PMID- 22847488
OWN - NLM
STAT- MEDLINE
DCOM- 20121003
LR  - 20211021
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 65
DP  - 2012 Jul 23
TI  - Analysis of pulmonary dendritic cell maturation and migration during allergic 
      airway inflammation.
PG  - e4014
LID - 4014 [pii]
LID - 10.3791/4014 [doi]
AB  - Dendritic cells (DCs) are the key players involved in initiation of adaptive 
      immune response by activating antigen-specific T cells. DCs are present in 
      peripheral tissues in steady state; however in response to antigen stimulation, 
      DCs take up the antigen and rapidly migrate to the draining lymph nodes where 
      they initiate T cell response against the antigen. Additionally, DCs also play a 
      key role in initiating autoimmune as well as allergic immune response. DCs play 
      an essential role in both initiation of immune response and induction of 
      tolerance in the setting of lung environment. Lung environment is largely 
      tolerogenic, owing to the exposure to vast array of environmental antigens. 
      However, in some individuals there is a break in tolerance, which leads to 
      induction of allergy and asthma. In this study, we describe a strategy, which can 
      be used to monitor airway DC maturation and migration in response to the antigen 
      used for sensitization. The measurement of airway DC maturation and migration 
      allows for assessment of the kinetics of immune response during airway allergic 
      inflammation and also assists in understanding the magnitude of the subsequent 
      immune response along with the underlying mechanisms. Our strategy is based on 
      the use of ovalbumin as a sensitizing agent. Ovalbumin-induced allergic asthma is 
      a widely used model to reproduce the airway eosinophilia, pulmonary inflammation 
      and elevated IgE levels found during asthma. After sensitization, mice are 
      challenged by intranasal delivery of FITC labeled ovalbumin, which allows for 
      specific labeling of airway DCs which uptake ovalbumin. Next, using several DC 
      specific markers, we can assess the maturation of these DCs and can also assess 
      their migration to the draining lymph nodes by employing flow cytometry.
FAU - Kushwah, Rahul
AU  - Kushwah R
AD  - Stem Cell and Cancer Research Institute, McMaster University, Hamilton.
FAU - Hu, Jim
AU  - Hu J
LA  - eng
GR  - Canadian Institutes of Health Research/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Video-Audio Media
DEP - 20120723
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 9006-59-1 (Ovalbumin)
SB  - IM
MH  - Animals
MH  - Asthma/immunology/*pathology
MH  - Cell Movement/*immunology
MH  - Dendritic Cells/immunology/*pathology
MH  - Flow Cytometry/methods
MH  - Lung/immunology/pathology
MH  - Lymph Nodes/immunology/pathology
MH  - Mice
MH  - Ovalbumin/immunology
PMC - PMC3476425
EDAT- 2012/08/01 06:00
MHDA- 2012/10/04 06:00
PMCR- 2014/07/23
CRDT- 2012/08/01 06:00
PHST- 2012/08/01 06:00 [entrez]
PHST- 2012/08/01 06:00 [pubmed]
PHST- 2012/10/04 06:00 [medline]
PHST- 2014/07/23 00:00 [pmc-release]
AID - 4014 [pii]
AID - 10.3791/4014 [doi]
PST - epublish
SO  - J Vis Exp. 2012 Jul 23;(65):e4014. doi: 10.3791/4014.