Identification of novel, serologically reactive recombinant Varicella-Zoster Virus (VZV) antigens by a systematic, genome-scale approach.

VIZOSO PINTO, M. G.; PFREPPER, K-I.; NOELTING, C.; JANKE, T.; LUEKING, A.; HAAS, J.; NITSCHKO, H.; JAEGER, G.; BAIKER, A.

Cernobbio, Como lake, Italy

Congreso; Fourth European Congress of Virology; 2010

Background: Varicella-Zoster virus (VZV) is a member of thealphaherpesvirus subfamily, causing chickenpox upon primary infection and shingles after reactivation from latency.Currently available serological tests to detect VZV-specific antibodies from patient samples are exclusively basedon antigens derived from VZV-infected cell culture and suffer from a number of limitations. Methods: All 71known VZV open reading frames (ORFs) were recombinatorially cloned into a customized, N-terminally His-taggedbacterial expression vector and analyzed for small scale recombinant protein expression and purification. All purifiedrecombinant VZV proteins were subsequently examined for their antigenic potential by serum profiling experimentsin a microarray format. All identified novel VZV antigens were rearranged in line assay format and validated. Results:Initial serum profiling experiments using purified, His-tagged VZV proteins and clinically defined VZV patient serain a microarray format revealed 6 putative candidate antigens, which exhibit reactivity with sera from patients withdifferent stages of VZV infection. These antigens (i.e. ORF1, ORF4, ORF14, ORF49, ORF62, and ORF68) wererearranged in a line assay format and validated with a number of defined serum samples. The results of theseexperiments confirmed the seroreactivity of the identified antigens and revealed the suitability of the recombinantline assay for VZV serodiagnostics. Future experiments will reveal whether this novel line assay may be utilized forthe diagnosis of distinct VZV-associated diseases and for the control of varicella vaccination response. Conclusion:We have developed a systematic, genome-scale screening approach for the identification of novel, serologicallyreactive recombinant antigens.