Optimized workflow for human PBMC multiomic immunosurveillance studies.

Area of Science:

• Immunology
• Molecular Biology
• Genomics

Background:

• Deep immune profiling is crucial for understanding human health and disease.
• Interpreting complex immunological data necessitates consistent laboratory processing to minimize batch-to-batch variation.

Purpose of the Study:

• To detail a robust pipeline for the comprehensive profiling of human peripheral blood mononuclear cells.
• To reduce batch effects and enhance data reproducibility in high-dimensional flow cytometry and single-cell RNA-seq.

Main Methods:

• Development of a standardized laboratory pipeline for processing human peripheral blood mononuclear cells.
• Application of high-dimensional flow cytometry for immune cell surface marker analysis.
• Implementation of single-cell RNA sequencing for transcriptomic profiling.

Main Results:

• The developed protocols significantly reduce batch effects in immunological data.
• The pipeline generates reproducible data, enhancing the reliability of experimental findings.
• Increased throughput was achieved, allowing for more extensive immune profiling studies.

Conclusions:

• This robust pipeline provides a standardized approach for deep immune profiling.
• The methods ensure data consistency and reproducibility, crucial for biological interpretation.
• The protocol is essential for advancing the understanding of the human immune system in various conditions.