TY - JOUR
T1 - A new approach for analyzing cellular infiltration during allergic airway inflammation
AU - Hoffmann, Peter R.
AU - Gurary, Alexandra
AU - Hoffmann, Fukun W.
AU - Jourdan-Le Saux, Claude
AU - Teeters, Kelsa
AU - Hashimoto, Ann C.
AU - Tam, Elizabeth K.
AU - Berry, Marla J.
N1 - Funding Information:
This publication was made possible by Grant Number G12 RR003061 and P20RR016467 from the National Center for Research Resources (NCRR), a component of the National Institutes of Health (NIH). Its contents are solely the responsibility of the authors and do not necessarily represent the official view of NCRR or NIH. This work was also supported by NIH award R01 DK40302, a Hawaiʻi Community Foundation Grant, and the American Lung Association of Hawaiʻi.
PY - 2007/12/1
Y1 - 2007/12/1
N2 - A mouse model for allergic airway inflammation involving ovalbumin (OVA) sensitization and challenge has been developed that reproduces hallmark features of human asthma and has provided valuable insight into the mechanisms by which this disease occurs. Cellular infiltrate in lungs of mice used in this model have conventionally been evaluated using histological examination of tissue sections and light microscopic analysis of lung lavage samples. As an alternative or complementary approach for characterizing cellular infiltrate, we developed a multicolor fluorescence-activated cell sorter (FACS) method involving the simultaneous detection of seven different markers on lung cell suspensions: CD4, CD8, B220, CD11b, Gr-1, CD49b, and Fce{open}RI. Only some of these cell types increased in OVA-challenged mice compared to PBS controls, including the CD4+, B220+, CD11b+, and Fce{open}RI+ groups. We also examined subpopulations of cells for coexpression of these markers and dissected heterogeneous populations as further evaluation procedures to characterize the cellular infiltrate resulting from OVA challenge. Finally, we combined FACS with real-time PCR to analyze certain cell types in terms of mRNA levels for factors involved in asthma, including GATA-3 and IL-1β. Overall, these FACS-based techniques provide a powerful approach for analyzing cellular profiles in lung tissue from mice used in the mouse model of asthma and may also prove valuable in evaluating cellular infiltrates for other models of inflammation and immune responses.
AB - A mouse model for allergic airway inflammation involving ovalbumin (OVA) sensitization and challenge has been developed that reproduces hallmark features of human asthma and has provided valuable insight into the mechanisms by which this disease occurs. Cellular infiltrate in lungs of mice used in this model have conventionally been evaluated using histological examination of tissue sections and light microscopic analysis of lung lavage samples. As an alternative or complementary approach for characterizing cellular infiltrate, we developed a multicolor fluorescence-activated cell sorter (FACS) method involving the simultaneous detection of seven different markers on lung cell suspensions: CD4, CD8, B220, CD11b, Gr-1, CD49b, and Fce{open}RI. Only some of these cell types increased in OVA-challenged mice compared to PBS controls, including the CD4+, B220+, CD11b+, and Fce{open}RI+ groups. We also examined subpopulations of cells for coexpression of these markers and dissected heterogeneous populations as further evaluation procedures to characterize the cellular infiltrate resulting from OVA challenge. Finally, we combined FACS with real-time PCR to analyze certain cell types in terms of mRNA levels for factors involved in asthma, including GATA-3 and IL-1β. Overall, these FACS-based techniques provide a powerful approach for analyzing cellular profiles in lung tissue from mice used in the mouse model of asthma and may also prove valuable in evaluating cellular infiltrates for other models of inflammation and immune responses.
KW - Airway inflammation
KW - Allergy
KW - Asthma
KW - Cellular infiltration
KW - FACS
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U2 - 10.1016/j.jim.2007.07.019
DO - 10.1016/j.jim.2007.07.019
M3 - Article
C2 - 17825315
AN - SCOPUS:35548956327
VL - 328
SP - 21
EP - 33
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 1-2
ER -