Back to Search
Start Over
Techniques for time-efficient isolation of human skin dendritic cell subsets and assessment of their antigen uptake capacity
- Source :
- Journal of Immunological Methods. 348:42-56
- Publication Year :
- 2009
- Publisher :
- Elsevier BV, 2009.
-
Abstract
- Dendritic cells (DCs) residing in skin are important sentinels for foreign antigens. Methods to facilitate studies of subsets of skin DCs are important to increase the understanding of various pathogens, allergens, topical treatments or vaccine components targeting the skin. In this study, we developed a new DC purification method using a skin graft mesher, clinically used for expansion of skin grafts, to accelerate processing of skin into nets that allowed efficient enzymatic disruption and single cell isolation. The reduction in processing time using the skin graft mesher enabled processing of larger skin samples and also limited the ex vivo handling of the specimens which is associated with maturation of DCs. In addition, a skin explant model to functionally monitor early events of antigen uptake by DC subsets in situ was developed. DCs isolated from epidermis represented a uniform CD1a(+) HLA-DR(+) CD11c(+) Langerin(+) DC-SIGN(-) DC-LAMP(int) DEC-205(int) Langerhans cell (LC) population whereas three subtypes of HLA-DR(+) CD11c(+) DCs were isolated from dermis based on their varying expression of CD1a. Epidermal LCs showed a significantly higher antigen uptake capacity of fluorescently-labelled ovalbumin (OVA) and dextran as compared to any of the dermal DC (dDC) subsets. In contrast, injection of antigen directly into skin explants followed by in situ imaging revealed that the majority of DCs with internalized antigen were localized in the dermis, likely as a consequence of the anatomical site for antigen delivery. These methods offer potency for various applications addressing antigen uptake, microbial DC interactions or other antigenic stimulation targeting the skin and can enhance our knowledge of basic DC biology in human skin.
- Subjects :
- Interferon Inducers
Langerhans cell
Langerin
Ovalbumin
Immunology
Population
Human skin
Cell Separation
Dermis
Antigen
medicine
Humans
Immunology and Allergy
Antigens
education
education.field_of_study
Interferon inducer
integumentary system
biology
Dextrans
Dendritic cell
Endocytosis
Hydrazines
Poly I-C
medicine.anatomical_structure
Epidermal Cells
Langerhans Cells
biology.protein
Female
Epidermis
Subjects
Details
- ISSN :
- 00221759
- Volume :
- 348
- Database :
- OpenAIRE
- Journal :
- Journal of Immunological Methods
- Accession number :
- edsair.doi.dedup.....2070c9eb36611e4d32c39734cdb0161f