Urinary interleukin-6 in acute Kawasaki disease

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URINARY INTERLEUKIN-6 IN ACUTE KAWASAKI DISEASE THE SIGNIFICANCE OF INFLAMATORY CYTOKINES A5 PREDITABLE FACTORS OF ANEURISMIN KAWASAKI SYNDROME

iazuhide Ohta, hkiko Seno, E i j i Kate, h k i h i r o Yaehie, Hidetoshi Seki, Toshio Miyawaki, Noberu Taniguchi

Takao Kohsaka, Hiroshi Miyasaka, Akio Nakamura, Noboru Kobayashi The Study Group of KS Pathogenesis In Kawasaki Syndrome(KS), to foresee the severe aneu rism is the most important problem. The KS patients with typical clinical findings were studied in 533 cases, statistically. Na, T-protein, neutrocyte count in early stage, and CRP, anemia( Hb, Ht, RBC), albumin, WBC, in aggravating stage are useful for predicting the severity of the disease. The infla~_matory cytokines (IL-6, TNF & IL-I ) were measured and investigated on its clinical significance. The serum samples of 408 from 128 patients in various stages (acute, aggravating and convalescent) were collected to assay IL-6 and TNF , while cultured mononuclear cells were used to measure IL-I . IL-I concentration in supernates in cell culture remain unchanged by clinical phase, but varied individually according to secretion amount and response to LPS immune complex stimulation. The majority of non-responders and high secretors of IL-I are grouped into the patients accompanied by aneurism. TNF in serum concentration is detected in one third of the patients by ELISA methods. The values didn'!t correlate with other laboratory values which influence the severity of disease. The serum concentration of IL-6 at early stage is correlative with CRP and other inflar,matoryl proteins, albumin, content of neutrophils in aggravating stage and febrile term which were found most important predictable factors. The cases with IL~6 value in early stage above 135ng/ml complised 90% of ANm plus AN1 groups. Cytokines values "especially 1L-6 are useful for predicting clinical courses and suggest the significant role of vascular permeability and formation of aneurism in KS patients.

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ANTI-NEU'I~OPHILCYTOPLASMICANTIBODIESIN KAWASAKIDISEASE Kazunari Kaneko, Vanita Shah, Gill Gaskin, Charles D. Pusey, Michael J. Dillon, Institute of Child Health and Royal Postgraduate Medical School, London,U.K. Anti-neutrophilcytoplasmicantibodies (ANCA) which are found in patients with systemic vasculitis have two major subtypes (p-ANCA and c-ANCA) according to the staining pattern by indirect immunoflourescencemicroscopy. The antigensagainst which p- and c-ANCA are directed have been considered to be myeloperoxidase (MPO) and 29kDa protein from azurophilic granules, respectively. Although it has been reported that ANCA are detected in patients with Kawasaki disease (KD), it is unclear which antigen ANCA are directed against. The aim of our study was to identify, if possible, the antigen for ANCA in KD. Nineteen patients and 12 healthy control children were enrolled. The ANCA directed against acid extracted granulocytes(crude antigens; A), MPO (B) and a granules (C) were measuredby ELISA. No. of Cases (%)

Type of antibodies A B C

12/19 (63.2%) 4119 (21.0%) 3/19 (15.8%)

(-) (+) (+)

(-) (-) (-)

(-) (-) (+)

These data show that anti.MPO antibodies are not detected in KD. However, anti-a granules antibodies are present in 15.8% of patients but the findingssuggest that other antigensare also involved.

Kawasaki disease (KD) is often presented with v a r i able degree of a s e p t i c pyuria and mild p r o t e i n u r i a during the acute phase, implying renal involvement in the u r i n e s from p a t i e n t s with acute KD to e l u c i d a t e the s i g n i f i c a n c e of IL-6 in the pathogenesis of renal lesions in KD. The s u b j e c t s included 16 p a t i e n t s with KD and 20 other age-matched f e b r i l e c o n t r o l s without abnormal u r i n a r y f i n d i n g s . IL-6 levels were d e t e r mined by a s e n s i t i v e sandwich ELISA assay (FUJIREBIO Inc., Tokyo). hlthongh increased serum IL-6 levels were seen in acute KD as well as f e b r i l e c o n t r o l s (23 to 557 and 6 to 970 pg/ml, r e s p e c t i v e l y ) , there was a marked d i f f e r e n c e between beth groups in u r i n a r y IL-6. In p a t i e n t s with acute KD, I b 6 levels in the urines were c o n s i s t e n t l y elevated (28 to 488 pg/mg-Cr), but u r i n a r y I L 6 l e v e l s in f e b r i l e c o n t r o l s remained low ( l e s s than 16 pg/mg-Cr). Urinary IL-6 levels returned to normal during the convalescent period of KD. In most p a t i e n t s of acute KD, u r i n a r y IL-6 levels were r e l a t e d to elevated levels of NAG and~2-microglobulin. The r e s u l t s support the notion that KD is complicated with some inflmmatory process i n the renal parenchyma, including the renal tubules. The assessment of u r i n a r y IL-6 may be a c l i n i c a l l y useful marker for diagnosis and monitoring of KD,

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AUTOANTIBODIESTO ENDOTHELIALCELLS AND NEUTROPHILCYTOPLASMICANTIGENSIN KAWASAKI DISEASE Kazunari Kaneko, Caroline O.S. Savage, Bruce E. Porringer, Vanita Shah, Jeremy D. Pearson, Michael J. Dillon, Institute of Child Health, Londonand ClinicalResearch Centre,Harrow, U.K. Anti-endothelialcell autoantibodies (AECA) have been found in a numberof presumed autoimmuneconditions. Recently AECA have also been reported in Kawasaki disease (KD). In addition, antineutrophil cytoplasmic antibodies (ANCA) which are known to be found in some patients with systemic vasculitis includingWegener's granulomatosisand microscopicpolyarteritishave also been reported in KD. The coexistence of these autoantibodies prompted us to investigate their relationship. Methods. Twenty acute phase KD patients and 18 healthy control children were studied. IgM-AECA and IgM-ANCA were measured by ELISA using human umbilical vein endothelial cells an~i disrupted granulocytes as antigens, respectively. Tiues were considered positive if greater than the control mean +3SD. Results. Titres of IgM-AECA were significantlygreater in patients than in controls. Positive titres were found in 13/20 (65%). Titres of IgM-ANCAwere also significantly greater in patients than in controls. Positive titres were found in 9/20 (45%) and all of them showed positive IgM-AECA. The correlationcoefficientbetweenIgM-AECAand IgM-ANCAwas 0.43 and statisticallysignificant(0.01