Mass Cytometry Workflow to Achieve High-Dimensional Immunophenotyping in Resource-Limited or Decentralized...

Area of Science:

• Immunology
• Biotechnology
• Clinical Research

Background:

• Regional and remote communities face higher disease burdens but are underrepresented in clinical studies due to logistical challenges.
• Immune phenotyping data from urban populations may not generalize to rural communities with unique environmental exposures.
• Existing research methods often require specialized equipment unavailable in resource-limited settings.

Purpose of the Study:

• To present a novel workflow for high-parameter mass cytometry studies adaptable to resource-limited settings.
• To enable the inclusion of geographically dispersed and underserved communities in immunological research.
• To facilitate scalable immune phenotyping by overcoming logistical barriers in sample collection and processing.

Main Methods:

• Whole blood (WB) or peripheral blood mononuclear cells (PBMCs) are collected, stained for surface antigens, and cryopreserved at the collection site.
• Samples are shipped to a central site for further processing: neutrophil depletion, fixation, barcoding, intracellular staining, and data acquisition.
• The workflow, particularly WB staining, avoids the need for specialized equipment like centrifuges, making it feasible in resource-limited environments.

Main Results:

• The workflow was successfully applied to analyze immune differences between lung cancer patients and healthy donors.
• Example data demonstrated the feasibility and application of the proposed method.
• The protocol includes detailed steps for data preprocessing and cleanup.

Conclusions:

• This workflow enhances access to clinical research for underserved populations.
• It enables the scalability of immune phenotyping studies across geographically dispersed clinical centers.
• This approach facilitates a more inclusive and representative understanding of immune responses in diverse communities.