Volume 5 Supplement 1

Proceedings of the 12-th International Conference on Malignancies in AIDS and Other Acquired Immunodeficiencies (ICMAOI)

Open Access

Development of a proteomic platform for EBV and KSHV serological screening

  • Leyao Wang1Email author,
  • Dasheng Zheng1,
  • Jian Zhu2,
  • Gangling Liao1,
  • Crystal Woodard2,
  • C-J Chiou1,
  • Gary Hayward1, 3,
  • Prashant Desai1, 3,
  • Richard Ambinder1, 3,
  • Heng Zhu2, 3 and
  • S Diane Hayward1, 3
Infectious Agents and Cancer20105(Suppl 1):A18

https://doi.org/10.1186/1750-9378-5-S1-A18

Published: 11 October 2010

In the context of HIV infection, there is an increased susceptibility to EBV- and KSHV-associated malignancies. Evaluating a possible role for specific humoral responses to individual viral proteins in the control of viral load and prevention of EBV or KSHV related disease development in this population has been hampered by the limited availability of suitable screening reagents.

We have developed a protein array platform that can be utilized for EBV and KSHV serological screening. The array consists of EBV and KSHV proteins printed onto glass slides. All eighty-four EBV open reading frames and eighty-six KSHV open reading frames were cloned into bacterial vectors and the inserts validated by DNA sequencing in both directions. Authenticated inserts were transferred to a yeast vector that expresses proteins as N-terminal GST-fusions. Seventy-nine EBV proteins and eighty-one KSHV proteins were successfully purified from yeast. First-generation EBV/KSHV protein arrays have been printed with these proteins plus a variety of control proteins that include GST, and human IgG, IgM, and IgA.

Glycosylation of viral proteins may affect immunogenicity. To evaluate this aspect, EBV and KSHV virion glycoproteins are being expressed in insect cells modified with the human glycosylation machinery. Once purified, these proteins will be added to the EBV/KSHV protein arrays.

The ability to compare serological responses to the complete repertoire of individual EBV and KSHV encoded proteins should provide new insight into B cell mediated immune regulation of these viruses.

Declarations

Acknowledgements

This article has been published as part of Infectious Agents and Cancer Volume 5 Supplement 1, 2010: Proceedings of the 12th International Conference on Malignancies in AIDS and Other Acquired Immunodeficiencies (ICMAOI).The full contents of the supplement are available online at http://www.biomedcentral.com/1750-9378/5?issue=S1.

Authors’ Affiliations

(1)
Department of Oncology, Johns Hopkins School of Medicine
(2)
Department of Pharmacology, Johns Hopkins School of Medicine
(3)
Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins School of Medicine

Copyright

© Wang et al; licensee BioMed Central Ltd. 2010

This article is published under license to BioMed Central Ltd.

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