Dry eye in systemic lupus erythematosus patients / Olho seco em pacientes com lúpus eritematoso sistêmico

Abstract Objective: To study the association of dry eye with lupus disease activity and cumulative damage. To verify if epidemiological, treatment and autoantibody profile of SLE (systemic Lupus erythematosus) patients influence the presence of dry eye. Methods: We studied 70 SLE patients for the presence of dry eye by Schirmer test, disease activity by SLEDAI (SLE-Disease activity index) and cumulative damage by SLICC/ACR DI (Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index). Patients were also submitted to the OSDI (Ocular Surface Disease Index) questionnaire. Epidemiological and treatment data and autoantibody profile were extracted from the charts. Results: Dry eye by Schirmer test was present in 51.4% of the sample. No association of the presence of dry eye with SLEDAI and SLICC DI were found (p = ns). Subjective symptoms of dry eye measured by OSDI showed a modest correlation with SLEDAI (Spearman rho = 0.32). Treatment profile did not influence in the presence of dry eye that was more common in older patients (p < 0.0001). Anti dsDNA had a negative association with the presence of positive Schirmer test (p = 0.0008). Conclusions: Dry eye detected by Schirmer test in SLE patients has no association with disease activity nor cumulative damage. Anti dsDNA seems to have a protective effect in this context.

Resumo Objetivos: Estudar a associação do olho seco com a atividade do lúpus eritematoso sistêmico (LES) e seus danos cumulativos. Verificar se o perfil epidemiológico, de tratamento e de auto anticorpos de pacientes com LES influencia a presença de olho seco. Métodos: Foram estudados 70 pacientes com LES para a presença de olho seco pelo teste de Schirmer, atividade da doença por SLEDAI (SLE Disease Activity Index) e dano cumulativo por SLICC/ACR DI (Clínicas Colaborativas Internacionais de Lúpus Eritematoso Sistêmico/American College of Rheumatology Damage Index). Os pacientes também foram submetidos ao questionário OSDI (índice de doenças da superfície ocular). Os dados epidemiológicos e de tratamento e o perfil de auto anticorpos foram extraídos dos prontuários. Resultados: Olho seco pelo teste de Schirmer esteve presente em 51,4% da amostra. Nenhuma associação da presença de olho seco com SLEDAI e SLICC/ACR DI foi encontrada (p = ns). Os sintomas subjetivos do olho seco medidos por OSDI mostraram uma correlação modesta com SLEDAI (Rho de Spearman = 0,32) . O perfil do tratamento não influenciou na presença de olho seco que era mais comum em uns pacientes mais idosos (p < 0, 1). Anti dsDNA teve uma associação negativa com a presença de teste positivo de Schirmer (p = 0, 8). Conclusões: Olho seco detectado pelo teste de Schirmer em pacientes com LES não tem associação com atividade da doença nem dano cumulativo. Anti dsDNA parece ter um efeito protetor neste contexto.

Evaluation of Anti-dsDNA Antibody Tests: Crithidia luciliae Immunofluorescence Test, Immunoblot, Enzyme-linked Immunosorbent Assay, Chemiluminescence Immunoassay / 대한진단검사의학회지

BACKGROUND: Anti-double stranded DNA antibody (anti-dsDNA) test is useful for the diagnosis and monitoring of systemic lupus erythematosus (SLE). Although several methods are available, none of them is completely satisfactory and differences among them have been reported. We evaluated the diagnostic performance of 6 commercial kits for anti-dsDNA detection. METHODS: A total of 142 sera (SLE [N=74], other systemic rheumatic diseases [N=50], other diseases [N=18]) were tested by 6 different assay kits using different antigenic sources of DNA: Crithidia luciliae immunofluorescence test (CLIFT), salmon testes (immunoblot, IB), human (ELISA I), salmon testes with nucleosome linker (ELISA II), plasmid (ELISA III), and synthetic oligonucleotides (chemiluminescence immunoassay, CLIA). RESULTS: With manufacturers' cut-off values, 6 test kits showed sensitivities of 55.4-91.9%. ELISA I had a greater sensitivity than the other five assays (P0.05). With cut-off values set at 95% of specificity, ELISA II had a higher sensitivity than ELISA III (63.5% vs. 41.9%, P<0.05). IB had poor concordance rates with other assays (42.0-65.0%). Pearson correlation coefficients among 4 quantitative assays were 0.667-0.798. CONCLUSIONS: Six different assays showed various performances depending on the methods and cut-off values used. Except IB, the other five assays can be used for the detection of anti-dsDNA.

Anti-double stranded antibody association with titers and fluorescence patterns of anti-nuclear antibody in systemic lupus erythematosus

Many laboratories do not test antinuclear antibodies [ANA] by indirect immune-fluorescence [IIF] in parallel with anti-double stranded [ds] DNA antibodies. This study attempts to investigate the legitimacy of such practice. A retrospective laboratory data analysis of simultaneous assessment of ANA and anti-dsDNA antibody results of 106 patients with either diagnosed or suspected systemic lupus erythematosus [SLE] was performed at King Khalid University Hospital, Riyadh, Kingdom of Saudi Arabia. The ANA was detected by IIF on HEp2 cells and anti-dsDNA antibodies were assessed by specific ELISA test. Among the patients, female preponderance [96.3%] was evident and a coarse speckled fluorescence pattern was commonly observed [60.4%]. There was almost no detection of anti-dsDNA antibodies up to an ANA titer of 1:320. Anti-dsDNA antibodies were often detected at ANA titers of 1:640 and beyond. Other patterns of fluorescence observed at ANA titers as low as 1:40 and at higher dilutions were, fine speckled [14.15%], homogeneous [9.4%], anti-mitochondrial [7.5%], ribosomal [4.7%], and nucleolar [3.8%]. Linear regression analysis revealed a statistically significant relationship [p=0.02] between ANA titers and anti-dsDNA antibodies only in the presence of a coarse speckled pattern. The rare occurrence of anti-dsDNA antibodies at clinically significant ANA titers associated with the coarse speckled pattern may mask the diagnosis of SLE. Similarly, the diagnosis of SLE may be overlooked if anti-dsDNA antibodies are not checked in the presence of clinically insignificant ANA titers associated with other patterns of fluorescence

Detection of Anti-ENA and anti-dsDNA Antibodies Using Line Immunoassay in Systemic Autoimmune Diseases / 대한진단검사의학회지

BACKGROUND: Detection of antibodies to extractable nuclear antigens (ENAs) and dsDNA is needed for the diagnosis of and predicting prognosis in systemic autoimmune diseases. Recently introduced line immunoassay (LIA) has the advantage of detecting several autoantibodies simultaneously, and we evaluated its usefulness in the diagnosis of autoimmune diseases in comparison with enzyme-linked immunosorbent assay (ELISA). METHODS: Samples were collected from 437 patients referred by rheumatologists. FANA (fluorescent antinuclear antibody) test and LIA for the detection of 13 different autoantibodies, including 6 ENAs and dsDNA were performed. LIA-positive samples for ENA or dsDNA antibodies were further tested with ELISA. Final diagnosis was made by rheumatologists according to the diagnostic criteria. Agreement of results between LIA and ELISA was analyzed in 53 selected patients with systemic autoimmune diseases. RESULTS: The LIA detected antibodies to ENA and dsDNA in 118 and 22 patients, respectively, and ELISA detected 70.3% (83/118) and 45.5% (10/22) of LIA positive samples. Especially, 60.2% (71/118) of patients with positive ENA antibody on LIA was diagnosed as systemic autoimmune diseases. Patients having strong FANA titer and homogenous/speckled pattern showed higher prevalence of autoantibodies, but a small proportion of FANA negative patients also showed positive reactivity (LIA 10.8%, ELISA 5.2%). LIA showed a good agreement with ELISA for the anti-ENA antibodies (> or =80%), and a lower agreement for the anti-dsDNA antibody (67.9%). CONCLUSIONS: LIA detecting several autoantibodies simultaneously might replace ELISA for anti-ENA antibodies, but not for anti-dsDNA antibodies. When LIA is performed considering clinical manifestations and FANA, it could contribute to the diagnosis of systemic autoimmune disease.

Kikuchi's disease associated with systemic lupus erythematosus.

A 25-year-old multiparous female presented with fever, joint pains, facial rash and lymphadenopathy of three months' duration. Lymph node biopsy revealed a diagnosis of Kikuchi's disease. She fulfilled seven out of the 11 ARA criteria for SLE. The association of Kikuchi's disease and SLE is rare.

Expression of anti-Z-DNA single chain antibody variable fragment on the filamentous phage surface

We describe the expression of an anti-Z-DNA single chain variable region antibody fragment (scFv) on a filamentous phage surface. Four vectors for phage display were constructed. Two of them are able to display multiple copies of the antibody fragment, and the others can be used to make monovalent libraries. The vectors use different promoter/leader sequences to direct the expression of the fused proteins. All were able to promote the assembly of fusion virion particles. In this paper we also show the affinity selection (biopanning) of those phage-antibodies based on the capacity of their products to recognize the antigen. We used biotinylated Z-DNA and the selection was performed in a solution phase fashion. The data presented here indicate that these vectors can be further used to construct anti-nucleic acid antibody fragment libraries that can be used to study the basis of nucleic acid-protein interaction and its role in autoimmunity mechanisms.

Hepatitis C virus genotyping by polymerase chain reaction and DNA enzyme immunoassay among Saudi patients in the Western Province, Saudi Arabia

The distribution of hepatitis C virus [HCV] genotypes in the Western Province of Saudi Arabia is unknown. The purpose of our study was to determine the prevalent HCV genotypes among HCV seropositive Saudi patients in the Western Province, and to study the relationship between types/subtypes, clinical status and liver histology. Patients and Serum samples were collected from 140 consecutive patients attending the Hepatology Clinic with varying grades of liver diseases, high alanine transferase [ALT] for >6 months, positive HCV, qualitative PCR, and who had had liver biopsy. HCV genotyping was determined on patients who had tested positive by both HCV enzyme immunoassay [EIA] and recombinant immunoblot assay [RIBA]. Of the 140 patients, 97 [69.2%] had genotype 4, 18 [12.8%] had genotype 1a, and 16 [11.4%] had genotype 1b. Genotypes 2b and 5 were found in two patients [1.4%] each, while 5 patients [3.6%] had mixed infections with genotypes 4 and 5. Of the 97 patients infected with genotype 4, 84 [86.6%] had chronic active hepatitis [CAH], two [2.1%] had CAH with active cirrhosis, 9 [9.3%] had cirrhosis and two [2.1%] had normal liver histology [NLH]. The most prevalent HCV genotype in the Western Province of Saudi Arabia was genotype 4 [69.2%]. Genotype 1b was encountered in 16 [11.4%] patients. For the first time, genotype 5 was identified in the Western Province of Saudi Arabia. Genotypes 1b and 4 were associated with different histological grades of liver disease

Anti-C1q antibody as a marker of disease activity in systemic lupus erythematosus.

The present study was conducted to examine the usefulness of anti-C1q antibody as a marker of disease activity in Indian patients with systemic lupus erythematosus (SLE). We standardized the assay for detection of IgG anti-C1q antibody using ELISA. The normal cut-off level was determined by testing 57 healthy, age and sex matched controls to be 53 units/m1 (mean +/- 2 SD). Patients with SEL (97 females and 13 males) were studied and the following parameters were obtained on all: SLE disease activity index (SLEDAI), anti-C1q, anti-ds DNA and C3. Correlations were tested between these parameters using Spearman's rank correlation coefficients. Anti-C1q was found positive in 66 (60%) patients while anti-ds DNA was found in 78 (71%). The positive predictive values of anti-C1q and anti-ds DNA for lupus nephritis were 59 and 61 per cent respectively. The titres of anti-C1q correlated positively with SLEDAI (P < 0.01) and anti-ds DNA (P < 0.01) and negatively with C3 levels (P < 0.001). No significant correlation was observed between anti-C1q positivity and any particular organ involvement. Similarly, no correlation was found between anti-C1q and proliferative lupus nephritis. Anti-C1q was found positive in 5 of 9 patients with moderate SLEDAI scores and negative for anti-ds DNA antibody. It is concluded that anti-C1q antibody can serve as a general marker for lupus activity, supplementing the currently used serum markers.

A radioimmunoassay method for detection of DNA based on chemical immobilization of anti-DNA antibody

High selectivity provided by biomolecules such as antibodies and enzymes has been exploited during the last two decades for development of biosensors. Of particular importance are efficient immobilization methods for biomolecules in order to preserve their biological activities. In this study, we have evaluated immobilization strategies for an anti-DNA antibody on a self-assembled monolayer of omega-functionalized thiols. The antibody was immobilized via peptide bond formation between the primary amines in the antibody and the carboxyl groups on the self-assembled monolayer. The peptide bond coupling was achieved by activating COOH groups on the surface through N-Hydroxysuccimide (NHS)-ester formation, followed by acylation of NH2 group in the antibody. DNA binding activity of the immobilized antibody was examined by counting beta emission from 35S-labeled DNA.

Extrachromosomal nucleic acids in bovine babesia

Two kinds of small extrachromosomal nucleic acid elements were found in the bovine babesias, Babesia bovis and B. bigemina. One element with an apparent size of 5.5 kilobase pairs (kbp) is a double stranded RNA related to virus like particles. Another molecule is a double stranded DNA with a molecular size of about 6.2 kbp. Southern blot comparison of restriction DNA fragments of the latter molecule, which is present in both B. bovis and B. bigemina is described

Analise citogenetica relacionada a idade e ao tratamento com drogas em macrofagos peritoneais e celulas da medula ossea de camundongos femeas Swiss hiperimunes intactos ou ooforectomizados / Citogenetic studies relationships to age and to tretment with drugs in peritoneal macrophages and bone marrow cells in the intact and ovarectomized female mouse Swiss hiperimunes

Estudos citogeneticos relacionados com a idade em camundongos Swiss femeas hiperimunes intactos mostraram aumento da frequencia de macrofgos peritoneais com alteracoes cromossomicas estruturais aos 6 meses e aumento progressivo da frequencia de macrofagos hiperdiploides dos 6 aos 15 meses de idade. Nos animais ooferectomizados, os macrofagos apresentaram estes aumentos aos 2 meses, 30 dias apos a ooferectomia, e dos 6 aos 18 meses, respectivamente. Os resultados evidenciam uma relacao entre alteracoes hormonais e aberracoes cromossomicas nos macrofagos peritoneais. Estas aberracoes cromossomicas nao foram observadas em celulas da medula ossea dos animais intactos e dos ooferectomizados, sugerindo respostas diferentes nos macrofagos peritoneais e seus precurssores na medula ossea.Os antioxidantes acido ascorbico e alfa-tocoferol protegeram os macrofagos peritoneais de camundongos ooforectomizados das aberracoes cromossomicas estruturais, evidenciando papel dos radicais livres no aparecimento das mesmas. Considerando que os estrogenos têm acao antioxidante na peroxidacao lipidica, que o tocoferol é um captador de radicais lipofilicos e que o ácido ascórbico pode ser um regenarador do tocoferol, é provável que as quebras verificadas nos cromossomos dos macrófagos peritoneais de camundongos ooforectomizados resultam de peridoxidacao lipidica. O metotrexato mostrou efeito sinergico com a ooforectomia em relacäo ás alteracöes cromossômicas, e teve este efeito parcialmente suprimido pelo ácido ascórbico, tanto nos animais intactos, como nos ooferectomizados, sugerindo que, além da inibicäo de reparo de DNA, a droga tenha possível acäo através dos radicais livres. O benzonidazol provocou um grande número de delecöes, sobretudo nos macrófagos peritoneais de animais ooferectomizados. Seu efeito foi antagonizado, em grande proporcäo, pelo tocoferol, o que corrobora os dados da literatura em relacäo á atuacäo do benzonidazol através da formacäo de radicais livres. Por serem mais suscetíveis ás alteracöes hormonais e aos tratamentos com drogas que as celulas da medula óssea, conclui-se que os macrófagos säo adequados para estudos dos fenômenos ligados ao envelhecimento, dos mecanismos de inducäo de aberracöes cromossomicas, dos efeitos de drogas e seu modo de acäo, constituindo, assim, importante modelo para investigacöes em citogenetica

Auto antibodies to nucleosides in systemic lupus erythematosus.

Serological studies in 110 patients with systemic lupus erythematosus (SLE) have shown that autoantibodies to DNA and RNA had subspecificity to adenosine (30.9%), cytidine (79%), guanosine (44.5%), thymidine (20%) and uracil (56.3%). It was also observed that DNA antibodies are heterogenous and that antibody with specificity for both the native confirmation as well as exposed nucleoside of the denatured molecule were present in sera of most of the patients with SLE. There was also alteration in the pattern of antibody to nucleoside in some patients who were treated with steroids or immunosuppressive drugs.

Naturally occurring anti-native DNA antibodies in SLE binding Z-DNA.

A high degree of specificity of circulating anti-DNA antibody for double stranded DNA in the sera of SLE patients was found, as compared to single stranded polymer. The antibody recognized brominated DNA, a polymer that appears to attain Z-conformation as indicated on the basis of UV absorption characteristics. The existence in native DNA of regions undergoing B----Z transition has been detected.

Systemic lupus erythematosus in Indian children.

Twenty cases of systemic lupus erythematosus (SLE) in prepubertal children (less than 14 years of age) were seen over a period of 14 years. The male:female ratio was 1:2.3, and the mean age at onset was 9.37 years. Fever with joint involvement was the commonest presenting manifestation (60%), followed by nephrotic syndrome (25%). Notable clinical features included a high incidence of renal involvement (75%), significant hypertension (45%) and reversibility of acute renal failure (2 cases). The other organs and systems involved included: mucocutaneous manifestations (60%), cardiovascular system (30%), respiratory system (25%), neuropsychiatric manifestations (45%), and anemia (75%). Raynaud's phenomenon and thrombocytopenia were rare while leucopenia was not seen in a single patient. Immunological abnormalities noted were 100% positivity for antinuclear antibodies, and 87.5 and 75% positivity for antibodies to double-stranded and single-stranded DNA, respectively. Hypocomplementemia was seen in 75% of patients tested.

Lúpus induzido por insoniazida / Lupus induced by isoniazide

Grande número de drogas tem sido responsabilizado pela induçäo de auto-anticorpos; algumas dessas drogas säo capazes de induzir quadro clínico de LES, embora geralmente sem envolvimento renal ou de sistema nervoso. Os autores investigam a freqüência de induçäo de lúpus pela isoniazida em 20 portadores de tuberculose após três a seis meses da introduçäo dessa medicaçäo. Nenhum paciente apresentou quadro clínico sugestivo de lúpus após introduçäo da isoniazida e apenas três pacientes (15%) mostraram positividade do FAN. A freqüência dos anticorpos anti-histona e antiDNA desnaturado foi maior após inicio da terapêutica. Esses dados sugerem que o uso de isoniazida näo estaria associado à induçäo de lúpus clínico, embora essa droga seja capaz de induzir o aparecimento de anticorpos antinucleares

Alteraciones inmunologicas en pacientes con actividad lúpica y compromiso renal / Immunological changes in patients with lupus activity and renal involvement

Una población de pacientes lúpidicos que se encontaban en distintos períodos de actividad y remisión fue evaluadas por parámetros convencionales de laboratorio y por la determinación de anticuerpos anti-ADN fijadores de complemento (anti-ADN FC) en virtud de su posible relación con el compromiso renal y actividad de la enfermedad. Los estudios convencionales de laboratorio confirmaron, como parámetro de elección para determinar el estado de actividad, el nivel de CH50, más aún existiendo compromiso renal. También se observó que los anticuerpos anti-ADN-hemaglutinantes permiten distinguir entre estados de actividad y remisión. Un menor valor discriminativo se adjudicaría a los niveles de complejos inmunes circulantes (CIC) y a los valores de vía alterna del complemento (VAC). Con respecto a los sueros anti-ADN FC positivos se encontró una relación con la actividad lúpica ni con el compromiso renal. Tampoco se observó que los sueros con bajo nivel de CH50 se relacionaran exclusivamente con la presencia de ati-ADN FC; parecería que algún otro mecanismo estaría involucrado en la patogenia de la nefritis lúpica

Anticuerpos ant-ADN y manifestaciones clínicas del lupus eritematoso sistémico / Anti DNA antibodies and clinical manifestations of systemic lupus erythematosus

Se determinó la presencia de anticuerpos anti-ADN/doble cadena en 73 pacientes con lupus eritematoso sistémico y en 20 pacientes con enfermedades reumáticas no inflamatorias, usando ADN marcado con 1**125 y precipitación con sulfato de amonio. Los pacientes testigo con enfermedades reumáticas no inflamatorias presentaron títulos promedio de 5.8 unidades (ámbito de 1 a 11). Cuarenta y siete pacientes (64%) con lupus eritematoso sistémico presentaron elevación de anticuerpos. En cada una de las manifestaciones clínicas evaluadas, los porcentajes de pacientes con elevación de títulos de anticuerpos anti-ADN fueron; manifestaciones cutáneas y de mucosa (72%), artritis (79%), nefropatía (67%), hipocomplementemia (77%), serositis (100%) y lupus del sistema nervioso central (100%). La elevación de anticuerpos anti-ADN/dc es un dato importante en la evaluación de actividad de la enfermedad, pero la existencia de cifras normales no descarta la presencia de dicha actividad