Cellular Immuno Assays and Phenotyping

Location: Thomas Building, DE-550
Contact phone: (206) 667-6455
Contact fax: (206) 667-6845
Contact e-mail: jcao@fredhutch.org

Lymphoproliferation assay

The Lymphoproliferation assay is the classic method for assessing lymphocyte response to stimulation. Lymphocytes proliferate after incubation in vitro with mitogens and specific antigens, or in a mixed lymphocyte reaction, which is then measured by 3H-thymidine incorporation. The rate of proliferation provides an indication for general lymphocyte activity or responsiveness to specific antigens. The Immune Monitoring resource has provided this assay for several clinical trials in accordance to specific study requirements.

Chromium-51 release assay

Cytotoxic T cells (CTL) and Natural Killer (NK) cells function as their names indicate by killing their target cells, which can be measured by the 51Cr - release assay. Specific target cells are loaded with Chromium-51, which is then released into the medium when the cells are lysed by specific CTL or NK cells. This functional assay is widely used in studies of antigen-specific CTL and NK activity.

Immune phenotyping by FACS analysis

Analyzing cell surface and intracellular proteins by labeling with fluorochrome-conjugated antibodies and flow cytometry is a common method in immunology. Flow cytometric analysis can be used to enumerate cell subsets according to their differential markers, activation, functional and other characteristics. Specific cell subsets can be sorted out from a large population of cells using a cell sorter. Antigen-specific T cells can be analyzed after labeling with specific pMHC tetramer. The Immune Monitoring lab provides cellular analysis by flow cytometry in accordance to specific study requirements. For example, we have provided complete peripheral lymphocyte phenotyping of B cells, T cells, Monocytes, NK and dendritic cells for the evaluation of immune reconstitution after stem cell transplant.

ELISPOT Assay

The Enzyme-linked ImmunoSpot (Elispot) assay is a high-throughput procedure to evaluate antigen-specific T cell responses on a single cell basis. In this assay, cells that are responsding to antigenic stimulation secrete cytokines which are captured locally by antibodies, and visualized by staining in the form of spots. Thus, precise enumeration of responsive cells can be made. The Immune Monitoring resource provides training and technical support for investigators performing this assay and using the Elispot reader. We also provide complete assays of clinical cell or blood samples.

For any of the above assays please use iLab to request the service.