RANTES and eotaxin enhance CD11b and CD18 expression on eosinophils from allergic patients with eosinophilia in the application of whole Blood flow cytometry analysis.
BACKGROUND
C-C chemokines and adhesion molecules expressed on eosinophils play an important role in the pathology of allergic inflammatory disease. C-C chemokines such as eotaxin or RANTES are involved in beta(2) integrin expression on purified eosinophils; so far we have no data on unpurified eosinophils in the peripheral blood. We measured beta(1) and beta(2) integrin activation after stimulation with eotaxin or RANTES in vitro using whole-blood flow-cytometric analysis.


METHODS
Heparinized whole blood obtained from allergic patients with eosinophilia or normal subjects was diluted with the same volume of RPMI 1640, and then cells were incubated in the presence or absence of PMA/ionomycin or chemokines for 45 min at 37 degrees C. After hemolyzation with lysing solution, expression of CD11b, CD11a, CD18 and CD49d on eosinophils was measured using flow cytometry.


RESULTS
The expression of CD11b, CD11a and CD18 in allergic patients was significantly higher than that in normal subjects. CD11b and CD18 expression showed a significant increase after stimulation with C-C chemokines, which was remarkable in allergic patients.


CONCLUSIONS
Eosinophils in the blood of allergic patients exhibited a higher expression of beta(2) integrins and were more sensitive to RANTES and eotaxin than those of normal subjects.
Chiba
Takahito
T
Department of Clinical and Laboratory Medicine, Akita University School of Medicine, Japan.
Kamada
Yumiko
Y
Saito
Norihiro
N
Oyamada
Hajime
H
Ueki
Shigeharu
S
Kobayashi
Yoshiki
Y
Kayaba
Hiroyuki
H
Chihara
Junichi
J
D000328:Adult; D040881:CD11a Antigen / Q000096:biosynthesis / Q000276:immunology; D039481:CD11b Antigen / Q000096:biosynthesis* / Q000276:immunology; D018821:CD18 Antigens / Q000096:biosynthesis* / Q000276:immunology; D054413:Chemokine CCL11; D018946:Chemokine CCL5 / Q000494:pharmacology*; D019742:Chemokines, CC / Q000494:pharmacology*; D004802:Eosinophilia / Q000097:blood / Q000276:immunology*; D004804:Eosinophils / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D006967:Hypersensitivity / Q000097:blood / Q000276:immunology*; D039641:Integrin beta Chains / Q000096:biosynthesis / Q000276:immunology; D015759:Ionomycin / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
C515456:CCL11 protein, human; D040881:CD11a Antigen; D039481:CD11b Antigen; D018821:CD18 Antigens; D054413:Chemokine CCL11; D018946:Chemokine CCL5; D019742:Chemokines, CC; D039641:Integrin beta Chains; D015759:Ionomycin; D013755:Tetradecanoylphorbol Acetate