Immunology Based Assays
The immune system is capable of mounting 2 types of responses which are separated temporally, but are linked functionally. The innate immune response which occurs during hours or a few days is usually the first line of defense against pathogens and tumors that threaten the host. This initial encounter results in activating the more elaborate components of the acquired or adaptive immune response which can take a few days to weeks to be mounted effectively and confer long lasting protective immunity against the specific pathogen. But the immune system is the proverbial double-edged sword, and while immune activation is the desired effect during a vaccination protocol or during infection, an out of control immune response that cannot be “reigned in” can lead to autoimmune disease. Not surprisingly, many recent drug development programs have focused on “immunomodulatory” drugs that can selectively target immune cell types to achieve the desired effect depending on the kind of condition being treated. There has also been growing interest in specifically targeting the innate immune system activation, which can then trigger the appropriate adaptive response downstream.
CDAS Immunology offers a broad range of in vitro and in vivo assays that allow investigators to evaluate their compounds with respect to their effect on the immune system. The molecular and cellular components of both types of immune responses can be measured qualitatively and quantitatively, thus allowing for a comprehensive immunological profile of the candidate drug, which can be then used to define efficacy.
In vitro cell-based assays:
CDAS utilizes state of the art technology including Luminex, Flow Cytometry, qPCR and Gene Arrays to help investigators define the molecular basis of drug action. Listed are some of the assays we perform routinely:
- Cell viability determination
- Identification of Toll-like receptor (TLR) agonists or antagonists by a NF-kB-Luciferase gene reporter assay in HEK293-TLR transfectants
- Activation or Inhibition assays that measure cytokines or chemokines by Luminex technology in culture supernatants from drug-treated cells
- Measurement of cell surface markers by multi-color Flow Cytometry
- Measurement of intracellular signaling phospho-proteins by Luminex technology and/or Western Blotting
- Determination of gene expression patterns by real time qPCR analysis and pathway-focused Gene Arrays
- T cell and B cell proliferation assays in response to specific peptide antigens, or mitogens (Con A, PHA) using tritiated thymidine incorporation
In vivo assays:
We can custom build in vivo assays to suit your specific needs. Some of the routine procedures are listed below:
- Immunization
- Antibody titer determinations by ELISA
- T cell activation assays (antigen-specific)
- Immunophenotyping - Enumeration of different cell types: T cells- CD4 and/or CD8, B cells, NK cells and Dendritic cells by Flow Cytometry
- Isolation of specific cell types by magnetic sorting (MACS or DYNAL) for subsequent gene profiling studies or ex vivo experimentation
