Association of Sendai virion envelope and a mouse surface membrane polypeptide on newly infected cells: lack of association with H-2K/D or alteration of viral immunogenicity

Journal Article · Fri Jan 01 04:00:00 EST 1982 · J. Immunol.; (United States)

OSTI ID:5503520

Zarling, D A ^[1]; Miskimen, J A; Fan, D P; Fujimoto, E K; Smith, P K

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1. Univ. of Minnesota, St. Paul

The reagent N-succinimidyl 4-azidophenyl-1,3'-dithiopropionate (SADP) was synthesized and then coupled to purified Sendai virions by the amino-reactive end of the SADP molecule. This SADP-coupled virus was fused into the membranes of surface radioiodinated P815 cells, and target structures were allowed to form. Next, the photosensitive group on SADP was activated with ultraviolet light to covalently couple the viral proteins to any neighboring cell surface proteins. The cellular neighbors were isolated from detergent extracts of membrane proteins after immunoprecipitation with antibody specific for Sendai virion proteins. The covalent cross-links between the nonradioactive Sendai proteins and the radioiodinated cellular polypeptide neighbors were broken, and the host cell polypeptides were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and detected by autoradiography. One of these neighboring cellular proteins had an apparent m.w. of 17,000, and none was found with the characteristic size and tryptic map of either the H-2K or D gene products. Thus, the H-2K or D proteins are unlikely to be SADP- detectable neighbors of Sendai viral antigens recognized by CTL. In further experiments, the complexes of Sendai virion proteins crosslinked to cellular polypeptide neighbors were isolated from the membrane of newly infected cells and were shown to be able to stimulate CTL in vitro with approximately the same efficiency as uncross-linked Sendai virion proteins. Thus, Sendai viral proteins in the membrane of newly infected cells do not appear to be in highly immunogenic complexes with either H-2K/D or any other cellular proteins.

• OSTI ID: 5503520
• Journal Information: J. Immunol.; (United States), Journal Name: J. Immunol.; (United States) Vol. 128:1; ISSN JOIMA
• Country of Publication: United States
• Language: English

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63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANTIGENS
BETA DECAY RADIOISOTOPES
BIOLOGICAL EFFECTS
BIOLOGICAL FUNCTIONS
BIOLOGICAL MATERIALS
BIOLOGICAL RADIATION EFFECTS
BLOOD
BLOOD CELLS
BODY FLUIDS
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL REACTIONS
CONNECTIVE TISSUE CELLS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
ELECTROMAGNETIC RADIATION
ELECTRON CAPTURE RADIOISOTOPES
FUNCTIONS
IMMUNOLOGY
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LABELLED COMPOUNDS
LEUKOCYTES
LYMPHOCYTES
MATERIALS
MEMBRANES
MICROORGANISMS
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
PARASITES
PEPTIDES
POLYMERIZATION
PROTEINS
RADIATION EFFECTS
RADIATIONS
RADIOISOTOPES
RECEPTORS
SOMATIC CELLS
TRACER TECHNIQUES
ULTRAVIOLET RADIATION
VIRUSES