Protocol for immunophenotyping out-of-hospital cardiac arrest patients

Kohei Yamada; Jaivardhan A Menon; Yaunghyun Kim; Changde Cheng; Wenan Chen; Jenny A Shih; Ana B Villasenor-Altamirano; Xiang Chen; Immunology of Cardiac Arrest Network (I-CAN); Tomoyoshi Tamura; Louis T Merriam; Edy Y Kim; Alexandra J Weissman

doi:10.1016/j.xpro.2024.102874

Protocol for immunophenotyping out-of-hospital cardiac arrest patients

STAR Protoc. 2024 Mar 15;5(1):102874. doi: 10.1016/j.xpro.2024.102874. Epub 2024 Feb 3.

Affiliations

¹ Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Harvard Medical School, Boston, MA 02115, USA. Electronic address: kyamada7@bwh.harvard.edu.
² Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Harvard Medical School, Boston, MA 02115, USA.
³ Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA.
⁴ Department of Computational Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
⁵ Center for Applied Bioinformatics, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
⁶ Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Harvard Medical School, Boston, MA 02115, USA; Department of Emergency and Critical Care Medicine, Keio University School of Medicine, Tokyo 160-8582, Japan.
⁷ Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Harvard Medical School, Boston, MA 02115, USA. Electronic address: ekim11@bwh.harvard.edu.
⁸ Department of Emergency Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. Electronic address: weissmanaj@upmc.edu.

Abstract

Immunophenotyping of out-of-hospital cardiac arrest (OHCA) patients is of increasing interest but has challenges. Here, we describe steps for the design of the clinical cohort, planning patient enrollment and sample collection, and ethical review of the study protocol. We detail procedures for blood sample collection and cryopreservation of peripheral blood mononuclear cells (PBMCs). We detail steps to modulate immune checkpoints in OHCA PBMC ex vivo. This protocol also has relevance for immunophenotyping other types of critical illness. For complete details on the use and execution of this protocol, please refer to Tamura et al. (2023).¹.

Keywords: Clinical Protocol; Flow Cytometry/Mass Cytometry; Immunology; RNA-seq; Single Cell.

MeSH terms

Cryopreservation
Humans
Immunophenotyping
Leukocytes, Mononuclear*
Out-of-Hospital Cardiac Arrest* / diagnosis