A novel TNFRSF1A splice mutation associated with increased nuclear factor kappaB (NF-kappaB) transcription factor activation in patients with tumour necrosis factor receptor associated periodic syndrome (TRAPS).

OBJECTIVE: To characterise and investigate the functional consequences of a novel TNFRSF1A splice site mutation causing tumour necrosis factor receptor associated periodic syndrome (TRAPS) in a 16-year-old male patient and his mother. METHODS: Mutational DNA screening was performed in the patient and his mother. Western blotting was used to analyse protein expression levels of TNFR1. A multiplex bead immunoassay was used to quantify serum levels of range of cytokines, and an ELISA-based transcription factor assay to measure nuclear factor (NF)-kappaB transactivation. Serum levels of soluble TNFR1 (sTNFR1) were measured by ELISA and fluorescence-activated cell sorting (FACS) analysis used to measure monocyte TNFR1 cell surface expression. RESULTS: A novel mutation, c.472+1G>A (C158delinsYERSSPEAKPSPHPRG), involving a splice site in intron 4 of TNFRSF1A, was found in the proband and affected mother leading to a 45 nucleotide insertion of intronic DNA into the mRNA, resulting in an in-frame insertion of 15 amino acids in the mature TNFR1 protein and a deletion of a cysteine residue C129 (158) in cysteine rich domain (CRD)3. The patients had reduced serum sTNFR1 and surface expression levels of TNFR1, with marked increases in pro- and anti-inflammatory cytokine. Their peripheral blood mononuclear cells (PBMC) had increased basal NF-kappaB activation compared with healthy controls and also had increased p50 nuclear expression following tumour necrosis factor (TNF) stimulation compared with PBMC from healthy controls, as well as T50M (T79M) and C88R (C117R) patients with TRAPS and patients with rheumatoid arthritis (RA). CONCLUSION: A novel, TRAPS causing, TNFRSF1A splice site mutation is associated with decreased sTNFR1 levels, cell surface and whole cell extract expression and increased NF-kappaB transcription factor activation.

Postpartum bilateral renal vein thrombosis in the primary antiphospholipid syndrome.

A young Caucasian woman developed bilateral deep vein thromboses (DVT) while taking the oral contraceptive (OC) in 1984. After a 9-month period of longterm anticoagulation therapy with warfarin, she experienced recurrent DVT 2 weeks following discontinuation of anticoagulation. She was then maintained with warfarin for the following 6 years. Elevated antibodies to cardiolipin (IgG aCL) were first detected in 1988. In 1990, warfarin was switched to aspirin and low molecular weight heparin as she wished to become pregnant. At 24 weeks she developed eclampsia and labor was induced. During the first postpartum week she experienced loin pain, associated with rapidly rising creatinine levels and features of a nephrotic syndrome. An abdominal ultrasound showed bilateral renal vein thrombosis with enlarged kidneys. She was treated with continuous ambulatory peritoneal dialysis (CAPD) for 4 months and her renal function is now recovering. Our patient is another example of the primary antiphospholipid syndrome.

A study of 100 high risk lupus pregnancies.

Certain subgroups of lupus patients and those with circulating antiphospholipid antibodies (aPL) in particular, suffer a high rate of fetal loss. Over the past 4 years, we have prospectively studied 100 pregnancies in patients with systemic lupus erythematosus (SLE) and primary antiphospholipid syndrome. In addition to conventional methods of monitoring SLE and fetal development, we have also used Doppler flow assessment of placental perfusion from the 14th wk of pregnancy onward. Patients with the antiphospholipid syndrome and previous history of thrombotic events were treated with daily heparin (10,000 IU) and low-dose aspirin (75 mg). Those without a history of thrombosis were treated with low-dose prednisolone, azathioprine, or hydroxychloroquine. Pregnancy loss was reduced from 81.3% in 101 previous pregnancies to 36.8% in 100 pregnancies managed by us. None of the patients who received hydroxychloroquine throughout the pregnancy presented fetal malformations. Careful management and close monitoring of the lupus pregnancy has substantially improved fetal outcome.

Early Doppler studies in lupus pregnancy.

The management of 56 pregnancies in 54 patients is presented, 52 with systemic lupus erythematosus and two patients with primary antiphospholipid syndrome. All underwent serial Doppler blood flow studies of the umbilical and uterine arteries from 14 weeks of gestation. Drug therapy was directed at disease activity and the fetus and mother monitored intensively to identify optimum time for delivery. End diastolic blood flow (EDF) studies were compared with anticardiolipin antibodies (aCL) and the lupus anticoagulant (LAC) to predict outcome of pregnancy in terms of mortality, Caesarian delivery, hypertension, and gestation. The absence of end-diastolic blood flow predicted Caesarian delivery more accurately. The presence of EDF and the absence of aCL was consistent with a normotensive pregnancy. The presence of LAC was the best of the three in predicting fetal demise.

Antiphospholipid syndrome: five year follow up.

Nineteen patients out of 250 subjects with antiphospholipid antibodies, who had initially presented to the lupus clinic at St Thomas's Hospital, London five or more years ago with a history of venous/arterial occlusions, were entered into the study. The patients were divided into two main groups: I those who remained well without any further thromboembolic complications (n = 10); II those who developed recurrent thrombotic events in the five year period (n = 9). The patients were followed up to determine the relation between the level or the isotype of the anticardiolipin antibodies, or both, to the recurrent thromboembolic events, and the effect of a variety of treatments (corticosteroids, immunosuppression, anticoagulation) in the prevention of further vascular occlusions. Lupus activity over the five year period varied considerably between the two groups--those in group I tending to be relatively inactive compared with those in group II. For some patients in group II thromboembolic events seemed to occur at the time of lupus activity. Antiphospholipid antibodies remained positive in all patients, the levels remaining fairly constant. Levels fell in only one patient in group I and in two in group II. Patients in group II had more systemic lupus erythematosus related disease than those in group I; most were receiving concomitant steroid and immunosuppressive therapy, but this did not seem to protect against the development of further occlusions. All patients were given anticoagulation treatment (warfarin/heparin) or salicylates (low dose aspirin 75 mg daily), or both. Patients with deep vein thromboses developed more complications during anticoagulation therapy than those with cerebrovascular symptoms. Problems in anticoagulation control and recurrent thromboses consequent on warfarin withdrawal despite the administration of subcutaneous heparin were responsible for complications in most patients in group II.

Antibodies to endothelial cells in systemic lupus erythematosus: a potential marker for nephritis and vasculitis.

Using an ELISA, anti-endothelial cell antibodies (AECA) have been found in sera obtained at the time of renal biopsy in 46 out of 57 patients (81%) with systemic lupus erythematosus (SLE) and nephritis (mean binding index (BI) = 84% +/- 52.8) compared with 22 out of 50 SLE patients (44%) without nephritis (mean BI = 45% +/- 35.9). Seventy normal human sera had a mean BI of 10% +/- 9.8. The highest levels were seen in patients with diffuse proliferative glomerulonephritis (WHO grade IV) and in patients with proteinuria and nephrotic syndrome. When the biopsies were assessed for activity and chronicity scores, AECA were associated with active renal lesions (P less than 0.001). AECA levels correlated with low complement levels but not with anti-DNA antibodies to extractable nuclear antigens (ENA), anti-cardiolipin or anti-neutrophil cytoplasmic antibodies. The presence of AECA conferred a positive predictive value of 0.68 for the presence of nephritis. Twenty-five patients had active vasculitis at the time of assay and the highest AECA values were seen in patients with both nephritis and vasculitis. No correlation was seen with serum immunoglobulin levels and immune complexes did not bind significantly to the endothelial surface. The possible role of these antibodies as a marker in lupus nephritis is discussed.

Antiendothelial cell antibodies detected by a cellular based ELISA in Kawasaki disease.

Kawasaki disease is an acute vasculitic illness of childhood associated with significant morbidity and mortality. A cellular based enzyme linked immunosorbent assay (ELISA) was used to demonstrate the presence of antiendothelial cell antibodies in sera from children with Kawasaki disease. Twenty one of 32 patients with Kawasaki disease had raised IgM antibody titres and four had raised IgG antiendothelial antibody titres. There was a significant difference in the IgM antiendothelial cell antibody titres when comparing the patients with normals and febrile controls. The antibody titre paralleled the disease activity in patients studied serially. There was no relative increase in binding of antiendothelial cell antibodies after cytokine stimulation. These findings may be of importance in further research into the understanding of mechanisms involved in this and other forms of vasculitis in man.

Myocardial infarction and antiphospholipid antibodies in SLE and related disorders.

The clinical and serological findings in 13 patients with myocardial infarction and antiphospholipid antibodies (the 'lupus anticoagulant', antibodies to cardiolipin, antibodies to phosphatidylethanolamine (one patient] seen by our unit and other units from 1984 to 1989, are presented (eight males and five females, ages ranging from 20 to 52 years). Five suffered myocardial infarction before the age of 30; four of these five were in their early 20s. Other risk factors such as excessive smoking (greater than 20 cigarettes a day) (two patients), long-term treatment with steroid (one) and use of oral contraceptives (one) were present. One patient had demonstrated a plasminogen activator deficiency and one a deficiency of protein C. Two patients developed myocardial infarction six to eight weeks after warfarin was discontinued for recurrent deep vein thrombosis. Six patients had SLE as defined by the revised 1982 criteria, three suffered from 'lupus-like' disease, while four patients conformed to a 'primary' antiphospholipid syndrome.

Ear, nose, and throat symptoms in subacute Wegener's granulomatosis.

The standard description of Wegener's granulomatosis emphasises renal failure and thus a distorted impression may be given. Subacute and even chronic cases occur, and in these patients the presentation is varied and often insidious, leading to delay in diagnosis. Twenty two such patients (13 women and nine men) with a mean age of 44 years were seen in our connective tissue disease clinic. The mean duration of symptoms before diagnosis was 3.6 years and the mean duration of disease 5.9 years (19 years in one patient). All patients had malaise and ear, nose, and throat symptoms, and most had joint pains. Impaired renal function was seen in seven patients only. Tissue biopsy was diagnostic in half of the patients, and appreciably high titres of antineutrophil cytoplasmic antibodies were detected in only nine of 18 patients in whom these were measured. The most useful investigations were neutrophil counts, chest radiographs, and computed tomography of the sinuses and orbits. The most effective treatment was with intravenous pulses of cyclophosphamide. No deaths occurred. At the time of writing two patients were in remission and no longer being treated and 18 patients were in partial remission on continued treatment. Patients with subacute forms of Wegener's granulomatosis present with a variety of clinical features and the insidious presentation often leads to delay in diagnosis. A history of ear, nose, and throat symptoms was universal in our patients.

Cerebrovascular disease and antiphospholipid antibodies in systemic lupus erythematosus, lupus-like disease, and the primary antiphospholipid syndrome.

PURPOSE AND PATIENTS AND METHODS: Antiphospholipid antibodies, lupus anticoagulant antibodies to cardiolipin, and a false-positive result on testing for syphilis have been linked to thrombotic vascular occlusions, particularly in patients with systemic lupus erythematosus (SLE) or lupus-like disease, i.e., patients not fulfilling four American Rheumatism Association criteria for the classification of SLE. The clinical and serologic features of 35 patients with cerebrovascular disease (strokes/transient ischemic attacks) who demonstrated antibodies to phospholipids are presented. Complete histories were obtained from all 35 patients, and all underwent routine physical examinations, radiography, electrocardiography, computed tomographic brain scanning, and immunologic studies. Psychometric tests were performed in nine patients. RESULTS: The strokes were often multiple and were followed by multi-infarct dementia in nine patients. Of particular interest were 10 patients in whom the presence of antiphospholipid antibodies was the major and often the sole immunologic disturbance present. Several of these patients were antinuclear antibody-negative, and the antinuclear antibodies, when present, were usually of a low titer (1:40 to 1:160). These patients conform to a group classified as having a primary antiphospholipid syndrome. CONCLUSION: Antiphospholipid antibodies are strongly associated with cerebrovascular occlusions in patients with SLE as well as in those with lupus-like disease and the primary antiphospholipid syndrome. All patients with any of these conditions who present with vascular events should be screened for these antibodies, as their occurrence may have a bearing on future therapy.

Anticardiolipin antibody levels in diabetic subjects with and without coronary artery disease.

Moderate (greater than 20 units) and high (greater than 80 units) IgG anticardiolipin antibody (aCL) titres are strongly predictive for recurrent thrombosis and early myocardial infarction in non-diabetic subjects. We have tested the hypothesis that the excess risk of myocardial infarction in diabetic subjects relates to the presence of aCL by measuring the frequency and titre of aCL in two groups of diabetic subjects and in 2500 healthy controls. One non-diabetic subject (0.04%) had low (5-20 units) IgG aCL titres. Seven out of 126 diabetics without cardiovascular disease (5.6%) and 9 out of 79 diabetics who were either myocardial infarction survivors or who had angiographically-proven coronary artery disease (11.4%) had low aCL titres (P less than 0.01 for comparison of either diabetic group with controls, and P less than 0.1 for comparison between diabetic groups). One subject in each diabetic group, but no non-diabetics, had moderate IgM aCL titres. No subjects had high aCL titres. Diabetics have an increased frequency of low aCL titres which may relate to macrovascular disease. Macrovascular disease in diabetics is not associated with moderate or high aCL titres.

Budd Chiari syndrome, visceral arterial occlusions, recurrent fetal loss and the "lupus anticoagulant" in systemic lupus erythematosus.

A 30-year-old Chinese woman had been diagnosed as having systemic lupus erythematosus (SLE) at age 25 after 4 spontaneous abortions, all between 8 and 12 weeks of gestation. A year later she developed mild jaundice and at laparotomy, nodular enlargement of the right lobe of the liver and splenomegaly were seen. She was found to have a "lupus anticoagulant" and false positive test for syphilis (VDRL), but persistently negative antibodies to cardiolipin. Angiography demonstrated occlusion and stenoses of visceral abdominal arteries and hepatic venography also showed evidence of hepatic venous occlusions. She subsequently developed thyrotoxicosis which was treated with carbimazole, followed by radioactive iodine. The SLE and lupus anticoagulant were treated with systemic steroids (prednisolone) and anticoagulation (warfarin).

Recurrent calcific periarthritis leading to erosive osteoarthritis.

Over a period of 31 years the patient described had recurrent attacks of acute calcific periarthritis at multiple sites. In the latter years of her follow-up she has developed an erosive polyarticular osteoarthritis at sites previously affected by calcific periarthritis and we speculate that there may be a pathogenetic link between these two conditions in some patients.

Diagnostic and therapeutic problems in two patients with antiphospholipid antibodies, heart valve lesions, and transient ischaemic attacks.

Two young women (aged 32 and 25 years) with systemic lupus erythematosus and heart valve lesions in association with antiphospholipid antibodies are presented. In addition to the presence of the 'lupus anticoagulant' and false positive Venereal Disease Research Laboratory (VDRL) tests, both patients had high levels of IgG anticardiolipin antibodies. The first patient additionally had contraceptive induced chorea, chorea gravidarum, seven miscarriages, livedo reticularis, pulmonary embolism, and thrombocytopenia and developed culture negative endocarditis as well as hypertension. The second patient, who had presented with hypertension, developed aortic and mitral regurgitation, suspected myocarditis, manifested transient ischaemic attacks, and responded well to anticoagulation and steroid treatment.