| Budget Amount *help |
¥11,000,000 (Direct Cost: ¥11,000,000)
Fiscal Year 1985: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1984: ¥10,000,000 (Direct Cost: ¥10,000,000)
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| Research Abstract |
All DNA segment of varicella zoster virus (VZV) except for terminal region were cloned with Ecoli vector (PBR 322, PVC 12). Variations in size of DNA segment were observed for HpaI-F,G K, EdoRI-P, KpnI-E, Sal I-E. The viable regions were investigated by Southerns hybridization with cloned VZV DNA and three variable regions (VRI, VRII, VRIII) could be mapped: VRI ( <+]-> 150 base pairs in variation, 0.16 map unit) VRII ( <+]-> 300bp 0.35, very unstable) VRIII( <+]-> 50bp, 0.95, relatively stable). No homology could be detected among DNAs of three variable regions. Many monoclonal antibodies against VZV glycoproteins(gp2, gp3, gp5) were isolated and used for analysis VZV glycoproteins. VZV gp3 was purified using monoclonal antibody to gp3 and monospecific antibody to gp3 was prepared in rabbit. While no neutralizing activity was found in monoclonal antibody against gp3, neutralizing activity was detected in monospecific antibody to gp3 indicating that gp3 is related to neutralization. Cross reactivity was found between VZV gp3 and HSV gB in immunofluorescent staing but there was no cross-reactivity in neutralization between then, which indicate that these two viruses have domain with common antigenicity but each has specific domain of neutralization.
A new method was devised to assay the potency of VZV skin test antigen (mainly gp5) prepared from culture fluid of VZV-infected human diploid cells. Human monoclonal antibody to VZV gp5 was adsorbed to wells of plates. Skin test antigen was added to it, and mouse-monoclonal antibody to gp5 was overlaied and antigen titer of skin test antigen was assay by ELISA. The titer thus obtained was well correlated with the skin test in imnuel individuals.
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