Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1991 Oct;29(10):2266–2272. doi: 10.1128/jcm.29.10.2266-2272.1991

Advantages of a human immunodeficiency virus type 1 (HIV-1) persistently infected HeLa T4+ cell line for HIV-1 indirect immunofluorescence serology.

B Forghani 1, J W Hurst 1, C S Chan 1
PMCID: PMC270311  PMID: 1939583

Abstract

A HeLa T4+ cell line persistently infected with human immunodeficiency virus type 1 (HIV-1) was used in an indirect immunofluorescent antibody assay (IFA) system to explore its potential suitability as an alternative source of viral antigen for confirmatory IFA in HIV serology. In a study of 121 serum samples chosen because they were reactive on repeat examination by enzyme immunoassay but nonspecific by IFA by using HIV-1-infected H9 cells (H9 IFA) or gave discrepant results by enzyme immunoassay and H9 IFA, the specificity and sensitivity of the HeLa T4+ IFA were comparable to those of Western blot (immunoblot), and identification of the true positive samples among these discrepant or nonspecific samples by HeLa T4+ IFA was approximately twice that by H9 IFA. The primary advantages of using the HeLa cell line rather than lymphoid cell lines in IFA are that cells can be grown as a monolayer and that the individual cells are much larger. The cell membrane, cytoplasm, and nucleus are easily discernible; this allows specific and nonspecific staining to be distinguished. At least eight different nonspecific nuclear and cytoplasmic staining patterns were identified in this study by using T4+ cells.

Full text

PDF
2266

Images in this article

Selected References

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

  1. Ascher M. S., Wilber J. C. Immunofluorescence for serodiagnosis of retrovirus infection. Arch Pathol Lab Med. 1990 Mar;114(3):246–248. [PubMed] [Google Scholar]
  2. Barré-Sinoussi F., Chermann J. C., Rey F., Nugeyre M. T., Chamaret S., Gruest J., Dauguet C., Axler-Blin C., Vézinet-Brun F., Rouzioux C. Isolation of a T-lymphotropic retrovirus from a patient at risk for acquired immune deficiency syndrome (AIDS). Science. 1983 May 20;220(4599):868–871. doi: 10.1126/science.6189183. [DOI] [PubMed] [Google Scholar]
  3. Biberfeld G., Bredberg-Rådén U., Böttiger B., Putkonen P. O., Blomberg J., Juto P., Wadell G. Blood donor sera with false-positive Western blot reactions to human immunodeficiency virus. Lancet. 1986 Aug 2;2(8501):289–290. doi: 10.1016/s0140-6736(86)92111-2. [DOI] [PubMed] [Google Scholar]
  4. Blomberg J., Klasse P. J. Specificities and sensitivities of three systems for determination of antibodies to human immunodeficiency virus by electrophoretic immunoblotting. J Clin Microbiol. 1988 Jan;26(1):106–110. doi: 10.1128/jcm.26.1.106-110.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blumberg R. S., Sandstrom E. G., Paradis T. J., Neumeyer D. N., Sarngadharan M. G., Hartshorn K. L., Byington R. E., Hirsch M. S., Schooley R. T. Detection of human T-cell lymphotropic virus type III-related antigens and anti-human T-cell lymphotropic virus type III antibodies by anticomplementary immunofluorescence. J Clin Microbiol. 1986 Jun;23(6):1072–1077. doi: 10.1128/jcm.23.6.1072-1077.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burke D. S., Brandt B. L., Redfield R. R., Lee T. H., Thorn R. M., Beltz G. A., Hung C. H. Diagnosis of human immunodeficiency virus infection by immunoassay using a molecularly cloned and expressed virus envelope polypeptide. Comparison to Western blot on 2707 consecutive serum samples. Ann Intern Med. 1987 May;106(5):671–676. doi: 10.7326/0003-4819-106-5-671. [DOI] [PubMed] [Google Scholar]
  7. Burke D. S., Brundage J. F., Redfield R. R., Damato J. J., Schable C. A., Putman P., Visintine R., Kim H. I. Measurement of the false positive rate in a screening program for human immunodeficiency virus infections. N Engl J Med. 1988 Oct 13;319(15):961–964. doi: 10.1056/NEJM198810133191501. [DOI] [PubMed] [Google Scholar]
  8. Carlson J. R., Bryant M. L., Hinrichs S. H., Yamamoto J. K., Levy N. B., Yee J., Higgins J., Levine A. M., Holland P., Gardner M. B. AIDS serology testing in low- and high-risk groups. JAMA. 1985 Jun 21;253(23):3405–3408. [PubMed] [Google Scholar]
  9. Carlson J. R., Yee J., Hinrichs S. H., Bryant M. L., Gardner M. B., Pedersen N. C. Comparison of indirect immunofluorescence and Western blot for detection of anti-human immunodeficiency virus antibodies. J Clin Microbiol. 1987 Mar;25(3):494–497. doi: 10.1128/jcm.25.3.494-497.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cheingsong-Popov R., Weiss R. A., Dalgleish A., Tedder R. S., Shanson D. C., Jeffries D. J., Ferns R. B., Briggs E. M., Weller I. V., Mitton S. Prevalence of antibody to human T-lymphotropic virus type III in AIDS and AIDS-risk patients in Britain. Lancet. 1984 Sep 1;2(8401):477–480. doi: 10.1016/s0140-6736(84)92562-5. [DOI] [PubMed] [Google Scholar]
  11. Chesebro B., Wehrly K. Development of a sensitive quantitative focal assay for human immunodeficiency virus infectivity. J Virol. 1988 Oct;62(10):3779–3788. doi: 10.1128/jvi.62.10.3779-3788.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Chesebro B., Wehrly K., Metcalf J., Griffin D. E. Use of a new CD4-positive HeLa cell clone for direct quantitation of infectious human immunodeficiency virus from blood cells of AIDS patients. J Infect Dis. 1991 Jan;163(1):64–70. doi: 10.1093/infdis/163.1.64. [DOI] [PubMed] [Google Scholar]
  13. Cooper D. A., Imrie A. A., Penny R. Antibody response to human immunodeficiency virus after primary infection. J Infect Dis. 1987 Jun;155(6):1113–1118. doi: 10.1093/infdis/155.6.1113. [DOI] [PubMed] [Google Scholar]
  14. Couroucé A. M., Muller J. Y., Richard D. False-positive Western blot reactions to human immunodeficiency virus in blood donors. Lancet. 1986 Oct 18;2(8512):921–922. doi: 10.1016/s0140-6736(86)90443-5. [DOI] [PubMed] [Google Scholar]
  15. Drabick J. J., Baker J. R., Jr HLA antigen contamination of commercial Western blot strips for detecting human immunodeficiency virus. J Infect Dis. 1989 Feb;159(2):357–358. doi: 10.1093/infdis/159.2.357. [DOI] [PubMed] [Google Scholar]
  16. Essex M., McLane M. F., Lee T. H., Falk L., Howe C. W., Mullins J. I., Cabradilla C., Francis D. P. Antibodies to cell membrane antigens associated with human T-cell leukemia virus in patients with AIDS. Science. 1983 May 20;220(4599):859–862. doi: 10.1126/science.6342136. [DOI] [PubMed] [Google Scholar]
  17. Forghani B., Schmidt N. J. Antigen requirements, sensitivity, and specificity of enzyme immunoassays for measles and rubella viral antibodies. J Clin Microbiol. 1979 Jun;9(6):657–664. doi: 10.1128/jcm.9.6.657-664.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Forghani B., Schmidt N. J., Dennis J. Antibody assays for varicella-zoster virus: comparison of enzyme immunoassay with neutralization, immune adherence hemagglutination, and complement fixation. J Clin Microbiol. 1978 Nov;8(5):545–552. doi: 10.1128/jcm.8.5.545-552.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Forghani B., Schmidt N. J. Humoral immune response to virions and dense bodies of human cytomegalovirus determined by enzyme immunofluorescence assay. J Med Virol. 1980;6(2):119–127. doi: 10.1002/jmv.1890060204. [DOI] [PubMed] [Google Scholar]
  20. Forghani B., Yu G. J., Hurst J. W. Comparison of biotinylated DNA and RNA probes for rapid detection of varicella-zoster virus genome by in situ hybridization. J Clin Microbiol. 1991 Mar;29(3):583–591. doi: 10.1128/jcm.29.3.583-591.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gallo D., Diggs J. L., Shell G. R., Dailey P. J., Hoffman M. N., Riggs J. L. Comparison of detection of antibody to the acquired immune deficiency syndrome virus by enzyme immunoassay, immunofluorescence, and Western blot methods. J Clin Microbiol. 1986 Jun;23(6):1049–1051. doi: 10.1128/jcm.23.6.1049-1051.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hausler W. J., Jr, Getchell J. P. Report of the Fourth Consensus Conference on HIV Testing sponsored by the Association of State and Territorial Public Health Laboratory Directors (ASTPHLD). Infect Control Hosp Epidemiol. 1989 Aug;10(8):354–367. doi: 10.1086/646045. [DOI] [PubMed] [Google Scholar]
  23. Imrie A. A., Kehrer S., Smith G. W., Penny R., Cooper D. A. Seroimmunology of AIDS retrovirus infection. I. Use of immunofluorescence assay to confirm sera with ELISA reactivity. Pathology. 1986 Oct;18(4):438–443. doi: 10.3109/00313028609087565. [DOI] [PubMed] [Google Scholar]
  24. Jackson J. B., Balfour H. H., Jr Practical diagnostic testing for human immunodeficiency virus. Clin Microbiol Rev. 1988 Jan;1(1):124–138. doi: 10.1128/cmr.1.1.124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Katchaki J. N., Siem T. H., Brouwer R., Nieste H. L. Anti-HTLV-III screening and false positivity. Lancet. 1986 Feb 22;1(8478):448–448. doi: 10.1016/s0140-6736(86)92408-6. [DOI] [PubMed] [Google Scholar]
  26. Kenrick K. G., Bolton W., Doran T. J. False-positive anti-HTLV-III assay results and HLA-DR4. Med J Aust. 1985 Jul 22;143(2):88–89. doi: 10.5694/j.1326-5377.1985.tb122812.x. [DOI] [PubMed] [Google Scholar]
  27. Kitchen L. W., Barin F., Sullivan J. L., McLane M. F., Brettler D. B., Levine P. H., Essex M. Aetiology of AIDS--antibodies to human T-cell leukaemia virus (type III) in haemophiliacs. Nature. 1984 Nov 22;312(5992):367–369. doi: 10.1038/312367a0. [DOI] [PubMed] [Google Scholar]
  28. Kvinesdal B. B., Nielsen C. M., Poulsen A. G., Højlyng N. Immunofluorescence assay for detection of antibodies to human immunodeficiency virus type 2. J Clin Microbiol. 1989 Nov;27(11):2502–2504. doi: 10.1128/jcm.27.11.2502-2504.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kühnl P., Seidl S., Holzberger G. HLA DR4 antibodies cause positive HTLV-III antibody ELISA results. Lancet. 1985 May 25;1(8439):1222–1223. doi: 10.1016/s0140-6736(85)92910-1. [DOI] [PubMed] [Google Scholar]
  30. Lennette E. T., Karpatkin S., Levy J. A. Indirect immunofluorescence assay for antibodies to human immunodeficiency virus. J Clin Microbiol. 1987 Feb;25(2):199–202. doi: 10.1128/jcm.25.2.199-202.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Maddon P. J., Dalgleish A. G., McDougal J. S., Clapham P. R., Weiss R. A., Axel R. The T4 gene encodes the AIDS virus receptor and is expressed in the immune system and the brain. Cell. 1986 Nov 7;47(3):333–348. doi: 10.1016/0092-8674(86)90590-8. [DOI] [PubMed] [Google Scholar]
  32. Maddon P. J., Littman D. R., Godfrey M., Maddon D. E., Chess L., Axel R. The isolation and nucleotide sequence of a cDNA encoding the T cell surface protein T4: a new member of the immunoglobulin gene family. Cell. 1985 Aug;42(1):93–104. doi: 10.1016/s0092-8674(85)80105-7. [DOI] [PubMed] [Google Scholar]
  33. Mahony J., Rosenthal K., Chernesky M., Castriciano S., Scheid E., Blajchman M., Harnish D. Agreement study between two laboratories of immunofluorescence as a confirmatory test for human immunodeficiency virus type 1 antibody screening. J Clin Microbiol. 1989 Jun;27(6):1234–1237. doi: 10.1128/jcm.27.6.1234-1237.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. McHugh T. M., Stites D. P., Casavant C. H., Carlson J. R., Yee J., McVay P. A., Busch M. P., Levy J. A. Evaluation of the indirect immunofluorescence assay as a confirmatory test for detecting antibodies to the human immunodeficiency virus. Diagn Immunol. 1986;4(5):233–240. [PubMed] [Google Scholar]
  35. Meyer K. B., Pauker S. G. Screening for HIV: can we afford the false positive rate? N Engl J Med. 1987 Jul 23;317(4):238–241. doi: 10.1056/NEJM198707233170410. [DOI] [PubMed] [Google Scholar]
  36. Morgan J., Tate R., Farr A. D., Urbaniak S. J. Potential source of error in HTLV-III antibody testing. Lancet. 1986 Mar 29;1(8483):739–740. doi: 10.1016/s0140-6736(86)91130-x. [DOI] [PubMed] [Google Scholar]
  37. Mortimer P. P., Parry J. V., Mortimer J. Y. Which anti-HTLV III/LAV assays for screening and confirmatory testing? Lancet. 1985 Oct 19;2(8460):873–877. doi: 10.1016/s0140-6736(85)90136-9. [DOI] [PubMed] [Google Scholar]
  38. Page M., Thorpe R. Controlling for false-positive antisera in human immunodeficiency virus immunoblotting. J Infect Dis. 1990 Jul;162(1):278–279. doi: 10.1093/infdis/162.1.278. [DOI] [PubMed] [Google Scholar]
  39. Roy S., Portnoy J., Wainberg M. A. Need for caution in interpretation of Western Blot tests for HIV. JAMA. 1987 Feb 27;257(8):1047–1047. [PubMed] [Google Scholar]
  40. Saag M. S., Britz J. Asymptomatic blood donor with a false positive HTLV-III Western blot. N Engl J Med. 1986 Jan 9;314(2):118–118. doi: 10.1056/nejm198601093140212. [DOI] [PubMed] [Google Scholar]
  41. Sandstrom E. G., Schooley R. T., Ho D. D., Byington R., Sarngadharan M. G., MacLane M. E., Essex M., Gallo R. C., Hirsch M. S. Detection of human anti-HTLV-III antibodies by indirect immunofluorescence using fixed cells. Transfusion. 1985 Jul-Aug;25(4):308–312. doi: 10.1046/j.1537-2995.1985.25485273806.x. [DOI] [PubMed] [Google Scholar]
  42. Schmidt B. L., Hutterer J., Kunz C., Gschnait F. Serologie der HIV-Infektion: Vergleich von indirekter Immunfluoreszenz, Western Blot und Enzym Immunoassay. Wien Med Wochenschr. 1988 Apr 30;138(8):174–180. [PubMed] [Google Scholar]
  43. Schochetman G., Epstein J. S., Zuck T. F. Serodiagnosis of infection with the AIDS virus and other human retroviruses. Annu Rev Microbiol. 1989;43:629–659. doi: 10.1146/annurev.mi.43.100189.003213. [DOI] [PubMed] [Google Scholar]
  44. Sligh J. M., Roodman S. T., Tsai C. C. Flow cytometric indirect immunofluorescence assay with high sensitivity and specificity for detection of antibodies to human immunodeficiency virus (HIV). Am J Clin Pathol. 1989 Feb;91(2):210–214. doi: 10.1093/ajcp/91.2.210. [DOI] [PubMed] [Google Scholar]
  45. Thorpe R., Brasher M. D., Bird C. R., Garrett A. J., Jacobs J. P., Minor P. D., Schild G. C. An improved immunoblotting procedure for the detection of antibodies against HIV. J Virol Methods. 1987 May;16(1-2):87–96. doi: 10.1016/0166-0934(87)90034-6. [DOI] [PubMed] [Google Scholar]
  46. Vercauteren G., van der Groen G., Piot P. Comparison of enzyme immunoassays and an immunofluorescence test for detection of antibody to human immunodeficiency virus in African sera. Eur J Clin Microbiol. 1987 Apr;6(2):132–135. doi: 10.1007/BF02018193. [DOI] [PubMed] [Google Scholar]
  47. van der Groen G., Vercauteren G., Piot P. Immunofluorescence tests for HIV antibody and their value as confirmatory tests. J Virol Methods. 1987 Aug;17(1-2):35–43. doi: 10.1016/0166-0934(87)90066-8. [DOI] [PubMed] [Google Scholar]

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

RESOURCES