegalic Inclusion Disease><Cytomegalovirus><Cytomegalovirus Infections><Cytomegalovirus Vaccines><DNA Viruses><Data><Development><Endothelial Cells><Envelope Protein><Environment><Enzyme Gene><Enzymes><Epithelial Cells><Epithelium><Exhibits><Fabaceae><Fibroblasts><GeneHomolog><Generations><Genes><Genetic><Genetic Alteration><Genetic Change><Genetic defect><Genetics-Mutagenesis><Glycans><Glycobiology><Glycocalyx><Glycoproteins><Goals><HCMV><Half-Cystine><Herpesviridae><Herpesviruses><Homolog><Homologous Gene><Homologue><Human><Immune><Immune Evasion><Immune Globulins><Immune response><Immunes><Immunity><Immunize><Immunoglobulins><Immunological response><Inclusion Disease><Infection><Ions><Knowledge><L-Cysteine><Lectin><Legumes><Leguminosae><Leguminoseae><Life><Ligand Binding Protein><Ligand Binding Protein Gene><Link><Lymphatic cell><Lymphocyte><Lymphocytic><Malignant Neoplasms><Malignant Tumor><Maps><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measures><Mediating><Membrane><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Messenger RNA><Metabolic Glycosylation><Metals><Mice><Mice Mammals><Modern Man><Modification><Molecular><Molecular Interaction><Murine><Mus><Mutagenesis><Mutagenesis Molecular Biology><Mutation><N-Acetylneuraminic Acids><N-Acylneuraminate Glycohydrolases><NRP2 Protein><Neuraminidase><Neurodevelopmental Disability><Neuropilin-2><Neutralization Tests><Neutralizing antibody assay><Npn-2 Protein><Oligosaccharide Sialidase><Oligosaccharides><Opportunistic Infections><Origin of Life><Ortholog><Orthologous Gene><PDGF alpha Receptor><PDGF receptor α><PDGF-R-alpha><PDGFR-α><PDGFRα><Pathway interactions><Patients><Pattern><Persons><Platelet-Derived Growth Factor Receptor Alpha Polypeptide><Platelet-Derived Growth Factor alpha Receptor><Play><Polysaccharides><Production><Protein Binding><Protein Glycosylation><Proteins><Proteomics><Quality Control><Reagent><Receptor Protein><Recombinants><Research><Resistance><Role><Salivary Gland Virus Disease><Salivary Gland Viruses><Sialic Acids><Sialidase><Sialyltransferases><Structural Models><Structure><Surface><Surface Proteins><Testing><Tissues><Tropism><Vaccines><Viral><Viral Envelope Proteins><Viral Gene Products><Viral Gene Proteins><Viral Proteins><Virion><Virus><Virus Particle><acute infection><adaptive immunity><aminoacid><analog><antibody neutralization test><antibody titering><biologic><biological adaptation to stress><bound protein><cell stress><cell type><chronic infection><congenital infection><cytomegalovirus group><developmental><env Antigens><env Gene Products><env Polyproteins><env Protein><exo alpha sialidase><genetic approach><genetic strategy><genome mutation><glycosylation><herpes virus><host response><hypoimmunity><immune deficiency><immune evasive><immune system response><immunization strategy><immunodeficiency><immunogen><immunogenicity><immunoresponse><kids><lymph cell><mRNA><malignancy><membrane structure><mouse model><murine model><mutant><neoplasm/cancer><neutralizing antibody><neutralizing antibody test><non-human primate><nonhuman primate><pathway><pea family><permanent hearing loss><persistent infection><pharmacologic><platelet-derived growth factor receptor α><polyclonal antibody><prevent><preventing><programs><protein structure><protein structures><proteins structure><reaction; crisis><receptor><resistant><response><sialic acid binding Ig-like lectin><sialylation><siglec><social role><stress response><stress; reaction><sugar><three dimensional><treatment strategy><vaccination strategy><virus host interaction><virus protein><youngster>