Flow cytometric assays
The laboratory currently performs cell surface phenotyping for a wide range of markers, apoptotic assays, and intracellular cytokine assays by flow cytometry for many clinical studies including the adult and pediatric AIDS Clinical Trials Group. Basic and specialized flow panels can be custom-made to meet investigator needs, and memory T cell and common variable immunodeficiency flow panels have been developed to identify children with immunodeficiency disorders.
Functional Analyses of Lymphocytes
The laboratory performs lymphocyte functional assays, such as antigen/mitogen proliferation assays and natural killer cell assays. A wide range of monokine, chemokine, and cytokine assays are routinely available as are tests for soluble cytokine receptors such as TNF-RII and IL-2R. Markers of cellular activation are also available and include beta-2-microglobulin, C-reactive protein and neopterin.
Other tests
Several endocrine tests such as cortisol, norepinephrine and epinephrine are available on serum and plasma samples. Salivary cortisol testing is also available.
HIV screening and western blot confirmation testing is performed on serum samples.
Sample Preparation and Storage
Separation and aliquoting of cells, serum and plasma for storage is provided. Long-term storage of peripheral blood mononuclear cells in liquid nitrogen freezers and storage of serum, plasma, saliva, urine and other body fluids is available at ultra-cold temperatures. Detailed records of specimen type, number, and location are computerized using repository management hardware and software that allows sample barcoding for rapid processing and storage/retrieval.
Quality Control Program
A major feature of services provided by the CIRL is the extensive quality control program. For natural killer cell and proliferation assays, day-to-day variability is evaluated throughout the week by parallel testing of samples obtained from healthy donors. The same donor is used throughout the week to ensure acceptable assay variability. Historical records are used to confirm long-term assay performance. The CIRL also participates in the NIH/NIAID/DAIDS quality assessment program for lymphocyte proliferation assays.
For cytokine and soluble marker/activation marker assays, approximately 10% of all samples tested in each run are re-assayed in a subsequent assay to ensure reproducible results. In addition, in-house control materials are included in each run to ensure acceptable long-term analytical performance. All immunoassays have been extensively evaluated for performance characteristics such as specimen stability, linearity, precision, lower limit of detection, and analyze interferences prior to use for unknown samples. The laboratory also subscribes to the CAP proficiency program for soluble serum markers such as beta-2 microglobulin.
For long-term storage of viable cells in liquid nitrogen, selected vials are thawed on a monthly basis and examined for viability and recovery as part of the quality control program.
