PMID: 8960094Nov 29, 1996Paper

A method to recover, enumerate and identify lymphomyeloid cells present in an inflammatory dermal site: a study in laboratory mice

Journal of Immunological Methods
Y BelkaidG Milon

Abstract

We describe a new method to recover and study cells present in the dermis of mouse ear at homeostasis or after intradermal injection of disturbing agents (lipopolysaccharide or Listeria monocytogenes). The ears either left untreated or inoculated were handled and processed as culture explants of the dorsal and ventral leaflets, their dermal sides being spread on a buffered medium. Within this medium emigrate/sediment, with different kinetics: neutrophils, mononuclear phagocytes, dendritic leucocytes, T lymphocytes expressing either gamma delta or alpha beta TCRs, and other minor subsets, the identification of which deserves more relevant reagents: they are likely to be NK, mast cells, eosinophils and their local progenitors. All the major subsets were identified through a combination of immunocytochemical and flow cytometry labeling. Two examples illustrating the advantages and limitations of this new method are given: either 1 microgram of LPS or 10(4) Listeria monocytogenes were injected within the ear 48, 24, 12, 6, 3 h before ear explant culture. This ear explant culture has been further compared to the ear sheet treatment with collagenase/disease for three cell populations, the epidermal dendritic leucocytes, the gamma del...Continue Reading

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Citations

Nov 11, 1999·Journal of Immunological Methods·G K DeKrey, R G Titus
Jan 14, 2012·Nature Protocols·Jackson LiangYao LiLai Guan Ng
May 23, 2007·The Journal of Experimental Medicine·Emmanuelle CoutanceauCaroline Demangel
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May 22, 2013·Journal of Immunological Methods·Mali CamberisGraham Le Gros
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Oct 9, 2002·The Journal of Immunology : Official Journal of the American Association of Immunologists·Takako HirataBruce Furie

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