The Cellular Immunity Core provides comprehensive flow cytometry, cell sorting, and cellular immune analysis support for the AIDS and Cancer Virus Program and for collaborative preclinical research studies and basic and translational research projects, using cutting-edge instrumentation, methods, and analysis software. Additionally, the core produces specialized reagents for the HIV/AIDS research community, which can be obtained at no charge. The core  provides flow cytometry protocols, consultations, and training to the research community.  

We focus on: 

  • Flow cytometry technologies, instrumentation, reagents, assays, and data analysis

  • Nonhuman primate SIV/AIDS model and cellular immunology 

  • Conventional and spectral flow cytometry protocol and antibody/reagent panel development 

  • Infectious sample cell sorting under biocontainment 

  • Flow cytometry, cell sorting, and data analysis software training 

  • SIV, HIV, and KSHV flow cytometry reagent development 

Monitoring immune system health and changes at the cellular level 

Measuring the quantity of specific immune system cells (known as “cell counts”) circulating in the body is an established and essential method for monitoring overall health in humans, especially when combating diseases like cancer and HIV infection.

Disease-causing agents like HIV and simian immunodeficiency virus, which cause AIDS in humans and nonhuman primates, respectively, directly target certain immune system cell populations. So, it is crucial to quantify these affected populations throughout the course of infection.

In support of the AIDS and Cancer Virus Program and collaborative preclinical studies, the core uses flow cytometry to detect and quantify most of the critical immune system cell populations in the blood stream, including those targeted by HIV/SIV. These immune cell population counts are used by principal investigators and other scientists to assess the effectiveness of established and experimental medications and treatment regimens and to alter treatment and study virus biology.  

Collaboration Opportunities

The Cellular Immunity Core offers services to external researchers. We consult with investigators to determine their needs and how flow cytometry could support their research goals, providing cutting-edge flow cytometry support, technical expertise, and complex data analysis. 

Our team provides immune system monitoring for internal and collaborative nonhuman primate studies, such as whole blood absolute cell counting, surface/intracellular marker phenotyping of blood and tissue suspensions, and intracellular cytokine staining to monitor functional immune responses. 

We perform single-cell and population sorting of infectious samples under biocontainment and provide novel nonhuman primate and SIV flow cytometry reagents to the AIDS and Cancer Virus Program and the greater AIDS research community.

Contact Mac Trubey for inquiries about collaborations and the use of the core’s services. 

Sharing expertise and access to novel equipment 

The core provides consultations, hands-on technical assistance, flow cytometry assay and reagent panel development, and instrument and data analysis software training to Frederick National Laboratory for Cancer Research intramural and extramural staff.

The core provides access to our well-maintained and calibrated analytical flow cytometers and biocontained cell sorters (training required before use). The core hosts the only accessible cell sorter on the Frederick National Laboratory campus that is equipped to sort infectious samples.  

Our capabilities and specializations

Streamlined absolute cell counting 

Semi-automated assay detects 15 distinct cellular markers and exogenous, infused fluorochrome-labeled cells and agents. It is used to monitor and quantify cell counts for 85+ unique leukocyte populations in a host of preclinical nonhuman primate studies. We perform absolute cell counting primarily with an Agilent NovoCyte Penteon analytical flow cytometer.

  • Advanced, 5-Laser, 27-color Agilent NovoCyte Penteon automated flow cytometer with Spectroflo software

  • Data analysis performed with De Novo Softwarer's FCS Express Version 7 program

  • All immune cell populations quantified as number of cells per microliter of whole blood

Infectious sample cell sorting 

Fluorescence-activated cell sorting (FACS) with extensive biocontainment systems and procedures, approved by our Institutional Biosafety Committee, curtail exposure to aerosols, which have the potential to contain infectious agents.

Real-time, full-spectrum analysis while sorting provides increased fluorochrome detection sensitivity and signal-to-noise.

  • Cutting-edge, 6-laser, 56-detector full-spectrum analysis, biocontained ThermoFisher Bigfoot infectious-sample cell sorter 

  • High-throughput single-cell, 96-/384-/1536-well plate, microscope slide and 10X Genomics chip sorting and simultaneous 6+ population sorting into various tubes and plates 

Spectral flow cytometry 

Spectral, or full-spectrum, analytical flow cytometers and cell sorters provide increased sensitivity and complexity analyses, enhance cost-efficiency, spare precious specimens and given their near-unlimited fluorochrome detection capability, support high-dimension cellular assessments.

  • State-of-the-art Cytek Aurora 5-laser automated spectral flow cytometer with a 64-detector APD array and 96-well plate/40-tube rack loader 

  • Increased marker detection only limited by available, laser-excitable fluorochromes 

  • Detailed cell population immunophenotyping with background autofluorescence subtraction to increase cellular resolution