Hamster polyomavirus-encoded proteins: gene cloning, heterologous expression and immunoreactivity.
The hamster polyomavirus (HaPV) is associated with spontaneously appearing skin epithelioma of the Syrian hamster Z3 strain. Virus particles prepared from the skin epithelioma cause lymphoma and leukemia when injected into newborn hamsters from a distinct Syrian hamster colony (HaP); in contrast to the skin epithelioma the hemopoietic tumors are virus free but accumulate viral DNA. To study the humoral immune response of HaPV-infected Z3 hamsters we produced recombinant HaPV proteins in Escherichia coli as beta-galactosidase-, TrpE- and dihydrofolate reductase-fusion proteins or as non-fused proteins. Recombinant plasmids carried segments of all putative early and late HaPV proteins. The recombinant proteins were detected in stained SDS polyacrylamide gels and in Western blots using monoclonal anti-TrpE and anti-beta-galactosidase antibodies and sera of HaPV-infected hamsters. Sera from HaPV-infected Z3 hamsters and crude lysates of all clones were applied to Western blots to characterize the humoral immune response in the animals. HaPV-specific antibodies were found to be directed against early protein segments translated from the first common exon and from the second unique exon of LT and MT, resp., as well as against the late proteins VP1 and VP2/3. The almost complete VP2 was recognized by all sera whereas VP1 was detected only by a half of the sera. Our data suggest the presence of at least 2 immunodominant regions in VP2, one in the C-terminal VP1 and at least 4 in early proteins.
Ulrich
R
R
Institut für Medizinische Virologie, Universitätsklinikum Charité der Humboldt-Universität, Berlin, Germany.
Sommerfeld
K
K
Schröder
A
A
Prokoph
H
H
Arnold
W
W
Krüger
D H
DH
Scherneck
S
S
D000818:Animals; D000917:Antibody Formation; D002277:Carcinoma / Q000662:veterinary / Q000821:virology; D003001:Cloning, Molecular; D006224:Cricetinae; D007938:Leukemia / Q000662:veterinary / Q000821:virology; D008223:Lymphoma / Q000662:veterinary / Q000821:virology; D008647:Mesocricetus; D011120:Polyomavirus / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry; D012878:Skin Neoplasms / Q000662:veterinary / Q000821:virology; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics*
D011994:Recombinant Proteins; D014764:Viral Proteins