A novel method for detection and ex vivo expansion of HIV type 1-specific cytolytic T lymphocytes

M. N. Lubaki, M. A. Egan, R. F. Siliciano, K. J. Weinhold, R. C. Bollinger

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Studies have shown that cytolytic T lymphocyte (CTL) responses may be critical to the clearance of the early viremia in acute HIV-1 infection. It is likely that these cells play an important role in prolonging the asymptomatic phase of the infection. Although HIV-1-specific CTL activity can be detected in direct assays of freshly isolated peripheral blood lymphocytes (PBL) from some infected individuals, this method fails to detect CTL that are present at low frequency and resting, memory CTL. For these reasons, direct CTL assays on PBL from seropositive individuals may underestimate the level of CTL immunity. As part of ongoing investigations of CTL activity in HIV-1-infected individuals, we developed a novel strategy for the detection and ex vivo expansion of HIV-1-specific CTL. This technique involves selective stimulation of PBL from seropositive individuals with autologous Epstein-Barr virus (EBV)-transformed, B-lymphoblastoid cell lines (B-LCL) infected with vaccinia vectors expressing various HIV-1 genes. Prior to their use for in vitro stimulation, B-LCL are treated with psoralen and UV light to inactivate vaccinia virus. After 1 week of stimulation, CTL activity in stimulated cultures is measured in a standard 51Cr release assay. This ex vivo expansion method can selectively increase the bulk culture CTL activity against env, gag and nef, even in some seropositive individuals with low CD4 counts and little evidence of HIV-1-specific CTL in assays of freshly isolated PBL. These expanded CTL are predominantly of the CD8+ phenotype. This method provides an efficient and sensitive method for the detection and ex vivo expansion of HIV-1-specific CTL from infected individuals at all stages of disease and may prove useful in following changes in CTL responses with disease progression and therapy. In addition, this method may allow for the ex vivo expansion of HIV-1-specific CTL.

Original languageEnglish (US)
Pages (from-to)1427-1431
Number of pages5
JournalAIDS Research and Human Retroviruses
Volume10
Issue number11
StatePublished - 1994
Externally publishedYes

Fingerprint

HIV-1
T-Lymphocytes
Lymphocytes
Ficusin
Vaccinia
Cell Line
Asymptomatic Infections
Vaccinia virus
Viremia
Ultraviolet Rays
CD4 Lymphocyte Count
Human Herpesvirus 4
HIV Infections
Disease Progression
Immunity
Phenotype

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Lubaki, M. N., Egan, M. A., Siliciano, R. F., Weinhold, K. J., & Bollinger, R. C. (1994). A novel method for detection and ex vivo expansion of HIV type 1-specific cytolytic T lymphocytes. AIDS Research and Human Retroviruses, 10(11), 1427-1431.

A novel method for detection and ex vivo expansion of HIV type 1-specific cytolytic T lymphocytes. / Lubaki, M. N.; Egan, M. A.; Siliciano, R. F.; Weinhold, K. J.; Bollinger, R. C.

In: AIDS Research and Human Retroviruses, Vol. 10, No. 11, 1994, p. 1427-1431.

Research output: Contribution to journalArticle

Lubaki, MN, Egan, MA, Siliciano, RF, Weinhold, KJ & Bollinger, RC 1994, 'A novel method for detection and ex vivo expansion of HIV type 1-specific cytolytic T lymphocytes', AIDS Research and Human Retroviruses, vol. 10, no. 11, pp. 1427-1431.
Lubaki, M. N. ; Egan, M. A. ; Siliciano, R. F. ; Weinhold, K. J. ; Bollinger, R. C. / A novel method for detection and ex vivo expansion of HIV type 1-specific cytolytic T lymphocytes. In: AIDS Research and Human Retroviruses. 1994 ; Vol. 10, No. 11. pp. 1427-1431.
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