Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1985 Jan;53(1):198–204. doi: 10.1128/jvi.53.1.198-204.1985

Detection of immunologically cross-reacting capsid protein of alphaviruses on the surfaces of infected L929 cells.

C Smith, J A Wolcott, C J Wust, A Brown
PMCID: PMC255008  PMID: 3965743

Abstract

Hyperimmune, but not normal immune, monospecific antiserum made to capsid protein of Sindbis virus (SIN) was found to cause cytolysis equally well of both SIN- and Semliki Forest virus-infected L929 cells in antibody-dependent, complement-mediated cytotoxicity assays. The cell surface reactivity of the hyperimmune antiserum was also demonstrated by solid-phase radioimmune assays with unfixed infected cells or infected cells fixed with low concentrations of glutaraldehyde (0.025%) before reactivity with antisera. Higher concentrations of glutaraldehyde lowered the sensitivity of detection. Purified SIN capsid protein specifically inhibited antibody-dependent, complement-mediated cytotoxicity by the monospecific anti-capsid protein serum on SIN- and Semliki Forest virus-infected target cells. That hyperimmune anti-SIN serum also cross-reacts with capsid protein on the surface of Semliki Forest virus-infected cells was suggested by the fact that capsid protein inhibited cross-cytolysis in the antibody-dependent, complement-mediated cytotoxicity assay. The latter antiserum was collected after repeated injections of purified virions over a 9-month period. The results suggest that hyperimmune monospecific antisera made to SIN capsid protein or hyperimmune antisera to SIN or Semliki Forest virions detect homologous and cross-reacting capsid protein determinants on the surface of infected cells.

Full text

PDF
198

Images in this article

201Image
on p.201

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aliperti G., Schlesinger M. J. Evidence for an autoprotease activity of sindbis virus capsid protein. Virology. 1978 Oct 15;90(2):366–369. doi: 10.1016/0042-6822(78)90321-5. [DOI] [PubMed] [Google Scholar]
  2. Bell J. R., Strauss E. G., Strauss J. H. Purification and amino acid compositions of the structural proteins of sindbis virus. Virology. 1979 Sep;97(2):287–294. doi: 10.1016/0042-6822(79)90340-4. [DOI] [PubMed] [Google Scholar]
  3. Bennink J. R., Yewdell J. W., Gerhard W. A viral polymerase involved in recognition of influenza virus-infected cells by a cytotoxic T-cell clone. Nature. 1982 Mar 4;296(5852):75–76. doi: 10.1038/296075a0. [DOI] [PubMed] [Google Scholar]
  4. Biddison W. E., Doherty P. C., Webster R. G. Antibody to influenza virus matrix protein detects a common antigen on the surface of cells infected with type A influenza viruses. J Exp Med. 1977 Sep 1;146(3):690–697. doi: 10.1084/jem.146.3.690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Braciale T. J. Immunologic recognition of influenza virus-infected cells. II. Expression of influenza A matrix protein on the infected cell surface and its role in recognition by cross-reactive cytotoxic T cells. J Exp Med. 1977 Sep 1;146(3):673–689. doi: 10.1084/jem.146.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  7. Cancedda R., Villa-Komaroff L., Lodish H. F., Schlesinger M. Initiation sites for translation of sindbis virus 42S and 26S messenger RNAs. Cell. 1975 Oct;6(2):215–222. doi: 10.1016/0092-8674(75)90012-4. [DOI] [PubMed] [Google Scholar]
  8. Clegg J. C., Chanas A. C., Gould E. A. Conformational changes in Sindbis virus E1 glycoprotein induced by monoclonal antibody binding. J Gen Virol. 1983 May;64(Pt 5):1121–1126. doi: 10.1099/0022-1317-64-5-1121. [DOI] [PubMed] [Google Scholar]
  9. Clegg J. C., Kennedy S. I. Initiation of synthesis of the structural proteins of Semliki Forest virus. J Mol Biol. 1975 Oct 5;97(4):401–411. doi: 10.1016/s0022-2836(75)80050-7. [DOI] [PubMed] [Google Scholar]
  10. Clegg J. C. Sequential translation of capsid and membrane protein genes of alphaviruses. Nature. 1975 Apr 3;254(5499):454–455. doi: 10.1038/254454a0. [DOI] [PubMed] [Google Scholar]
  11. Effros R. B., Doherty P. C., Gerhard W., Bennink J. Generation of both cross-reactive and virus-specific T-cell populations after immunization with serologically distinct influenza A viruses. J Exp Med. 1977 Mar 1;145(3):557–568. doi: 10.1084/jem.145.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garoff H., Frischauf A. M., Simons K., Lehrach H., Delius H. Nucleotide sequence of cdna coding for Semliki Forest virus membrane glycoproteins. Nature. 1980 Nov 20;288(5788):236–241. doi: 10.1038/288236a0. [DOI] [PubMed] [Google Scholar]
  13. Garoff H., Frischauf A. M., Simons K., Lehrach H., Delius H. The capsid protein of Semliki Forest virus has clusters of basic amino acids and prolines in its amino-terminal region. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6376–6380. doi: 10.1073/pnas.77.11.6376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Garoff H., Simons K., Renkonen O. Isolation and characterization of the membrane proteins of Semliki Forest virus. Virology. 1974 Oct;61(2):493–504. doi: 10.1016/0042-6822(74)90285-2. [DOI] [PubMed] [Google Scholar]
  15. Gates D., Brown A., Wust C. J. Comparison of specific and cross-reactive antigens of alphaviruses on virions and infected cells. Infect Immun. 1982 Jan;35(1):248–255. doi: 10.1128/iai.35.1.248-255.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Glanville N., Ranki M., Morser J., Käriäinen L., Smith A. E. Initiation of translation directed by 42S and 26S RNAs from Semliki Forest virus in vitro. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3059–3063. doi: 10.1073/pnas.73.9.3059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Grant J. P., Bigner D. D., Fischinger P. J., Bolognesi D. P. Expression of murine leukemia virus structural antigens on the surface of chemically induced murine sarcomas. Proc Natl Acad Sci U S A. 1974 Dec;71(12):5037–5041. doi: 10.1073/pnas.71.12.5037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. HUNTER W. M., GREENWOOD F. C. Preparation of iodine-131 labelled human growth hormone of high specific activity. Nature. 1962 May 5;194:495–496. doi: 10.1038/194495a0. [DOI] [PubMed] [Google Scholar]
  19. Hackett C. J., Askonas B. A., Webster R. G., Van Wyke K. Monoclonal antibodies to influenza matrix protein: detection of low levels of matrix protein on abortively infected cells. J Gen Virol. 1980 Apr;47(2):497–501. doi: 10.1099/0022-1317-47-2-497. [DOI] [PubMed] [Google Scholar]
  20. Holloway P. W. A simple procedure for removal of Triton X-100 from protein samples. Anal Biochem. 1973 May;53(1):304–308. doi: 10.1016/0003-2697(73)90436-3. [DOI] [PubMed] [Google Scholar]
  21. Hunsmann G., Claviez M., Moennig V., Schwarz H., Schäfer W. Properties of mouse leukemia viruses. X. Occurrence of viral structural antigens on the cell surface as revealed by a cytotoxicity test. Virology. 1976 Jan;69(1):157–168. doi: 10.1016/0042-6822(76)90203-8. [DOI] [PubMed] [Google Scholar]
  22. Kaluza G. Effect of impaired glycosylation on the biosynthesis of Semliki forest virus glycoproteins. J Virol. 1975 Sep;16(3):602–612. doi: 10.1128/jvi.16.3.602-612.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kaluza G., Rott R., Schwarz R. T. Carbohydrate-induced conformational changes of Semliki forest virus glycoproteins determine antigenicity. Virology. 1980 Apr 30;102(2):286–299. doi: 10.1016/0042-6822(80)90096-3. [DOI] [PubMed] [Google Scholar]
  24. Kees U., Krammer P. H. Most influenza A virus-specific memory cytotoxic T lymphocytes react with antigenic epitopes associated with internal virus determinants. J Exp Med. 1984 Feb 1;159(2):365–377. doi: 10.1084/jem.159.2.365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. Lerner R. A. Tapping the immunological repertoire to produce antibodies of predetermined specificity. Nature. 1982 Oct 14;299(5884):593–596. doi: 10.1038/299592a0. [DOI] [PubMed] [Google Scholar]
  28. Nowinski R. C., Watson A. Immune response of the mouse to the major core protein (p30) of ecotropic leukemia viruses. J Immunol. 1976 Aug;117(2):693–696. [PubMed] [Google Scholar]
  29. Powell K. L., Courtney R. J. Polypeptide synthesized in herpes simplex virus type 2-infected HEp-2 cells. Virology. 1975 Jul;66(1):217–228. doi: 10.1016/0042-6822(75)90192-0. [DOI] [PubMed] [Google Scholar]
  30. Rice C. M., Strauss J. H. Association of sindbis virion glycoproteins and their precursors. J Mol Biol. 1982 Jan 15;154(2):325–348. doi: 10.1016/0022-2836(82)90067-5. [DOI] [PubMed] [Google Scholar]
  31. Rice C. M., Strauss J. H. Nucleotide sequence of the 26S mRNA of Sindbis virus and deduced sequence of the encoded virus structural proteins. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2062–2066. doi: 10.1073/pnas.78.4.2062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rice C. M., Strauss J. H. Synthesis, cleavage and sequence analysis of DNA complementary to the 26 S messenger RNA of Sindbis virus. J Mol Biol. 1981 Aug 15;150(3):315–340. doi: 10.1016/0022-2836(81)90550-7. [DOI] [PubMed] [Google Scholar]
  33. Scheefers H., Scheefers-Borchel U., Edwards J., Brown D. T. Distribution of virus structural proteins and protein-protein interactions in plasma membrane of baby hamster kidney cells infected with Sindbis or vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7277–7281. doi: 10.1073/pnas.77.12.7277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shellam G. R., Knight R. A., Mitchison N. A., Gorczynski R. M., Maoz A. The specificity of effector T cells activated by tumours induced by murine oncornaviruses. Transplant Rev. 1976;29:249–276. doi: 10.1111/j.1600-065x.1976.tb00204.x. [DOI] [PubMed] [Google Scholar]
  35. Simmons D. T., Strauss J. H. Replication of Sindbis virus. I. Relative size and genetic content of 26 s and 49 s RNA. J Mol Biol. 1972 Nov 28;71(3):599–613. [PubMed] [Google Scholar]
  36. Smith J. F., Brown D. T. Envelopments of Sindbis virus: synthesis and organization of proteins in cells infected with wild type and maturation-defective mutants. J Virol. 1977 Jun;22(3):662–678. doi: 10.1128/jvi.22.3.662-678.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Virelizier J. L., Allison A. C., Oxford J. S., Schild G. C. Early presence of ribonucleoprotein antigen on surface of influenza virus-infected cells. Nature. 1977 Mar 3;266(5597):52–54. doi: 10.1038/266052a0. [DOI] [PubMed] [Google Scholar]
  39. Watanabe H., Mackenzie J. S. The detection of influenza A virus antigens in cultured cells by enzyme-linked immunosorbent assay. Arch Virol. 1981;67(1):31–43. doi: 10.1007/BF01314599. [DOI] [PubMed] [Google Scholar]
  40. Welch W. J., Sefton B. M. Characterization of a small, nonstructural viral polypeptide present late during infection of BHK cells by Semliki Forest virus. J Virol. 1980 Jan;33(1):230–237. doi: 10.1128/jvi.33.1.230-237.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Wirth D. F., Katz F., Small B., Lodish H. F. How a single Sindbis virus mRNA directs the synthesis of one soluble protein and two integral membrane glycoproteins. Cell. 1977 Feb;10(2):253–263. doi: 10.1016/0092-8674(77)90219-7. [DOI] [PubMed] [Google Scholar]
  42. Wolcott J. A., Gates D. W., Wust C. J., Brown A. Cross-reactive, cell-associated antigen on L929 cells infected with temperature-sensitive mutants of sindbis virus. Infect Immun. 1982 May;36(2):704–709. doi: 10.1128/iai.36.2.704-709.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wolcott J. A., Wust C. J., Brown A. Identification of immunologically cross-reactive proteins of Sindbis virus: evidence for unique conformation of E1 glycoprotein from infected cells. J Virol. 1984 Feb;49(2):379–385. doi: 10.1128/jvi.49.2.379-385.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wolcott J. A., Wust C. J., Brown A. Immunization with one alphavirus cross-primes cellular and humoral immune responses to a second alphavirus. J Immunol. 1982 Sep;129(3):1267–1271. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES