Skip to main content

Advertisement

SpringerLink
Log in

Immunogenicity analysis of prokaryotic expression products of Kaposi’s sarcoma associated herpesvirus orf65

  • Published:
Virologica Sinica

Abstract

To purify the protein encoding the small capsid protein (SCP) of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi’s sarcoma associated-herpesvirus (KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf65 was induced by isopropyl-β-D-thiogalactopyranoside (IPTG) and the fusion protein was purified by chromatography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and showed that 9 kDa was the expected size of the purified orf65 protein. The antiserum was produced in rabbit which was immunized by purified orf65 protein. An ELISA assay was established to analyze the immunogenicity of the purified orf65 protein. The ELISA analysis demonstrated that orf65 protein has strong immune activity, and the immune activity of polyclonal antibody against orf65 was more than 4 fold higher than that in the serum of the non-immunized rabbit. These results demonstrate that purified orf65 protein has very strong immunogenicity and can be used in screening KSHV infection in the general population using ELISA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Belec L, Cancre N, Hallouin M C, et al. 1998. High prevalence in Central Africa of blood donors who are potentially infectious for human herpesvirus 8. Transfusion, 38(8): 771–775.

    Article  PubMed  CAS  Google Scholar 

  2. Cesarman E, Chang Y, Moore P S, et al. 1995. Kaposi’s sarcoma-associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lymphomas. N Engl J Med, 332(18):1186–1191.

    Article  PubMed  CAS  Google Scholar 

  3. Chang Y, Cesarman E, Pessin M S, et al. 1994. Identification of herpesvirus-like DNA sequences in AIDS-associated Kaposi’s sarcoma. Science, 266(5192): 1865–1869.

    Article  PubMed  CAS  Google Scholar 

  4. Engels E A, Sinclair M D, Biggar R J, et al. 2000. Latent class analysis of human herpesvirus 8 assay performance and infection prevalence in sub-saharan Africa and Malta. Int J Cancer, 88(6): 1003–1008.

    Article  PubMed  CAS  Google Scholar 

  5. Engels E A, Whitby D, Goebel P B, et al. 2000. Identifying human herpesvirus 8 infection: performance characteristics of serologic assays. J Acquir Immune Defic Syndr, 23(4): 346–354.

    PubMed  CAS  Google Scholar 

  6. Fujii T, Taguchi H, Katano H, et al. 1999. Seroprevalence of human herpesvirus 8 in human immuno-deficiency virus 1-positive and human immunodeficiency virus 1-negative populations in Japan. J Med Virol, 57(2): 159–162.

    Article  PubMed  CAS  Google Scholar 

  7. Lin S F, Sun R, Heston L, et al. 1997. Identification, expression, and immunogenicity of Kaposi’s sarcoma-associated herpesvirus-encoded small viral capsid antigen. J Virol, 71(4): 3069–3076.

    PubMed  CAS  Google Scholar 

  8. Melbye M, Cook P M, Hjalgrim H, et al. 1998. Risk factors for Kaposi’s-sarcoma-associated herpesvirus (KSHV/ HHV-8) seropositivity in a cohort of homosexual men, 1981–1996. Int J Cancer, 77(4): 543–548.

    Article  PubMed  CAS  Google Scholar 

  9. Russo J J, Bohenzky R A, Chien M C, et al. 1996. Nucleotide sequence of the Kaposi sarcoma-associated herpesvirus (HHV8). Proc Natl Acad Sci USA, 93(25): 14862–14867.

    Article  PubMed  CAS  Google Scholar 

  10. Schulz T F. 1998. Kaposi’s sarcoma-associated herpesvirus (human herpesvirus-8). J Gen Virol, 79( Pt 7): 1573–1591.

    PubMed  CAS  Google Scholar 

  11. Simpson G R, Schulz T F, Whitby D, et al. 1996. Prevalence of Kaposi’s sarcoma associated herpesvirus infection measured by antibodies to recombinant capsid protein and latent immunofluorescence antigen. Lancet, 348(9035): 1133–1138.

    Article  PubMed  CAS  Google Scholar 

  12. Wang L. 2007. Pathogenesis and associated Diseases of Kaposi’s Sarcoma-associated Herpesvirus. Virol Sin, 22(3): 248–255..

    Article  CAS  Google Scholar 

  13. Whitby D, Luppi M, Barozzi P, et al. 1998. Human herpesvirus 8 seroprevalence in blood donors and lymphoma patients from different regions of Italy. J Natl Cancer, Inst90(5): 395–397.

    Article  Google Scholar 

  14. Zeng Y, Zhang X, Huang Z, et al. 2007. Intracellular Tat of human immunodeficiency virus type 1 activates lytic cycle replication of Kaposi’s sarcoma-associated herpes-virus: role of JAK/STAT signaling. J Virol, 81(5): 2401–2417.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, China

    Bi-shi Fu, Bao-lin Li & Lin-ding Wang

  2. Graduate School of the Chinese Academy of Sciences, Beijing, 100039, China

    Bi-shi Fu & Bao-lin Li

Authors
  1. Bi-shi Fu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bao-lin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lin-ding Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lin-ding Wang.

Additional information

Foundation items: Supported by the Knowledge Innovation Program of the Chinese Academy of Sciences (0702121Y-J1)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fu, Bs., Li, Bl. & Wang, Ld. Immunogenicity analysis of prokaryotic expression products of Kaposi’s sarcoma associated herpesvirus orf65. Virol. Sin. 23, 196–202 (2008). https://doi.org/10.1007/s12250-008-2925-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12250-008-2925-z

Key words

CLC number

Search

Navigation