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CD4 and CD8 expression by dendritic cell subtypes in mouse thymus and spleen
D Vremec1, J Pooley, H Hochrein
1The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria, Australia.
Insights
Researchers identified three mature dendritic cell (DC) types in mouse spleens, including a novel CD4+ population. Thymic DCs showed artifacts, with chimeric mice revealing antigen uptake from thymocytes.
Area of Science:
- Immunology
- Cell Biology
- Developmental Biology
Background:
- Dendritic cells (DCs) are crucial immune regulators.
- Understanding DC heterogeneity in spleen and thymus is vital for immunology.
- Previous studies identified some DC subsets, but comprehensive characterization is ongoing.
Purpose of the Study:
- To characterize dendritic cell (DC) populations in mouse spleen and thymus.
- To investigate the expression of CD4 and CD8 markers on these DCs.
- To identify potential artifacts in DC marker analysis and understand thymic DC interactions.
Main Methods:
- Flow cytometry was used to analyze spleen and thymus dendritic cells (DCs) for CD4 and CD8 expression.
- Careful methods were employed to prevent selective extraction or depletion of DC subpopulations.
- Chimeric mice were constructed using bone marrow from wild-type and CD4/CD8-deficient mice to study antigen pickup.
Main Results:
- Three major DC types were identified in the mouse spleen: CD4+8- DEC-205low CD11bhigh, CD4-8- DEC-205low CD11bhigh, and CD4-8alphaalpha+ DEC-205high CD11blow.
- The CD4 and CD8alpha expression on splenic DCs were confirmed to be intrinsic.
- Two DC types were found in the thymus, differing in CD8alphaalpha expression, with evidence of artifactual marker expression and significant thymic DC antigen uptake from thymocytes in chimeric mice.
Conclusions:
- Mouse spleen contains at least three mature DC subsets with distinct phenotypes.
- Thymic DC analysis requires careful consideration of artifacts, and thymic DCs can acquire antigens from thymocytes.
- This study refines the understanding of DC heterogeneity and function in immune surveillance and T cell development.
Abstract:
The dendritic cells (DC) of mouse spleen and thymus were examined for expression of CD4 and CD8. Provided care was taken to avoid selective extraction or selective depletion of DC subpopulations, three main types of DC were detected in mouse spleen: a major new population of CD4+8- DEC-205low CD11bhigh DC, together with the previously described CD4-8- DEC-205low CD11bhigh DC and CD4-8alphaalpha+ DEC-205high CD11blow DC. The CD4 on the surface of the CD4+ splenic DC subpopulation was produced by the DC themselves, and CD4 RNA transcripts were present. Likewise, the CD8alpha on the surface of the splenic CD8+ DC was shown to be a product of the DC themselves, in agreement with earlier evidence. All three spleen DC types would be considered as mature, based on expression of CD80, CD86, and CD40 as well as on T cell stimulating function. Mouse thymuses appeared to contain two DC types; both were DEC-205highCD11blow, but they differed in the level of CD8alphaalpha expression. However, as well as this authenticated marker expression, immunofluorescent staining was also found to reflect a series of artifacts, due to the autofluorescence of contaminating cells and due to pickup of CD4 and CD8alphabeta. By constructing mice chimeric for the hemopoietic lineages using mixtures of wild-type bone marrow with CD4null or CD8alphanull bone marrow, a marked pickup by thymic DC of Ags derived from thymocytes was demonstrated.
