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Published on: September 16, 2012
Screening of the HLDA9 panel on peripheral blood dendritic cell populations
Yitian Ding1, Xinsheng Ju, Maryam Azlan
1Mater Medical Research Institute, Aubigny Place, South Brisbane, Queensland, Australia.
Insights
Dendritic cells (DCs) are crucial for T cell responses. This study screened DC populations using flow cytometry, revealing similarities between myeloid DC subsets and an immature state in peripheral blood DCs.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are vital leukocytes for initiating T lymphocyte responses.
- DCs are identified as Lineage-negative, HLA-DR-positive blood cells.
- DCs can be subdivided into CD11c(+) myeloid DCs and CD11c(-) plasmacytoid DCs (pDCs).
Purpose of the Study:
- To characterize human blood dendritic cell populations.
- To investigate cell surface antigen expression on different DC subsets.
- To identify potential cross-reactivity with B lymphocyte antigens.
Main Methods:
- Flow cytometry analysis of fresh human blood DC populations.
- Utilized the HLDA9 panel of 63 directly labeled monoclonal antibodies (mAbs).
- Screened for mAb binding to Lin(-)HLA-DR(+) DC populations and their subsets.
Main Results:
- 23 mAbs did not bind to Lin(-)HLA-DR(+) DCs.
- 10 mAbs bound to all four DC populations analyzed.
- 8 mAbs bound to the three CD11c(+) myeloid DC populations, but none bound exclusively to pDCs.
- Some expected mAbs failed to bind DC populations, indicating unexpected antigen expression patterns.
Conclusions:
- The study identified distinct binding patterns of mAbs across different DC subsets.
- Highlighted similarities between CD11c(+) DC subsets.
- Suggested a relatively immature state of peripheral blood dendritic cells.
- Provided insights into DC heterogeneity and antigen expression for improved characterization.
Abstract:
Dendritic cells (DC) are a heterogeneous population of bone marrow derived leucocytes that are essential in the initiation of primary T lymphocyte responses. DC are identified as Lineage negative, HLA-DR(+) blood cells that can be further subdivided by CD11c to distinguish CD11c(+) DC and the CD11c(-) plasmacytoid DC. Plasmacytoid DC are the primary IFNα producing cells and express CD303, CD304 and CD123. The CD11c(+) myeloid DC can be divided into populations by CD1c, CD16 and CD141 expression. Despite DC being a functionally unique population, they share many cell surface antigens with myeloid lineage cells and B lymphocytes. We used flow cytometry to screen fresh human blood DC populations with the HLDA9 panel of 63 directly labelled mAb which included mAb specific for a number of B lymphocyte antigens. Of this panel, 23 mAb did not bind Lin(-)HLA-DR(+) DC and 10 bound all four populations. Eight mAb bound to the three CD11c(+) DC populations whilst no mAb tested bound to only pDC. Some of the mAb expected to bind to DC populations failed in this analysis. Overall, this screening highlighted similarities between the CD11c(+) DC subsets and the relatively immature state of peripheral blood DC.
