Related Experiment Video
Updated: May 14, 2026
Precision-cut Mouse Lung Slices to Visualize Live Pulmonary Dendritic Cells
Published on: April 5, 2017
Visualization of intrapulmonary lymph vessels in healthy and inflamed murine lung using CD90/Thy-1 as a marker
Sarah Kretschmer1, Ina Dethlefsen, Stefanie Hagner-Benes
1Institut für Anatomie, Zentrum für medizinische Struktur- und Zellbiologie, Universität zu Lübeck, Lübeck, Germany.
Insights
CD90/Thy-1 is a reliable marker for identifying lymphatic vessels in mouse lungs. This method helps visualize lung lymphatic anatomy in healthy and inflamed states, aiding immune cell studies.
Area of Science:
- Immunology
- Vascular Biology
- Pulmonary Medicine
Background:
- Lung lymphatic vessels are crucial for fluid balance and immune cell transport.
- Identifying murine lung lymphatics is challenging due to marker overlap with other cell types.
- CD90/Thy-1 expression on lymphatic endothelium offers a potential solution for specific identification.
Purpose of the Study:
- To evaluate CD90/Thy-1 as a specific marker for murine pulmonary lymphatic vessels.
- To characterize the anatomical routes of lung lymphatic vessels under normal and inflammatory conditions.
Main Methods:
- Immunohistochemistry using anti-CD90.2/Thy-1.2 antibody on precision-cut lung slices.
- Electron microscopy and cross-reactivity tests to confirm specificity.
- Double-labeling with α-smooth muscle actin to identify lymphatic exit routes.
- Analysis of T cell distribution following house dust mite challenge.
Main Results:
- CD90/Thy-1 labeled a vascular network distinct from blood vessels in murine lungs.
- Antibody specificity was confirmed via electron microscopy and FVB mouse controls.
- Two distinct lymphatic exit routes from the lung were identified.
- Inflammation induced by house dust mite altered T cell distribution, including around arteries lacking lymphatics.
Conclusions:
- CD90/Thy-1 is a suitable marker for identifying murine lung lymphatic endothelial cells.
- This technique enables visualization of lung lymphatic anatomy in health and disease.
- Findings provide insights into immune cell trafficking in the lung during inflammation.
Background:
Lymphatic vessels play a pivotal role in fluid drainage and egress of immune cells from the lung. However, examining murine lung lymphatics is hampered by the expression of classical lymph endothelial markers on other cell types, which hinders the unambiguous identification of lymphatics. The expression of CD90/Thy-1 on lymph endothelium was recently described and we therefore examined its suitability to identify murine pulmonary lymph vessels under healthy and inflammatory conditions.
Methodology/Principal Findings:
Immunohistochemistry with a monoclonal antibody against CD90.2/Thy-1.2 on 200 µm thick precision cut lung slices labeled a vascular network that was distinct from blood vessels. Preembedding immunostaining and electron microscopy verified that the anti-CD90.2/Thy-1.2 antibody labeled lymphatic endothelium. Absence of staining in CD90.1/Thy-1.1 expressing FVB mice indicated that CD90/Thy-1 was expressed on lymph endothelium and labeling was not due to antibody cross reactivity. Double-labeling immunohistochemistry for CD90/Thy-1 and α-smooth muscle actin identified two routes for lymph vessel exit from the murine lung. One started in the parenchyma or around veins and left via venous blood vessels. The other began in the space around airways or in the space between airways and pulmonary arteries and left via the main bronchi. As expected from the pulmonary distribution of lymph vessels, intranasal application of house dust mite led to accumulation of T cells around veins and in the connective tissue between airways and pulmonary arteries. Surprisingly, increased numbers of T cells were also detected around intraacinar arteries that lack lymph vessels. This arterial T cell sheath extended to the pulmonary arteries where lymph vessels were located.
Conclusions/Significance:
These results indicate that CD90/Thy-1 is expressed on lymphatic endothelial cells and represents a suitable marker for murine lung lymph vessels. Combining CD90/Thy-1 labeling with precision cut lung slices allows visualizing the anatomy of the lymphatic system in normal and inflamed conditions.
