1887

Abstract

Our earlier identification of the non-structural protein 3 (NS3) of Japanese encephalitis virus (JEV) as a dominant CD4 as well as CD8 T cell-eliciting antigen in a healthy JEV-endemic cohort with a wide HLA distribution implied the presence of several epitopes dispersed over the length of the protein. Use of various truncated versions of NS3 in lymphocyte stimulation and interferon (IFN)- secretion assays revealed that amino acids (aa) 193–324 of NS3 were comparable with, if not superior to, the full-length protein in evoking Th1 responses. The potential of this 14·4 kDa stretch to stimulate IFN- production from both subtypes of T cells in a manner qualitatively and quantitatively similar to the 68 kDa parent protein suggested the presence within it of both class I and II epitopes and demonstrated that the entire immunogenicity of NS3 was focused on aa 193–324. Interestingly, this segment contained five of the eight helicase motifs of NS3. Analysis of variability of the NS3 protein sequence across 16 JEV isolates revealed complete identity of aa 219–318, which is contained within the above segment, suggesting that NS3-specific epitopes tend to cluster in relatively conserved regions that harbour functionally critical domains of the protein.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.19698-0
2004-05-01
2019-11-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/85/5/vir851131.html?itemId=/content/journal/jgv/10.1099/vir.0.19698-0&mimeType=html&fmt=ahah

References

  1. Aihara, H., Takasaki, T., Matsutani, T., Suzuki, R. & Kurane, I. ( 1998; ). Establishment and characterization of Japanese encephalitis virus-specific, human CD4+ T-cell clones: flavivirus cross-reactivity, protein recognition, and cytotoxic activity. J Virol 72, 8032–8036.
    [Google Scholar]
  2. Becker-Hapak, M., McAllister, S. S. & Dowdy, S. F. ( 2001; ). TAT-mediated protein transduction into mammalian cells. Methods 24, 247–256.[CrossRef]
    [Google Scholar]
  3. Billoir, F., de Chesse, R., Tolou, H., de Micco, P., Gould, E. A. & de Lamballerie, X. ( 2000; ). Phylogeny of the genus Flavivirus using complete coding sequences of arthropod-borne viruses and viruses with no known vector. J Gen Virol 81, 781–790.
    [Google Scholar]
  4. Co, M. D., Terajima, M., Cruz, J., Ennis, F. A. & Rothman, A. L. ( 2002; ). Human cytotoxic T lymphocyte responses to live attenuated 17D yellow fever vaccine: identification of HLA-B35-restricted CTL epitopes on nonstructural proteins NS1, NS2b, NS3, and the structural protein E. Virology 293, 151–163.[CrossRef]
    [Google Scholar]
  5. Desai, A., Ravi, V., Chandramuki, A. & Gourie-Devi, M. ( 1995; ). Proliferative response of human peripheral blood mononuclear cells to Japanese encephalitis virus. Microbiol Immunol 39, 269–273.[CrossRef]
    [Google Scholar]
  6. Diepolder, H., Gerlach, J., Zachoval, R. & 9 other authors ( 1997; ). Immunodominant CD4+ T-cell epitope within nonstructural protein 3 in acute hepatitis C virus infection. J Virol 71, 6011–6019.
    [Google Scholar]
  7. Hall, T. ( 1999; ). Bioedit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41, 95–98.
    [Google Scholar]
  8. Konishi, E., Kurane, I., Mason, P. W., Innis, B. L. & Ennis, F. A. ( 1995; ). Japanese encephalitis virus-specific proliferative responses of human peripheral blood T lymphocytes. Am J Trop Med Hyg 53, 278–283.
    [Google Scholar]
  9. Ku, C. C., King, C. C., Lin, C. Y., Hsu, H. C., Chen, L. Y., Yueh, Y. Y. & Chang, G. J. ( 1994; ). Homologous and heterologous neutralization antibody responses after immunization with Japanese encephalitis vaccine among Taiwan children. J Med Virol 44, 122–131.[CrossRef]
    [Google Scholar]
  10. Kumar, P., Amara, R. R., Challu, V. K., Chadda, V. K. & Satchidanandam, V. ( 2003a; ). The Apa protein of Mycobacterium tuberculosis stimulates gamma interferon-secreting CD4+ and CD8+ T cells from purified protein derivative-positive individuals and affords protection in a guinea pig model. Infect Immun 71, 1929–1937.[CrossRef]
    [Google Scholar]
  11. Kumar, P., Uchil, P. D., Sulochana, P., Nirmala, G., Chandrashekar, R., Haridattatreya, M. & Satchidanandam, V. ( 2003b; ). Screening for T cell-eliciting proteins of Japanese encephalitis virus in a healthy JE-endemic human cohort using recombinant baculovirus-infected insect cell preparations. Arch Virol 148, 1569–1591.[CrossRef]
    [Google Scholar]
  12. Kumar, P., Krishna, V., Sulochana, P., Nirmala, G., Haridattatreya, M. & Satchidanandam, V. ( 2004a; ). Cell mediated immune responses in healthy children with a history of Japanese encephalitis virus subclinical infection: analysis of CD4+ and CD8+ T cell target specificities by intracellular delivery of viral proteins using the HIV Tat protein transduction domain. J Gen Virol 85, 471–482.[CrossRef]
    [Google Scholar]
  13. Kumar, P., Sulochana, P., Nirmala, G., Haridattatreya, M. & Satchidanandam, V. ( 2004b; ). Impaired Th1 function of NS3-specific T cells in Japanese encephalitis patients with neurological sequelae. J Infect Dis (in press).
    [Google Scholar]
  14. Kurane, I., Dai, L. C., Livingston, P. G., Reed, E. & Ennis, F. A. ( 1993; ). Definition of an HLA-DPw2-restricted epitope on NS3, recognized by a dengue virus serotype-cross-reactive human CD4+ CD8 cytotoxic T-cell clone. J Virol 67, 6285–6288.
    [Google Scholar]
  15. Kurane, I., Okamoto, Y., Dai, L. C., Zeng, L. L., Brinton, M. A. & Ennis, F. A. ( 1995; ). Flavivirus-cross-reactive, HLA-DR15-restricted epitope on NS3 recognized by human CD4+ CD8 cytotoxic T lymphocyte clones. J Gen Virol 76, 2243–2249.[CrossRef]
    [Google Scholar]
  16. Kurane, I., Zeng, L., Brinton, M. A. & Ennis, F. A. ( 1998; ). Definition of an epitope on NS3 recognized by human CD4+ cytotoxic T lymphocyte clones cross-reactive for dengue virus types 2, 3, and 4. Virology 240, 169–174.[CrossRef]
    [Google Scholar]
  17. Lamonaca, V., Missale, G., Urbani, S. & 10 other authors ( 1999; ). Conserved hepatitis C virus sequences are highly immunogenic for CD4+ T cells: implications for vaccine development. Hepatology 30, 1088–1098.[CrossRef]
    [Google Scholar]
  18. Li, W. H., Tanimura, M. & Sharp, P. M. ( 1988; ). Rates and dates of divergence between AIDS virus nucleotide sequences. Mol Biol Evol 5, 313–330.
    [Google Scholar]
  19. Lin, C. & Kim, J. L. ( 1999; ). Structure-based mutagenesis study of hepatitis C virus NS3 helicase. J Virol 73, 8798–8807.
    [Google Scholar]
  20. Lobigs, M., Arthur, C. E., Mullbacher, A. & Blanden, R. V. ( 1994; ). The flavivirus nonstructural protein NS3 is a dominant source of cytotoxic T cell peptide determinants. Virology 202, 195–201.[CrossRef]
    [Google Scholar]
  21. Loke, H., Bethell, D. B., Phuong, C. X., Dung, M., Schneider, J., White, N. J., Day, N. P., Farrar, J. & Hill, A. V. ( 2001; ). Strong HLA class I-restricted T cell responses in dengue hemorrhagic fever: a double-edged sword? J Infect Dis 184, 1369–1373.[CrossRef]
    [Google Scholar]
  22. Mackenzie, J. S., Chua, K. B., Daniels, P. W. & 18 other authors ( 2001; ). Emerging viral diseases of southeast Asia and the western Pacific. Emerg Infect Dis 7, 497–504.[CrossRef]
    [Google Scholar]
  23. Mackenzie, J. S., Barrett, A. D. T. & Deubel, V. ( 2002; ). Japanese Encephalitis and West Nile Viruses. Current Topics in Microbiology and Immunology, vol. 267. Edited by R. W. Compans, M. D. Cooper, H. Koprowski, M. B. A. Oldstone, M. Potter & P. K. Vogt. New York: Springer.
  24. Mathew, A., Kurane, I., Rothman, A. L., Zeng, L. L., Brinton, M. A. & Ennis, F. A. ( 1996; ). Dominant recognition by human CD8+ cytotoxic T lymphocytes of dengue virus nonstructural proteins NS3 and NS1.2a. J Clin Invest 98, 1684–1691.[CrossRef]
    [Google Scholar]
  25. Mathew, A., Kurane, I., Green, S. & 7 other authors ( 1998; ). Predominance of HLA-restricted cytotoxic T-lymphocyte responses to serotype-cross-reactive epitopes on nonstructural proteins following natural secondary dengue virus infection. J Virol 72, 3999–4004.
    [Google Scholar]
  26. Matusan, A. E., Pryor, M. J., Davidson, A. D. & Wright, P. J. ( 2001; ). Mutagenesis of the dengue virus type 2 NS3 protein within and outside helicase motifs: effects on enzyme activity and virus replication. J Virol 75, 9633–9643.[CrossRef]
    [Google Scholar]
  27. Monath, P. T. ( 1999; ). Yellow fever. In Vaccines, 3rd edn, pp. 815–879. Edited by S. A. Plotkins & W. A. Orenstein. Philadelphia, PA: W. B. Saunders.
  28. Okamoto, Y., Kurane, I., Leporati, A. M. & Ennis, F. A. ( 1998; ). Definition of the region on NS3 which contains multiple epitopes recognized by dengue virus serotype-cross-reactive and flavivirus-cross-reactive, HLA-DPw2-restricted CD4+ T cell clones. J Gen Virol 79, 697–704.
    [Google Scholar]
  29. Pan, C. H., Chen, H. W., Huang, H. W. & Tao, M. H. ( 2001; ). Protective mechanisms induced by a Japanese encephalitis virus DNA vaccine: requirement for antibody but not CD8+ cytotoxic T-cell responses. J Virol 75, 11457–11463.[CrossRef]
    [Google Scholar]
  30. Rao-Bhau, L. N., Singh, G., Gowal, D., Saxena, S. N., Kobayashi, M., Oya, A. & Yoshioka, I. ( 1988; ). Safety and efficacy of Japanese encephalitis vaccine produced in India. Indian J Med Res 88, 301–307.
    [Google Scholar]
  31. Regner, M., Lobigs, M., Blanden, R. V., Milburn, P. & Mullbacher, A. ( 2001; ). Antiviral cytotoxic T cells cross-reactively recognize disparate peptide determinants from related viruses but ignore more similar self- and foreign determinants. J Immunol 166, 3820–3828.[CrossRef]
    [Google Scholar]
  32. Rice, C. M. & Lindenbach, B. D. ( 2001; ). Flaviviridae. In Fields Virology, 4th edn, pp. 991–1041. Edited by D. M. Knipe & P. M. Howley. Philadelphia, PA: Lippincott Williams & Wilkins.
  33. Thimme, R., Chang, K.-M., Pemberton, J., Sette, A. & Chisari, F. V. ( 2001; ). Degenerate immunogenicity of an HLA-A2-restricted hepatitis B virus nucleocapsid cytotoxic T-lymphocyte epitope that is also presented by HLA-B51. J Virol 75, 3984–3987.[CrossRef]
    [Google Scholar]
  34. Ting, S. H., See, E., Tan, H. C., Lee, M. A. & Ooi, E. E. ( 2001; ). Development of a simplified assay for the detection of neutralizing antibodies to Japanese encephalitis virus. J Virol Methods 93, 43–47.[CrossRef]
    [Google Scholar]
  35. Tomiyama, H., Yamada, N., Komatsu, H., Hirayama, K. & Takiguchi, M. ( 2000; ). A single CTL clone can recognize a naturally processed HIV-1 epitope presented by two different HLA class I molecules. Eur J Immunol 30, 2521–2530.[CrossRef]
    [Google Scholar]
  36. Tsai, T. F., Chang, G. J. & Yu, Y. X. ( 1999; ). Japanese encephalitis vaccines. In Vaccines, 3rd edn, pp. 672–710. Edited by S. A. Plotkins & W. A. Orenstein. Philadelphia, PA: W. B. Saunders.
  37. Yusim, K., Kesmir, C., Gaschen, B., Addo, M. M., Altfeld, M., Brunak, S., Chigaev, A., Detours, V. & Korber, B. T. ( 2002; ). Clustering patterns of cytotoxic T-lymphocyte epitopes in human immunodeficiency virus type 1 (HIV-1) proteins reveal imprints of immune evasion on HIV-1 global variation. J Virol 76, 8757–8768.[CrossRef]
    [Google Scholar]
  38. Zanotto, P. M., Gould, E. A., Gao, G. F., Harvey, P. H. & Holmes, E. C. ( 1996; ). Population dynamics of flaviviruses revealed by molecular phylogenies. Proc Natl Acad Sci U S A 93, 548–553.[CrossRef]
    [Google Scholar]
  39. Zeng, L., Kurane, I., Okamoto, Y., Ennis, F. A. & Brinton, M. A. ( 1996; ). Identification of amino acids involved in recognition by dengue virus NS3-specific, HLA-DR15-restricted cytotoxic CD4+ T-cell clones. J Virol 70, 3108–3117.
    [Google Scholar]
  40. Zivna, I., Green, S., Vaughn, D. W., Kalayanarooj, S., Stephens, H. A., Chandanayingyong, D., Nisalak, A., Ennis, F. A. & Rothman, A. L. ( 2002; ). T cell responses to an HLA-B*07-restricted epitope on the dengue NS3 protein correlate with disease severity. J Immunol 168, 5959–5965.[CrossRef]
    [Google Scholar]
  41. Zivny, J., DeFronzo, M., Jarry, W., Jameson, J., Cruz, J., Ennis, F. A. & Rothman, A. L. ( 1999; ). Partial agonist effect influences the CTL response to a heterologous dengue virus serotype. J Immunol 163, 2754–2760.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.19698-0
Loading
/content/journal/jgv/10.1099/vir.0.19698-0
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error