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Abstract

Human cytomegalovirus (HCMV) gpUL18 is a HLA class I (HLA-I) homologue with high affinity for the inhibitory receptor LIR-1/ILT2. The previously described 67 kDa form of gpUL18 is shown here to be sensitive to endoglycosidase-H (EndoH). A novel form of gpUL18 with a molecular mass of ∼160 kDa and resistance to EndoH was identified in cells infected with HCMV strain AD169 or the low passage HCMV isolates Merlin and Toledo. The 67 kDa EndoH-sensitive gpUL18 glycoform was detected earlier in a productive infection (from 24 h post-infection) than the slower-migrating EndoH-resistant glycoform (from 72 h post-infection). Deletion of the US2–US11 region from the HCMV genome was associated with a substantial up-regulation of endogenous HLA-I in infected cells, but had no obvious effect on the gpUL18 expression pattern. Vaccinia virus and adenovirus vectors were used to further analyse gpUL18 expression. Depending on the delivery vector system, differences in the electrophoretic motility of the EndoH-resistant >105 kDa form of gpUL18, but not the EndoH-sensitive 67 kDa form, were observed; post-translational modification of the higher molecular mass glycoform appears to be influenced by active virus infection and vector delivery. The EndoH-sensitive 67 kDa gpUL18 had a rapid turnover, while the maturation to the EndoH-resistant >105 kDa form was relatively slow and inefficient. However, synthesis of the EndoH-resistant >105 kDa form was enhanced with elevated levels of -microglobulin. When expressed by using an adenovirus vector, both the EndoH-sensitive 67 kDa and the EndoH-resistant >105 kDa gpUL18 forms could be detected on the cell surface.

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2005-11-01
2019-10-20
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References

  1. Abenes, G., Chan, K., Lee, M., Haghjoo, E., Zhu, J., Zhou, T., Zhan, X. & Liu, F. ( 2004; ). Murine cytomegalovirus with a transposon insertional mutation at open reading frame m155 is deficient in growth and virulence in mice. J Virol 78, 6891–6899.[CrossRef]
    [Google Scholar]
  2. Akter, P., Cunningham, C., McSharry, B. P. & 8 other authors ( 2003; ). Two novel spliced genes in human cytomegalovirus. J Gen Virol 84, 1117–1122.[CrossRef]
    [Google Scholar]
  3. Arase, H., Mocarski, E. S., Campbell, A. E., Hill, A. B. & Lanier, L. L. ( 2002; ). Direct recognition of cytomegalovirus by activating and inhibitory NK cell receptors. Science 296, 1323–1326.[CrossRef]
    [Google Scholar]
  4. Beck, S. & Barrell, B. G. ( 1988; ). Human cytomegalovirus encodes a glycoprotein homologous to MHC class-I antigens. Nature 331, 269–272.[CrossRef]
    [Google Scholar]
  5. Benedict, C. A., Butrovich, K. D., Lurain, N. S., Corbeil, J., Rooney, I., Schneider, P., Tschopp, J. & Ware, C. F. ( 1999; ). Cutting edge: a novel viral TNF receptor superfamily member in virulent strains of human cytomegalovirus. J Immunol 162, 6967–6970.
    [Google Scholar]
  6. Berg, L., Riise, G. C., Cosman, D., Bergström, T., Olofsson, S., Kärre, K. & Carbone, E. ( 2003; ). LIR-1 expression on lymphocytes, and cytomegalovirus disease in lung-transplant recipients. Lancet 361, 1099–1101.[CrossRef]
    [Google Scholar]
  7. Biron, C. A., Byron, K. S. & Sullivan, J. L. ( 1989; ). Severe herpesvirus infections in an adolescent without natural killer cells. N Engl J Med 320, 1731–1735.[CrossRef]
    [Google Scholar]
  8. Browne, H., Smith, G., Beck, S. & Minson, T. ( 1990; ). A complex between the MHC class I homologue encoded by human cytomegalovirus and β 2 microglobulin. Nature 347, 770–772.[CrossRef]
    [Google Scholar]
  9. Browne, H., Churcher, M. & Minson, T. ( 1992; ). Construction and characterization of a human cytomegalovirus mutant with the UL18 (class I homolog) gene deleted. J Virol 66, 6784–6787.
    [Google Scholar]
  10. Cebulla, C. M., Miller, D. M., Knight, D. A., Briggs, B. R., McGaughy, V. & Sedmak, D. D. ( 2000; ). Cytomegalovirus induces sialyl Lewisx and Lewisx on human endothelial cells. Transplantation 69, 1202–1209.[CrossRef]
    [Google Scholar]
  11. Chapman, T. L., Heikeman, A. P. & Bjorkman, P. J. ( 1999; ). The inhibitory receptor LIR-1 uses a common binding interaction to recognize class I MHC molecules and the viral homolog UL18. Immunity 11, 603–613.[CrossRef]
    [Google Scholar]
  12. Chee, M. S., Bankier, A. T., Beck, S. & 7 other authors ( 1990; ). Analysis of the protein-coding content of the sequence of human cytomegalovirus strain AD169. Curr Top Microbiol Immunol 154, 125–169.
    [Google Scholar]
  13. Child, S. J., Hakki, M., De Niro, K. L. & Geballe, A. P. ( 2004; ). Evasion of cellular antiviral responses by human cytomegalovirus TRS1 and IRS1. J Virol 78, 197–205.[CrossRef]
    [Google Scholar]
  14. Cosman, D., Fanger, N., Borges, L., Kubin, M., Chin, W., Peterson, L. & Hsu, M. L. ( 1997; ). A novel immunoglobulin superfamily receptor for cellular and viral MHC class I molecules. Immunity 7, 273–282.[CrossRef]
    [Google Scholar]
  15. Cosman, D., Mullberg, J., Sutherland, C. L., Chin, W., Armitage, R., Fanslow, W., Kubin, M. & Chalupny, N. J. ( 2001; ). ULBPs, novel MHC class I-related molecules, bind to CMV glycoprotein UL16 and stimulate NK cytotoxicity through the NKG2D receptor. Immunity 14, 123–133.[CrossRef]
    [Google Scholar]
  16. Davis, D. M., Reyburn, H. T., Pazmany, L., Chiu, I., Mandelboim, O. & Strominger, J. L. ( 1997; ). Impaired spontaneous endocytosis of HLA-G. Eur J Immunol 27, 2714–2719.[CrossRef]
    [Google Scholar]
  17. Dolan, A., Cunningham, C., Hector, R. D. & 12 other authors ( 2004; ). Genetic content of wild type human cytomegalovirus. J Gen Virol 85, 1301–1312.[CrossRef]
    [Google Scholar]
  18. Fahnestock, M. L., Johnson, J. L., Feldman, R. M., Neveu, J. M., Lane, W. S. & Bjorkman, P. J. ( 1995; ). The MHC class I homolog encoded by human cytomegalovirus binds endogenous peptides. Immunity 3, 583–590.[CrossRef]
    [Google Scholar]
  19. Farrell, H. E., Vally, H., Lynch, D. M., Fleming, P., Shellam, G. R., Scalzo, A. A. & Davis-Poynter, N. J. ( 1997; ). Inhibition of natural killer cells by a cytomegalovirus MHC class I homologue in vivo. Nature 386, 510–514.[CrossRef]
    [Google Scholar]
  20. Gazit, R., Garty, B. Z., Monselise, Y. & 9 other authors ( 2004; ). Expression of KIR2DL1 on the entire NK cell population: a possible novel immunodeficiency syndrome. Blood 103, 1965–1966.[CrossRef]
    [Google Scholar]
  21. Jones, T. R., Hanson, L. K., Sun, L., Slater, J. S., Stenberg, R. M. & Campbell, A. E. ( 1995; ). Multiple independent loci within the human cytomegalovirus unique short region down-regulate expression of major histocompatibility complex class I heavy chains. J Virol 69, 4830–4841.
    [Google Scholar]
  22. Keller, R., Peitchel, R., Goldman, J. N. & Goldman, M. ( 1976; ). An IgG-Fc receptor induced in cytomegalovirus-infected human fibroblasts. J Immunol 116, 772–777.
    [Google Scholar]
  23. Kim, J. S., Choi, S. E., Yun, I. H. & 8 other authors ( 2004; ). Human cytomegalovirus UL18 alleviated human NK-mediated swine endothelial cell lysis. Biochem Biophys Res Commun 315, 144–150.[CrossRef]
    [Google Scholar]
  24. Kotenko, S. V., Saccani, S., Izotova, L. S., Mirochnitchenko, O. V. & Pestka, S. ( 2000; ). Human cytomegalovirus harbors its own unique IL-10 homolog (cmvIL-10). Proc Natl Acad Sci U S A 97, 1695–1700.[CrossRef]
    [Google Scholar]
  25. Leong, C. C., Chapman, T. L., Bjorkman, P. J., Formankova, D., Mocarski, E. S., Phillips, J. H. & Lanier, L. L. ( 1998; ). Modulation of natural killer cell cytotoxicity in human cytomegalovirus infection: the role of endogenous class I major histocompatibility complex and a viral class I homolog. J Exp Med 187, 1681–1687.[CrossRef]
    [Google Scholar]
  26. Lepin, E. J., Bastin, J. M., Allan, D. S. & 8 other authors ( 2000; ). Functional characterization of HLA-F and binding of HLA-F tetramers to ILT2 and ILT4 receptors. Eur J Immunol 30, 3552–3561.[CrossRef]
    [Google Scholar]
  27. Lin, G., Simmons, G., Pohlmann, S. & 8 other authors ( 2003; ). Differential N-linked glycosylation of human immunodeficiency virus and Ebola virus envelope glycoproteins modulates interactions with DC-SIGN and DC-SIGNR. J Virol 77, 1337–1346.[CrossRef]
    [Google Scholar]
  28. Llano, M., Guma, M., Ortega, M., Angulo, A. & Lopez-Botet, M. ( 2003; ). Differential effects of US2, US6 and US11 human cytomegalovirus proteins on HLA class Ia and HLA-E expression: impact on target susceptibility to NK cell subsets. Eur J Immunol 33, 2744–2754.[CrossRef]
    [Google Scholar]
  29. Markine-Goriaynoff, N., Gillet, L., Van Etten, J. L., Korres, H., Verma, N. & Vanderplasschen, A. ( 2004; ). Glycosyltransferases encoded by viruses. J Gen Virol 85, 2741–2754.[CrossRef]
    [Google Scholar]
  30. McGrory, W. J., Bautista, D. S. & Graham, F. L. ( 1988; ). A simple technique for the rescue of early region I mutations into infectious human adenovirus type 5. Virology 163, 614–617.[CrossRef]
    [Google Scholar]
  31. Neote, K., DiGregorio, D., Mak, J. Y., Horuk, R. & Schall, T. J. ( 1993; ). Molecular cloning, functional expression, and signaling characteristics of a C-C chemokine receptor. Cell 72, 415–425.[CrossRef]
    [Google Scholar]
  32. Odeberg, J., Cerboni, C., Browne, H., Karre, K., Moller, E., Carbone, E. & Soderberg-Naucler, C. ( 2002; ). Human cytomegalovirus (HCMV)-infected endothelial cells and macrophages are less susceptible to natural killer lysis independent of the downregulation of classical HLA class I molecules or expression of the HCMV class I homologue, UL18. Scand J Immunol 55, 149–161.[CrossRef]
    [Google Scholar]
  33. Park, B., Oh, H., Lee, S., Song, Y., Shin, J., Sung, Y. C., Hwang, S.-Y. & Ahn, K. ( 2002; ). The MHC class I homolog of human cytomegalovirus is resistant to down-regulation mediated by the unique short region protein (US)2, US3, US6, and US11 gene products. J Immunol 168, 3464–3469.[CrossRef]
    [Google Scholar]
  34. Penfold, M. E. T., Dairaghi, D. J., Duke, G. M., Saederup, N., Mocarski, E. S., Kemble, G. W. & Schall, T. J. ( 1999; ). Cytomegalovirus encodes a potent α chemokine. Proc Natl Acad Sci U S A 96, 9839–9844.[CrossRef]
    [Google Scholar]
  35. Reyburn, H. T., Mandelboim, O., Valés-Gómez, M., Davis, D. M., Pazmany, L. & Strominger, J. L. ( 1997; ). The class I MHC homologue of human cytomegalovirus inhibits attack by natural killer cells. Nature 386, 514–517.[CrossRef]
    [Google Scholar]
  36. Saverino, D., Ghiotto, F., Merlo, A. & 12 other authors ( 2004; ). Specific recognition of the viral protein UL18 by CD85j/LIR-1/ILT2 on CD8+ T cells mediates the non-MHC-restricted lysis of human cytomegalovirus-infected cells. J Immunol 172, 5629–5637.[CrossRef]
    [Google Scholar]
  37. Shiroishi, M., Tsumoto, K., Amano, K. & 11 other authors ( 2003; ). Human inhibitory receptors Ig-like transcript 2 (ILT2) and ILT4 compete with CD8 for MHC class I binding and bind preferentially to HLA-G. Proc Natl Acad Sci U S A 100, 8856–8861.[CrossRef]
    [Google Scholar]
  38. Spaete, R. R. & Mocarski, E. S. ( 1987; ). Insertion and deletion mutagenesis of the human cytomegalovirus genome. Proc Natl Acad Sci U S A 84, 7213–7217.[CrossRef]
    [Google Scholar]
  39. Tomasec, P., Braud, V. M., Rickards, C. & 7 other authors ( 2000; ). Surface expression of HLA-E, an inhibitor of natural killer cells, enhanced by human cytomegalovirus gpUL40. Science 287, 1031.[CrossRef]
    [Google Scholar]
  40. Tomasec, P., Wang, E. C. Y., Davison, A. J. & 10 other authors ( 2005; ). Downregulation of natural killer cell-activating ligand CD155 by human cytomegalovirus UL141. Nat Immunol 6, 181–188.
    [Google Scholar]
  41. Tortorella, D., Gewurz, B., Schust, D., Furman, M. & Ploegh, H. ( 2000; ). Down-regulation of MHC class I antigen presentation by HCMV; lessons for tumor immunology. Immunol Invest 29, 97–100.[CrossRef]
    [Google Scholar]
  42. Valés-Gómez, M., Shiroishi, M., Maenaka, K. & Reyburn, H. T. ( 2005; ). Genetic variability of the major histocompatibility complex class I homologue encoded by human cytomegalovirus leads to differential binding to the inhibitory receptor ILT2. J Virol 79, 2251–2260.[CrossRef]
    [Google Scholar]
  43. Varnum, S. M., Streblow, D. N., Monroe, M. E. & 11 other authors ( 2004; ). Identification of proteins in human cytomegalovirus (HCMV) particles: the HCMV proteome. J Virol 78, 10960–10966.[CrossRef]
    [Google Scholar]
  44. Wilkinson, G. W. & Akrigg, A. ( 1992; ). Constitutive and enhanced expression from the CMV major IE promoter in a defective adenovirus vector. Nucleic Acids Res 20, 2233–2239.[CrossRef]
    [Google Scholar]
  45. Willcox, B. E., Thomas, L. M. & Bjorkman, P. J. ( 2003; ). Crystal structure of HLA-A2 bound to LIR-1, a host and viral major histocompatibility complex receptor. Nat Immunol 4, 913–919.
    [Google Scholar]
  46. Young, N. T., Uhrberg, M., Phillips, J. H., Lanier, L. L. & Parham, P. ( 2001; ). Differential expression of leukocyte receptor complex-encoded Ig-like receptors correlates with the transition from effector to memory CTL. J Immunol 166, 3933–3941.[CrossRef]
    [Google Scholar]
  47. Zhu, H., Shen, Y. & Shenk, T. ( 1995; ). Human cytomegalovirus IE1 and IE2 proteins block apoptosis. J Virol 69, 7960–7970.
    [Google Scholar]
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vol. , part 11, pp. 2999 - 3008

IMAGE

Relative mobility of highly glycosylated forms of UL18. HFFFs were infected with HCMV AD169 or RAd-UL18 and analysed by Western blotting with anti-UL18 mAb. UL18 detected during productive HCMV infection had slower mobility during SDS-PAGE.

IMAGE

Amino acid sequence alignment of UL18-encoding sequences from various HCMV strains. The UL18-encoding gene from strain Toledo and clinical isolates Merlin, 6397 and 3157 were sequenced, and the translated sequences were aligned with the prototype strain AD169 sequence (GenBank accession no. P08560) using Megalign (DNASTAR). Differences to the AD169 strain UL18-encoding sequence in the other strains are boxed in grey. The GenBank accession no. for clinical isolate 6397 is bankit736177, and for clinical isolate 3157 is bankit748724. The UL18 sequences of strain Toledo (AAS48934) and Merlin (YP_081477) were identical to those determined and deposited independently in GenBank, thus the existing accession numbers are cited here.

IMAGE

Amino acid sequence alignment of UL18 from various HCMV strains. The DNA sequences of UL18 genes for the GenBank-deposited strain sequences Towne (accession no. AC146851), FIX (AC146907), PH (AC146904) and TR (AC146906) were translated and aligned using Megalign (DNASTAR). Differences to the AD169 UL18 sequence (P08560) are boxed in grey.

IMAGE

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