PMID- 19576898
OWN - NLM
STAT- MEDLINE
DCOM- 20090921
LR  - 20181201
IS  - 1872-7905 (Electronic)
IS  - 0022-1759 (Linking)
VI  - 348
IP  - 1-2
DP  - 2009 Aug 31
TI  - Techniques for time-efficient isolation of human skin dendritic cell subsets and 
      assessment of their antigen uptake capacity.
PG  - 42-56
LID - 10.1016/j.jim.2009.06.012 [doi]
AB  - 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.
FAU - Bond, Emily
AU  - Bond E
AD  - Center for Infectious Medicine, Department of Medicine, Karolinska Institutet, 
      Stockholm, Sweden.
FAU - Adams, William C
AU  - Adams WC
FAU - Smed-Sorensen, Anna
AU  - Smed-Sorensen A
FAU - Sandgren, Kerrie J
AU  - Sandgren KJ
FAU - Perbeck, Leif
AU  - Perbeck L
FAU - Hofmann, Anette
AU  - Hofmann A
FAU - Andersson, Jan
AU  - Andersson J
FAU - Lore, Karin
AU  - Lore K
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090702
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Alexa 488 hydrazide)
RN  - 0 (Antigens)
RN  - 0 (Dextrans)
RN  - 0 (Hydrazines)
RN  - 0 (Interferon Inducers)
RN  - 9006-59-1 (Ovalbumin)
RN  - O84C90HH2L (Poly I-C)
SB  - IM
MH  - Antigens/*immunology
MH  - Cell Separation/*methods
MH  - Dermis/cytology/immunology
MH  - Dextrans/pharmacology
MH  - Endocytosis
MH  - Epidermal Cells
MH  - Epidermis/immunology
MH  - Female
MH  - Humans
MH  - Hydrazines/pharmacology
MH  - Interferon Inducers/pharmacology
MH  - Langerhans Cells/cytology/*immunology
MH  - Ovalbumin/immunology
MH  - Poly I-C/pharmacology
EDAT- 2009/07/07 09:00
MHDA- 2009/09/22 06:00
CRDT- 2009/07/07 09:00
PHST- 2008/12/18 00:00 [received]
PHST- 2009/06/22 00:00 [revised]
PHST- 2009/06/24 00:00 [accepted]
PHST- 2009/07/07 09:00 [entrez]
PHST- 2009/07/07 09:00 [pubmed]
PHST- 2009/09/22 06:00 [medline]
AID - S0022-1759(09)00199-9 [pii]
AID - 10.1016/j.jim.2009.06.012 [doi]
PST - ppublish
SO  - J Immunol Methods. 2009 Aug 31;348(1-2):42-56. doi: 10.1016/j.jim.2009.06.012. 
      Epub 2009 Jul 2.