Deep Immune Status Monitoring with Multi-color Flow Cytometry

Weiwei Wang ,Guang Fan ,Lisong Shen

doi:10.33696/haematology.2.039

Abstract

With the development of immune therapy, the role of immune monitoring during treatments has been emphasized. Flow cytometry is the best method for immune status monitoring because of its characteristic of multi-parameter analysis

Highlights

The white blood cell (WBC) count and lymphocyte percentage were obtained from SysmexKX-21N, which data were used to prepare single cell suspension with cell count in the range of 1 to 2 × 106/μL for antibody staining
The major immune cell populations detected by 29 antibody panel included T cell subsets (CD3+ total T, CD4+Th, CD8+Tc, Treg, CD8hi T, CD8dim T, αβTCR cells, γδTCR cells, naïve T and memory T cells), T cell activation markers and one immune checkpoint PD1(CD279), B cell subsets (CD5+B1, switched memory, non-switched, naïve B, CD27-IgD- B cells, and the light chain Kappa or Lambda), neutrophils, basophils, four monocytic cell subsets (CD13+CD14hi CD16- Mo1, CD13+CD14lowCD16+ Mo2, CD13+CD14hiCD16+ Mo3, and CD13+CD14lowCD16Mo4), dendritic cells, four NK cell subsets (CD16+CD56- NK1, CD16+CD56+ NK2, CD16-CD56low NK3, CD16low/-CD56hi NK4), and NKT (CD3+CD16+or CD3+CD56+)
Combined immunodeficiency (SCID); lymphocytosis with increased percentage of double negative T-cells (DNT) and DNT αβ-T-cells is the diagnostic criterion of autoimmune lymphoproliferative syndrome (ALPS); quantitative assessment of B cell populations and subpopulations is useful for the diagnosis of X-linked agammaglobulinemia (XLA) and Common variable immunodeficiency (CVID)

Summary

Introduction

The white blood cell (WBC) count and lymphocyte percentage were obtained from SysmexKX-21N, which data were used to prepare single cell suspension with cell count in the range of 1 to 2 × 106/μL for antibody staining. The major immune cell populations detected by 29 antibody panel included T cell subsets (CD3+ total T, CD4+Th, CD8+Tc, Treg, CD8hi T, CD8dim T, αβTCR cells, γδTCR cells, naïve T and memory T cells), T cell activation markers (percentage and MFI of CD25, CD69 and HLA-DR) and one immune checkpoint PD1(CD279), B cell subsets (CD5+B1, switched memory, non-switched, naïve B, CD27-IgD- B cells, and the light chain Kappa or Lambda), neutrophils, basophils, four monocytic cell subsets (CD13+CD14hi CD16- Mo1, CD13+CD14lowCD16+ Mo2, CD13+CD14hiCD16+ Mo3, and CD13+CD14lowCD16Mo4), dendritic cells (pDCs and mDCs), four NK cell subsets (CD16+CD56- NK1, CD16+CD56+ NK2, CD16-CD56low NK3, CD16low/-CD56hi NK4), and NKT (CD3+CD16+or CD3+CD56+). These panels of antibodies had been applied to monitor immune status (percentage and absolute number) in total 303 clinical cases with various diseases before or after treatments, such as leukemia, lymphoma, cancers, immune deficiencies, and autoimmune diseases.

Results

Conclusion