Development of Immunological Reagents for the Identification of New World Monkey Biomarkers

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PROJECT SUMMARY This R24 OD030215 Administrative Supplement request seeks funds for the acquisition of a Cytek Northern Lights spectral flow cytometer. The requested instrument is equipped with three excitation laser lines, 16 detectors per laser, an automatic sample loader, and basic start-up Training Kit; the instrument will be housed in the Center for Sciences and Innovation (CSI) building of Trinity University in San Antonio, Texas. The goal of the “New World Monkey Immunoreagent Resource” parent award is to improve the translational value of the New World Monkey (NWM) species marmoset (MAR), squirrel monkey (SQM), and owl monkey (OLM) by generating novel monoclonal antibodies and assays able to identify important biomarkers of inflammation and metabolism, for which there is scarcity of reagents. The need for this instrument is based on the following reasons: 1) It will significantly improve the development of the immunological assays proposed in Specific Aim 2 of the parent award; these flow cytometry assays apply to six of the ten selected targets (IL-4, IL-6, IL-10, IP-10, granzyme B, and CD69) and will expand the limited assay scope caused by the use of antiquated instrumentation. 2) It will increase the rigor and reproducibility of these assays by concentrating assay development for all three NWM species in one laboratory; administrative changes in one of our collaborating institutions reduces access to more modern instruments and limits assays to just one species. 3) It will generate deeper scientific output from the limited number of cells that can be recovered from these small monkeys; spectral flow cytometry allows the use of common fluorescent dyes that are incompatible in conventional flow cytometry and can use archived spectral data, which reduces the number of cells used for control tubes. 4) It will generate new avenues for training undergraduate students and technical personnel in the practical understanding of spectral flow cytometry, a technique that has revolutionized immunophenotyping; presence of this instrument at Trinity University will give access to novel technologies to upper division Biology students. The capacity of the spectral flow cytometer to detect a large number of markers in a single cell will increase the ability to identify multiple cell subsets in small blood or tissue samples, which is critical for improving the translational value of these small body NWM species. Institutional support is provided for the long-term maintenance of this equipment.