Optimal conditions for recovery of the human immunodeficiency virus from peripheral blood mononuclear cells

Castro, Blesila A.; Weiss, Carol D.; Wiviott, Lory D.; Levy, Jay A.

doi:10.1128/jcm.26.11.2371-2376.1988

Cited by 105 publications

(34 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Viral Phenotypic Analysis Virus was isolated from both patients' peripheral blood mononuclear cells (PBMCs) by cocultivation with phytohemagglutinin (PHA)-stimulated PBMC (stimulated for 2 days) from seronegative blood donors as previously described [Castro et al, 1988]. HIV-1 strains obtained from initial culture were propagated by shortterm (7-10 days) passage and then 10 4 TCID 50 of each of the sequential viruses from the couple were used in infecting 2 × 10 6 cells of selected continuous cell lines (human T-cell lines: Molt-4 clone 8 provided by M. Hayami, CEMSS provided by P. Nara, and MT-2 cells provided by M. Koot).…”

Section: Methodsmentioning

confidence: 99%

Virologic, immunologic, and clinical follow-up of a couple infected by the human immunodeficiency virus type one, group O

et al. 1997

View full text Add to dashboard Cite

The pathogenic course (virologic, immunologic, and clinical changes) of infection due to human immunodeficiency virus type one (HIV-1) group O viruses is unknown at present. To address this issue, serial HIV-1 isolates from a married couple (patients A and B) infected with a group O virus were analyzed to determine the temporal association between disease status and alterations in several parameters including plasma viral burden as measured by semiquantitative polymerase chain reaction, changes in CD4+ T cells, presence of neutralizing antibodies, and the ability to induce syncytia on the MT2 cells. For patient A who has been asymptomatic for at least 8 years, both the absence of syncytium-inducing (SI) variants and the presence of autologous and heterologous neutralizing antibodies correlated with a clinically healthier status. In contrast, a switch from NSI to SI variants was observed in patient B in 1990, followed by an expanded in vitro host range, increased viral burden, and a sharp decrease in CD4+ T cells 4 years later. Moreover, plasma obtained from this patient uniformly failed to neutralize both autologous and heterologous viruses. These observations in patient B correlated with a slightly unfavorable clinical status. Based on our preliminary results, it appears that the pathogenic course of infections due to group O viruses is similar to that reported previously for infections due to group M viruses.

show abstract

Section: Methodsmentioning

confidence: 99%

Virologic, immunologic, and clinical follow-up of a couple infected by the human immunodeficiency virus type one, group O

et al. 1997

View full text Add to dashboard Cite

show abstract

“…This is accomplished using T-cell growth factor (11), now called interleukin-2 (IL-2). Although numerous articles have described optimal conditions for HIV isolation (1,(5)(6)(7), experimental studies on the influence of the IL-2 concentration on HIV isolation from plasma are lacking. The concentrations currently used range between 5% (10 U/ml) (2,6) and 10% (20 U/ml) (1, 5) IL-2/ml culture medium.…”

mentioning

confidence: 99%

Influence of Exogenous Interleukin‐2 Concentration on the Isolation of Human Immunodeficiency Virus

Zöllner

Laufs

1995

Microbiology and Immunology

View full text Add to dashboard Cite

A significant increase (P=0.015) in the HIV isolation rate from plasma samples was achieved by use of 10 U/ml exogenous interleukin‐2 compared to 20 U/ml. The sensitivity rose from 0% to 29% on patients negative for p24 core antigen (P=0.031) and from 71% to 86% in patients positive for p24 core antigen in plasma (P>0.05). Titration of infectious HIV revealed that 10 U/ml interleukin‐2 is the optimal concentration to isolate low numbers of infectious particles of HIV.

show abstract

“…Briefly, p24 antigen was quantitated using a commercial ELISA kit (p24 Antigen Capture ELISA, Abbott Diagnostics Division), following the instructions of the manufacturer. Samples were assayed for reverse transcriptase activity, as previously described (Castro et al, 1988). 2.6. Bestatin-mediated inhibition of LAPase and protein kinase C (PKC) acti6ities in HIV infected and control cells.…”

Section: Effect Of Bestatin On P24 Antigen Le6els and Re6erse Transcrmentioning

confidence: 99%

Bestatin-mediated inhibition of leucine aminopeptidase may hinder HIV infection

et al. 1997

View full text Add to dashboard Cite

Bestatin, an inhibitor of leucine aminopeptidase (LAPase), significantly decreased HIV infection as reflected by a reduced number of positive immunofluorescent cells, p24 levels, reverse transcriptase activity and the number of proviral copies found in Bestatin-treated cells. Cellular and extracellular LAPase activity in infected cells was higher than the LAPase activity found in uninfected cells. However, cellular and extracellular LAPase activity as well as total protein kinase C activity was lower in Bestatin-treated cells. Conversely, the incubation of human lymphocytic HUT78 cells with LAPase promotes HIV infectivity. The possible role of LAPase in the pathophysiology of HIV was assessed by determining LAPase serum levels in HIV infected patients. LAPase activity levels were three orders of magnitude greater in sera obtained from HIV patients than those detected in sera of uninfected individuals. Although Bestatin reduced HIV infection, a moderate decrease in the reverse transcriptase activity of chronically-infected H9 human T-lymphocytic cells was observed. Based on the higher levels of LAPase present in the serum of HIV patients and on the combined inhibitory effect of Bestatin on LAPase and on protein kinase C activities, we suggest that LAPase may play an important role in the early events of HIV infection such as viral entry.

show abstract

Optimal conditions for recovery of the human immunodeficiency virus from peripheral blood mononuclear cells

Cited by 105 publications

References 21 publications

Virologic, immunologic, and clinical follow-up of a couple infected by the human immunodeficiency virus type one, group O

Virologic, immunologic, and clinical follow-up of a couple infected by the human immunodeficiency virus type one, group O

Influence of Exogenous Interleukin‐2 Concentration on the Isolation of Human Immunodeficiency Virus

Bestatin-mediated inhibition of leucine aminopeptidase may hinder HIV infection

Contact Info

Product

Resources

About