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Penn Center for AIDS Research

Immunology Core Services

Mission | Personnel | Services | Contact Info

The Immunology Core provides a wide range of services and reagents to benefit the immunological research of the Penn CFAR Community. Services are provided by facilities in both the University of Pennsylvania and the Children's Hospital of Philadelphia Research Institute and are housed at several sites. Services include:

  • Primary Cell Purification (Riley Lab, 8th Floor Similow Center for Translational Research)
    • Freshly isolated primary PBMCs, CD4, CD8, T cells, NK, monocytes and B cells
    • CCR5 deficient T cells and monocytes from Δ32 donors
  • Standard Immunological Assays (12th Floor Abramson Research Center, CHOP)
    • Cell Based Assays
    • Flow Based Assays and Services
  • Cellular, Humoral and Molecular Immunology assays (HIC, 410 & 704 Stellar Chance)
    • Specimen Processing, Banking and Sample Shipment
    • Luminex Assays
    • Flow Cytometry
    • ELISPOT
    • ELISA
    • Sequencing of Human and Murine IgH rearrangement
    • Sequencing of Human TCR Vβ Rearrangements
    • Amplification and cloning of Ig rearrangements
    • Data analysis pipeline for repertoire studies
    • Customized data analysis
    • Scientific Consultation and Assay Development
  • BSL-3 Cell Sorting Facility
  • Small Animal Model of HIV-1 Infection

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Primary Cell Purification

Cell PurityWithin the last several years there has been a greatly increased demand for primary cells in both HIV and immunological research. There are multiple reasons for this. There has been a shift in the field from the use of cell lines to primary cells for research. For example, during the transition from primary cell to transformed cell line, critical signal transduction pathways are altered, which complicates the extrapolation of data to primary cells. Moreover, certain cell types, such as dendritic cells (DC), do not have cell lines that faithfully model their activities, necessitating the use of primary cells. Lastly, certain experiments require cells with a particular genotype or HIV status. Transformed cell lines are popular because they are inexpensive, require little expertise to use, and are easy to propagate; the major goal of this Immunology Core service is to make primary cells as accessible and easy to use as transformed cells.

Dr. Riley and his lab have been purifying primary human cells for research since 1995 and distributing to others as a function of the CFAR Immunology Core since 2003. For the most part, investigators prefer cells that have been purified via a negative selection process, which leaves them as minimally manipulated as possible. Cells are obtained from healthy donors by leukapharesis, and depending on users' need, are separated into subsets by negative selection using Rosette-Sep platform developed by STEMCELL Technologies. Cells are generally provided fresh to investigators or, for those who require large batches of single-donor cells to be used at different time points, can be provided as frozen aliquots. The cell purity is always greater than 90% and is routinely greater than 95%. During the time Core E has served the Penn CFAR community, we have isolated cells from greater than 600 apheresis products; a sample summary for one such product is shown in Table 1. Several investigators have requested purified cells from HIV-1 infected donors. This can be set up on a case by case basis. These cells are only available to investigators who have specifically requested cells from HIV-1 infected donors.

Table 1. Typical Cell Request

Cell Type

# Requested

Purity

CD4 T cell

350 million

95%

CD8 T cell

100 million

97%

Monocytes

350 million

95%

PBMC

200 million

N/A

CD8-depleted PBMC

100 million

98%

This is an ideal core function because the effort required to isolate 10 million cells is the same as that to isolate one billion cells. Rather than having all laboratories who wish to work with primary cells obtain their own IRB-approved protocol, recruit donors, and develop the expertise to purify cells, the Immunology Core performs all of these functions, and since the entire apheresis product is utilized, the price per cell is much less than if the researchers were to obtain their own cells. A major part of the appeal and popularity of this service is that it removes the regulatory paperwork required to work with primary cells: The Immunology Core maintains the IRB-approved protocol and the individual core users receipt of these cells is considered secondary use of de-identified human specimens which is not considered human use by both NIH and our IRB. This recent clarification of what human use is removes a significant burden to use primary human cells to study HIV-1. We have found that the decreased regulatory burden often makes it more attractive for labs to carry out even pilot experiments with primary human cells, and not resort to cell lines. We do maintain an inventory of frozen cells so please inquire if you would prefer frozen cells.

Donors are not tested for any infectious agents. So as with all blood borne products, users are encouraged to follow universal precautions.

Ordering information:
Order cells at the Human Immunology Core
See when cells will be available at the Apheresis Research Donor Calendar
Follow order status on Twitter via @ImmuneCellsRUs    

Primary Contact:
James L. Riley, Ph.D.
rileyj@exchange.upenn.edu
215-573-6792