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Published on: November 4, 2016
The expression of mouse CLEC-2 on leucocyte subsets varies according to their anatomical location and inflammatory
Kate L Lowe1, Leyre Navarro-Núñez1, Cécile Bénézech2
1Centre for Cardiovascular Sciences, University of Birmingham, Birmingham, UK.
Insights
This study clarifies C-type lectin-like receptor 2 (CLEC-2) expression, finding it absent on resting dendritic cells (DCs) and upregulated on activated DCs during inflammation. Peripheral blood B lymphocytes present exogenous CLEC-2, which is lost upon entering lymphoid organs.
Area of Science:
- Immunology
- Cell Biology
Background:
- C-type lectin-like receptor 2 (CLEC-2) expression has been reported on myeloid cells and dendritic cells (DCs).
- Previous studies lacked CLEC-2-deficient mice and relied on single antibody clones, leading to potential inaccuracies.
Purpose of the Study:
- To accurately investigate CLEC-2 surface expression on hematopoietic cells.
- To clarify CLEC-2 expression patterns in resting and activated immune cells, particularly DCs and B lymphocytes.
Main Methods:
- Generated CLEC-2-deficient adult mice using two independent approaches.
- Employed two anti-mouse CLEC-2 antibody clones for surface expression analysis.
- Analyzed hematopoietic cells from peripheral blood and secondary lymphoid organs.
Main Results:
- Ruled out constitutive CLEC-2 expression on resting dendritic cells (DCs).
- Showed CLEC-2 upregulation on activated DCs in mesenteric lymph nodes during LPS-induced inflammation.
- Demonstrated peripheral blood B lymphocytes present exogenous CLEC-2.
- Observed loss of CLEC-2 on circulating B lymphocytes and myeloid cells upon entering secondary lymphoid organs.
Conclusions:
- CLEC-2 is not constitutively expressed on resting DCs.
- CLEC-2 expression is dynamically regulated during inflammation and cell trafficking.
- Peripheral B lymphocytes acquire exogenous CLEC-2, which is subsequently lost in lymphoid organs.
Abstract:
Expression of mouse C-type lectin-like receptor 2 (CLEC-2) has been reported on circulating CD11b(high) Gr-1(high) myeloid cells and dendritic cells (DCs) under basal conditions, as well as on a variety of leucocyte subsets following inflammatory stimuli or in vitro cell culture. However, previous studies assessing CLEC-2 expression failed to use CLEC-2-deficient mice as negative controls and instead relied heavily on single antibody clones. Here, we generated CLEC-2-deficient adult mice using two independent approaches and employed two anti-mouse CLEC-2 antibody clones to investigate surface expression on hematopoietic cells from peripheral blood and secondary lymphoid organs. We rule out constitutive CLEC-2 expression on resting DCs and show that CLEC-2 is upregulated in response to LPS-induced systemic inflammation in a small subset of activated DCs isolated from the mesenteric lymph nodes but not the spleen. Moreover, we demonstrate for the first time that peripheral blood B lymphocytes present exogenously derived CLEC-2 and suggest that both circulating B lymphocytes and CD11b(high) Gr-1(high) myeloid cells lose CLEC-2 following entry into secondary lymphoid organs. These results have significant implications for our understanding of CLEC-2 physiological functions.
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