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Human cytomegalovirus glycoprotein O as a new drug target and subunit vaccine candidate
| Details |
Inventors: Compton, Teresa; Huber, Mary T.;
Assignee: Wisconsin Alumni Research Foundation (Madison, WI)
Primary Examiner: Housel; James
Assistant Examiner: Li; Bao Qun
Attorney, Agent or Firm: Quarles & Brady LLP
A method of designing a new anti-CMV drug is disclosed. In one embodiment, the invention comprises (a) analyzing the binding of glycoprotein O to a glycoprotein O receptor and (b) designing a candidate drug that would competitively interfere with glycoprotein O binding to glycoprotein O receptor and (c) showing that the candidate drug competitively inhibits glycoprotein O binding to glycoprotein O receptor. A method of screening anti-CMV drugs, a vaccine effective to diminish CMV infection, and a method of diminishing CMV infection are also disclosed. |
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DETAILED DESCRIPTION OF THE DRAWINGS FIG.
1 is a diagram of HCMV entry into human fibroblasts.
FIG.
2 is a model of the tripartite gCIII complex in the HCMV envelope.
FIG.
3 is the predicted amino acid sequence of gO.
DETAILED DESCRIPTION OF THE INVENTION A.
In General Discovery of Glycoprotein O One envelope complex of CMV known to be required for entry of the virus is the glycoprotein H (gH)-containing complex.
Homologs of gH are found in all herpes viruses and are generally complexed with a second gene product, glycoprotein L (gL).
We initially attempted to reconstitute the gH complex of CMV from recombinant sources by co-expression of gH and gL in a baculovirus system.
However, these efforts were unsuccessful.
Using antibodies specific for gH, we found that the gH complex of CMV likely contained a third viral component (Huber, M.
T.
and T.
Compton, J.
Virol.
71:5391-8, 1997).
The genome of a laboratory strain of CMV is completely deduced (Chee, M.
S.
, et al.
, Curr.
Top.
Microbiol.
Immunol.
154(125):125-69, 1990), and analysis of open reading frames encoded in the genome revealed that as many as 55 of the 208 genes of CMV potentially encode envelope glycoproteins.
To date, however, only 5 genes are linked to defined envelope protein components.
To determine which gene may encode the third gH-complex component, we subjected a preparation of purified protein to microsequence analysis.
Our analysis of the results of the protein sequence allowed us to identify this protein as the product of a particular gene (UL74), one of the predicted envelope glycoproteins.
We designated the UL74 gene product as glycoprotein O (gO) (Huber, M.
T.
and T.
Compton, J.
Virol.
72:8191-97, 1998).
Potential Function of gO To determine whether gO was important in the lifecycle of CMV, we produced a high-titer antiserum.
This antiserum is monospecific for gO and is capable of precipitating gO and its complexing partners, gH and gL (Huber, M.
T.
and T.
Compton, supra, 1998) in CMV-infected cells (Huber, M.
T.
and T
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