Interferon-gamma pretreatment of peripheral blood mononuclear cells partially restores defective cytokine production in children with atopic dermatitis

Interferon-gamma (IFN-γ) is considered an important determinant of the balance between T-helper type 1 and 2 cytokines and has been used experimentally for the treatment of atopic dermatitis. However, contrasting results have been reported relative to the Th-UTh-2 cytokine profile in atopic patients. In this study, we examined cytokine production by polyclonally activated peripheral blood mononuclear cells (PBMC) from children with atopic dermatitis, and assessed the influence of in vitro IFN-γ pretreatment on these cells. A fraction of PBMC isolated from children with severe atopic dermatitis, as well as from age-matched controls, was initially exposed to IFN-γ. After washing, both treated and untreated cells were then put into culture either alone or with the addition of phytohemagglutinin (PHA) or phorbol myristate acetate (PMA) plus ionomycin. IL-4, IL-5, IL-10 and IFN-γ production were measured in the supernatants using commercially available ELISAs. PBMC from atopic patients produced more IL-4 (P = 0.04) and IL-10 (P = 0.03) and less IFN-γ (P = 0.01) than controls, when stimulated with PHA. Interestingly, in PMA + ionomycin stimulated cultures, the atopic cytokine profile was different with more IL-5 (P = 0.0068) and less IFN-γ production (P = 0.00046) than the control group. When cells were pretreated with IFN-γ, there were no significant differences between patients and controls. PBMC from children with atopic dermatitis show alterations in cytokine production, compatible in general terms with the Th-l/ Th-2 model. Exposure of PBMC to IFN-γ before activation results in a reduction of these differences, so that cytokine production becomes similar in the atopic and normal groups.     

Defective Leukocyte Interferon Response in Children with Recurrent Infections Accompanied by Arthralgia

ABSTRACT. Children with recurrent and/or unusually severe infections were investigated for possible defects in the interferon (IFN)-natural killer (NK) cell system. Two series, each of 13 children, were examined, one in 1982 and one in 1983. Healthy children, seven in 1982 and eight in 1983, served as controls. Peripheral blood mononuclear leukocytes were examined for IFN production induced by the IFN-α inducers Sendai virus and Escherichia coli and by the IFN-γ inducers Concanavalin A and Lens culinaris lectin. None of these inducers discriminated patients from controls. However, the bacteria Staphylococcus aureus Cowan I (SACol), inducers of atypical IFN in null lymphocytes, yielded significantly lower IFN production in infection-prone children than in controls, particularly in children with recurrent infections accompanied by arthralgia. No differences in basal NK activity or in the in vitro enhancement of such activity by IFN-α were found between patients and controls.     

Interferon-γ production by cord-blood mononuclear cells is reduced in newborns with a family history of atopic disease and is independent from cord blood IgE-levels

For newborn children both elevated serum IgE levels in the cord blood and a positive family history of atopic disease have been shown to be risk factors for the manifestation of atopic diseases. In adult patients with atopic dermatitis, in vitro interferon-γ (IFN-γ) production is reduced and a negative correlation with serum IgE levels has been shown. We have now raised the question if newborn infants at risk for the development of atopic disease have similar abnormalities of cytokine production at birth. In vitro production of interleukin 2, interleukin 6 and interferon-γ by peripheral blood mononuclear cells was measured in 53 newborns: 21 had cord blood IgE levels above 0. 9 kU/1, 21 had a positive family history, 7 had both elevated IgE and a positive family history; 18 newborns with no identitiable risk for atopic disease served as controls. Umbilical cord blood mononuclear cells were stimulated with PHA or monoclonal antibody OKT3. In vitro production of interleukin 2 and 6 was comparable in all groups. Compared to controls IFN-γ production of peripheral mononuclear cells (PBMC) from newborns with elevated cord blood IgE was not different, but PMBC from newborns with a familial risk showed a significant decrease in PHA induced IFN-γ production (p < 0.005, U-test). No correlation between umbilical cord blood IgE and diminished IFN-γ production was found in newborns with or without a positive family history. We conclude that immunoregulatory abnormalities in newborns of atopic families are detectable already at birth and are unrelated to cord blood IgE.     

Ontogeny of T-helper 1 and T-helper 2 cytokine production in childhood

Compared to adults, infants and young children demonstrate differences in their immune response, indicating that there is maturation or change over time and it is probable that this may be reflected in cytokine production. Cytokine responses have been demonstrated to be different in atopic and non-atopic individuals. In this study, we examined T-helper 1 (Th1) (interferon-γ[IFN-γ]) and T-helper 2 (Th2) (interleukin [IL]-4, IL-5, and IL-13) cytokine release from atopic and non-atopic children in response to the staphylococcal superantigen, staphylococcal enterotoxin B (SEB). In non-atopic and atopic children, IFN-γ, IL-4, and IL-5 release was significantly related to age. Non-atopic children younger than 2 years of age were found to have significantly reduced Th2 (IL-4, IL-5, and IL-13) responses when compared with older, non-atopic children. Atopic children had a reduced IFN-γ response when compared with non-atopics in early childhood; however, the decreased IFN-γ response seen in early childhood did not persist after 10 years. These age-related changes in cytokine production provide further support for the concept that cytokine deviations may determine the natural history of atopic disease during early childhood. In addition, the present study indicates the necessity of age-matched controls when examining children for both Th1 (IFN-γ) and Th2 (IL-4) cytokine release.     

Comparison of cytokine production in blood cell cultures of healthy children and adults

The production of the cytokines IL-l-α, IL-l-β, IL-2, TNF-α, and IFN-γ was measured by a sensitive immunological assay in stimulated whole blood cell cultures from 52 healthy children (33 aged from 1 to 9 years and 19 aged between 10 and 17 years) and 67 healthy adults. When the higher absolute mononuclear cell counts in the peripheral blood samples of the children were taken into account, the relative production of all measured cytokines was lower in the cell cultures of the children than of the adults. In the group of the younger children (< 10 years) the differences were significant for all measured cytokines. In the group of older children (> = 10 years) the values were higher than in the younger children but lower than in adults. The findings indicate that the cellular immunological competence is or can be reduced in children and adolescents, particularly young children below 10 years of age. There seems to be a gradual development of cytokine production during childhood.     

Plasma cytokine profiles in patients with celiac disease and selective IgA deficiency

Celiac disease (CD) and selective IgA deficiency (IgAD) are frequently associated, and share the same genetic background. The aim of the present study was to evaluate both Type 1 and 2 plasma cytokine levels in CD and in CD-IgAD. IL-2, TNF-α, IL-10, IL-4 and IL-13 plasma levels were measured both at diagnosis and after a gluten-free diet (GFD) in 32 CD patients, in 27 CD-IgAD patients and in 30 healthy controls. IFN-γ levels were significantly higher in CD and CD-IgAD than in controls, TNF-α displayed significantly higher levels in CD-IgAD when compared both with controls and with CD, and IL-2 was in CD-IgAD significantly increased respect to controls. Kinetics of the Type 1 cytokine plasma levels did not show a clear relationship with the GFD in both groups of CD patients, and particularly in those with IgAD. IL-4 and IL-13, both at diagnosis and after a GFD, were not significantly different in controls and in celiac patients (with and without IgAD). IL-10, whose production is stimulated by the TNF-α, had significantly higher plasma levels in CD-IgAD, but not in CD patients, with a significant decrease after a GFD. CD and especially CD-IgAD patients display persistently higher pro-inflammatory cytokine levels, suggesting a persistent state of activation of pro-phlogistic signals in CD, particularly when IgAD coexists. Serial measurement of serum IL-10 may be an adjunctive evaluating criterion in the follow-up of CD-IgAD patients.     

Evidence of Circulating Interferon-γ in Newly Diagnosed Diabetic Children

ABSTRACT. Acid-stable interferons (IFN-α or IFN-β) are produced by nucleated cells infected by virus, while acid-labile interferon (IFN-γ) is synthesized by activated T-lymphocytes. IFN-γ or an atypical form of acid-labile IFN-α have been observed in the circulation of some patients with autoimmune disease. It is believed that autoimmunity and/or viral infections are involved in the pathogenesis of insulin dependent diabetes mellitus. We therefore examined the sera of newly diagnosed diabetic children for the presence of virus-induced or acid-labile IFN. Significant IFN levels (≥8 U/ml) were observed in 11 of 29 patients when compared to 31 healthy children in the same age range. The inhibitor is characterized by species specificity, acid-lability and neutralization with anti-serum for IFN-γ.     

Effects of breast milk from allergic and non-allergic mothers on mitogen- and allergen-induced cytokine production

Breast milk contains several components that provide specific immunity and affect the maturation of the infant's immune system. The aim of this study was to analyze the effects of breast milk, on mitogen- and allergen-induced cytokine production from cord blood mononuclear cells (CBMC), and if those effects differ between allergic and non-allergic mothers. The cells were incubated for 96 h with phytohemagglutinin (PHA), ovalbumin or cat dander in the presence of various dilutions of colostrum. Colostrum inhibited both mitogen- and cat-induced IFN-γ and mitogen-induced interleukin-4 (IL-4) production. The inhibition on IFN-γ production was to some extent caused by TGF-β, as the effect was modified when an anti-TGF-β antibody was added to the cultures. In contrast, colostrum enhanced allergen-induced production of the Th2-like cytokines IL-5 and IL-13, and this was accompanied with increased production of IL-10. No differences were found between allergic and non-allergic mothers. The inhibitory effect of breast milk on IFN-γ production, which was partly due to the high levels of TGF-β, together with the enhancing effect on IL-10 secretion, confirm that breast milk is anti-inflammatory. Although the production of IL-5 and IL-13 was enhanced by colostrum, this was accompanied with an increased production of IL-10. Together with the high levels of TGF-β in breast milk and inhibitory effect of colostrum on IL-4 production, this suggests a possible mechanism whereby breast-feeding may protect against the development of allergy. Despite differences in the composition of breast milk between allergic and non-allergic mothers, the effects of breast milk on cytokine production from CBMC were independent of the atopic status of the mothers.     

Cytokine profiles in 1-yr-old very low-birth-weight infants after enteral glutamine supplementation in the neonatal period

In a previous study, we found that glutamine-enriched enteral nutrition in 102 very low-birth-weight (VLBW) infants decreased both the incidence of serious neonatal infections and atopic dermatitis during the first year of life. The aims of this follow-up study were to determine whether these beneficial effects are attended by changes in Th₁ and Th₂ cytokine profiles at age 1 yr. Furthermore, we studied changes in cytokine profiles during the first year of life in these VLBW infants. In total, 89 infants were eligible for the follow-up study (12 died, 1 exclusion due to a chromosomal abnormality). Th₁ (IFN-γ, TNF- α and IL-2) and Th₂ cytokine (IL-10, IL-5, and IL-4) profiles following in vitro whole blood stimulation were measured at 1 yr. Cytokine profiles were measured in 59/89 (66%) infants. Glutamine-enriched enteral nutrition in neonatal period did not influence cytokine profiles at 1 yr. Cytokine profiles were not different in infants with and without allergic or infectious diseases. The beneficial effect of glutamine-enriched enteral nutrition on the incidence of serious neonatal infections and atopic dermatitis during the first year of life is not related to changes in the Th₁ and Th₂ cytokine profiles. Both Th₁ and Th₂ cytokine profiles increased during the first year of life in this cohort of VLBW infants.     

Interleukin 6, but not tumour necrosis factor-α, is a good predictor of severe infection in febrile neutropenic and non-neutropenic children with malignancy

Objective : Interleukin-6 (IL6), tumor necrosis factor-a (TNF-a) and interferon-gamma (IFN-7) are important mediators of the inflammatory response in human infection. The aim of this study was to determine the relationship between serum levels of IL6, TNF-α, IFN-γ and CRP in febrile children with malignant disease, and relate these levels to aetiology of fever, presence of neutropenia and the effect of untreated malignancy. Methods: 110 febrile episodes in 70 children with malignant disease were included. Cytokine analyses were performed with sensitive immunoradiometric methods using double monoclonal antibodies. Results: IL6 had a sensitivity of 74% in detecting sepsis in children with fever and malignant disease. This sensitivity was not influenced by the presence of neutropenia or newly diagnosed malignancy. A positive correlation between IL6 and the CRP levels on the following day was observed ( r = 53). TNF-α was elevated in 22% of the episodes and mean levels were significantly higher in untreated malignancy but lower in neutropenic patients. IFN-γ was elevated in 18% of cases and correlated strongly with mean TNF-a levels. Conclusions: IL6 is a sensitive and early predictor of bacterial infection in both neutropenic and non-neutropenic febrile children with malignancy. It is more sensitive than CRP in detecting sepsis, but the predictive value is too low to allow IL6 levels to influence initial treatment decisions in patients with granulocytopenia. TNF-α production seems to be impaired in neutropenic children and serum TNF-α cannot be employed as an indicator of bacterial infection.     

Comparison of cytokine responses in nasopharyngeal aspirates from children with viral lower respiratory tract infections

Aim: To determine whether nasopharyngeal aspirates (NPAs) cytokine response is different according to the causative viruses in children with lower respiratory tract infections (LRTI).
Methods: NPAs from 277 children with LRTI caused by respiratory virus were evaluated. Based on the proven viral agents, LRTI patients were divided into four groups. Levels of IL-4, IL-5 and IFN-γ were determined by ELISA.
Results: Patients with influenza virus infection demonstrated significantly lower IL-4 and IL-5 levels than those with other three groups. Patients with respiratory syncytial virus (RSV) infection showed an increase in production of IL-4 and IL-5, and a decrease in the IFN-γ level when compared to patients with influenza virus infection. Interestingly, a similar Th2 response was seen in patients with parainfluenza virus or adenovirus infection.
Conclusion: These results demonstrate that respiratory viruses can induce different local cytokine responses. However, Th2 biased responses are not unique for RSV but seem to be predominant in respiratory viruses of young children.     

T-cell signal transduction in children with cow's milk allergy – increased MAP kinase activation in patients with acute symptoms of cow's milk allergy

The precise immune mechanisms behind cow's milk allergy (CMA) are still unknown. Previously, the production of the cytokines TNF-α and IFN-γ in T cells from children with CMA has been shown to be decreased, and the production of IL-4 has been shown to be increased when compared to healthy children. As these aberrations in cytokine production may be associated with disturbances in cellular function, we investigated whether T-cell signal transduction is abnormal in children with CMA. For this purpose we evaluated the activation of the MAP kinase Erk2. Thirty-nine infants were included in the study. Of those with CMA, 13 had acute symptoms and 9 were free of symptoms due to a successful elimination diet at the time of the study. To activate T cells and to stimulate MAP kinase phosphorylation, peripheral blood mononuclear cells (PBMC) were incubated with Concanavalin A (ConA). The change in MAP kinase phosphorylation was measured by Western blotting. The increase in MAP kinase phosphorylation after stimulation with ConA for 5 min was significantly higher in cells from patients with acute symptoms of CMA than in cells from CMA patients free of symptoms or cells from healthy children. A time-course experiment showed that the change in MAP kinase phosphorylation was still increasing after 10 min incubation in cells from patients with acute symptoms of CMA. The increased MAP kinase activation was found to correlate positively with non-IgE mediated CMA in patients with acute symptoms of CMA.     

In vitro cytokine production and phenotype expression by blood mononuclear cellsfrom umbilical cords, children and adults

Age related differences in immunological reactions include variations in the in vitro functions of blood mononuclear cells (MNC). In an attempt to understand the mechanism behind these differences we examined age related differences in the phenotype profiles of MNC in parallel with the in vitro production of interleukin IL-6, tumour necrosis factor alpha (TNFα) and in-terferon gamma (IFNg) in neonates, children and adults. In cultures without added polyclonal activators IL-6 and TNFα levels in children were 3–6 times higher than those of umbilical cords and adults. However, using optimal in vitro stimulation ( E. coli lipopolysaccharide (LPS), phytohaemmag-glutinin or pokeweed mitogen (PWM)) no significant differences in the levels of these cytokines were observed. The levels of IFNg in PWM driven cultures followed a different pattern with comparable levels in children and adults, and unmeasurable levels in cord blood MNC. Flow cytometry analysis of the phenotypic distribution of MNC revealed age related differences in the expression of CD3, CD4, CD8, CD14, CD19, CD45RA, CD45RO, CD2, LFA-1, ICAM-1 and LFA-3. Correlation studies did not indicate that the observed differences in cytokine production could be ascribed to differences in the frequency of monocytes, T cells or B cells. The TNFα levels in suboptimally stimulated cultures correlated negatively with the expression of LFA-3 and positively with CD45RA. while IFNg correlated positively with CD2, LFA-1, CD45R0 and CD8.
In conclusion, the study provides evidence of age related differences in the production of TNFα, IL-6 and IFNg among neonates, children and adults. These differences may to some extent be caused by differences in the expression of cell surface molecules involved in cellular interactions and signalling.     

Cow's milk protein-specific T-helper type I/II cytokine responses in infants with necrotizing enterocolitis

Enteral feeding, in particular with formula feeds, is associated with necrotizing enterocolitis (NEC). In this study, we have examined, in the systemic and mucosal immune compartments, for evidence of bovine milk antigen sensitization in infants with NEC. Eleven newborns with Bell's staging 2–3 NEC [median post-conceptional age 31 wk (range 27–41 wk)], 21 neonatal controls [33 (28–40) wk] and 15 infants undergoing intestinal resection or mucosal biopsy for non-inflammatory conditions [39 (34–42) wk] were studied. Spontaneous and antigen or mitogen elicited interferon-γ (IFN-γ) [T-helper type I (Th1)], interleukin (IL)-4 and IL-5 [T-helper type II (Th2)] responses were enumerated using single-cell enzyme-linked immunospot (ELISPOT) assay in peripheral blood (PBMC) or lamina propria mononuclear cells. NEC infants, compared with controls, showed a significant elevation in baseline PBMC cytokine secreting cells, vigorous mitogen responses (20- to 120-fold increase) for IFN-γ, IL-4 and IL-5 (p < 0.001), strong responses to beta-lactoglobulin (βlg) (IFN-γ > IL-4/IL-5, p ≤ 0.001), and somewhat smaller casein responses. Similarly, in the lamina propria, a small but significant increase in spontaneous cytokine-secreting cells was detected in NEC infants (p < 0.01), with an IFN-γ/IL-4 predominant phytohemagglutinin (PHA)/concanavalin-A (ConA) response. Three of nine NEC infants (but no controls) also showed a positive ELISPOT response to βlg (IFN-γ only) but none to casein. We have thus demonstrated significant cow's milk protein (CMP) sensitization in NEC, at least in the systemic compartment (mixed Th1/Th2), with minimal mucosal activation in some cases. These novel findings provide a potential mechanism for a direct contributory role of CMP in the pathogenesis of NEC.     

Method of birth alters interferon-gamma and interleukin-12 production by cord blood mononuclear cells

Many uncertainties exist regarding the capability of cord blood mononuclear cells (CBMC) to produce cytokines. A number of conflicting reports led us to examine the effects of method of birth on CBMC production of interferon-gamma (IFN-γ), interleukin-4 (IL-4) and interleukin-12 (IL-12). While constitutive production of IL-4 was found in both vaginally and cesarean-delivered infants, constitutive IFN-γ or IL-12 production was found in neither. CBMC from vaginally delivered infants responded to stimulation with concanavalin A/phorbol 12-myristate 13-acetate (Con A/PMA), phytohemagglutinin (PHA), and lipopolysaccharide (LPS) with significantly higher levels of IFN-γ than CBMC from unlabored cesarean section (CS) infants. Production of IL-12 was increased in the vaginally delivered group in response to LPS and PHA but not to ConA/PMA. In contrast, mode of delivery was not associated with differences in IL-4 production. These results indicate that mode of delivery significantly alters the capability of CBMC to produce some cytokines and therefore should be taken into account in interpreting fetal/neonatal mononuclear cell function studies.     

Intracellular production of IL-2, IL-4, IFN-γ, and TNF-α by peripheral blood CD3+ and CD4+ T cells in children with atopic dermatitis

The role of the type-2 T helper (Th2) cell-mediated immune response in the immunopathogenesis of atopic dermatitis (AD) is well documented. Whether polarized immunoresponse is confined to antigen-specific T cells or is distributed among all T cell subsets is still controversial. We investigated frequencies of interleukin-2 (IL-2), IL-4, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) producing CD3⁺ and CD4⁺ T cells in peripheral blood from children with atopic dermatitis and healthy subjects with and without in vitro stimulation. Children with severe AD had a significantly lower percentage of CD4⁺ T cells spontaneously expressing IL-4 compared with healthy controls (p <0.01). Polyclonal stimulation significantly increased cytokine production in both AD patients and healthy individuals. Frequencies of CD3⁺ and CD4⁺ producing IL-2, IL-4, IFN-γ, and TNF-α after in vitro stimulation with phorbol-12-myristate 13-acetate (PMA) + ionomycin were comparable in the AD and control groups. In response to PMA/ionomycin, children with AD and asthma symptoms had a significantly lower percentage of CD3⁺ T cells producing TNF-α. We failed to demonstrate evidence of an imbalance with respect to type-2 cytokine productions in children with AD. Comparable induction of Th1 and Th2 cytokines in polyclonally stimulated peripheral CD3⁺ and CD4⁺T cells from AD patients and controls puts into question the polarized Th2 immune response as a general characteristic of T cells in children with atopic dermatitis.     

Allergen-induced cytokine secretion in atopic and non-atopic asthmatic children

Atopic asthma is characterized by excessive T helper 2 (Th2)-like immunity to allergens in the bronchial mucosa. The Th2-cytokine interleukin (IL)-4 induces IgE production, while the Th2-cytokine IL-5 promotes eosinophilic inflammation in the airways of asthmatics. Most asthmatics are atopic, but a subgroup is non-atopic. We hypothesize that allergen-induced Th2, particularly IL-5, responses can be observed in peripheral blood in both atopic and non-atopic asthmatic children but not in healthy control children. The aim of the present study was to determine IL-4, IL-5, IL-9, IL-10, IL-13 and IFN-γ secretion induced from peripheral blood mononuclear cells (PBMC) by a broad panel of inhalant allergens (timothy, cat, birch, dog and house dust mite) in asthmatic children with and without sensitization. The study included 13 atopic asthmatic, 5 non-atopic asthmatic, and 12 non-atopic non-asthmatic children. PBMC were stimulated with allergens and cytokine production was measured with enzyme-linked immunosorbent assay (ELISA). Higher levels of cat and dog antigen-induced IL-5 release were more commonly observed in both atopic and non-atopic asthmatics than in controls. Children with atopic, but not non-atopic, asthma produced higher levels of allergen-induced IL-4 and IL-9 than controls. Non-atopic asthmatics produced more IL-10 than atopic asthmatics after cat stimulation. High levels of eosinophilia-associated IL-5 responses are induced by cat and dog allergen in both atopic and non-atopic asthmatic children. The Th2 cytokines IL-4 and IL-9 were associated only with atopic asthma, probably due to their IgE-inducing properties.     

Macrophage inflammatory protein‐1α and RANTES are present in nasal secretions during ongoing upper respiratory tract infection

Marcophage inflammatory protein‐1α (MIP‐1α) and RANTES (regulated upon activation, normal T‐cell expressed and secreted) were measured by enzyme‐linked immunosorbent assay (ELISA) from virus‐infected respiratory cell culture supernatants and from 100 nasal wash samples obtained from patients aged 8 d to 10 yr. The results of the nasal wash samples were analyzed in relation to the etiology of the viral infection. In vitro , respiratory syncytial virus (RSV) induced the production of MIP‐1α, while both RSV and adenovirus were associated with the production of RANTES. Both MIP‐1α and RANTES were detected in nasopharyngeal secretions from pediatric patients with acute upper respiratory tract RSV, adenovirus, influenza, and parainfluenza virus infection (p<0.001 by Fisher's exact test). As both of these chemokines have potent effects on the recruitment and degranulation of eosinophils and basophils, further understanding of their role in upper respiratory tract infections may provide valuble insights into the immunopathogenesis of respiratory viral infections.     

Effects of immunoglobulin and gamma-interferon on the production of tumour necrosis factor-α and interleukin-1β by peripheral blood monocytes in the acute phase of Kawasaki disease

Abstract In order to study the in vitro effects of intact immunoglobulin (Ig) and gamma-interferon (INF-) in patients with Kawasaki disease, the production of tumour necrosis factor- (TNF-) and interleukin-1 (IL-1) was measured in peripheral blood monocytes (PBM) both before and after intravenous immunoglobulin (IVIG) therapy. Spontaneous production of TNF- and IL-1 both before and after IVIG therapy was significantly higher than in healthy controls. Intact Ig enhanced in vitro the production of TNF- and IL-1 both before and after IVIG therapy approximately 3–4 times as compared to the spontaneous production. INF- did not affect the production of the two cytokines. Ig enhanced IL-1 mRNA expression in PBM of KD by 3–8 times more than that of spontaneous production.Conclusion These results suggest that: (1) the mechanism of action of IVIG therapy in KD is not to cut down the production of inflammatory cytokines such as TNF- and IL-1 andf that (2) the changes of these cytokine levels may be related to the clinical effectiveness of high dose IVIG.