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A Simple and Efficient Method for Testing Immunomodulatory Agents for Generation of Tolerogenic Dendritic Cells from Human CD14+ Monocytes
Published on: April 11, 2025
Human spleen contains different subsets of dendritic cells and regulatory T lymphocytes
M M Velásquez-Lopera1, L A Correa, L F García
1Grupo de Inmunología Celular e Inmunogenética, Sección de Dermatología, Universidad de Antioquia, Medellín, Colombia.
Insights
This study quantifies dendritic cells (DCs) and regulatory T cells (Tregs) in human spleens. Findings reveal their distribution and phenotype, offering insights for future immunotherapies.
Area of Science:
- Immunology
- Cell Biology
Background:
- Limited understanding of dendritic cell (DC) and regulatory T cell (Treg) presence in human lymphoid organs.
- Most data on these cells derived from circulating populations.
Purpose of the Study:
- To determine the frequency, phenotype, and location of DCs and Tregs within human spleens.
- To explore the potential of spleen cells in transplantation and autoimmune disease treatment.
Main Methods:
- Flow cytometry analysis of spleen cells from deceased organ donors.
- Identification of conventional/myeloid DCs (cDCs), plasmacytoid DCs (pDCs), and Tregs using specific cell surface markers.
- Assessment of cell distribution within splenic microenvironments.
Main Results:
- Conventional/myeloid DCs constituted approximately 2-4% of total spleen cells, with distinct mature and immature populations.
- Plasmacytoid DCs represented about 0.3% of spleen cells.
- CD4+CD25high Tregs were found at 0.2% of spleen lymphocytes, with FOXP3+ cells in T- and B-cell areas. Cells expressing cDC and pDC markers (BDCA-1, BDCA-3, BDCA-2, BDCA-4) were frequent.
- DCs and Tregs were localized to specific splenic regions, including the subcapsular zone, T-cell areas, and B-cell follicles.
Conclusions:
- The study provides a detailed characterization of DC and Treg populations in the human spleen.
- Advancements in cell manipulation techniques could enable the use of donor spleen cells for inducing allograft tolerance or treating autoimmune conditions.
Abstract:
Most knowledge about dendritic cells (DCs) and regulatory T cells in humans has been gathered from circulating cells but little is known about their frequency and distribution in lymphoid organs. This report shows the frequency, phenotype and location of DCs and regulatory T cells in deceased organ donors' spleens. As determined by flow cytometry, conventional/myeloid DCs (cDCs) CD11c(high)HLA-DR(+)CD123(-/low) were 2.3 +/- 0.9% and LIN(-) HLA-DR(+)CD11c(high) 2.1 +/- 0.3% of total spleen cells. Mature CD11c(high)HLA-DR(+)CD83(+) were 1.5 +/- 0.8% and 1.0 +/- 1.6% immature CD11c(high)HLA-DR(+)CD83(-) cDC. There were 0.3 +/- 0.3% plasmacytoid DCs (pDC) CD11c(-/low)HLA-DR(+)CD123(high) and 0.3 +/- 0.1% LIN(-)HLA-DR(+)CD123(high). Cells expressing cDCs markers, BDCA-1 and BDCA-3, and pDCs markers BDCA-2 and BDCA-4 were observed in higher frequencies than DCs with other phenotypes evaluated. CD11c(+), CD123(+) and CD83(+) cells were located in subcapsular zone, T cells areas and B-cell follicles. CD4(+)CD25(high) Tregs were 0.2 +/- 0.2% and CD8(+)CD28(-) comprised 11.5 +/- 8.1% of spleen lymphocytes. FOXP3(+) cells were found in T- and B-cell areas. The improvement in cell separation, manipulation and expansion techniques, will facilitate the manipulation of donor spleen cells as a part of protocols for induction and maintenance of allograft tolerance or treatment of autoimmune diseases.
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