Murine leukemia virus infectious centers are dependent on the rate of virus production by infected cells

Chesebro, Bruce; Wehrly, Kathy; Watson, Kenneth F.; Chesebro, Kevin

doi:10.1016/0042-6822(78)90239-8

Cited by 30 publications

(41 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Stock preparations of HIV-1 LAI containing approximately 10 6 infectious units͞ml were titered by syncytium induction (14). HIV was added at a multiplicity of 0.01 to 1-ml aliquots of medium containing 2 ϫ 10 5 CEM cells that were previously incubated for 1 hr with or without different deoxynucleoside analogs.…”

Section: Methodsmentioning

confidence: 99%

Lethal mutagenesis of HIV with mutagenic nucleoside analogs

Loeb

Essigmann

Kazazi

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

338

289

View full text Add to dashboard Cite

The human immunodeficiency virus (HIV) replicates its genome and mutates at exceptionally high rates. As a result, the virus is able to evade immunological and chemical antiviral agents. We tested the hypothesis that a further increase in the mutation rate by promutagenic nucleoside analogs would abolish viral replication. We evaluated deoxynucleoside analogs for lack of toxicity to human cells, incorporation by HIV reverse transcriptase, resistance to repair when incorporated into the DNA strand of an RNA⅐DNA hybrid, and mispairing at high frequency. Among the candidates tested, 5-hydroxydeoxycytidine (5-OH-dC) fulfilled these criteria. In seven of nine experiments, the presence of this analog resulted in the loss of viral replicative potential after 9-24 sequential passages of HIV in human CEM cells.

show abstract

Section: Methodsmentioning

confidence: 99%

Lethal mutagenesis of HIV with mutagenic nucleoside analogs

Loeb

Essigmann

Kazazi

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

338

289

View full text Add to dashboard Cite

show abstract

“…As an indicator of cell proliferation, absorbancy was measured at 490 nm, using a 96-well plate reader (Spectra Fluor, Tecan, Research Triangle Park, NC), Cells and Antibodies for Fluorescence-activated Cell Sorter-Sup-T1, a non-Hodgkin's T-cell lymphoma cell line (43), and the same cells stably infected with HIV type 1, III-B (A1953), were kindly provided by Dr. J. Hoxie. The mouse monoclonal antibody 902, specific for gp120 of HIV-1 III-B (44,45), was used to differentiate infected versus noninfected cells. Mouse sera were tested with dilutions ranging between 1:10 and 1:100.…”

Section: Methodsmentioning

confidence: 99%

Directing the Immune Response to Carbohydrate Antigens

Cunto-Amesty¹,

Dam²,

Luo³

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

Peptide mimetics may substitute for carbohydrate antigens in vaccine design applications. At present, the structural and immunological aspects of antigenic mimicry, which translate into immunologic mimicry, as well as the functional correlates of each, are unknown. In contrast to screening peptide display libraries, we demonstrate the feasibility of a structure-assisted vaccine design approach to identify functional mimeotopes. By using concanavalin A (ConA), as a recognition template, peptide mimetics reactive with ConA were identified. Designed peptides were observed to compete with synthetic carbohydrate probes for ConA binding, as demonstrated by enzyme-linked immunosorbent assay and isothermal titration calorimetry (ITC) analysis. ITC measurements indicate that a multivalent form of one particular mimetic binds to ConA with similar affinity as does trimannoside. Splenocytes from mimeotope-immunized mice display a peptide-specific cellular response, confirming a T-cell-dependent nature for the mimetic. As ConA binds to the Envelope protein of the human immunodeficiency virus, type 1 (HIV-1), we observed that mimeotope-induced serum also binds to HIV-1-infected cells, as assessed by flow cytometry, and could neutralize T-cell line adapted HIV-1 isolates in vitro, albeit at low titers. These studies emphasize that mimicry is based more upon functional rather than structural determinants that regulate mimeotope-induced T-dependent antibody responses to polysaccharide and emphasize that rational approaches can be employed to develop further vaccine candidates.

show abstract

“…CEM-GFP cells were infected with HIV-1 LAI strain at a MOI of 0.001TCID 50 per milliliter. At days 1 to 6 after infection, 2 ϫ 10 5 cells were harvested from the culture, centrifuged 2 min at 2,000 ϫ g, and incubated for 30 min at 4ЊC in the presence of 1 g͞ml of mouse anti-gp120 mAbs (Hybridoma 902 recognizing the V3 loop; obtained through the AIDS Research and Reference Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, from Bruce Chesebro) (19,20). The cells then were washed three times with PBS containing 0.2% BSA and 0.1% Na-azide, resuspended with 20 l of a 1:50 diluted anti-mouse IgG labeled with phycoerythrin (Boehringer-Mannheim), and incubated 30 min at 4ЊC.…”

mentioning

confidence: 99%

A new reporter cell line to monitor HIV infection and drug susceptibilityin vitro

Gervaix

West

Leoni

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

230

168

View full text Add to dashboard Cite

Determination of HIV infectivity in vitro and its inhibition by antiretroviral drugs by monitoring reduction of production of p24 antigen is expensive and time consuming. Such assays also do not allow accurate quantitation of the number of infected cells over time. To develop a simple, rapid, and direct method for monitoring HIV infection, we generated a stable T-cell line (CEM) containing a plasmid encoding the green f luorescent protein (humanized S65T GFP) driven by the HIV-1 long terminal repeat. Clones were selected that displayed low constitutive background f luorescence, but a high level of GFP expression upon infection with HIV. HIV-1 infection induced a 100-to 1,000-fold increase in relative f luorescence of cells over 2 to 4 days as monitored by f luorescence microscopy, cytof luorimetry, and f low cytometry. Addition of inhibitors of reverse transcriptase, protease, and other targets at different multiplicities of infection permitted the accurate determination of drug susceptibility. This technique also permitted quantitation of infectivity of viral preparations by assessment of number of cells infected in the first round of infection. In conclusion, the CEM-GFP reporter cell line provides a simple, rapid, and direct method for monitoring HIV infectivity titers and antiretroviral drug susceptibility of syncytium-inducing strains.

show abstract

Murine leukemia virus infectious centers are dependent on the rate of virus production by infected cells

Cited by 30 publications

References 8 publications

Lethal mutagenesis of HIV with mutagenic nucleoside analogs

Lethal mutagenesis of HIV with mutagenic nucleoside analogs

Directing the Immune Response to Carbohydrate Antigens

A new reporter cell line to monitor HIV infection and drug susceptibilityin vitro

Contact Info

Product

Resources

About