Objetivo: Ao nosso conhecimento, não há estudos na literatura avaliando simultaneamente um grande número de autoanticorpos órgãoespecíficos, bem como a prevalência de doenças autoimunes órgãoespecíficas em populações com lúpus eritematoso sistêmico juvenil (LESJ) e dermatomiosite juvenil (DMJ). Portanto, o objetivo deste estudo foi avaliar autoanticorpos e doenças autoimunes órgão-específicas em pacientes com LESJ e DMJ. Métodos: Quarenta e um pacientes com LESJ e 41 com DMJ foram investigados para os autoanticorpos séricos associados com hepatite autoimune, cirrose biliar primária, diabetes melito tipo 1 (DM1), tireoidite autoimune, gastrite autoimune e doença celíaca. Pacientes com positividade para anticorpos órgão-específicos foram avaliados para a presença das respectivas doenças autoimunes órgão-específicas. Resultados: A média de idade ao diagnóstico foi significativamente maior em pacientes com LESJ em comparação com DMJ (10,3 ± 3,4 vs. 7,3 ± 3,1 anos, p=0,0001), enquanto a média de duração da doença foi similar em ambos os grupos (p=0,92). As freqüências de autoanticorpos órgão-específicos foram semelhantes nos pacientes com LESJ e DMJ (p>0,05). Notavelmente, uma alta prevalência de autoanticorpos relacionados a tireoidite autoimune e DM1 e foi observada em ambos os grupos (20% vs. 15%, p=0,77 e 24% vs. 15%, p=0,41, respectivamente). A elevada freqüência de fator antinúcleo - FAN (93% vs. 59%, p=0,0006), anti-DNA (61% vs. 2%, p<0,0001), anti-Ro (35% vs. 0%, p<0,0001 ), anti-Sm (p=0,01), anti-RNP (p=0,02), anti-La (p=0,03) e aCL IgG (p=0,001) foram observadas em pacientes com LESJ em comparação com DMJ. Doenças autoimunes órgão-específicas foram evidenciadas apenas em pacientes com LESJ (24% vs. 0%, p=0,13). Dois pacientes com LESJ apresentavam DM1 associada com tireoidite de Hashimoto e um terceiro paciente apresentava tireoidite subclínica. Outro paciente com LESJ preenchia diagnóstico de doença celíaca com base em anemia por deficiência de ferro, a presença de anticorpo anti-endomísio, biópsia duodenal compatível com doença celíaca e resposta a dieta livre de glúten. Conclusão: Doenças órgão-específicas foram observadas apenas em pacientes com LESJ e exigiram tratamento específico. A presença destes anticorpos sugere a avaliação de doenças órgão-específicas e um acompanhamento rigoroso destes pacientes / Objective: To our knowledge, no study has assessed simultaneously a large number of organ-specific autoantibodies, as well as the prevalence of organ-specific autoimmune diseases in juvenile systemic lupus erythematosus (JSLE) and juvenile dermatomyositis (JDM) populations. Therefore, the purpose of this study was to evaluate organ-specific autoantibodies and autoimmune diseases in JSLE and JDM patients. Methods: Forty-one JSLE and 41 JDM patients were investigated for serum autoantibodies associated with autoimmune hepatitis, primary biliary cirrhosis, type 1 diabetes mellitus (T1DM), autoimmune thyroiditis, autoimmune gastritis and celiac disease. Patients with positive organspecific antibodies were assessed for the presence of the respective organ-specific autoimmune diseases. Results: Mean age at diagnosis was significantly higher in JSLE compared to JDM patients (10.3±3.4 vs. 7.3±3.1years, p=0.0001), whereas the mean disease duration was similar in both groups (p=0.92). The frequencies of organ-specific autoantibodies were similar in JSLE and JDM patients (p>0.05). Of note, a high prevalence of autoantibodies related to T1DM and autoimmune thyroiditis were observed in both groups (20% vs. 15%, p=0.77 and 24% vs. 15%, p=0.41; respectively). Higher frequencies of antinuclear antibody - ANA (93% vs. 59%, p=0.0006), anti-dsDNA (61% vs. 2%, p<0.0001), anti-Ro (35% vs. 0%, p<0.0001), anti-Sm (p=0.01), anti-RNP (p=0.02), anti-La (p=0.03) and IgG aCL (p=0.001) were observed in JSLE compared to JDM patients. Organ-specific autoimmune diseases were evidenced only in JSLE patients (24% vs. 0%, p=0.13). Two JSLE patients had T1DM associated with Hashimoto thyroiditis and another had subclinical thyroiditis. Another JSLE patient had celiac disease diagnosis based on iron deficiency anaemia, presence of anti-endomysial antibody, duodenal biopsy compatible to celiac disease and response to a gluten-free diet. Conclusion: Organ-specific diseases were observed solely in JSLE patients and required specific therapy. The presence of these antibodies recommends the evaluation of organ-specific diseases and a rigorous follow-up of these patients

O Lúpus Eritematoso Sistêmico (LES) é uma doença inflamatória crônica autoimune que apresenta uma ampla variedade de manifestações clínicas e anormalidades imunológicas, afetando principalmente mulheres. É caracterizado pela perturbação da homeostase imunológica, que envolve a indução e produção de autoanticorpos, bem como pela formação e deposição de complexos imunes, que conduzem a uma intensa resposta inflamatória e dano tecidual. Há evidências de que fatores imunológicos, ambientais, hormonais e genéticos estão implicados na patogênese da doença. Genes e proteínas envolvidas na metabolização/detoxificação de xenobióticos são frequentemente utilizados como marcadores de susceptibilidade para o desenvolvimento de doenças, cuja etiologia está relacionada à exposição a fatores de risco ambientais. Enzimas do Citocromo P450 (CYP) são as principais responsáveis pela fase I de detoxificação, na qual ativam o xenobiótico, tornando-o mais eletrofílico e, desta forma, mais reativo. As Glutationa S-transferases (GST) são enzimas detoxificantes de fase II e normalmente conjugam a glutationa reduzida com uma variedade de compostos eletrofílicos, como espécies reativas de oxigênio, facilitando a excreção de produtos tóxicos. Polimorfismos nos genes CYP e GST são capazes de alterar a expressão e a atividade catalítica das enzimas, sendo responsáveis por diferenças interindividuais quanto à capacidade de biotransformação de xenobióticos. O objetivo do nosso trabalho foi avaliar a influência de três polimorfismos GST (GSTM1 nulo, GSTT1 nulo, e GSTP1*Val) e dois polimorfismos CYP (CYP1A1*2C e CYP2E1*5B) na predisposição ao LES em uma amostra de 370 pacientes com LES e 329 doadores de sangue saudáveis provenientes da região sul do Brasil. Os polimorfismos CYP foram genotipados por PCR-RFLP, enquanto que os polimorfismos GST foram genotipados por PCR multiplex (GSTM1 nulo, GSTT1 nulo) e PCR-RFLP para GSTP1. As freqüências alélicas e genotípicas foram comparadas entre pacientes e controles usando o teste de Qui-Quadrado ou o teste Exato de Fisher. As análises foram realizadas subdividindo os indivíduos de acordo com sua origem étnica. Entre os indivíduos Euro-descendentes, observou-se uma menor freqüência de genótipos heterozigotos GSTP1*Val em pacientes com LES em comparação aos controles (p=0,0047; OR 0,63 CI 95% 0,43 – 0,93 em relação a GSTP1*Ile/Ile e OR 0,49 CI 95% 0,26 – 0,92 em relação a GSTP1*Val/Val). No grupo Afro-descendente, houve tendência a uma maior freqüência do alelo GSTP1*Val em pacientes quando comparados aos controles (p=0,061). O alelo CYP2E1*5B foi significativamente mais freqüente nos pacientes do que em controles (p=0,038; OR 2,69 CI 95% 1,00 – 8,42). Não foi observada qualquer implicação clínica dos polimorfismos CYP e GST nos pacientes com LES. Nossos dados sugerem um papel protetor do genótipo heterozigoto GSTP1*105Ile/Val em Euro-descendentes e uma possível influência do alelo CYP2E1*5B na susceptibilidade ao LES entre Afro-descendentes. / Systemic lupus erythematosus (SLE) is an autoimmune chronic inflammatory disease that presents a variety of clinical manifestations and immunological abnormalities, particularly affecting women. It is characterized by disruption of immunologic homeostasis, which results in the induction and production of autoantibodies, as well as the formation and deposition of immune complexes, leading to an intense inflammatory response and tissue damage. There is evidence that immunological, environmental, hormonal and genetic factors are involved in the pathogenesis of the disease. Genes and proteins involved in metabolism/detoxification of xenobiotics are often used as markers of susceptibility to the development of diseases whose etiology is related to exposure to environmental risk factors. Cytochrome P450 (CYP) enzymes are primarily responsible for phase I detoxification, in which activate the xenobiotic, making it more electrophilic and thus more reactive. The Glutathione S-transferases (GST) are phase II detoxifying enzymes and usually conjugate reduced glutathione with a variety of electrophilic compounds, such as reactive oxygen species, facilitating the excretion of toxic products. Polymorphisms in the CYP and GST genes can alter the expression and catalytic activity of enzymes, being responsible for interindividual differences regarding the capacity of xenobiotics biotransformation. The aim of our study was to evaluate the influence of three GST polymorphisms (GSTM1 null, GSTT1 null and GSTP1*Val) and two CYP polymorphisms (CYP1A1*2C and CYP2E1*5B) in SLE predisposition in a sample of 370 SLE patients and 329 healthy blood donors, both from southern Brazil. The CYP polymorphisms were genotyped by PCR-RFLP, while the GST polymorphisms were genotyped by multiplex PCR and PCR-RFLP for GSTP1. Allelic and genotypic frequencies were compared between patients and controls using the Chi-square test or Fisher´s exact test. Analyses were performed subdividing the individuals according to their ethnic origin. Among European-derived individuals, it was observed a lower frequency of GSTP1*Val heterozygous genotypes in SLE patients compared to controls (p = 0.0047; OR 0.63 CI 95% 0.43 - 0.93 in relation to GSTP1*Ile/Ile) and (OR 0.49 95% CI 0.26 - 0.92 in relation to GSTP1*Val/Val). In African-derived group, there was a trend to a higher frequency of GSTP1*Val allele in patients when compared to controls (p=0.061). The CYP2E1*5B allele was significantly more frequent in patients than controls (p=0.038, OR 2.69 95% CI 1.00 - 8.42). We did not observe any clinical implication of the CYP and GST polymorphisms in patients with SLE. Our data suggest a protective role of the GSTP1*105Ile/Val heterozygous genotype in European-derived and a possible influence of the CYP2E1*5B allele in SLE susceptibility among African-derived.

This research study explored the experiences of 19 women who had been diagnosed with, or were still seeking the diagnosis of SLE (lupus) and their decisions regarding support group participation. The aim of this study was to evaluate the variety of factors influencing their choices in types and sources of support, their coping strategies and the reasons behind their decisions to either choose or not choose lupus support groups as a viable support resource. Those women identified as support groups attendees recalled a more emotion-focused response to their diagnosis and showed stronger reliance on seeking emotional forms of support. Conversely, those women who chose not to participate in groups (non-attendees) utilized more problem-focused strategies when they received their news of the illness and indicated more reliance on instrumental forms of support. Additionally, the women who do not attend support groups did not seem to have more social support from outside sources, but did show a tendency to utilize relationship-focused coping more than other forms of coping strategies overall. Both groups showed a heavy reliance on their medical providers for both emotional and instrumental forms of support suggesting this source as an important factor in individual choices in coping strategies and support sources. The decisions to attend or not attend differed only in the strategies they relied on and specific group structure, timing and locations. The results of this study supports earlier research in the types and sources of social support used in adapting to a chronic illness. This study also encourages incorporating individual support services through medical providers and the development of programs that acknowledge individual coping and support needs.

Bibliography: leaves 99-106.

<p>Single nucleotide polymorphisms (SNPs) are the most common form of genetic variation. We developed a novel multiplexed method for SNP genotyping based on four-color fluorophore tag-microarray minisequencing. This method allows simultaneous genotyping of 80 samples and up to 200 SNPs in any allele combination. In study I we set up the method for a panel of SNPs from genes in the type I interferon system, and applied it in study III. In study II we used the technique to genotype SNPs from the coding region of the mitochondrial genome. A panel of 150 SNPs was genotyped in 265 individuals representing nine different populations. We demonstrated that the multiplexed SNP genotyping method for mitochondrial DNA increases the power of forensic identification in combination with sequencing of the hypervariable region of mitochondrial DNA. </p><p>In study III we performed a genetic association study of SNPs in genes related to the type I Interferon system in Systemic Lupus Erythematosus (SLE). SLE is a chronic autoimmune inflammatory disease with a complex etiology. The SNPs were genotyped in DNA samples from Swedish, Finnish, and Icelandic patients with SLE, unaffected family members, and unrelated controls. The analysis identified SNPs in two genes, the tyrosine kinase 2 (TYK2) and interferon regulatory factor 5 (IRF5) genes that are highly associated with SLE with p-values <10^-7 for joint linkage and association. </p><p>Study IV describes the analysis of the TYK2 and IRF5 SNPs in a large Rheumatoid Arthritis (RA) sample cohort. We found that SNPs in the IRF5 gene were significantly associated with RA with a p-value = 0.00008. In contrast, we did not detect an association with SNPs in the TYK2 gene. These findings demonstrate that SLE and RA may have a common genetic background in the case of IRF5, while the TYK2 variants appear to be unique for SLE. </p>

Single nucleotide polymorphisms (SNPs) are the most common form of genetic variation. We developed a novel multiplexed method for SNP genotyping based on four-color fluorophore tag-microarray minisequencing. This method allows simultaneous genotyping of 80 samples and up to 200 SNPs in any allele combination. In study I we set up the method for a panel of SNPs from genes in the type I interferon system, and applied it in study III. In study II we used the technique to genotype SNPs from the coding region of the mitochondrial genome. A panel of 150 SNPs was genotyped in 265 individuals representing nine different populations. We demonstrated that the multiplexed SNP genotyping method for mitochondrial DNA increases the power of forensic identification in combination with sequencing of the hypervariable region of mitochondrial DNA. In study III we performed a genetic association study of SNPs in genes related to the type I Interferon system in Systemic Lupus Erythematosus (SLE). SLE is a chronic autoimmune inflammatory disease with a complex etiology. The SNPs were genotyped in DNA samples from Swedish, Finnish, and Icelandic patients with SLE, unaffected family members, and unrelated controls. The analysis identified SNPs in two genes, the tyrosine kinase 2 (TYK2) and interferon regulatory factor 5 (IRF5) genes that are highly associated with SLE with p-values &lt;10-7 for joint linkage and association. Study IV describes the analysis of the TYK2 and IRF5 SNPs in a large Rheumatoid Arthritis (RA) sample cohort. We found that SNPs in the IRF5 gene were significantly associated with RA with a p-value = 0.00008. In contrast, we did not detect an association with SNPs in the TYK2 gene. These findings demonstrate that SLE and RA may have a common genetic background in the case of IRF5, while the TYK2 variants appear to be unique for SLE.

Pre-eclampsia is associated with increased risk of cardiovascular disease (CVD) later in life. The ‘gold standard’ for estimating cardiovascular risk - ultrasound assessment of the common carotid artery intima-media thickness (CCA-IMT) - does not convincingly demonstrate this increased risk. The aim of this thesis was to examine whether high-frequency (22 MHz) ultrasound assessment of the individual CCA intima and media layers and calculation of the intima/media (I/M) ratio - can indicate the increased cardiovascular risk after pre-eclampsia. After validation of the method in premenopausal women with systemic lupus erythematosus (SLE) who have a recognized increased risk of CVD, women during and after normal and preeclamptic pregnancies were investigated. Assessment of the individual artery wall layers reliably demonstrated the increased cardiovascular risk in premenopausal women with SLE, while CCA-IMT did not. The artery wall layer dimensions in women with SLE were comparable to those of postmenopausal women without SLE and were 30 years older. Among the women with normal pregnancies negative changes to the artery wall later on in the pregnancy were seen in those with lower serum estradiol, older age, higher body mass index or higher blood pressure early in the pregnancy. About one year postpartum, both the mean intima thickness and the I/M ratio had improved, compared to values during pregnancy. These findings support the theory that normal pregnancy is a stress on the vascular system. Women who developed pre-eclampsia (mean age 31 years) had thicker intima layers, thinner media layers and higher I/M ratios, both at diagnosis and one year postpartum, than women with normal pregnancies, indicating increased cardiovascular risk. Women with a history of severe pre-eclampsia (mean age 44 years; mean 11 years since the last delivery) had thicker intima layers and higher I/M ratios than women with a history of normal pregnancies, indicating long-standing negative vascular effects. Assessment of individual CCA wall layers, but not of CCA-IMT, provided clear evidence of the well-known increased cardiovascular risk in women with SLE or pre-eclampsia. The method has the potential to become an important tool in reducing cardiovascular morbidity and mortality in these women through early diagnosis and intervention.

Systemic Lupus Erythematosus (SLE) is a complex autoimmune disease. HTLV-1 Related Endogenous Sequence (HRES-1), a human endogenous retrovirus, produces 2 retroviral-like Gag capsid proteins (p8 and p15) that share significant sequence homology to the U1-subunit of the small ribonucleoprotein complex (U1sn-RNP), an autoantigen of lupus. The central hypothesis is that molecular mimicry between HRES-1 and U1sn-RNP serves as a priming event in SLE via the production of cross-reactive autoantibodies. Anti-HRES-1/U1sn-RNP serological responses in subjects with SLE and comparison populations were characterized. An overlapping peptide set mapping the HRES-1 p8 and p15 proteins was used. SLE subjects produce IgG to several regions of HRES-1. Healthy subjects or those with RA, HIV-1 infection, or HTLV-1-infection produced no significant anti-HRES-1 IgG. Anti-HRES-1 antibodies deposited in the kidneys of patients with SLE glomerulonephritis were identified. Our data suggests that HRES-1 plays a role in SLE by means of a molecular mimicry mechanism with U1sn-RNP.

Systemic lupus erythematosus (SLE) is an autoimmune, inflammatory disease characterized by autoantibody production and an activated type I interferon system. Cardiovascular disease (CVD) is as a major cause of morbidity and mortality. The aim of this thesis was to identify genetic risk factors for CVD in SLE. The role of T cells in regulation of the interferon-α (IFNα) production by plasmacytoid dendritic cells (pDCs) was also investigated.    In paper I, a thicker intima, thinner media and increased intima/media ratio was found in young premenopausal women with SLE compared to healthy controls indicating increased cardiovascular risk. As traditional ultrasound assessment of the common carotid intima-media thickness (CCA-IMT) in SLE has given conflicting results separate measurement of the intima and media can be a useful tool to identify SLE patients at increased risk of CVD.    In paper II, an association was demonstrated in SLE between a STAT4 risk allele and ischemic cerebrovascular disease and presence of anti-phospholipid antibodies (aPL). The association remained after adjustment for traditional CVD risk factors. A possible mechanism for this association is that the risk allele leads to increased production of aPL, which promotes thromboembolism.    In paper III, a genetic locus in IRF8 was identified to be associated to coronary heart disease (CHD) in SLE. The association remained after adjustment of other CHD risk factors.  Patients with the IRF8 risk variant had increased CCA-IMT, more carotid plaques and reduced frequency of circulating B cells. Weaker binding of nuclear protein to the risk allele was demonstrated, suggesting a regulatory function of the IRF8 risk variant.    In paper IV, activated T cells were found to strongly enhance the IFNα production by pDC stimulated with RNA-containing immune complexes via GM-CSF and IL-3. Activated SLE T cells enhanced the IFNα production to the same extent as T cells from healthy controls. This finding together with previous observations in SLE of increased levels of GM-CSF and IL-3 suggests that T cells contribute to the activated type I interferon system in SLE.    In conclusion, this thesis demonstrates that genetic predisposition is important for CVD in SLE and describes a new role for T cells in the pathogenesis of SLE.

Chilblain lupus erythematosus is a rare form of cutaneous lupus erythematosus characterized by bluish red infiltrates in acral locations of the body mostly affecting middle-aged women. We recently described a familial form of chilblain lupus manifesting in early childhood caused by a heterozygous mutation in the TREX1 gene, which encodes a 3′-5′ DNA exonuclease. Thus, familial chilblain lupus represents the first monogenic form of cutaneous lupus erythematosus. Here we describe the unusual clinical course of this newly defined genodermatosis in an 18-year-old female member of the family in which familial chilblain lupus was originally described. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.