Increased expression of high affinity IgE (FcepsilonRI) receptor-alpha chain mRNA and protein-bearing eosinophils in human allergen-induced atopic asthma.
FcepsilonRI receptors play an important role in allergen-induced mediator release and antigen presentation by mast cells, basophils, and monocyte/macrophages in atopic disorders. The expression of FcepsilonRI by tissue eosinophils in atopic asthma after allergen challenge has not been established. For this reason we attempted to identify mRNA and protein product + FcepsilonRIalpha eosinophils in cytospins made from bronchoalveolar lavage (BAL) from atopic asthmatics (n = 9) and nonatopic normal subjects (n = 4) 24 h after segmental challenge with allergen or diluent. Messenger RNA for FcepsilonRIalpha was determined using in situ hybridization and FcepsilonRIalpha protein expression by immunocytochemistry using a mouse monoclonal antibody 22E7. Colocalization of FcepsilonRIalpha receptors to eosinophils was performed using chromotrope 2R. When compared with a control challenge, segmental challenge with Dermatophagoides pteronyssinus induced significant BAL eosinophilia (p = 0.007). The total number of BAL FcepsilonRIalpha mRNA and protein-positive cells also increased in asthmatics, median values 2 (0.7-7.2) and 11.5 (0.6-65.0) x 10(6) cells (p = 0.02) and 0 (0-0.3 x 10(6)) and 3.1 x 10(6) (0.45 - 162.5 x 10(6)) cells (p = 0.007), respectively, for mRNA and protein. Net increases in FcepsilonRIalpha+ cells correlated with the net increases in BAL eosinophils (r = 0.98, p = 0.0001 for mRNA and r = 0.72, p = 0.02 for protein). Colocalization studies with chromotrope 2R revealed that only 4% of FcepsilonRIalpha+ cells were eosinophils after control challenge and, in contrast, 85 to 95% of FcepsilonRIalpha+ cells were eosinophils after allergen. There were no differences in the numbers of FcepsilonRIalpha+ cells or eosinophils in normal control subjects. Our results demonstrated that local endobronchial allergen provocation in atopic asthmatics results in increased synthesis and expression of FcepsilonRIalpha predominantly on BAL eosinophils.
Rajakulasingam
K
K
Upper Respiratory Medicine and Allergy and Clinical Immunology, Imperial College School of Medicine at the National Heart and Lung Institute, Dovehouse Street, London, United Kingdom.
Till
S
S
Ying
S
S
Humbert
M
M
Barkans
J
J
Sullivan
M
M
Meng
Q
Q
Corrigan
C J
CJ
Bungre
J
J
Grant
J A
JA
Kay
A B
AB
Durham
S R
SR
D000328:Adult; D000911:Antibodies, Monoclonal; D001249:Asthma / Q000276:immunology*; D001985:Bronchial Provocation Tests; D001992:Bronchoalveolar Lavage Fluid; D001999:Bronchoscopy; D002452:Cell Count; D004804:Eosinophils / Q000276:immunology*; D005260:Female; D006801:Humans; D007141:Immunoglobulin Fc Fragments; D007137:Immunoglobulin alpha-Chains; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008214:Lymphocytes / Q000276:immunology; D008264:Macrophages / Q000276:immunology; D008297:Male; D009504:Neutrophils / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism*; D017455:Receptors, IgE / Q000096:biosynthesis*
D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D007137:Immunoglobulin alpha-Chains; D012333:RNA, Messenger; D017455:Receptors, IgE