EXPERIMENTAL VIROLOGY CORE SERVICES

The Experimental Virology Core of the UMMS Center for AIDS Research offers numerous services at highly competitive prices. We foster an environment of collaborative and innovative AIDS research for UMass- and non-UMass investigators to work on independent and collaborative projects that are related to HIV and new emerging infectious human pathogens.

Training Services

  • The training session begins with an interview with the Core Director who will establish the baseline level of competence and knowledge of the potential users
  • Review the NIH guidelines for safe-handling of pathogenic agents
  • The individual is provided an orientation "walk-through" to get acquainted with the day-to-day working practices
  • The individual is briefed regarding the accidental pathogen exposure reporting system
  • A hard copy of the biocontainment procedures (Standard Operating Procedures) to be followed while using the facility is provided to the individual
  • The individual is also required to watch a video presentation of the safe practices in handling pathogens like HIV
    • Periodic presentation of this video (or updated versions) along with a review of the biocontainment procedures is scheduled to serve as a refresher course for the users of the Core D facility.

Maintenance and other appropriate (technical) personnel will work in the lab under the supervision of the Core Director.

Research Services

Elutriation Services

  • Offer monocytes/macrophages and matching PBMCs

Molecular and Virology Assays

  • Infectivity, tropism and phenotype assays
  • Develop assays and perform molecular, virology and diagnostic assays

Cells and Virus Services

  • Isolate PBMC from normal human donor blood and provide activated or non-activated primary PBMC for viral infectivity studies
  • Provide cell lines and T-cells (including enriched CD4 or CD8 T-cells) for infectivity and tropism studies
  • Provide macrophages free of T-cell contamination using immunobeads or other improved cell-separation techniques (from monocytes derived from elutriated PBMC) and autologous PBMC for infectivity and tropism studies
    • HLA-typed on request
  • The Core D facility provides phenotypically and genotypically characterized HIV-1 viral stocks
  • Transfection, Generate Pseudovirus and Labeled Virus
  • Provide well-characterized viral stocks of primary isolates and functional, biological

Qualitative and Quantitative quantification of virus

  • Instruments
  • Perform molecular detection/diagnostic assays including qualitative and quantitative RNA and DNA-PCR for viral genomes; p24 antigen ELISA
  • Perform Taqman- & LightCycler-based PCR assays for real-time detection and quantitation of viral DNA in clinical samples & for LTR-circle assay
  • Detect and quantitate HIV RNA and DNA in Whole Blood and DBS, EBV, CMV
  • Identify low levels of viral replication and loads
  • Determine HAART-induced viral load changes and resistance
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    Phenotypic & Genotypic characterizations of virus

     

    • Titer virus in appropriate host cells
    • Characterize the macrophage-tropism
    • Single genome amplification (SGA) of env genes from plasma and cells
    • Full-length sequencing of env gene
    • Determine expression of functional env protein
    • Generae env+-pseudotyped viruses
    • Determine the infectivity of the virus in CD4 T-cells
    • Determine the entry phenotype & kinetics and replication kinetics and receptor-tropism in primary T-cells, T-cell lines, and cell lines co-expressing CD4 and chemokine receptors
    • Determine Neutralizing and Inhibitory titers to huMab and entry-inhibitors
    • Determine the entry phenotype & kinetics
    • Identify and/or monitor the efficacy of potential anti-HIV compounds/drugs targeted to structural or regulatory gene functions using cell-culture systems
    • Participate in Quality Control & Certification programs related to cell-culture and PCR diagnostics and monitoring

    Repository Services

    • Maintain a repository of cell lines (also with indicator read-out genes) routinely used for viral phenotype characterization for immediate availability to investigators
    • Maintain a repository of titered viral stocks (primary isolates, lab-adapted and reference virus, pseudotyped and env+-pseudotyped viruses)
    • Maintain a repository of HIV- wild type and mutant plasmids & cell-culture reagents
    • Maintain vast repository of primary donor PBMC, and cell lines

    Fostering Interaction, Collaboration and Outreach Services

    • Training and Mentoring -- Biosafety procedures; Assays
    • Training in use of HIV and Blood-Borne Pathogens
    • Lab orientation and training for UMMS community, neighboring schools and international programs
    • Certificate Programs  (NIH, CLIA)
    • Qualitative and Quantitative PCR
    • Qualitative Co-culture Assays
    • Support of Pilot Projects (UMMS/National/International)
    • Help non-HIV researchers obtain HIV grants by providing free resources
    • Provide resources to investigators in non-AIDS research (Emerging diseases)
    • Seminars and Lab Meetings

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