Increased levels of IL-5 positive peripheral blood eosinophils and lymphocytes in mild asthmatics after allergen inhalation provocation


Halldén Department of Laboratory Medicine, Division of Clinical Immunology, Karolinska Hospital, S-171 76 Stockholm, Sweden.



In allergic inflammation eosinophils and TH2-like lymphocytes are supposed to be the major effector cells and considered to contribute as cellular source of the key cytokine interleukin (IL)-5.


The purpose of this study was to enable detection of IL-5 containing leucocytes and to investigate whether the number of these cells in the blood circulation differed between healthy and asthmatics before and after allergen provocation.


The distribution of intracellular IL-5 in human peripheral blood eosinophils (PBE) and lymphocytes (PBL) has been investigated using fixation and cell membrane permeabilization with octyl-glucopyranoside, the FOG-method, and flow cytometry. The intracellular staining was performed on leucocytes without any prior purification and in vitro stimulation. The specificity of IL-5 binding to intracellular compartment of both PBE and PBL was confirmed by complete inhibition with human recombinant IL-5.


Preformed intracellular IL-5 was detected in the main population of PBE (> 70%) in both healthy individuals and asymptomatic patients. Moreover, preformed intracellular IL-5 was also detected in 4.8% and 2.4% of PBL from healthy individuals and asymptomatic patients, respectively. There was a correlation between the absolute number of PBE and IL-5 positive PBE. In patients with pollen-related asthma, the number of IL-5 positive PBE and PBL increased significantly 24 h after an allergen inhalation provocation (P < 0.05). In the healthy control group no differences regarding IL-5 positive PBE and PBL were obtained pre- and post-allergen challenge.


In patients with mild allergic asthma, but not in healthy individuals, allergen provocation induces an increased absolute number of IL-5 positive PBE and PBL. The reason for the relatively high number of IL-5 positive PBL is unclear, but a plausible explanation might be that other lymphocyte subsets besides CD4+ TH2 can produce IL-5. However, enumeration of IL-5 positive leucocytes may be used as an activity marker and also be a useful tool in monitoring the inflammation in asthma.