Rhesus Macaque CODEX Multiplexed Immunohistochemistry Panel for Studying Immune Responses During Ebola Infection.

Non-human primate (NHP) animal models are an integral part of the drug research and development process. For some biothreat pathogens, animal model challenge studies may offer the only possibility to evaluate medical countermeasure efficacy. A thorough understanding of host immune responses in such NHP models is therefore vital. However, applying antibody-based immune characterization techniques to NHP models requires extensive reagent development for species compatibility. In the case of studies involving high consequence pathogens, further optimization for use of inactivated samples may be required. Here, we describe the first optimized CO-Detection by indEXing (CODEX) multiplexed tissue imaging antibody panel for deep profiling of spatially resolved single-cell immune responses in rhesus macaques. This 21-marker panel is composed of a set of 18 antibodies that stratify major immune cell types along with a set three Ebola virus (EBOV)-specific antibodies. We validated these two sets of markers using immunohistochemistry and CODEX in fully inactivated Formalin-Fixed Paraffin-Embedded (FFPE) tissues from mock and EBOV challenged macaques respectively and provide an efficient framework for orthogonal validation of multiple antibody clones using CODEX multiplexed tissue imaging. We also provide the antibody clones and oligonucleotide tag sequences as a valuable resource for other researchers to recreate this reagent set for future studies of tissue immune responses to EBOV infection and other diseases.

Informations de copyright

Copyright © 2021 Jiang, Mukherjee, Bennett, Chen, Logue, Dighero-Kemp, Kurtz, Adams, Phillips, Schürch, Goltsev, Hickey, McCaffrey, Delmastro, Chu, Reader, Keesler, Galván, Zlobec, Van Rompay, Liu, Hensley, Nolan and McIlwain.

Déclaration de conflit d'intérêts

GN and YG are co-founders and stockholders of Akoya Biosciences, Inc. GN, YG, and DM are inventors on CODEX-related patents US9909167 and US9909167B2, respectively. CS is a scientific advisor to Enable Medicine, Inc. CS, DP, and GN are inventors on a patent application related to CODEX (PCT/US2021/016641). EFM has previously consulted for Ionpath, Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.