Measurement of Surface Antigen by Specific Bacterial Adherence and Scanning Electron Microscopy (SABA/SEM) in Cells Infected by Vesiculovirus ts Mutants
Temperature-sensitive (ts) mutants of the rhabdoviruses vesicular stomatitis virus and Chandipura virus have been used to measure the appearance of virus antigen on the surface of infected cells by the technique of surface analysis by bacterial adherence and scanning electron microscopy (SABA/SEM). The number of staphylococci specifically adhering to antiserum-treated infected PTK-2 or BSC-1 cells at permissive (31 °C) and restrictive (39 °C) temperatures was followed in time-course experiments and a close correspondence was observed between the proportion of staphylococci bound at 39 °C and the known phenotypic properties of the ts mutants. Virus surface antigen was undetected in cells infected by transcription- and replication-defective ts mutants with thermolabile L proteins under restrictive conditions up to an input multiplicity of infection of 50, and in cells infected by a replication-defective NS protein mutant. Some surface antigen was detected late in infection in PTK-2 cells infected by a replication-defective N protein mutant. Surface antigen accumulated normally in maturation-defective mutants with lesions in envelope proteins. These results establish the suitability of the SABA/SEM technique for quantitative estimation of virus antigen on the surface of infected cells.
EmersonS. U.,
YuY. E.1975; Both NS and L proteins are required for in vitro RNA synthesis by vesicular stomatitis virus. Journal of Virology 15:1348–1356
EvansD.,
PringleC. R.,
SzilagyiJ. F.1979; Temperature-sensitive mutants of complementation group E of vesicular stomatitis virus New Jersey serotype possess altered NS polypeptides. Journal of Virology 31:325–333
GadkariD. A.,
PringleC. R.1980; Temperature-sensitive mutants of Chandipura virus. II. Phenotypic characteristics of the six complementation groups. Journal of Virology 33:107–114
HuntD. M.,
EmersonS. U.,
WagnerR. R.1976; RNA~ temperature-sensitive mutants of vesicular stomatitis virus: L-protein thermosensitivity accounts for transcriptase restriction of group I mutants. Journal of Virology 18:596–603
LesnawJ. A.,
ReichmannM. E.1975; RNA synthesis by temperature-sensitive mutants of vesicular stomatitis virus, New Jersey serotype. Virology 63:492–504
ParryJ. E.,
ShirodariaP. V.,
PringleC. R.1979; Pneumoviruses: the cell surface of lytically and persistently infected cells. Journal of General Virology 44:479–491
PringleC. R.,
ParryJ. E.1980; Location and quantitation of antigen on the surface of virus-infected cells by specific bacterial adherence and scanning electron microscopy. Journal of Virological Methods 1:61–75
PringleC. R.,
SzilagyiJ. F.1980; Gene assignment and complementation group. In The Rhabdoviruses vol 2: pp 141–161 Edited by
BishopD. H. L.
Boca Raton: CRC Press;
PringleC. R.,
DuncanI. B.,
StevensonM.1971; Isolation and characterisation of temperature-sensitive mutants of vesicular stomatitis virus, New Jersey serotype. Journal of Virology 8:836–841
PringleC. R.,
DevineV.,
WilkieM.,
PrestonC. M.,
DolanA.,
MogeochD. J.1981; Enhanced mutability associated with a temperature-sensitive mutant of vesicular stomatitis virus. Journal of Virology 39:377–389
RepikP.,
FlamandA.,
BishopD. H. L.1976; Synthesis of RNA by mutants of vesicular stomatitis virus (Indiana serotype) and the ability of wild-type VSV New Jersey to complement the VSV Indiana ts GI-114 transcription defect. Journal of Virology 20:157–169
SzilagyiJ. F.,
PringleC. R.1972; Effect of temperature-sensitive mutations on the virion-associated RNA transcriptase of vesicular stomatitis virus. Journal of Molecular Biology 71:281–291
SzilagyiJ. F.,
PringleC. R.1979; Effect of temperature-sensitive mutation on activity of the RNA transcriptase of vesicular stomatitis virus New Jersey. Journal of Virology 30:692–700
WestmorelandD.,
St jeorS.,
RappF.1976; The development by cytomegalovirus-infected cells of binding affinity for normal human immunoglobulin. Journal of Immunology 116:1566–1570
Measurement of Surface Antigen by Specific Bacterial Adherence and Scanning Electron Microscopy (SABA/SEM) in Cells Infected by Vesiculovirus ts Mutants