The mission of the USC Arnold and Mabel Beckman Center for Immune Monitoring is to provide the USC community and external customers with the highest quality support for clinical trials for immunotherapy of human cancers; facilitate basic, translational, and clinical research with an immunological monitoring component; and provide immunological assay training and education. The IM core provides quality assessments of immunological function necessary for the study of pathogenesis, treatment, and prevention of cancer and other human diseases.
Supporting clinical trials for immunotherapy of human cancer:
For physicians involved in clinical trials for immunotherapy of human cancer, the Beckman Center for Immune Monitoring Core offers several assays to assess patient immune responses. Such assays provide a highly sensitive and quantitative means of measuring the efficacy of an experimental treatment as a surrogate endpoint or as a correlative to clinical responses. It can help establish proof of principle that a treatment works, and suggest changes to adapt a standard treatment protocol to individual patient needs. Besides the Gold Standard of Immune monitoring: the ELISPOT assay for detection of cytokines released by activated T cells, we also perform intracellular cytokine staining and can detect tumor specific T cells by MHC tetramer analysis. Helper and killer T cells can be separated, and proliferative and cytotoxic responses are quantified in radioactive assays. The monitoring protocol of choice is tailor made to fit the needs and the possibilities of each treatment protocol. The core offers freezing and storage capacity for clinical samples and full analytical support for monitoring assays. It is also possible to send in samples for post-assay analysis using our state-of-the-art ELISPOT reader.
Facilitating research with an immune monitoring component:
For researchers within travel distance of our facility, we offer equipment and technical assistance for performance and/or analysis of ELISPOT assays, flow cytometry-based assays, radioisotope-based proliferation and cytotoxicity assays, multiplexing ELISA assays, and molecular-based quantitative PCR assays. Our lab has equipment for controlled rate freezing and storage of cryopreserved peripheral blood cells, tissue culture facilities and an ELISPOT reader to analyze your 96 well plates. Assays and technical support are also available to support translational projects that utilize in-vitro systems and in vivo animal models.
This assay is used to quantify the number of cells in your sample secreting a particular cytokine. It is most commonly used to estimate the proportion of T cells reactive with a particular antigen, since responder cells will secrete IFN-g when encountering their antigen. The IMC can currently evaluate T cell responses to either recall antigens to monitor general immune responsiveness or to a specific antigen of your choice using overlapping sets of peptides. The assay can be performed using whole blood samples taken from cancer patients before, during, and after treatment by immunotherapy. Antibody-coated culture wells are used to plate the responder cells. After in vitro stimulation, captured cytokine is detected by another, enzymatically labeled antibody. Colored spots in the well represent individual cytokine secreting cells. In dual color ELISPOT assays, two different cytokines are detected simultaneously using separate enzymatic labels. The IMC can perform the ELISPOT assay for you, or you can come in with your stained plates to assess the spot number using our state-of-the-art Zeiss ELISPOT analyzer.
Characterization of immune cell activation and effector phenotypes is an important measure of responses to immunotherapy. Flow cytometry is used to detect and quantify cells stained with fluorescently labeled antibodies. Markers of cellular differentiation and homing status can also be measured. Another method that requires flow cytometric analysis and is frequently used in immune monitoring is intracellular cytokine staining, based on the principle, that immunocytes will generate cytokines upon activation. Thus, we can look for T cells responsive to a peptide of interest by quantification of cytokine producing cells before and after addition of the antigen. In this case we are looking for cytokine molecules that have not yet been secreted by the cells. General markers of immune activation can also be used to assess immune responses in patients.
Cytokine measurements are performed either through traditional ELISA assays or on a Bio-Rad Bio-plex System using the Luminex xMAP® technology. The Luminex xMAP® technology allows for the measurement of up to 100 distinct proteins through a suspension bead array that employs a series of distinct color-coded microspheres to simultaneously detect multiple soluble analytes from a single serum, plasma, tissue culture supernatant, or other bodily fluid sample. Biomarker panels for obesity, cardiovascular disease, cancer, endocrinology, cell signaling, bone metabolism, and immunoglobulin isotyping are also available in addition to a wide selection of cytokines and chemokines. The system uses very small sample volumes and delivers fast and cost-effective results when analyzing multiple proteins. Please contact us to discuss which cytokines can be measured simultaneously and for recommendations for sample preparation.
MHC Tetramer analysis is used to quantify the number of T cells within a given sample that can recognize a particular antigenic epitope based upon expression of a particular T cell receptor. Antigenic epitopes, particularly those presented in the context of HLA-A2 molecules, are rapidly being identified. Using antigenic eptides bound to four (MHC tetramer) or five (MHC pentamer) copies of the HLA molecule linked to a fluorescent marker, a tool has been created to label T cells responsive to an antigen of interest within your sample. Fluorescently-labeled tetramer-positive cells are detected and quantified using flow cytometry. Cells can also be labeled with fluorescent antibodies to cell surface markers to identify phenotypic markers.
A very convenient service of the Immune Monitoring Core is the standardized processing and isolation of peripheral blood lymphocytes or serum from tubes of blood or bags from apheresed normal donors or patients. For some studies, isolation and freezing of PBMC is performed for analysis at a later date, for example, when multiple time points are collected from one patient over the course of a trial. For other studies, fresh PBMC are used in functional, descriptive, or molecular assays. PBMC are stored in the vapor phase of large capacity liquid nitrogen tanks. Serum or plasma samples are stored in -80° C ultra-low freezers.
T cell proliferation in response to an antigen of interest is considered a functional marker for immune reactivity. We measure proliferation by traditional tritiated thymidine incorporation assays, where radiolabeled nucleotides are built into newly synthesized DNA in the S phase of mitosis. Effector functions are measured by looking for actual cytotoxic activity towards target cells of interest. This important parameter of immune reactivity provides a close measure of tumor shrinking ability in cancer patients. Release of chromium by dying radiolabeled target cells serves as a measure of cytotoxicity, as mediated by specific CD8 T cells within your sample. CFSE dye-based proliferation assays can also be performed in conjunction with flow cytometry-based immunophenotyping.
Messenger RNA extracted from tumor biopsies can be analyzed by quantitative RT-PCR for specific transcripts, such as cytokines, cellular markers, or regulatory T cell markers.
Immune Monitoring Core Facility has not received any reviews.
Immune Monitoring Core Facility has not received any endorsements.