Immunological profile of peripheral blood lymphocytes and monocytes/macrophages in Kawasaki disease

Abstract

Kawasaki disease (KD) is an acute illness of early childhood characterized by prolonged fever, diffuse mucosal inflammation, indurative oedema of the hands and feet, a polymorphous skin rash and nonsuppurative lymphadenopathy. The histopathological findings in KD comprise panvasculitis with endothelial necrosis, and the infiltration of mononuclear cells into small and medium-sized blood vessels. The levels of many proinflammatory cytokines, chemokines and adhesion molecules can be elevated in sera from children with KD at the acute stage. Although many immunological studies on KD involving peripheral blood have been reported, the data obtained remain controversial. This review focuses on the immune response of peripheral blood lymphocytes and monocytes/macrophages during acute KD.

Fig. 1

NF-κB activation in peripheral blood CD3+ T cells and CD14 + monocytes/macrophages of a 2-month-old boy with KD. (a) Nuclear extracts were harvested from CD14+ monocytes/macrophages or CD3+ T cells. The nuclear extracts were used as the sample for Western blotting because activated NF-κB existed in the nucleus. Rabbit polyclonal antibodies against NF-κB-p65 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) was used as the primary antibodies.Western blot analysis demonstrated that intranuclear amount of NF-κB was increased in CD14 + monocytes/macrophages and CD3+ T cells at the acute stage compared with that at the convalescent stage. (b–e) Whole blood was labelled with phycoerythrin-conjugated with anti-CD14 moclonal antibodies and peridinin chlorophyll protein-conjugated anti-CD3 monoclonal antibodies and then permeabilized in 4% paraformaldehyde in phosphate-buffered saline, pH 7·2, containing 0·1% saponin and 10 mm HEPES. The cells were then labelled with a mouse anti-NF-κB (nuclear-localized signal) antibody (IgG3; Boehringer Mannheim, Mannheim, Germany). The mouse anti-NF-κB (nuclear-localized signal) antibody recognizes an epitope overlapping the nuclear location signal of NF-κB-p65 and therefore selectively recognizes the activated form of NF-κB. The cells were then labelled with a FITC-conjugated rat antimouse IgG3 monoclonal antibody (Pharmingen, San Diego, CA, USA). Immunofluorescence was analysed with a FACScan flow cytometer equipped with CellQuest software (Becton-Dickinson Biosciences, San Jose, CA, USA). The percentages of cells with intranuclear NF-κB in CD14+ monocytes/macrophages and CD3+ T cells by flow cytometric analysis are indicated. (b) CD3+ T cells at the acute stage; (c) CD3+ T cells at the convalescent stage; (d) CD14 + monocytes/macrophages at the acute stage; (e) CD14 + monocytes/macrophages at the convalescent stage.