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Visualizing Lymph Node Structure and Cellular Localization using Ex-Vivo Confocal Microscopy
Published on: August 9, 2019
The cervical lymph node preparation: a novel approach to study lymphocyte homing by intravital microscopy
R Schramm1, H-J Schäfers, Y Harder
1Institute for Clinical & Experimental Surgery, Department of Thoracic and Cardiovascular Surgery, University Hospitals of Saarland, Kirrberger Strasse, 66421 Homburg/Saar, Germany. reneschramm@hotmail.com
Insights
Researchers developed a novel mouse cervical lymph node preparation to study lymphocyte homing in vivo. This method enhances understanding of adaptive immune responses and lymphocyte interactions with high endothelial venules (HEVs).
Area of Science:
- Immunology
- Cell Biology
- Physiology
Background:
- Lymphocyte recirculation is crucial for adaptive immunity.
- Lymphocytes migrate into lymphoid organs via high endothelial venules (HEVs).
- In vivo study of lymphocyte homing requires advanced technical approaches.
Purpose of the Study:
- To develop a novel in vivo method for studying lymphocyte homing.
- To analyze lymphocyte interactions with HEV endothelium in the cervical lymph node.
- To investigate the role of L-selectin and LFA-1 in lymphocyte migration.
Main Methods:
- Creation of a mouse cervical lymph node preparation.
- Intravital fluorescence microscopy for analyzing hemodynamics and cell interactions.
- Assessment of lymphocyte rolling and firm adhesion under different conditions.
Main Results:
- The novel preparation enabled detailed in vivo analysis of lymphocyte-HEV interactions.
- L-selectin blockade significantly inhibited lymphocyte rolling (92%) and firm adhesion (66%).
- LFA-1 deficiency reduced lymphocyte firm adhesion by 70%.
- Afferent lymphatic transport from the oral mucosa was visualized.
Conclusions:
- A novel technical tool for in vivo analysis of adaptive immune responses was developed.
- The study confirms the critical roles of L-selectin and LFA-1 in lymphocyte homing.
- The method provides insights into immune cell trafficking and interactions within lymph nodes.
Objective:
Lymphocyte recirculation constitutes an integral part of the adaptive immune system. Blood-borne lymphocytes migrate into secondary lymphoid organs, crossing the vascular wall of site-specific high endothelial venules (HEVs). We created a preparation of the cervical lymph node in mice to study lymphocyte homing in vivo.
Methods And Results:
Our novel approach allowed the detailed analysis of hemodynamics and lymphocyte-HEV endothelium interactions by means of intravital fluorescence microscopy. We confirm the key roles of L-selectin and LFA-1 for lymphocyte homing. Blockade of L-selectin function inhibited lymphocyte rolling and firm adhesion by 92% and 66%. In LFA-1-deficient mice, lymphocyte firm adhesion was reduced by 70%. In addition to the microcirculation studies, the cervical lymph node preparation allowed for visualization of afferent lymphatic transport, which is mainly derived from the oral mucosa.
Conclusion:
This study reports a novel technical tool for the detailed in vivo analysis of adaptive immune responses.
