HIV-1 isolation from plasma specimens

Costa, C. L.; Morgado, M. G.; Santos, Valdiléa Gonçalves Veloso dos; Bongertz, Vera

doi:10.1590/s0074-02761996000600017

Cited by 4 publications

(1 citation statement)

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“…Viruses were cultured in a two-step procedure termed the small and large-scale cultures. The protocol utilized from culture was adapted from previously published methods (Albert et al, 1987; Falk et al, 1987; Costa et al, 1996; Aguado-Martinez et al, 2009). Briefly, cryopreserved PBMCs pooled from five donors, termed feeder cells, were activated with α-CD28 at 100ng/mL (BD Biosciences, San Jose, CA) and α-CD3 at 50ng/mL (eBioscience, San Diego, CA) three days prior to the start of culture.…”

Section: Methodsmentioning

confidence: 99%

Development of a contemporary globally diverse HIV viral panel by the EQAPOL program

Sánchez

DeMarco

Hora

et al. 2014

Journal of Immunological Methods

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The significant diversity among HIV-1 variants poses serious challenges for vaccine development and for developing sensitive assays for screening, surveillance, diagnosis and clinical management. Recognizing a need to develop a panel of HIV representing the current genetic and geographic diversity NIH/NIAID contracted the External Quality Assurance Program Oversight Laboratory (EQAPOL) to isolate, characterize and establish panels of HIV-1 strains representing global diverse subtypes and circulating recombinant forms (CRFs), and to make them available to the research community. HIV-positive plasma specimens and previously established isolates were collected through a variety of collaborations with a preference for samples from acutely/recently infected persons diagnosed since 2009. Source specimens were cultured to high-titer/high-volume using well-characterized cryopreserved PBMCs from healthy donors. Panel samples were stored as neat culture supernatant or diluted into defibrinated plasma. Characterization for the final expanded virus stocks included viral load, p24 antigen, infectivity (TCID), sterility, coreceptor usage, and near full-length genome sequencing. Viruses are made available to approved, interested laboratories using an online ordering application. The current EQAPOL Viral Diversity panel includes over 101 viral specimens representing 6 subtypes (A, B, C, D, F, and G), 2 sub-subtypes (F1 and F2), 7 CRFs (01, 02, 04, 14, 22, 24, and 47), 19 URFs and 3 group O viruses from 22 countries. The EQAPOL Viral Diversity panel is an invaluable collection of well-characterized reagents that are available to the scientific community, including researchers, epidemiologists, and commercial manufacturers of diagnostics and pharmaceuticals to support HIV research and diagnostic and vaccine development.

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Section: Methodsmentioning

confidence: 99%

Development of a contemporary globally diverse HIV viral panel by the EQAPOL program

Sánchez

DeMarco

Hora

et al. 2014

Journal of Immunological Methods

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Sequence analysis of HIV-1 isolates from Guinea-Bissau: selection of vaccine epitopes relevant in both West African and European countries

Vinner

Holmgren

Jensen

et al. 2011

Apmis

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For a CD8 epitope-based vaccine to match different geographic locations, the targeted epitopes for cytotoxic T-lymphocytes (CTLs) must be present in the local circulating HIV-1 strains. Secondly, the vaccine epitopes should match the host population HLA types. We characterized two new HIV-1 isolates from Guinea-Bissau. Also, we have identified 15 subdominant CD8 epitopes representing common HLA super-types theoretically covering most HLA alleles in any population. Herein we demonstrate that the selected vaccine epitopes are well conserved and simultaneously present in sequences from West Africa and Denmark. Use of the selected epitopes will likely ensure ≥10 immune targets in the majority of candidates for experimental therapeutic vaccination in both geographic regions. Our results warrant testing of the selected vaccine epitopes in both geographic locations.

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Effect of Complement on HIV-2 Plasma Antiviral Activity Is Intratype Specific and Potent

Şahin

Holmgren

Sheik-Khalil

et al. 2013

J Virol

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Human immunodeficiency virus type 2 (HIV-2)-infected individuals develop immunodeficiency with a considerable delay andtransmit the virus at rates lower than HIV-1-infected persons. Conceivably, comparative studies on the immune responsiveness of HIV-1-and HIV-2-infected hosts may help to explain the differences in pathogenesis and transmission between the two types of infection. Previous studies have shown that the neutralizing antibody response is more potent and broader in HIV-2 than in HIV-1 infection. In the present study, we have examined further the function of the humoral immune response and studied the effect of complement on the antiviral activity of plasma from singly HIV-1-or HIV-2-infected individuals, as well as HIV-1/ HIV-2 dually infected individuals. The neutralization and antibody-dependent complement-mediated inactivation of HIV-1 and HIV-2 isolates were tested in a plaque reduction assay using U87.CD4.CCR5 cells. The results showed that the addition of complement increased intratype antiviral activities of both HIV-1 and HIV-2 plasma samples, although the complement effect was more pronounced with HIV-2 than HIV-1 plasma. Using an area-under-the-curve (AUC)-based readout, multivariate statistical analysis confirmed that the type of HIV infection was independently associated with the magnitude of the complement effect. The analyses carried out with purified IgG indicated that the complement effect was largely exerted through the classical complement pathway involving IgG in both HIV-1 and HIV-2 infections. In summary, these findings suggest that antibody binding to HIV-2 structures facilitates the efficient use of complement and thereby may be one factor contributing to a strong antiviral activity present in HIV-2 infection.

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HIV-1 isolation from plasma specimens

Cited by 4 publications

References 0 publications

Development of a contemporary globally diverse HIV viral panel by the EQAPOL program

Development of a contemporary globally diverse HIV viral panel by the EQAPOL program

Sequence analysis of HIV-1 isolates from Guinea-Bissau: selection of vaccine epitopes relevant in both West African and European countries

Effect of Complement on HIV-2 Plasma Antiviral Activity Is Intratype Specific and Potent

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