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

Frontiers in Immunology

卷:12

J. Rachel Reader¹  Rebekah I. Keesler¹  Koen K. A. Van Rompay¹  Sizun Jiang²  Christian M. Schürch³  Garry P. Nolan⁴  Nilanjan Mukherjee⁴  Erin F. McCaffrey⁴  John W. Hickey⁴  Han Chen⁴  Yury Goltsev⁴  Pauline Chu⁴  Alea Delmastro⁴  David R. McIlwain⁴  Darci Phillips⁴  José A. Galván⁵  Inti Zlobec⁵  Richard S. Bennett⁶  James Logue⁶  Lisa E. Hensley⁶  Bonnie Dighero-Kemp⁶  Ricky Adams⁶  Jonathan R. Kurtz⁶  David X. Liu⁶ 

[1] California National Primate Research Center, University of California, Davis, CA, United States;

[2] Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Boston, MA, United States;

[3] Department of Pathology and Neuropathology, University Hospital and Comprehensive Cancer Center Tübingen, Tübingen, Germany;

[4] Department of Pathology, Stanford University School of Medicine, Stanford, CA, United States;

[5] Institute of Pathology, University of Bern, Bern, Switzerland;

[6] Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Frederick, MD, United States;

关键词: codex;    EBOV (Ebola virus);    rhesus macaque (Macaca mulatta tcheliensis);    NHP (non-human primate);    Spatial biology;    multiplexed immunofluorescencence and immunohistochemistry;   

DOI  :  10.3389/fimmu.2021.729845

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown