Analysis of dendritic cell trafficking using EGFP-transgenic mice.
Dendritic cells (DCs) are professional antigen presenting cells, well equipped to initiate an immune response. For effective induction of an immune response, DC should migrate from the periphery to the lymph node to present the antigen to T lymphocytes. Currently, tumor-antigen loaded DCs are used in clinical vaccination trials in cancer patients. To investigate the migratory capacity of DC in vivo, a variety of fluorescent and radioactive labels have been used. Here we introduce a novel tool to study DC migration in vivo: DCs generated from enhanced green fluorescent protein (EGFP)-transgenic mice. DC from EGFP-transgenic mice display typical DC behavior and can be matured without affecting their autofluorescence in vitro. In addition, the continuously produced cytoplasmic EGFP in living cells functions as a viability marker, since EGFP released from dying cells does not stain DC from C57Bl/6 mice upon coculture. In vivo migration studies using EGFP-DC and indium-111-labeled DC were performed to determine the efficiency of i.d. versus s.c. administered DC to reach the draining lymph node. The analysis demonstrates that i.d. injection increases the amount of EGFP-DC/indium-111-labeled DC in the lymph node compared to s.c. injection. Subsequent quantitative, phenotypical and ultrastuctural analysis demonstrate that DC generated from EGFP-transgenic mice are well suited to study the migratory behavior, distribution and phenotype of DC in vivo.
Eggert
Andreas A O
AA
Tumor Immunology Laboratory, University Medical Center Nijmegen St. Radboud, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands.
van der Voort
Robbert
R
Torensma
Ruurd
R
Moulin
Veronique
V
Boerman
Otto C
OC
Oyen
Wim J G
WJ
Punt
C J A
CJ
Diepstra
Heleen
H
de Boer
Annemiek J
AJ
Figdor
Carl G
CG
Adema
Gosse J
GJ
D000818:Animals; D000911:Antibodies, Monoclonal; D002465:Cell Movement; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology / Q000502:physiology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D049452:Green Fluorescent Proteins; D007279:Injections, Subcutaneous; D015847:Interleukin-4 / Q000494:pharmacology; D007668:Kidney / Q000276:immunology; D008164:Luminescent Proteins / Q000032:analysis; D008198:Lymph Nodes / Q000276:immunology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010641:Phenotype; D011994:Recombinant Proteins; D013154:Spleen / Q000276:immunology
D000911:Antibodies, Monoclonal; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D015847:Interleukin-4; D016178:Granulocyte-Macrophage Colony-Stimulating Factor