Minor displacements in the insertion site provoke major differences in the induction of antibody responses by chimeric parvovirus-like particles.

An antigen-delivery system based on hybrid virus-like particles (VLPs) formed by the self-assembly of the capsid VP2 protein of canine parvovirus (CPV) and expressing foreign peptides was investigated. In this report, we have studied the effects of inserting the poliovirus C3:B epitope in the four loops and the C terminus of the CPV VP2 on the particle structure and immunogenicity. Epitope insertions in the four loops allowed the recovery of capsids in all of the mutants. However, only insertions of the C3:B epitope in VP2 residue 225 of the loop 2 were able to elicit a significant anti-peptide antibody response, but not poliovirus-neutralizing antibodies, probably because residue 225 is located in an small depression of the surface. To fine modulate the insertion site in loop 2, a cassette-mutagenesis was carried out to insert the epitope in adjacent positions 226, 227, and 228. The epitope C3:B inserted into these positions was well recognized by the specific monoclonal antibody C3 by immunoelectron microscopy. BALB/c mice immunized with these chimeric C3:B CPV:VLPs were able to elicit an strong neutralizing antibody response (>3 log(10) units) against poliovirus type 1 (Mahoney strain). Therefore, minor displacements in the insertion place cause dramatic changes in the accessibility of the epitope and the induction of antibody responses.

[1]  H. A. George,et al.  Continuous culture study of the expression of hepatitis B surface antigen and its self‐assembly into virus‐like particles in Saccharomyces cerevisiae , 2000, Biotechnology and bioengineering.

[2]  C. Parrish,et al.  Canine parvovirus host range is determined by the specific conformation of an additional region of the capsid , 1997, Journal of virology.

[3]  C. Leclerc,et al.  Recombinant parvovirus-like particles as an antigen carrier: a novel nonreplicative exogenous antigen to elicit protective antiviral cytotoxic T cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. Rossmann,et al.  Structural analysis of a mutation in canine parvovirus which controls antigenicity and host range. , 1996, Virology.

[5]  J. Casal,et al.  Identification of domains in canine parvovirus VP2 essential for the assembly of virus-like particles , 1996, Journal of virology.

[6]  C. Leclerc,et al.  Immunogenicity of poliovirus B and T cell epitopes presented by hybrid porcine parvovirus particles. , 1995, The Journal of general virology.

[7]  John E. Johnson,et al.  Eukaryotic viral expression systems for polypeptides , 1995 .

[8]  D. Tresnan,et al.  Analysis of the cell and erythrocyte binding activities of the dimple and canyon regions of the canine parvovirus capsid. , 1995, Virology.

[9]  John E. Johnson,et al.  Development of cowpea mosaic virus as a high-yielding system for the presentation of foreign peptides. , 1994, Virology.

[10]  M G Rossmann,et al.  The structure of a neutralized virus: canine parvovirus complexed with neutralizing antibody fragment. , 1994, Structure.

[11]  J. Van lent,et al.  Chimeric parvovirus B19 capsids for the presentation of foreign epitopes. , 1994, Virology.

[12]  M. Rossmann,et al.  The canine parvovirus empty capsid structure. , 1993, Journal of molecular biology.

[13]  R. Meloen,et al.  Topographical analysis of canine parvovirus virions and recombinant VP2 capsids. , 1993, The Journal of general virology.

[14]  M. S. Chapman,et al.  Structure, sequence, and function correlations among parvoviruses. , 1993, Virology.

[15]  R. Meloen,et al.  B-cell epitopes of canine parvovirus: distribution on the primary structure and exposure on the viral surface , 1993, Journal of virology.

[16]  C. Parrish,et al.  Multiple amino acids in the capsid structure of canine parvovirus coordinately determine the canine host range and specific antigenic and hemagglutination properties , 1992, Journal of virology.

[17]  E. Cortés,et al.  Recombinant vaccine for canine parvovirus in dogs , 1992, Journal of virology.

[18]  J. Garcia,et al.  Fine mapping of canine parvovirus B cell epitopes. , 1991, The Journal of general virology.

[19]  M S Chapman,et al.  The three-dimensional structure of canine parvovirus and its functional implications. , 1991, Science.

[20]  K. Matsubara,et al.  The essential region for assembly and particle formation in hepatitis B virus surface antigen produced in yeast cells. , 1990, Gene.

[21]  Y. Matsuura,et al.  Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. , 1987, The Journal of general virology.

[22]  J. Slot,et al.  A new method of preparing gold probes for multiple-labeling cytochemistry. , 1985, European journal of cell biology.

[23]  I. Singer,et al.  Canine parvovirus: a biochemical and ultrastructural characterization. , 1982, The Journal of general virology.

[24]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[25]  C. Parrish,et al.  Two dominant neutralizing antigenic determinants of canine parvovirus are found on the threefold spike of the virus capsid. , 1994, Virology.

[26]  M. Summers,et al.  A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedures. , 1987 .

[27]  P. Tattersall,et al.  The autonomously replicating parvoviruses of vertebrates. , 1987, Advances in virus research.

[28]  R. Crainic,et al.  Identification and characterization of a continuous neutralization epitope (C3) present on type 1 poliovirus. , 1987, Progress in medical virology. Fortschritte der medizinischen Virusforschung. Progres en virologie medicale.