Skip to main content
NCBI home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1994 Aug;32(8):1870–1875. doi: 10.1128/jcm.32.8.1870-1875.1994

Differentiation between human immunodeficiency virus type 1 (HIV-1) and HIV-2 isolates by nonradioisotopic reverse transcriptase-typing assay.

T Urabe 1, K Sano 1, T Nakano 1, F Odawara 1, M H Lee 1, T Otake 1, S Okubo 1, M Hayami 1, H Misaki 1, M Baba 1, et al.
PMCID: PMC263894  PMID: 7527425

Abstract

We tested whether human immunodeficiency virus type 1 (HIV-1) could be differentiated from HIV-2 by a reverse transcriptase (RT)-typing assay that measured the reduction of enzyme activity owing to specific antibody. RT-inhibiting antibody was examined for HIV type specificity by a new nonradioisotopic RT assay. Antibodies from four rabbits immunized with recombinant HIV-1 RT and from 23 HIV-1-seropositive individuals all specifically inhibited the enzyme activities of two HIV-1 strains (LAV-1 and GH-3), three zidovudine-resistant HIV-1 mutants, and a recombinant HIV-1 RT. However, none of these antisera affected the activities of six HIV-2 strains (GH-1, GH-2, GH-4, GH-5, GH-6, LAV-2ROD), Rous-associated virus type 2, and DNA polymerase I from Escherichia coli. In contrast, HIV-2 antibody from a rabbit immunized with disrupted GH-1 virions blocked the enzyme activities of the six HIV-2 strains but not those of the three HIV-1 strains, Rous-associated virus type 2, or DNA polymerase I. These results indicate that the antigenic domains of HIV-1 and HIV-2 RTs recognized by their inhibiting antibodies are distinct from each other and are highly conserved. Clinical HIV isolates from 18 HIV-1-seropositive individuals and 3 HIV-2-seropositive Ghanaian individuals were identified as HIV-1 and HIV-2, respectively, by the nonradioisotopic RT-typing assay.

Full text

PDF
1870

Images in this article

1871Image
on p.1871

Selected References

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

  1. Advani M., Imagawa D. T., Lee M. H., Sano K., Morales F., Mitsuyasu R. T., Detels R. Cross-sectional study of reverse transcriptase-inhibiting antibody as a marker of acquired immune deficiency syndrome. J Clin Microbiol. 1989 Jul;27(7):1453–1455. doi: 10.1128/jcm.27.7.1453-1455.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allan J. S., Coligan J. E., Lee T. H., Barin F., Kanki P. J., M'Boup S., McLane M. F., Groopman J. E., Essex M. Immunogenic nature of a Pol gene product of HTLV-III/LAV. Blood. 1987 Jan;69(1):331–333. [PubMed] [Google Scholar]
  3. Baba M., De Clercq E., Tanaka H., Ubasawa M., Takashima H., Sekiya K., Nitta I., Umezu K., Nakashima H., Mori S. Potent and selective inhibition of human immunodeficiency virus type 1 (HIV-1) by 5-ethyl-6-phenylthiouracil derivatives through their interaction with the HIV-1 reverse transcriptase. Proc Natl Acad Sci U S A. 1991 Mar 15;88(6):2356–2360. doi: 10.1073/pnas.88.6.2356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. De Cock K. M., Odehouri K., Colebunders R. L., Adjorlolo G., Lafontaine M. F., Porter A., Gnaore E., Diaby L., Moreau J., Heyward W. L. A comparison of HIV-1 and HIV-2 infections in hospitalized patients in Abidjan, Côte d'Ivoire. AIDS. 1990 May;4(5):443–448. doi: 10.1097/00002030-199005000-00010. [DOI] [PubMed] [Google Scholar]
  5. DeVico A. L., Veronese F. D., Lee S. L., Gallo R. C., Sarngadharan M. G. High prevalence of serum antibodies to reverse transcriptase in HIV-1-infected individuals. AIDS Res Hum Retroviruses. 1988 Feb;4(1):17–22. doi: 10.1089/aid.1988.4.17. [DOI] [PubMed] [Google Scholar]
  6. Gelderblom H. R., Hausmann E. H., Ozel M., Pauli G., Koch M. A. Fine structure of human immunodeficiency virus (HIV) and immunolocalization of structural proteins. Virology. 1987 Jan;156(1):171–176. doi: 10.1016/0042-6822(87)90449-1. [DOI] [PubMed] [Google Scholar]
  7. Hollis V. W., Jr, Aoki T., Barrera O., Oldstone M. B., Dixon F. J. Detection of naturally occurring antibodies to RNA-dependent DNA polymerase of murine leukemia virus in kidney eluates of AKR mice. J Virol. 1974 Feb;13(2):448–454. doi: 10.1128/jvi.13.2.448-454.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ishikawa K., Tsujimoto H., Nakai M., Mingle J. A., Osei-Kwasi M., Aggrey S. E., Nettey V. B., Afoakwa S. N., Fukasawa M., Kodama T. Isolation and characterization of HIV-2 from an AIDS patient in Ghana. AIDS. 1988 Oct;2(5):383–388. doi: 10.1097/00002030-198810000-00009. [DOI] [PubMed] [Google Scholar]
  9. Kawamura M., Katahira J., Fukasawa M., Sakuragi J., Ishikawa K., Nakai M., Mingle J. A., Osei-Kwasi M., Netty V. B., Akari H. Isolation and characterization of a highly divergent HIV-2[GH-2]: generation of an infectious molecular clone and functional analysis of its rev-responsive element in response to primate retrovirus transactivators (Rev and Rex). Virology. 1992 Jun;188(2):850–853. doi: 10.1016/0042-6822(92)90540-6. [DOI] [PubMed] [Google Scholar]
  10. Kohlstaedt L. A., Wang J., Friedman J. M., Rice P. A., Steitz T. A. Crystal structure at 3.5 A resolution of HIV-1 reverse transcriptase complexed with an inhibitor. Science. 1992 Jun 26;256(5065):1783–1790. doi: 10.1126/science.1377403. [DOI] [PubMed] [Google Scholar]
  11. Kornfeld H., Riedel N., Viglianti G. A., Hirsch V., Mullins J. I. Cloning of HTLV-4 and its relation to simian and human immunodeficiency viruses. Nature. 1987 Apr 9;326(6113):610–613. doi: 10.1038/326610a0. [DOI] [PubMed] [Google Scholar]
  12. Larder B. A., Darby G., Richman D. D. HIV with reduced sensitivity to zidovudine (AZT) isolated during prolonged therapy. Science. 1989 Mar 31;243(4899):1731–1734. doi: 10.1126/science.2467383. [DOI] [PubMed] [Google Scholar]
  13. Larder B. A., Kemp S. D. Multiple mutations in HIV-1 reverse transcriptase confer high-level resistance to zidovudine (AZT). Science. 1989 Dec 1;246(4934):1155–1158. doi: 10.1126/science.2479983. [DOI] [PubMed] [Google Scholar]
  14. Laurence J., Saunders A., Kulkosky J. Characterization and clinical association of antibody inhibitory to HIV reverse transcriptase activity. Science. 1987 Mar 20;235(4795):1501–1504. doi: 10.1126/science.2435004. [DOI] [PubMed] [Google Scholar]
  15. Marlink R. G., Ricard D., M'Boup S., Kanki P. J., Romet-Lemonne J. L., N'Doye I., Diop K., Simpson M. A., Greco F., Chou M. J. Clinical, hematologic, and immunologic cross-sectional evaluation of individuals exposed to human immunodeficiency virus type-2 (HIV-2). AIDS Res Hum Retroviruses. 1988 Apr;4(2):137–148. doi: 10.1089/aid.1988.4.137. [DOI] [PubMed] [Google Scholar]
  16. McDougal J. S., Cort S. P., Kennedy M. S., Cabridilla C. D., Feorino P. M., Francis D. P., Hicks D., Kalyanaraman V. S., Martin L. S. Immunoassay for the detection and quantitation of infectious human retrovirus, lymphadenopathy-associated virus (LAV). J Immunol Methods. 1985 Jan 21;76(1):171–183. doi: 10.1016/0022-1759(85)90489-2. [DOI] [PubMed] [Google Scholar]
  17. Müller B., Restle T., Kühnel H., Goody R. S. Expression of the heterodimeric form of human immunodeficiency virus type 2 reverse transcriptase in Escherichia coli and characterization of the enzyme. J Biol Chem. 1991 Aug 5;266(22):14709–14713. [PubMed] [Google Scholar]
  18. Otake T., Mori H., Morimoto M., Ueba N., Kunita N., Sano K., Nakai M., Okubo S., Yasunaga K., Nagao N. [HIV isolation and clinical markers on the seropositive subjects]. Kansenshogaku Zasshi. 1990 Oct;64(10):1287–1294. doi: 10.11150/kansenshogakuzasshi1970.64.1287. [DOI] [PubMed] [Google Scholar]
  19. Pauwels R., Andries K., Desmyter J., Schols D., Kukla M. J., Breslin H. J., Raeymaeckers A., Van Gelder J., Woestenborghs R., Heykants J. Potent and selective inhibition of HIV-1 replication in vitro by a novel series of TIBO derivatives. Nature. 1990 Feb 1;343(6257):470–474. doi: 10.1038/343470a0. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. Rayfield M., De Cock K., Heyward W., Goldstein L., Krebs J., Kwok S., Lee S., McCormick J., Moreau J. M., Odehouri K. Mixed human immunodeficiency virus (HIV) infection in an individual: demonstration of both HIV type 1 and type 2 proviral sequences by using polymerase chain reaction. J Infect Dis. 1988 Dec;158(6):1170–1176. doi: 10.1093/infdis/158.6.1170. [DOI] [PubMed] [Google Scholar]
  22. Rho H. M., Poiesz B., Ruscetti F. W., Gallo R. C. Characterization of the reverse transcriptase from a new retrovirus (HTLV) produced by a human cutaneous T-cell lymphoma cell line. Virology. 1981 Jul 15;112(1):355–360. doi: 10.1016/0042-6822(81)90642-5. [DOI] [PubMed] [Google Scholar]
  23. Robert-Gurof M., Gallo R. C. Type-specific binding antibody to baboon endogenous virus (M7) reverse transcriptase. J Gen Virol. 1979 Apr;43(1):241–246. doi: 10.1099/0022-1317-43-1-241. [DOI] [PubMed] [Google Scholar]
  24. Saitoh A., Iwasaki H., Nakata A., Adachi A., Shinagawa H. Overproduction of human immunodeficiency virus type I reverse transcriptase in Escherichia coli and purification of the enzyme. Microbiol Immunol. 1990;34(6):509–521. doi: 10.1111/j.1348-0421.1990.tb03168.x. [DOI] [PubMed] [Google Scholar]
  25. Sano K., Lee M. H., Morales F., Nishanian P., Fahey J., Detels R., Imagawa D. T. Antibody that inhibits human immunodeficiency virus reverse transcriptase and association with inability to isolate virus. J Clin Microbiol. 1987 Dec;25(12):2415–2417. doi: 10.1128/jcm.25.12.2415-2417.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sarngadharan M. G., Popovic M., Bruch L., Schüpbach J., Gallo R. C. Antibodies reactive with human T-lymphotropic retroviruses (HTLV-III) in the serum of patients with AIDS. Science. 1984 May 4;224(4648):506–508. doi: 10.1126/science.6324345. [DOI] [PubMed] [Google Scholar]
  27. Schüpbach J., Popovic M., Gilden R. V., Gonda M. A., Sarngadharan M. G., Gallo R. C. Serological analysis of a subgroup of human T-lymphotropic retroviruses (HTLV-III) associated with AIDS. Science. 1984 May 4;224(4648):503–505. doi: 10.1126/science.6200937. [DOI] [PubMed] [Google Scholar]
  28. Snowden W., Coughlan N., Tisdale M., Stammers D. K. Isolation and characterization of monoclonal antibodies raised against the reverse transcriptase of human immunodeficiency virus type 2 and cross-reactivity with that of type 1. J Acquir Immune Defic Syndr. 1993 Nov;6(11):1187–1193. [PubMed] [Google Scholar]
  29. Urabe T., Sano K., Tanno M., Mizoguchi J., Otani M., Lee M. H., Takasaki T., Kusakabe H., Imagawa D. T., Nakai M. A non-radioisotopic reverse transcriptase assay using biotin-11-deoxyuridinetriphosphate on primer-immobilized microtiter plates. J Virol Methods. 1992 Nov;40(2):145–154. doi: 10.1016/0166-0934(92)90063-j. [DOI] [PubMed] [Google Scholar]
  30. Wolfs T. F., Geelen J. L., Schellekens H., Barin F., Dekker J. T., Goudsmit J. Serum capacity to inhibit reverse transcriptase in vitro distinguishes HIV-1 infection from HIV-2 or SIV infection. AIDS Res Hum Retroviruses. 1989 Oct;5(5):535–540. doi: 10.1089/aid.1989.5.535. [DOI] [PubMed] [Google Scholar]
  31. Wuu K. D., Graves D. C., Ferrer J. F. Inhibition of the reverse transcriptase of bovine leukemia virus by antibody in sera from leukemic cattle and immunological characterization of the enzyme. Cancer Res. 1977 May;37(5):1438–1442. [PubMed] [Google Scholar]
  32. di Marzo Veronese F., Copeland T. D., DeVico A. L., Rahman R., Oroszlan S., Gallo R. C., Sarngadharan M. G. Characterization of highly immunogenic p66/p51 as the reverse transcriptase of HTLV-III/LAV. Science. 1986 Mar 14;231(4743):1289–1291. doi: 10.1126/science.2418504. [DOI] [PubMed] [Google Scholar]

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

RESOURCES