The cervical lymph node preparation: a novel approach to study lymphocyte homing by intravital microscopy.
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.


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.


CONCLUSIONS
This study reports a novel technical tool for the detailed in vivo analysis of adaptive immune responses.
Schramm
R
R
Institute for Clinical & Experimental Surgery, Department of Thoracic and Cardiovascular Surgery, University Hospitals of Saarland, Kirrberger Strasse, 66421 Homburg/Saar, Germany. reneschramm@hotmail.com
Schäfers
H-J
HJ
Harder
Y
Y
Schmits
R
R
Thorlacius
H
H
Menger
M D
MD
D000818:Animals; D002448:Cell Adhesion; D002465:Cell Movement; D002574:Cervical Vertebrae / Q000276:immunology*; D042783:Endothelial Cells / Q000166:cytology; D019041:L-Selectin / Q000378:metabolism; D007958:Leukocyte Count; D008198:Lymph Nodes / Q000166:cytology* / Q000276:immunology*; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008856:Microscopy, Fluorescence; D012867:Skin / Q000166:cytology
D016169:Lymphocyte Function-Associated Antigen-1; D019041:L-Selectin