Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1989 Sep;63(9):4064–4068. doi: 10.1128/jvi.63.9.4064-4068.1989

Use of TrpE/Gag fusion proteins to characterize immunoreactive domains on the human immunodeficiency virus type 1 core protein.

M G Windheuser 1, G E Tegtmeier 1, C Wood 1
PMCID: PMC251008  PMID: 2474676

Abstract

The human immunodeficiency virus (HIV) p24 core protein is one of the most immunogenic of HIV structural proteins. Infected individuals develop high titers of antibodies against p24 early in infection, which makes anti-p24 antibodies important serological markers. However, despite the clinical importance of the anti-p24 response, no systematic study to characterize the antigenic domains on the p24 protein has been reported. We report here on the use of 12 overlapping fragments of the HIV type 1 p24 protein, synthesized in bacteria as TrpE/Gag fusion proteins, to identify at least two and possibly three antigenic domains on the p24 protein. In addition, we note that different HIV-seropositive sera exhibited different patterns of reactivity with the p24 domains presented on our fusion proteins.

Full text

PDF
4064

Images in this article

4065Image
on p.4065
4066Image
on p.4066

Selected References

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

  1. Biggar R. J., Melbye M., Ebbesen P., Alexander S., Nielsen J. O., Sarin P., Faber V. Variation in human T lymphotropic virus III (HTLV-III) antibodies in homosexual men: decline before onset of illness related to acquired immune deficiency syndrome (AIDS). Br Med J (Clin Res Ed) 1985 Oct 12;291(6501):997–998. doi: 10.1136/bmj.291.6501.997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cao Y. Z., Valentine F., Hojvat S., Allain J. P., Rubinstein P., Mirabile M., Czelusniak S., Leuther M., Baker L., Friedman-Kien A. E. Detection of HIV antigen and specific antibodies to HIV core and envelope proteins in sera of patients with HIV infection. Blood. 1987 Aug;70(2):575–578. [PubMed] [Google Scholar]
  4. Casey J. M., Kim Y., Andersen P. R., Watson K. F., Fox J. L., Devare S. G. Human T-cell lymphotropic virus type III: immunologic characterization and primary structure analysis of the major internal protein, p24. J Virol. 1985 Aug;55(2):417–423. doi: 10.1128/jvi.55.2.417-423.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Debouck C., Gorniak J. G., Strickler J. E., Meek T. D., Metcalf B. W., Rosenberg M. Human immunodeficiency virus protease expressed in Escherichia coli exhibits autoprocessing and specific maturation of the gag precursor. Proc Natl Acad Sci U S A. 1987 Dec;84(24):8903–8906. doi: 10.1073/pnas.84.24.8903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Henikoff S. Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol. 1987;155:156–165. doi: 10.1016/0076-6879(87)55014-5. [DOI] [PubMed] [Google Scholar]
  7. Kanner S. B., Cheng-Mayer C., Geffin R. B., Parks W. P., Beltz G. A., Arthur L. O., Samuel K. P., Papas T. S. Human retroviral env and gag polypeptides: serologic assays to measure infection. J Immunol. 1986 Jul 15;137(2):674–678. [PubMed] [Google Scholar]
  8. Kappes J. C., Morrow C. D., Lee S. W., Jameson B. A., Kent S. B., Hood L. E., Shaw G. M., Hahn B. H. Identification of a novel retroviral gene unique to human immunodeficiency virus type 2 and simian immunodeficiency virus SIVMAC. J Virol. 1988 Sep;62(9):3501–3505. doi: 10.1128/jvi.62.9.3501-3505.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kitchen L., Malone G., Orgad S., Barin F., Zaizov R., Ramot B., Gazit E., Kreiss J., Leal M., Wichmann I. Viral envelope protein of HTLV-III is the major target antigen for antibodies in hemophiliac patients. J Infect Dis. 1986 Apr;153(4):788–790. doi: 10.1093/infdis/153.4.788. [DOI] [PubMed] [Google Scholar]
  10. Kleid D. G., Yansura D., Small B., Dowbenko D., Moore D. M., Grubman M. J., McKercher P. D., Morgan D. O., Robertson B. H., Bachrach H. L. Cloned viral protein vaccine for foot-and-mouth disease: responses in cattle and swine. Science. 1981 Dec 4;214(4525):1125–1129. doi: 10.1126/science.6272395. [DOI] [PubMed] [Google Scholar]
  11. Kraft R., Tardiff J., Krauter K. S., Leinwand L. A. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544-6, 549. [PubMed] [Google Scholar]
  12. Marcus-Sekura C. J., Woerner A. M., Klutch M., Quinnan G. V., Jr Reactivity of an HIV gag gene polypeptide expressed in E. coli with sera from AIDS patients and monoclonal antibodies to gag. Biochim Biophys Acta. 1988 Feb 28;949(2):213–223. doi: 10.1016/0167-4781(88)90085-1. [DOI] [PubMed] [Google Scholar]
  13. McDougal J. S., Kennedy M. S., Nicholson J. K., Spira T. J., Jaffe H. W., Kaplan J. E., Fishbein D. B., O'Malley P., Aloisio C. H., Black C. M. Antibody response to human immunodeficiency virus in homosexual men. Relation of antibody specificity, titer, and isotype to clinical status, severity of immunodeficiency, and disease progression. J Clin Invest. 1987 Aug;80(2):316–324. doi: 10.1172/JCI113075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mervis R. J., Ahmad N., Lillehoj E. P., Raum M. G., Salazar F. H., Chan H. W., Venkatesan S. The gag gene products of human immunodeficiency virus type 1: alignment within the gag open reading frame, identification of posttranslational modifications, and evidence for alternative gag precursors. J Virol. 1988 Nov;62(11):3993–4002. doi: 10.1128/jvi.62.11.3993-4002.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pal R., Gallo R. C., Sarngadharan M. G. Processing of the structural proteins of human immunodeficiency virus type 1 in the presence of monensin and cerulenin. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9283–9286. doi: 10.1073/pnas.85.23.9283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pedersen C., Nielsen C. M., Vestergaard B. F., Gerstoft J., Krogsgaard K., Nielsen J. O. Temporal relation of antigenaemia and loss of antibodies to core antigens to development of clinical disease in HIV infection. Br Med J (Clin Res Ed) 1987 Sep 5;295(6598):567–569. doi: 10.1136/bmj.295.6598.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ratner L., Haseltine W., Patarca R., Livak K. J., Starcich B., Josephs S. F., Doran E. R., Rafalski J. A., Whitehorn E. A., Baumeister K. Complete nucleotide sequence of the AIDS virus, HTLV-III. Nature. 1985 Jan 24;313(6000):277–284. doi: 10.1038/313277a0. [DOI] [PubMed] [Google Scholar]
  18. Robey W. G., Safai B., Oroszlan S., Arthur L. O., Gonda M. A., Gallo R. C., Fischinger P. J. Characterization of envelope and core structural gene products of HTLV-III with sera from AIDS patients. Science. 1985 May 3;228(4699):593–595. doi: 10.1126/science.2984774. [DOI] [PubMed] [Google Scholar]
  19. Sanchez-Pescador R., Power M. D., Barr P. J., Steimer K. S., Stempien M. M., Brown-Shimer S. L., Gee W. W., Renard A., Randolph A., Levy J. A. Nucleotide sequence and expression of an AIDS-associated retrovirus (ARV-2). Science. 1985 Feb 1;227(4686):484–492. doi: 10.1126/science.2578227. [DOI] [PubMed] [Google Scholar]
  20. Schüpbach J., Haller O., Vogt M., Lüthy R., Joller H., Oelz O., Popovic M., Sarngadharan M. G., Gallo R. C. Antibodies to HTLV-III in Swiss patients with AIDS and pre-AIDS and in groups at risk for AIDS. N Engl J Med. 1985 Jan 31;312(5):265–270. doi: 10.1056/NEJM198501313120502. [DOI] [PubMed] [Google Scholar]
  21. Shoeman R. L., Young D., Pottathil R., Victor J., Conroy R. R., Crowl R. M., Coleman T., Heimer E., Lai C. Y., Ganguly K. Comparison of recombinant human immunodeficiency virus gag precursor and gag/env fusion proteins and a synthetic env peptide as diagnostic reagents. Anal Biochem. 1987 Mar;161(2):370–379. doi: 10.1016/0003-2697(87)90465-9. [DOI] [PubMed] [Google Scholar]
  22. Spindler K. R., Rosser D. S., Berk A. J. Analysis of adenovirus transforming proteins from early regions 1A and 1B with antisera to inducible fusion antigens produced in Escherichia coli. J Virol. 1984 Jan;49(1):132–141. doi: 10.1128/jvi.49.1.132-141.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Windheuser M. G., Wood C. Characterization of immunoreactive epitopes of the HIV-1 p41 envelope protein using fusion proteins synthesized in Escherichia coli. Gene. 1988 Apr 15;64(1):107–119. doi: 10.1016/0378-1119(88)90485-4. [DOI] [PubMed] [Google Scholar]
  24. de Wolf F., Lange J. M., Houweling J. T., Coutinho R. A., Schellekens P. T., van der Noordaa J., Goudsmit J. Numbers of CD4+ cells and the levels of core antigens of and antibodies to the human immunodeficiency virus as predictors of AIDS among seropositive homosexual men. J Infect Dis. 1988 Sep;158(3):615–622. doi: 10.1093/infdis/158.3.615. [DOI] [PubMed] [Google Scholar]

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

RESOURCES