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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Oct;86(19):7532–7536. doi: 10.1073/pnas.86.19.7532

In vitro synthesis of human immunodeficiency virus-specific antibodies in peripheral blood lymphocytes of infants.

S Pahwa 1, N Chirmule 1, C Leombruno 1, W Lim 1, R Harper 1, R Bhalla 1, R Pahwa 1, R P Nelson 1, R A Good 1
PMCID: PMC298099  PMID: 2798424

Abstract

An assay system was developed for the analysis of antibodies secreted in vitro against human immunodeficiency virus (HIV) by cultured peripheral blood lymphocytes of HIV-infected individuals. Cultures of peripheral blood lymphocytes were established with medium alone or with medium containing Epstein-Barr virus (EBV) or pokeweed mitogen. HIV antibodies were determined by an ELISA performed with commercial kits in which a whole virus extract served as antigen. Optimal antibody secretion was detected in 7-day peripheral blood lymphocyte cultures to which EBV had been added to provide polyclonal B-cell activation. Pokeweed mitogen-induced antibody secretion and spontaneous antibody secretion were less consistent. With EBV as a stimulus, the sensitivity and specificity of this assay for determining HIV infection status were each 100% in adults. When the assay was applied to infants and children, 23 of 24 symptomatic HIV-seropositive children (class P-2) and 11 of 33 asymptomatic seropositive infants aged less than or equal to 15 months (class P-0) tested positive for EBV-induced antibody secretion. Six of the 11 P-0 patients who tested positive have progressed to develop symptomatic disease, while the remainder are still seropositive at ages 2-15 months. Of the infants who were negative in this assay, all have remained asymptomatic. Treatment with 3'-azido-3'-deoxythymidine in infected adults and children has resulted in transient suppression of the in vitro antibody response in some instances. Thus EBV-induced synthesis of HIV-specific antibodies in vitro is a sensitive and specific indicator of HIV infection and is of help in determining infection status of asymptomatic seropositive infants who are classified as having "indeterminate" infection.

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Selected References

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  1. Amadori A., De Rossi A., Faulkner-Valle G. P., Chieco-Bianchi L. Spontaneous in vitro production of virus-specific antibody by lymphocytes from HIV-infected subjects. Clin Immunol Immunopathol. 1988 Mar;46(3):342–351. doi: 10.1016/0090-1229(88)90053-0. [DOI] [PubMed] [Google Scholar]
  2. Amadori A., de Rossi A., Giaquinto C., Faulkner-Valle G., Zacchello F., Chieco-Bianchi L. In-vitro production of HIV-specific antibody in children at risk of AIDS. Lancet. 1988 Apr 16;1(8590):852–854. doi: 10.1016/s0140-6736(88)91603-0. [DOI] [PubMed] [Google Scholar]
  3. Birx D. L., Redfield R. R., Tosato G. Defective regulation of Epstein-Barr virus infection in patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related disorders. N Engl J Med. 1986 Apr 3;314(14):874–879. doi: 10.1056/NEJM198604033141403. [DOI] [PubMed] [Google Scholar]
  4. Fauci A. S. The human immunodeficiency virus: infectivity and mechanisms of pathogenesis. Science. 1988 Feb 5;239(4840):617–622. doi: 10.1126/science.3277274. [DOI] [PubMed] [Google Scholar]
  5. Gaetano C., Scano G., Carbonari M., Giannini G., Mezzaroma I., Aiuti F., Marolla L., Casadei A. M., Carapella E. Delayed and defective anti-HIV IgM response in infants. Lancet. 1987 Mar 14;1(8533):631–631. doi: 10.1016/s0140-6736(87)90272-8. [DOI] [PubMed] [Google Scholar]
  6. Goudsmit J., Lange J. M., Paul D. A., Dawson G. J. Antigenemia and antibody titers to core and envelope antigens in AIDS, AIDS-related complex, and subclinical human immunodeficiency virus infection. J Infect Dis. 1987 Mar;155(3):558–560. doi: 10.1093/infdis/155.3.558. [DOI] [PubMed] [Google Scholar]
  7. Harnish D. G., Hammerberg O., Walker I. R., Rosenthal K. L. Early detection of HIV infection in a newborn. N Engl J Med. 1987 Jan 29;316(5):272–273. doi: 10.1056/NEJM198701293160510. [DOI] [PubMed] [Google Scholar]
  8. Johnson J. P., Nair P., Alexander S. Early diagnosis of HIV infection in the neonate. N Engl J Med. 1987 Jan 29;316(5):273–274. doi: 10.1056/NEJM198701293160511. [DOI] [PubMed] [Google Scholar]
  9. Lane H. C., Masur H., Edgar L. C., Whalen G., Rook A. H., Fauci A. S. Abnormalities of B-cell activation and immunoregulation in patients with the acquired immunodeficiency syndrome. N Engl J Med. 1983 Aug 25;309(8):453–458. doi: 10.1056/NEJM198308253090803. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Ou C. Y., Kwok S., Mitchell S. W., Mack D. H., Sninsky J. J., Krebs J. W., Feorino P., Warfield D., Schochetman G. DNA amplification for direct detection of HIV-1 in DNA of peripheral blood mononuclear cells. Science. 1988 Jan 15;239(4837):295–297. doi: 10.1126/science.3336784. [DOI] [PubMed] [Google Scholar]
  12. Pahwa S. G., Good R. A., Hoffmann M. K. The role of mitogens and antigens in the generation of antibody-producing human B lymphocytes. J Clin Immunol. 1981 Apr;1(2):101–105. doi: 10.1007/BF00915386. [DOI] [PubMed] [Google Scholar]
  13. Pahwa S. G., Quilop M. T., Lange M., Pahwa R. N., Grieco M. H. Defective B-lymphocyte function in homosexual men in relation to the acquired immunodeficiency syndrome. Ann Intern Med. 1984 Dec;101(6):757–763. doi: 10.7326/0003-4819-101-6-757. [DOI] [PubMed] [Google Scholar]
  14. Pahwa S., Pahwa R., Good R. A., Gallo R. C., Saxinger C. Stimulatory and inhibitory influences of human immunodeficiency virus on normal B lymphocytes. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9124–9128. doi: 10.1073/pnas.83.23.9124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Popovic M., Sarngadharan M. G., Read E., Gallo R. C. Detection, isolation, and continuous production of cytopathic retroviruses (HTLV-III) from patients with AIDS and pre-AIDS. Science. 1984 May 4;224(4648):497–500. doi: 10.1126/science.6200935. [DOI] [PubMed] [Google Scholar]
  16. Slade H. B., Lopatin D. E., Schwartz S. A. Measurement of immunoglobulin concentration in cell culture supernates by computer-assisted ELISA. J Immunol Methods. 1986 Nov 20;94(1-2):169–179. doi: 10.1016/0022-1759(86)90230-9. [DOI] [PubMed] [Google Scholar]
  17. Slade H. B., Pica R. V., Pahwa S. G. Detection of HIV-specific antibodies in infancy by isoelectric focusing and affinity immunoblotting. J Infect Dis. 1989 Jul;160(1):126–130. doi: 10.1093/infdis/160.1.126. [DOI] [PubMed] [Google Scholar]
  18. Taswell C. Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis. J Immunol. 1981 Apr;126(4):1614–1619. [PubMed] [Google Scholar]
  19. Teeuwsen V. J., Logtenberg T., Siebelink K. H., Lange J. M., Goudsmit J., Uytdehaag F. G., Osterhaus A. D. Analysis of the antigen- and mitogen-induced differentiation of B lymphocytes from asymptomatic human immunodeficiency virus-seropositive male homosexuals. Discrepancy between T cell-dependent and T cell-independent activation. J Immunol. 1987 Nov 1;139(9):2929–2935. [PubMed] [Google Scholar]
  20. Tosato G., Magrath I. T., Koski I. R., Dooley N. J., Blaese R. M. B cell differentiation and immunoregulatory T cell function in human cord blood lymphocytes. J Clin Invest. 1980 Aug;66(2):383–388. doi: 10.1172/JCI109867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tovo P. A., Gabiano C., Riva C., Palomba E., Valpreda A., Musso A. Specific antibody and virus antigen expression in congenital HIV infection. Lancet. 1987 May 23;1(8543):1201–1201. doi: 10.1016/s0140-6736(87)92166-0. [DOI] [PubMed] [Google Scholar]
  22. Turner D. A. An intuitive approach to receiver operating characteristic curve analysis. J Nucl Med. 1978 Feb;19(2):213–220. [PubMed] [Google Scholar]
  23. Yarchoan R., Tosato G., Blaese R. M., Simon R. M., Nelson D. L. Limiting dilution analysis of Epstein-Barr virus-induced immunoglobulin production by human B cells. J Exp Med. 1983 Jan 1;157(1):1–14. doi: 10.1084/jem.157.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]

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