Henryka Brózik

Distribution and clinical significance of blood dendritic cells in children with juvenile idiopathic arthritis

Background: A role for dendritic cells (DC) in the development of adult rheumatoid arthritis has been suggested. To date, this problem has been poorly explored in juvenile idiopathic arthritis (JIA). Objective: To analyse distribution and maturation status of blood DC (BDC) in JIA. Methods: Absolute BDC counts were assessed by the “single platform” method in peripheral blood (PB) of 47 untreated children with JIA and 32 healthy controls. Moreover, BDC were investigated in JIA synovial fluid (SF). When the panel of monoclonal antibodies against BDC antigens (BDCA) was used, three BDC subpopulations were determined: myeloid type 1 (mDC1; BDCA-1+/HLA-DR+/CD19−), myeloid type 2 (mDC2; BDCA-3+/HLA-DR+/CD14−) and plasmacytoid (pDC; BDCA-2+/HLA-DR+/CD123+). Results: A considerable deficiency of all subtypes of BDC was found in the PB of children with JIA. BDC counts in JIA SF were significantly higher than in PB both from children with JIA (p<0.001) and healthy children (p<0.001). SF BDC, especially mDC1 and mDC2 subtypes, had significantly higher expression of maturation markers (CD40, CD80, CD86 or CD83 antigens) than those from PB. A smaller number of PB BDC at diagnosis correlated significantly with poor response to treatment. Conclusions: A deficiency of BDC in PB is accompanied by enrichment of SF with those cells. Probably, circulating BDC migrate to joints where they undergo maturation and help to mediate and maintain the local immune response. Interestingly, the level of PD BDC deficiency seems to influence the outcome in children with JIA.

Apoptosis of peripheral blood lymphocytes in patients with juvenile idiopathic arthritis

Background: Recent data suggested that abnormalities in mechanisms regulating apoptosis may have a role in the development of the rheumatoid process. Objective: To evaluate different aspects of apoptosis in children with juvenile idiopathic arthritis (JIA). Methods: The frequency of TUNEL positive peripheral blood (PB) lymphocytes (apoptotic index (AI)), as well as serum CD95 (APO1/Fas) antigen expression and serum levels of sFas and interleukin 15 (IL15), were examined in 44 cases of JIA. Results were correlated with type of onset, activity of JIA, and acute phase indicators. Results: The AI of lymphocytes was significantly higher in patients with JIA than in controls (p=0.020). The mean AI of lymphocytes was increased in JIA with systemic type of onset and high activity (p=0.001). Moreover, IL15 levels in systemic disease were higher than in controls (p=0.012). An increased AI correlated with raised IL15 (p=0.046), erythrocyte sedimentation rate (p=0.005) and C reactive protein (CRP; p=0.017). Additionally, correlation was found between IL15 and CRP levels (p=0.039). CD95 and sFas levels were unchanged compared with controls. Conclusion: PB lymphocytes of children with JIA have an increased tendency to undergo apoptosis. The degree of apoptosis depends on the type of onset and activity of JIA and correlates with serum levels of IL15. Further studies are needed to explain whether this is an epiphenomenon of the disease activity or is related to the pathogenesis of JIA.

HPTLC screening assay for urinary cotinine as biomarker of environmental tobacco smoke exposure among male adolescents

For selective screening determination of urinary cotinine, i.e. (S)-1-methyl-5-(3-pyridyl)-2-pyrrolidinone, the major metabolite of nicotine, the high-performance thin-layer chromatographic (HPTLC) method have been proposed. Prior the final HPTLC analysis the procedure required a solid-phase extraction (SPE) of cotinine from collected urine samples with 1-methyl-2-pyrrolidinone as an internal standard. Densitometrical quantitation of cotinine on the chromatograms have been performed with a 16-grayscale scanner using the specialized software implemented on a desktop microcomputer. The lower detection limit of cotinine was 6 microg/l allowing the method to be applied for the measurement a concentration of this compound in urine samples collected from 35 elementary schoolboys exposed on both moderate and/or significant ETS. The mean recovery of cotinine from urine samples was 93%. The mean intra-day accuracy for the analysis of cotinine in range 6-750 microg/l. including four paralell measurements, was 2.9 %. The results of cotinine measurements by proposed SPE-HPTLC procedure were used in the pilot studies for assessment of hazard from home ETS on the health status of elementary schoolboys, especially an increased risk for infectious respiratory track diseases.

Anticitrullinated Protein Antibodies and Radiological Progression in Juvenile Idiopathic Arthritis

Objective. The aim of the study was to investigate whether determination of anticitrullinated protein antibodies (ACPA) provides predictive information on severity of disease course and joint destruction in children with juvenile idiopathic arthritis (JIA). Methods. Sera from 74 children with JIA were examined for ACPA using the ELISA test. To assess joint destruction, plain radiographs of both hands were scored twice according to the Steinbrocker scale: at the beginning of observation and after 8.9 to 15.2 months (median 11.5 months) of the followup. Correlations between ACPA serum levels and the disease characteristics (type of JIA onset, disease activity, disease duration, radiological status) were investigated. Results. Twenty-six out of 74 examined children with JIA (35.0%) were ACPA-positive [> 5 relative units (RU)/ml]. ACPA were present in all types of JIA onset, including 36.6% of children with early stage JIA (disease duration < 6 months). All of the IgM-rheumatoid factor (RF)-positive children with polyarticular type of JIA onset were simultaneously positive for ACPA. ACPA levels correlated positively with disease activity at the beginning of the study (rho = 0.7196; p < 0.0001) and after followup (rho = 0.2485; p = 0.0486). Disease duration did not significantly affect ACPA serum levels. ACPA levels correlated positively with radiological joint destruction in children with JIA, both at the beginning of the study (rho = 0.4599; p = 0.0004) and after the followup period (rho = 0.5523; p < 0.0001). Conclusion. ACPA were superior to IgM-RF in diagnosing JIA and provided predictive information on severity of disease course and radiological outcome.

Relationship between impaired apoptosis of lymphocytes and distribution of dendritic cells in peripheral blood and synovial fluid of children with juvenile idiopathic arthritis

Introduction:The pathogenesis of juvenile idiopathic arthritis (JIA) is not fully understood. Recently the present authors described disturbed apoptosis of JIA lymphocytes in both peripheral blood (PB) and synovial fluid (SF) as well as an abnormal distribution of blood dendritic cells (BDCs) between the PB and SF in this disease. Possible relationships between these events during the development of JIA process are assessed here.Materials and Methods:Lymphocyte apoptosis and BDC counts were assessed in the PB and SF of untreated JIA children. Lymphocyte apoptosis was analyzed by the Annexin-V/propydium iodide assay. Total DC (TDC) number was based on the sum of three BDC subpopulations determined using a panel of monoclonal antibodies against BDC antigens (BDCA): myeloid type 1 (mDC1, BDCA-1+/HLA-DR+/CD19-), myeloid type 2 (mDC2, BDCA-3+/HLA-DR+/CD14-), and plasmacytoid (pDC, BDCA-2+/HLA-DR+/CD123+). Cells were enumerated by the flow cytometric “single-platform” method. The concentration of tumor necrosis factor (TNF)-α and the distribution of particular lymphocyte subtypes in both PB and SF were also investigated.Results:There was significant positive correlation between apoptosis of PB lymphocytes and SF TDC count (p=0.002) as well as SF TNF-α concentration (p=0.007). SF TNF-α levels also correlated with SF TDC count (p=0.003). Moreover, JIA SF was distinctly enriched with CD4+ and CD8+ T lymphocytes and included CD4+/CD25high cells as well. There was significant positive correlation between the number of CD4+/CD25high cells and SF JIA BDC count (p=0.015).Conclusions:These data suggest a possible link between impaired apoptosis of PB/SF lymphocytes and increased recruitment of PB BDCs to SF and other elements of the immune system in JIA, including regulatory CD4+/CD25high cells.

Kompleksowa ocena zaburzeń funkcji rozkurczowej lewej komory u dzieci z młodzieńczym idiopatycznym zapaleniem stawów

Do najczestszych zapalnych ukladowych chorob tkanki lącznej u dzieci nalezy mlodziencze idiopatyczne zapalenie stawow (MIZS). U dzieci z MIZS do objecia procesem chorobowym serca dochodzi u 30–80% przypadkow, natomiast objawy kliniczne wystepują u 3–9% chorych. Celem pracy byla ocena zaburzen funkcji rozkurczowej lewej komory u pacjentow z MIZS. Metody Badaniem objeto 50 dzieci z rozpoznanym wcześniej MIZS. Grupe kontrolną stanowilo 20 zdrowych dzieci. U kazdego pacjenta wykonano badanie echokardiograficzne z oceną funkcji skurczowej i rozkurczowej serca (MPI, E/A, VpMV, AEF). Wyniki Statystycznie istotną roznice stwierdzono pomiedzy wartościami wskaźnika E/A (1,456 ± 0,353 i 1,864 ± 0,385). Wartości fali A w grupach wyniosly średnio 0,6 ± 0,131 m/s i 0,48 ± 0,078 m/s. Wartośc MPI w grupie badanej wyniosla średnio 0,476 ± 0,13. W grupie kontrolnej średnia wartośc MPI wyniosla 0,411 ± 0,075. Średnia wartośc AEF wyniosla 12,09 ± 6,04 kdyn i 6,17 ± 1,9 kdyn. Wartośc VpMV wyniosla średnio 0,7 ± 0,14 m/s i 0,81 ± 0,13 m/s. Wartości te statystycznie istotnie roznily sie miedzy sobą. Nie odnotowano statystycznie istotnej korelacji pomiedzy czasem trwania choroby a ktorymkolwiek ze wskaźnikow opisujących zaburzenia relaksacji lewej komory. Wnioski 1. U pacjentow z MIZS stwierdzono zaburzenia funkcji rozkurczowej lewej komory. 2. W ocenie zaburzen funkcji rozkurczowej lewej komory u pacjentow z MIZS wskazane jest zastosowanie kilku parametrow. Na szczegolną uwage, ze wzgledu na niezaleznośc od innych czynnikow, zaslugują MPI oraz VpMV. 3. Sila wyrzutu przedsionka (AEF) jest wartościowym parametrem oceniającym funkcje rozkurczową lewej komory.