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Human Immunology Core

Director: Eline Luning Prak, MD, PhD

The Human Immunology Core (HIC) in the Abramson Cancer Center (ACC) is one of the first and most active immune monitoring core facilities in the United States. The HIC mission is to provide wet bench expertise for the incorporation of the newest technologies into Phase I/II clinical trials using validated immunology assays, while also offering expert scientific and technical consultation to investigators conducting translational research. Services provided by the HIC are scientific consultation, sample processing, viable cell preservation and storage, standard, customized and novel assays in cellular and molecular immunology, data analysis, and human cell products. The HIC can perform studies on a pilot research basis or at the standard of Good Laboratory Practices, as needed. The Scientific Director of the HIC is Dr. Eline T. Luning Prak, Professor of Pathology and Laboratory Medicine, who is a clinical pathologist with 20 years of clinical lab experience and research expertise in B lymphocyte biology and immune repertoire profiling.

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Major Services

Human Cell Products: More than 1,500 cell product orders were filled in 2019, highlighting the importance of this service. Due to rising demand and volume, the Human Immunology Core (HIC) was able to lower prices by 10% or more for most cell products (freshly isolated cells). These prices are at least nine times lower than the most frequently used commercial vendor. Recently, Path BioResource, our administrative support team, developed an online tool that enables investigators to request products and view requests from other labs and allows HIC lab staff to better prioritize requests based upon investigator needs.

Wet Bench Support for Clinical Trials and Translational Immunology Studies: Primary and back-up processors are scheduled to accommodate variable sample processing workload throughout the week. There is a call schedule to accommodate samples arriving after hours. A manager is on call weekends and holidays. Staff are externally qualified for sample processing work and follow study-specific standard operating procedures (SOPs). The HIC has 63 SOPs that cover everything from processing and inventory management to freezer temperature monitoring and data back-up. Enhancements during the funding period include extensive validation of all HIC sample processing SOPs, detailed comparisons of different tube types and sample volumes, and studies on sample age effects on PBMC viability, recovery, and functional competence. In addition, the HIC began transferring new processing projects into a Laboratory Information Management System (LIMS; LabVantage) and is continuing to work with information services to modify LIMS data entry fields to adapt to different sample processing workflows and investigator needs for specific projects.

Perform Validated Immunology Assays With Rigorous Data Analysis: HIC staff are internally and (for some projects) externally qualified to perform flow cytometry, ELISAs, Luminex assays, ELISPOTs, and molecular immunology assays. During the project period, the HIC established a digital ELISA service and analyzed over 2,000 samples in 16 different assays after acquiring a Quanterix Simoa analyzer in 2016. The HIC also performs high-throughput sequencing of antibody and T cell receptor gene rearrangements and served as a beta test site for the 10X Genomics Human 5’ RNAseq + IgH/IgL kit. Assays are fully customizable. During the project period, a large number of different immune repertoire profiling assays were developed and optimized. These assays can be performed on fixed or fresh tissues or cells, on DNA or RNA, with or without molecular barcoding, with bulk or single cells. All assays utilize the HIC's fully integrated in-house data analysis and visualization tools.

For cell products and sample processing, the HIC has six biosafety cabinets, three large liquid nitrogen freezers and five -80 freezers for short-term sample storage. All freezers are on an electronic monitoring system. Enhancements during the funding period include acquisition of a Beckman Coulter ViCell for cell counting, a Zeiss AxioVert inverted microscope, three new Sorvall centrifuges, and an MVE Cryocart. For cellular immunology assays, the HIC relies on instruments in the Flow Cytometry and Cell Sorting Facility with access to a 10-color flow cytometer in the same building as our sample processing facilities for efficient, real-time analysis of clinical trial samples. The Core also acquired a BMG CLARIOstar microplate reader for ELISAs and a Quanterix Simoa analyzer for digital ELISA. The HIC has a CTL ImmunoSpot S3 ELISPOT reader and FlexMAP 3D and Bio-Plex 200 Luminex instruments. For molecular immunology assays, the Core has four Peltier type thermal cyclers, a Nanodrop reader for nucleic acid quantification, a BioRad Gel Documentation system, a Chromium Controller (10X Genomics) for single cell work (acquired during the most recent funding period), and an Illumina MiSeq instrument. HIC staff have access to instruments in other PSOM core facilities including an Agilent Bioanalyzer, and Illumina NovaSeq and NextSeq sequencers. For processing large datasets, a Dell with a 12-core Xeon processor and 128 Gb of memory is available. All data are stored on a dedicated 200 Tb Synology network attached storage device configured with RAID 5 for data protection. All data are also continuously backed up off-site. The HIC also uses two dedicated servers for data storage and (redundantly) backs up sample processing data.

For more information, please visit the Core's website: https://pathbio.med.upenn.edu/hic/site