Abstract for Grants
The City of Hope Clinical Immunobiology Correlative Studies laboratory (CICSL) is a Shared Resource for the City of Hope Comprehensive Cancer Center. The laboratory occupies approximately 1000 square feet of laboratory space in the Shapiro building (rooms 1002, 1044, and 1045). This space includes a 275 square foot tissue culture area, a 275 square foot area dedicated to qPCR and ELISA, a 250 square foot room dedicated to liquid nitrogen, -80°, and -20° C storage, and a 200 square foot room housing dedicated to Luminex equipment. CICSL laboratory operations are guided by GLP principles of operation, with all possible equipment 21 CFR part 11 compliant.
 
The CICSL includes state-of-the-art equipment for monitoring immune responses to cancer therapy. Specific equipment includes a fully automated microplate based multi-detection reader capable of reading fluorescence intensity, time-resolved fluorescence, high-performance luminescence (flash and glow), and absorbance, a Luminex FLEXMAP 3D bead array reader, a Luminex BioPlex™-HTX bead array reader, two MJR Chromo 4™ Q-PCR machine, and a Guava PCA-96 analytical cell counter and analyzer. The CICSL additionally makes use of access to instruments within the City of Hope Analytical Flow Cytometry Core, including a Beckman Coulter Gallios three-laser flow cytometer capable of evaluating fluorescence in ten distinct colors, and analytical cell sorters.
 
Current assays within the capabilities of the laboratory include:
 
  • Flow Cytometry: Characterization of immune cell activation/differentiation/homing status; analysis of immune cell effector phenotypes and functionality (ICC, CD107); absolute count determination and quantification of immune cell subsets.
 
  • Molecular: quantitative real-time PCR-based quantification of immune cell subsets (including Treg cells) and mRNA levels for immune effector genes; qPCR-based quantification of adoptively transferred cells; Vβ spectratyping.
 
  • Biochemical: Quantification of multiple biomarkers and analytes in serum, plasma, tissue samples and cell culture medium using Luminex bead array and ELISA platforms. These analytes include cytokines and chemokines, growth factors, endocrine markers and hormones, cardiovascular markers, bone metabolism markers, cell signaling molecules, and cancer markers.
 
  • Cell-based: Blood processing; PBMC/serum/plasma isolation; freezing, and storage; CD8 and NK cytolysis bioassays.