Age and gender leucocytes variances and references values generated using the standardized ONE-Study protocol.

Area of Science:

• Immunology
• Biotechnology

Background:

• Flow cytometry is crucial for analyzing immune cell changes but suffers from variability and errors, hindering cross-laboratory reproducibility.
• Standardization of flow cytometry is essential for reliable clinical trial data and patient monitoring.

Purpose of the Study:

• To validate a standardized whole blood flow cytometry protocol for profiling immune cell subsets.
• To identify age- and gender-dependent differences in immune cell populations.
• To establish a standardized reference cohort for clinical trials.

Main Methods:

• Analysis of whole blood samples from two cohorts (n=52 and n=46, ages 20-84) using standardized "ONE Study" panels.
• Profiling of >30 surface markers to enumerate >50 leukocyte subsets.
• Validation of findings by a different operator on the second cohort.

Main Results:

• Significant age-dependent changes observed, including increased activated/differentiated CD4(+) and CD8(+) T cells, memory Tregs, and decreased MDC2 and Marginal Zone B cells.
• Distinct age-related immune cell dynamics between males and females, suggesting faster immunosenescence in males.
• Standardized approach successfully validated age-dependent changes in a separate cohort.

Conclusions:

• The developed strategy standardizes whole blood flow cytometry, enabling reproducible analysis of immune cell subsets.
• Generated reproducible reference ranges account for age- and gender-specific immune variations.
• This standardization is vital for enhanced patient monitoring and personalized therapeutic strategies.