Our Immunomonitoring laboratory provides highly specialized immunoassay services, providing support for the development of our vaccines and immunotherapy products in human clinical trials. Or lab utilizes the following techniques to dissect the immune response in cancer and HIV patients to their disease and the effect our products have on changing the immune response. We also study the immune response in healthy individuals. This information is used to optimize our clinical protocols by selecting patients most likely to respond to immunotherapy and determining whether a patient that received immunotherapy has a response that correlates with a positive outcome, such as increase in survival. The data from our immunomonitoring laboratory also assists us in improving our products and optimizing the dose, route of administration and frequency of administration of our current products in clinical trials.
The enzyme-linked immunoassay (ELISA) is used to quantify proteins, peptides, and antibodies. We use ELISA to detect levels of cytokines or other immune markers in serum, plasma, or cell culture supernatants, which provide valuable information on the type of immune response elicited by our vaccines and immunotherapy.
When it is desired to measure multiple cytokines or chemokines in the plasma, the multiplex assay is used as an alternative to the ELISA as it is able to detect multiple signaling agents in a single sample, which saves on the volume of blood necessary to be drawn on each patient.
The Enzyme-Linked ImmunoSpot (ELISpot) Assay measures the frequency of antigen-specific cytokine-secreting cells at the single-cell level. The ELISpot involves the culture of immune cells (e.g. peripheral blood mononuclear cells) with antigen(s) of interest, such as chaperone proteins from tumor lysate as tumor antigens or lysate of AlloStim for alloantigens and the capture of a secreted cytokine of interest immediately after secretion. The limit of detection can be as low as one cell in 100,000. Each spot formed on the plate membrane represents one cytokine-secreting cell. This allows for enumeration of the spot forming cells (SFC) using our ELISpot plate reader and associated software. The ELISpot assay provides detail of the T cell-mediated immunity (Th1 and Th2) at a single-cell level. We use the ELISpot assay to monitor the effects that our immunotherapy products have on tumor-specific and antigen-specific immunity and altering the Th1/Th2 balance.
Our expertise in ELISpot and ELISA assays is complemented by our experienced flow cytometry team. Flow cytometry is capable of providing detailed information on immune cell populations by detecting the expression of specific cell surface markers. These cell surface markers can identify cell populations of interest and their activation status, and other characteristics which may change in response to a vaccine or immunotherapy. Flow cytometry is also used for intracellular cytokine staining to determine signaling pathways (e.g. STAT4, STAT 6), apoptosis evaluation, phagocytic and oxidative burst activity, and also to measure cell proliferation and viability. The data provided by flow cytometry provides a comprehensive profile of the immune response.
Real-time PCR is a tool for detecting and quantifying expression profiling of selected genes within immune cells. This tool combined with internal cytokine staining is used to analyze signaling pathways from surface receptor to gene transcription level in the nucleus. Cytokines mRNA quantification together with NKΚB and nuclear activation of is used in immunological research to dissect the early steps of immune responses or pathophysiological pathways.