The effect of fresh lymphocytes on increased sensitivity of hiv-1 isolation: A multicenter study

Homayoon Farzadegan, David Imagawa, Phalguni Gupta, Moon H. Lee, Lisa Paula Jacobson, Alfred Saah, Kathie Grovit, Charles R. Rinaldo, B. Frank Polk

Research output: Contribution to journalArticle

Abstract

A multicenter study was undertaken to determine the sensitivity and reproducibility of markers for human immunodeficiency virus type 1 (HIV-1) viral growth and the effect of various preparations of lymphocytes on the sensitivity of standard and routinely used procedures for HIV-1 isolation. In phase 1, cocultivated culture supernatants obtained from 10 HIV-1 cultures were transported to three Multicenter AIDS Cohort Study (MACS) Virology Laboratories. Three commercial HIV-p24 antigen capture (AC) tests and two reverse transcriptase (RT) assays were used to ascertain the replication of HIV-1. The Du Pont and Abbott AC assays were found to be most sensitive (85-100%), and the RT assay with 24-h incubation period had comparable sensitivity (75-100%). In phase II, the sensitivity of standard cocultivation procedure for HIV-1 isolation was compared using freshly phytohemaggluti-nin-P (PHA-P)-stimulated, stimulated-frozen, and frozen-thawed and then stimulated normal human peripheral blood mononuclear cells (PBMCs) as co-cultivating cells. Blood samples from 13 HIV-1 infected individuals with various CD4+ cell counts were cocultivated in each of the three MACS laboratories using one of the aforementioned normal PBMCs. The PHA-P-stimulated fresh normal PBMC showed a maximum isolation rate of 100% (13 of 13) with an average of 8 days to positivity. This rate of isolation was significantly greater than other rates using any one of the other PBMC preparations. These findings demonstrated that the use of freshly PH A-P stimulated PBMCs maximized HIV-1 isolation from blood when a sensitive HIV-1 p24 AC assay or RT assay with overnight incubation is employed for the detection of HIV in culture supernatant.

Original languageEnglish (US)
Pages (from-to)981-986
Number of pages6
JournalJournal of Acquired Immune Deficiency Syndromes
Volume3
Issue number10
StatePublished - 1990

Fingerprint

Multicenter Studies
HIV-1
Lymphocytes
Blood Cells
RNA-Directed DNA Polymerase
Acquired Immunodeficiency Syndrome
Cohort Studies
HIV Core Protein p24
Antigens
Virology
CD4 Lymphocyte Count
Coculture Techniques
HIV
Growth

Keywords

  • HIV-1
  • Isolation procedures
  • Lymphocytes

ASJC Scopus subject areas

  • Infectious Diseases
  • Pharmacology (medical)
  • Immunology and Allergy
  • Virology

Cite this

The effect of fresh lymphocytes on increased sensitivity of hiv-1 isolation : A multicenter study. / Farzadegan, Homayoon; Imagawa, David; Gupta, Phalguni; Lee, Moon H.; Jacobson, Lisa Paula; Saah, Alfred; Grovit, Kathie; Rinaldo, Charles R.; Polk, B. Frank.

In: Journal of Acquired Immune Deficiency Syndromes, Vol. 3, No. 10, 1990, p. 981-986.

Research output: Contribution to journalArticle

Farzadegan, H, Imagawa, D, Gupta, P, Lee, MH, Jacobson, LP, Saah, A, Grovit, K, Rinaldo, CR & Polk, BF 1990, 'The effect of fresh lymphocytes on increased sensitivity of hiv-1 isolation: A multicenter study', Journal of Acquired Immune Deficiency Syndromes, vol. 3, no. 10, pp. 981-986.
Farzadegan, Homayoon ; Imagawa, David ; Gupta, Phalguni ; Lee, Moon H. ; Jacobson, Lisa Paula ; Saah, Alfred ; Grovit, Kathie ; Rinaldo, Charles R. ; Polk, B. Frank. / The effect of fresh lymphocytes on increased sensitivity of hiv-1 isolation : A multicenter study. In: Journal of Acquired Immune Deficiency Syndromes. 1990 ; Vol. 3, No. 10. pp. 981-986.
@article{4a0b9ad22f7f4d48b111289ee8711aaa,
title = "The effect of fresh lymphocytes on increased sensitivity of hiv-1 isolation: A multicenter study",
abstract = "A multicenter study was undertaken to determine the sensitivity and reproducibility of markers for human immunodeficiency virus type 1 (HIV-1) viral growth and the effect of various preparations of lymphocytes on the sensitivity of standard and routinely used procedures for HIV-1 isolation. In phase 1, cocultivated culture supernatants obtained from 10 HIV-1 cultures were transported to three Multicenter AIDS Cohort Study (MACS) Virology Laboratories. Three commercial HIV-p24 antigen capture (AC) tests and two reverse transcriptase (RT) assays were used to ascertain the replication of HIV-1. The Du Pont and Abbott AC assays were found to be most sensitive (85-100{\%}), and the RT assay with 24-h incubation period had comparable sensitivity (75-100{\%}). In phase II, the sensitivity of standard cocultivation procedure for HIV-1 isolation was compared using freshly phytohemaggluti-nin-P (PHA-P)-stimulated, stimulated-frozen, and frozen-thawed and then stimulated normal human peripheral blood mononuclear cells (PBMCs) as co-cultivating cells. Blood samples from 13 HIV-1 infected individuals with various CD4+ cell counts were cocultivated in each of the three MACS laboratories using one of the aforementioned normal PBMCs. The PHA-P-stimulated fresh normal PBMC showed a maximum isolation rate of 100{\%} (13 of 13) with an average of 8 days to positivity. This rate of isolation was significantly greater than other rates using any one of the other PBMC preparations. These findings demonstrated that the use of freshly PH A-P stimulated PBMCs maximized HIV-1 isolation from blood when a sensitive HIV-1 p24 AC assay or RT assay with overnight incubation is employed for the detection of HIV in culture supernatant.",
keywords = "HIV-1, Isolation procedures, Lymphocytes",
author = "Homayoon Farzadegan and David Imagawa and Phalguni Gupta and Lee, {Moon H.} and Jacobson, {Lisa Paula} and Alfred Saah and Kathie Grovit and Rinaldo, {Charles R.} and Polk, {B. Frank}",
year = "1990",
language = "English (US)",
volume = "3",
pages = "981--986",
journal = "Journal of Acquired Immune Deficiency Syndromes",
issn = "1525-4135",
publisher = "Lippincott Williams and Wilkins",
number = "10",

}

TY - JOUR

T1 - The effect of fresh lymphocytes on increased sensitivity of hiv-1 isolation

T2 - A multicenter study

AU - Farzadegan, Homayoon

AU - Imagawa, David

AU - Gupta, Phalguni

AU - Lee, Moon H.

AU - Jacobson, Lisa Paula

AU - Saah, Alfred

AU - Grovit, Kathie

AU - Rinaldo, Charles R.

AU - Polk, B. Frank

PY - 1990

Y1 - 1990

N2 - A multicenter study was undertaken to determine the sensitivity and reproducibility of markers for human immunodeficiency virus type 1 (HIV-1) viral growth and the effect of various preparations of lymphocytes on the sensitivity of standard and routinely used procedures for HIV-1 isolation. In phase 1, cocultivated culture supernatants obtained from 10 HIV-1 cultures were transported to three Multicenter AIDS Cohort Study (MACS) Virology Laboratories. Three commercial HIV-p24 antigen capture (AC) tests and two reverse transcriptase (RT) assays were used to ascertain the replication of HIV-1. The Du Pont and Abbott AC assays were found to be most sensitive (85-100%), and the RT assay with 24-h incubation period had comparable sensitivity (75-100%). In phase II, the sensitivity of standard cocultivation procedure for HIV-1 isolation was compared using freshly phytohemaggluti-nin-P (PHA-P)-stimulated, stimulated-frozen, and frozen-thawed and then stimulated normal human peripheral blood mononuclear cells (PBMCs) as co-cultivating cells. Blood samples from 13 HIV-1 infected individuals with various CD4+ cell counts were cocultivated in each of the three MACS laboratories using one of the aforementioned normal PBMCs. The PHA-P-stimulated fresh normal PBMC showed a maximum isolation rate of 100% (13 of 13) with an average of 8 days to positivity. This rate of isolation was significantly greater than other rates using any one of the other PBMC preparations. These findings demonstrated that the use of freshly PH A-P stimulated PBMCs maximized HIV-1 isolation from blood when a sensitive HIV-1 p24 AC assay or RT assay with overnight incubation is employed for the detection of HIV in culture supernatant.

AB - A multicenter study was undertaken to determine the sensitivity and reproducibility of markers for human immunodeficiency virus type 1 (HIV-1) viral growth and the effect of various preparations of lymphocytes on the sensitivity of standard and routinely used procedures for HIV-1 isolation. In phase 1, cocultivated culture supernatants obtained from 10 HIV-1 cultures were transported to three Multicenter AIDS Cohort Study (MACS) Virology Laboratories. Three commercial HIV-p24 antigen capture (AC) tests and two reverse transcriptase (RT) assays were used to ascertain the replication of HIV-1. The Du Pont and Abbott AC assays were found to be most sensitive (85-100%), and the RT assay with 24-h incubation period had comparable sensitivity (75-100%). In phase II, the sensitivity of standard cocultivation procedure for HIV-1 isolation was compared using freshly phytohemaggluti-nin-P (PHA-P)-stimulated, stimulated-frozen, and frozen-thawed and then stimulated normal human peripheral blood mononuclear cells (PBMCs) as co-cultivating cells. Blood samples from 13 HIV-1 infected individuals with various CD4+ cell counts were cocultivated in each of the three MACS laboratories using one of the aforementioned normal PBMCs. The PHA-P-stimulated fresh normal PBMC showed a maximum isolation rate of 100% (13 of 13) with an average of 8 days to positivity. This rate of isolation was significantly greater than other rates using any one of the other PBMC preparations. These findings demonstrated that the use of freshly PH A-P stimulated PBMCs maximized HIV-1 isolation from blood when a sensitive HIV-1 p24 AC assay or RT assay with overnight incubation is employed for the detection of HIV in culture supernatant.

KW - HIV-1

KW - Isolation procedures

KW - Lymphocytes

UR - http://www.scopus.com/inward/record.url?scp=0025114216&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025114216&partnerID=8YFLogxK

M3 - Article

C2 - 2118952

AN - SCOPUS:0025114216

VL - 3

SP - 981

EP - 986

JO - Journal of Acquired Immune Deficiency Syndromes

JF - Journal of Acquired Immune Deficiency Syndromes

SN - 1525-4135

IS - 10

ER -