Identification of cytokine and adhesion molecule mRNA in murine lung tissue and isolated T cells and eosinophils by semi-quantitative reverse transcriptase-polymerase chain reaction.
We have used a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) assay to detect the expression of mRNA for inflammatory cytokines, integrins and selectins in murine lung tissue, and T cells and eosinophils isolated from lung and bronchoalveolar lavage (BAL) fluid in an in vivo model of ovalbumin (OA)-induced airway inflammation. RNA was isolated from whole lung tissue at 1, 6, 24, 48, 72 h, and 7 days after OA inhalation. mRNA for the Th2 cytokines, IL-4, -5, -6, -10 and -13 in OA-sensitized mice were significantly elevated compared with non-sensitized mice. IL-2, TNF-beta, and eotaxin mRNA were also increased, but IFN-gamma mRNA was not. P- and E-selectin mRNA levels were also enhanced in lung tissue between 6 and 72 h after challenge. Lung T cells were isolated by cell sorting with a flow cytometer at 3, 12, 24, 48 and 72 h after challenge. mRNA levels for IL-5 and -10 were greater in T cells from OA-sensitized and -challenged mice than controls at 24 h. BAL fluid from OA-sensitized and -challenged mice also had significantly higher IL-5 levels than controls. BAL fluid T cells and eosinophils were obtained at 48 and 72 h after aerosol challenge and were purified by cell sorting. Messenger RNA for IL-1 alpha, -2, -4, -5, -10, IFN-gamma, and beta 1 were detected in T cells at both time points. Transcripts for IL-1 alpha, -4, -5, -13, TNF-alpha and beta, and alpha 4, beta 1 and beta 7 were also identified in BAL eosinophils. These data show that in addition to murine lung T cells, airway eosinophils may also contribute to the inflammatory response by their ability to express mRNA for cytokines and integrins.
Krzesicki
R F
RF
Cell Biology and Inflammation Research, Pharmacia and Upjohn, Incorporated, Kalamazoo, Michigan 49001, USA.
Winterrowd
G E
GE
Brashler
J R
JR
Hatfield
C A
CA
Griffin
R L
RL
Fidler
S F
SF
Kolbasa
K P
KP
Shull
K L
KL
Richards
I M
IM
Chin
J E
JE
D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry / Q000166:cytology; D015815:Cell Adhesion Molecules / Q000235:genetics*; D002469:Cell Separation; D054413:Chemokine CCL11; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil / Q000235:genetics; D016207:Cytokines / Q000235:genetics* / Q000378:metabolism; D019040:E-Selectin / Q000235:genetics; D004804:Eosinophils / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D005260:Female; D007249:Inflammation / Q000378:metabolism; D039441:Integrin alpha4; D039641:Integrin beta Chains; D019012:Integrin beta1 / Q000235:genetics; D016023:Integrins / Q000235:genetics; D007371:Interferon-gamma / Q000235:genetics; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism; D018793:Interleukin-13 / Q000235:genetics / Q000378:metabolism; D007376:Interleukin-2 / Q000235:genetics; D015847:Interleukin-4 / Q000235:genetics / Q000378:metabolism; D015848:Interleukin-5 / Q000235:genetics / Q000378:metabolism; D008168:Lung / Q000166:cytology*; D008233:Lymphotoxin-alpha / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D019007:P-Selectin / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis; D013601:T-Lymphocytes / Q000737:chemistry* / Q000166:cytology / Q000276:immunology
D015703:Antigens, CD; C515457:Ccl11 protein, mouse; D015815:Cell Adhesion Molecules; D054413:Chemokine CCL11; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil; D016207:Cytokines; D019040:E-Selectin; D039641:Integrin beta Chains; D019012:Integrin beta1; D016023:Integrins; D018793:Interleukin-13; D007376:Interleukin-2; D015848:Interleukin-5; D008233:Lymphotoxin-alpha; D019007:P-Selectin; D012333:RNA, Messenger; C091368:integrin beta7; D016753:Interleukin-10; D039441:Integrin alpha4; D015847:Interleukin-4; D007371:Interferon-gamma