The clinical relevance of serum antinuclear antibodies in Japanese patients with systemic sclerosis.

BACKGROUND: Systemic sclerosis (SSc) is a connective tissue disorder with excessive fibrosis of the skin and various internal organs. Although SSc is a heterogeneous disease, it has been reported that the particular antinuclear antibodies (ANA) are often indicative of clinical features, disease course and overall severity. OBJECTIVE: To clarify the association of clinical and prognostic features with serum ANA in Japanese patients with SSc. METHODS: We studied 203 Japanese patients diagnosed with SSc, who visited our hospital during the period 1983-2005. Six SSc-related ANA were identified using indirect immunofluorescence, double immunodiffusion and immunoprecipitation assays. RESULTS: Patients with SSc were classified into six ANA-based subgroups and a group without ANA. As expected, antitopoisomerase I antibody (Ab, n=64), anti-RNA polymerases (RNAP) Ab (n=12) and anti-U3 RNP Ab (n=5) were associated with diffuse cutaneous SSc, whereas anticentromere Ab (ACA, n=75), anti-Th/To Ab (n=7) and anti-U1 RNP Ab (n=10) were frequently detected in patients with limited cutaneous SSc. Clinical features of the ANA-negative group (n=10) were heterogeneous. Consistent with previous findings in Caucasian and/or black African patients, antitopoisomerase I Ab was associated with the involvement of vascular and pulmonary fibrosis, leading to decreased survival rate. However, no patients with anti-RNAP Ab developed renal crisis and the frequency of isolated pulmonary hypertension in patients with ACA, anti-Th/To Ab or anti-U3 RNP Ab was similar to that in other ANA-based subgroups. CONCLUSION: These results indicate that the clinical relevance of SSc-related ANA in Japanese patients differs in some aspects from that in Caucasian and/or black African patients.

Anti-phosphatidylserine-prothrombin complex antibodies in patients with localized scleroderma.

OBJECTIVE: Although some antiphospholipid antibodies (Abs) are found in patients with localized scleroderma (LSc), Ab against phosphatidylserine-prothrombin complex (PS/PT) has not been examined. We investigated anti-PS/PT Ab levels in patients with LSc. METHODS: IgG anti-PS/PT Ab levels in serum samples taken from patients with LSc (n = 42) were measured using ELISA. RESULTS: IgG anti-PS/PT Ab was detected in 17% of the LSc patients, while it was not detected in any normal controls (n = 32) or psoriasis vulgaris (n = 25), and this frequency was similar to that of systemic sclerosis (17%, n = 41). Among 3 LSc subgroups, generalized morphea, the severest form of LSc, had a frequency (27%) comparable with that of systemic lupus erythematosus (32%, n = 25). Among 7 LSc patients with anti-PS/PT Ab, 2 developed symptomatic thromboembolism (A 70-year-old man developed deep vein thrombosis and pulmonary infarction, although he was negative for other antiphospholipid Abs. A 6-year-old boy positive for lupus anticoagulant had cerebral infarction). By contrast, symptomatic thromboembolism was not detected in 35 LSc patients without anti-PS/PT Ab. CONCLUSION: Patients with LSc, especially generalized morphea, exhibit anti-PS/PT Ab at a frequency comparable with collagen diseases such as systemic sclerosis and systemic lupus erythematosis. Examination of this Ab may be useful to recognize the risk of thromboembolism in patients with LSc.

Autoantibody against matrix metalloproteinase-3 in patients with systemic sclerosis.

Systemic sclerosis (SSc) is characterized by multi-organ fibrosis with an autoimmune background. Although autoantibodies are detected frequently in SSc patients, the role of autoantibody in the development of fibrosis remains unknown. Connective tissue homeostasis is a balance between the synthesis and degradation of the extracellular matrix (ECM); ECM degradation is regulated mainly by matrix metalloproteinases (MMPs). Anti-MMP-1 antibody is suggested to inhibit MMP-1 and be involved in the development of the fibrosis in SSc. However, the accumulation of various ECM components in the tissue of SSc cannot be explained by the anti-MMP-1 antibody alone. In this study, we examined the presence or levels of antibody to MMP-3, a protein which degrades various ECM components relevant to SSc fibrosis. Enzyme-linked immunosorbent assay (ELISA) using human recombinant MMP-3 revealed that IgG anti-MMP-3 autoantibody levels were elevated significantly in the sera from SSc patients, but not in patients with active systemic lupus erythematosus or dermatomyositis. IgG and IgM anti-MMP-3 antibody levels were significantly higher in diffuse cutaneous SSc, a severe form, than those in limited cutaneous SSc. Consistently, IgG anti-MMP-3 antibody levels correlated significantly with fibrosis of the skin, lung and renal blood vessels. The presence of IgG anti-MMP-3 autoantibody in sera from SSc patients was confirmed by immunoblotting analysis. Remarkably, MMP-3 activity was inhibited by IgG anti-MMP-3 antibody. These results suggest that anti-MMP-3 antibody is a serological marker that reflects the severity of SSc and also suggest that it may contribute to the development of fibrosis by inhibiting MMP-3 activity and reducing the ECM turnover.

Anti-agalactosyl IgG antibodies in sera from patients with systemic sclerosis.

OBJECTIVE: To determine the prevalence and clinical correlations of anti-agalactosyl IgG antibodies (anti-AG IgG) in patients with systemic sclerosis (SSc). METHODS: Serum samples from patients with limited cutaneous SSc (lSSc; n = 49), diffuse cutaneous SSc (dSSc; n = 21), rheumatoid arthritis (RA; n = 10), systemic lupus erythematosus (SLE; n = 20), and healthy individuals (n = 20) were examined by lectin-enzyme immunoassay using human agalactosyl IgG as antigen. RESULTS: Anti-AG IgG were detected in 52 (74%) of 70 patients with SSc, which was much higher than the frequency of rheumatoid factor positivity in SSc (16%). Levels of anti-agalactosyl IgG antibodies were significantly higher than in healthy controls or patients with SLE, but lower than patients with RA. Levels of anti-AG IgG in patients with dSSc were significantly higher than in lSSc. SSc patients with anti-topoisomerase I antibodies had significantly higher levels of anti-AG IgG than SSc patients with anticentromere antibodies. Concerning clinical correlation, patients with pulmonary fibrosis showed elevated levels of anti-AG IgG compared to those without pulmonary fibrosis. Patients with decreased %VC or %DLCO showed increased levels of anti-AG IgG. Elevated levels of anti-AG IgG were associated with the presence of contracture of phalanges or cutaneous calcinosis, but not the presence of arthritis/arthralgia. CONCLUSION: The results suggest that anti-agalactosyl IgG antibody is frequently detected in SSc and is a serological indicator for more severe SSc.

Delayed functional recovery by vincristine after sciatic nerve crush injury: a mouse model of vincristine neurotoxicity.

Neuropathy is the dose-limiting side effect of vincristine (VCR) in cancer therapy. However, no simple experimental model has yet been reported. Here, we present a simple experimental model of VCR neurotoxicity using a mouse sciatic nerve crush model, which allows evaluation within a few weeks. VCR administered intravenously once on the day after the crush lesion caused a dose-dependent delay of the recovery of motor and sensory functions. The minimal dose required to cause the delay was 0.25 mg/kg, which corresponded to five times the usual clinical dose for man and was far less than the reported doses required to cause functional impairment in intact animals. The model would be useful not only for the development of new drugs but also for the estimation of the drug interactions in combination cancer therapy.

Renal vascular damage in Japanese patients with systemic sclerosis.

OBJECTIVE: To evaluate renal vascular damage in Japanese patients with systemic sclerosis (SSc) by colour-flow Doppler ultrasonography. METHODS: The pulsatility index (PI) was measured in renal interlobar and segmental arteries by colour-flow Doppler ultrasonography. RESULTS: PI values of interlobar arteries were increased in SSc patients (n=53) with normal renal function compared with healthy persons (n=16), systemic lupus erythematosus patients (n=12) and dermatomyositis patients (n=3). SSc patients with elevated PI levels had digital pitting scar, short sublingual frenulum, contracture of phalanges, pulmonary fibrosis, decreased per cent vital capacity, heart involvement, positivity for anti-topoisomerase I antibody, and elevated C-reactive protein more frequently than those with normal PI levels. CONCLUSION: Although renal crisis is rare in Japanese SSc patients, our study suggests that latent and subclinical renal damage exists in these patients.

Survival activity of troglitazone in rat motoneurones.

Troglitazone (TGZ), an antidiabetic drug that improves insulin-resistance in the peripheral tissues, was tested for neurotrophic activity in motoneurones and other neurones in culture. In rat motoneurones, TGZ had a remarkable effect on survival, which was comparable or superior to that of brain-derived neurotrophic factor, a known potent neurotrophic factor for rat motoneurones. However, TGZ did not promote the survival of sensory, sympathetic, septal or hippocampal neurones. The effect of TGZ on motoneurones was additive to that of insulin-like growth factor-I and both activities were inhibited by phosphatidylinositol 3-kinase (PI3-kinase) inhibitors, wortmannin and LY294002, suggesting the involvement of the activation of PI3-kinase in the activity of TGZ. Pioglitazone, another antidiabetic drug structurally similar to TGZ, did not show any activity, indicating that the agonistic activity of TGZ for peroxisome proliferator-activated receptor-gamma is not involved in the survival activity. Chromanol, an antioxidant moiety of TGZ, showed little or no survival activity. These results indicate specific neurotrophic activity of TGZ for motoneurones through the activation of PI3-kinase and support the applicability of TGZ for the treatment of motor neurone diseases such as amyotrophic lateral sclerosis.

Diastolic dysfunction in patients with systemic sclerosis detected by gated myocardial perfusion SPECT: an early sign of cardiac involvement.

UNLABELLED: Diagnosis of cardiac involvement is important for the management of patients with systemic sclerosis (SSc). This study was undertaken to determine the significance of gated myocardial perfusion SPECT in patients with SSc and whether diastolic function measured by gated SPECT is an early sign of cardiac complications. METHODS: Thirty-four patients with SSc and 16 control patients were studied using exercise nongated and resting gated myocardial perfusion SPECT. The SSc was classified by the modified Rodnan total skin score (TSS) into high-TSS (score > or = 10; n = 18) and low-TSS (score < 10; n = 16) groups. Gated SPECT was performed using 99mTc-methoxyisobutylisonitrile with 16 frames per cardiac cycle and quantitatively analyzed by QGS software and Fourier filtering of the volume curve. The parameters of ejection fraction (EF), peak filling rate (PFR), one-third mean filling rate, and time to PFR (TPFR) were calculated. RESULTS: A slight perfusion abnormality was observed in four and five patients in the low-TSS and high-TSS groups, respectively (not statistically significant). A decreased resting EF less than 55% was found in no and two patients in the low-TSS and high-TSS groups, respectively. TPFR was 166 +/- 22, 168 +/- 38, and 216 +/- 82 ms (P = 0.05, high-TSS group versus low-TSS group; P = 0.04, control group versus high-TSS group) and TPFR/R-R interval was 0.18 +/- 0.02, 0.19 +/- 0.04, and 0.26 +/- 0.09 (P = 0.01, high-TSS group versus low-TSS group; P = 0.005, control group versus high-TSS group) for the control, low-TSS, and high-TSS groups, respectively. CONCLUSION: Diastolic function can be evaluated by gated myocardial perfusion SPECT. Significant diastolic abnormalities were shown even in patients with normal perfusion and systolic function and were related to the severity of SSc.

Levels of interleukin 12, a cytokine of type 1 helper T cells, are elevated in sera from patients with systemic sclerosis.

OBJECTIVE: To determine serum levels and spontaneous production by peripheral blood mononuclear cells (PBMC) of interleukin 12 (IL-12), a potent inducer of type 1 helper (Th1) T cells, in patients with systemic sclerosis (SSc). METHODS: Serum IL-12 levels and spontaneous production levels of IL-12 in culture supernatants from PBMC were examined by ELISA. Serum levels of IL-4, IL-6. IL-10, and IL-13 and production levels by PBMC of IL-6 and IL-10 were also examined by ELISA. Renal vascular damage was determined as a pulsatility index (PI) by color flow Doppler ultrasonography of kidneys. RESULTS: Serum IL-12 levels were significantly elevated in patients with SSc (n = 62) compared with healthy controls (n = 20). Similarly, spontaneous production levels of IL-12 by PBMC in patients with SSc (n = 47) were higher than those in controls (n = 20). Serum IL-12 levels did not correlate with serum levels of any Th2-type cytokines such as IL-4, IL-6, IL-10, and IL-13. However, spontaneous production levels of IL-10 by PBMC significantly correlated with serum IL-12 levels in patients. Patients with elevated serum IL-12 levels had the increased PI values more frequently than those with normal IL-12 levels. Further, serum levels of IL-12 and production levels of IL-12 by PBMC correlated significantly with the PI values in patients with SSc. CONCLUSION: These results suggest that the increased levels of IL-12 may relate to the activation of Th1 cells in SSc and that IL- 12 overproduction may be associated with renal vascular damage.

Elevated serum KL-6 levels in patients with systemic sclerosis: association with the severity of pulmonary fibrosis.

BACKGROUND: Serum KL-6 has been suggested to be a useful marker for the evaluation of interstitial lung disease activity. OBJECTIVE: To determine the correlation between serum KL-6 levels and pulmonary fibrosis in patients with systemic sclerosis (SSc). METHODS: Serum samples from patients with limited cutaneous SSc (lSSc; n = 19), diffuse cutaneous SSc (dSSc; n = 26) and normal individuals (n = 15) were examined by ELISA. RESULTS: Serum KL-6 levels in SSc patients were significantly higher than those in normal controls. KL-6 levels in dSSc patients were significantly elevated compared with those in lSSc patients. Elevated KL-6 levels were associated with the presence of pulmonary fibrosis in SSc patients or dSSc patients. Furthermore, KL-6 levels inversely correlated with percentages of diffusion capacity of carbon monoxide and vital capacity in SSc patients or dSSc patients. CONCLUSION: KL-6 may be a simple, serologic indicator for the severity of pulmonary fibrosis in SSc.

Urticarial vasculitis in systemic lupus erythematosus: fair response to prednisolone/dapsone and persistent hypocomplementemia.

Two cases of urticarial vasculitis (UV) accompanying systemic lupus erythematosus (SLE) are reported. Both patients developed characteristic wheal and purpuric lesions of UV followed by pigmentation, and histological examination revealed leucocytoclastic vasculitis. Although oral prednisolone was beneficial for the systemic symptoms and various serological abnormalities, one patient needed dapsone and the other needed dapsone and cyclophosphamide to control the UV. In both patients, hypocomplementemia with no evidence of congenital complement deficiency or complement consumption persisted even after all other laboratory data and symptoms improved.

Biosynthesis and accumulation of microgram quantities of chlorophyll by developing chloroplasts in vitro.

Developing chloroplasts were incubated under conditions previously shown to induce protochlorophyll and chlorophyll biosynthesis, as well as chloroplast maintenance and partial differentiation in vitro. In the presence of air, delta-aminolevulinic acid, coenzyme A, glutathione, potassium phosphate, methyl alcohol, magnesium, nicotinamide adenine dinucleotide, and adenosine triphosphate, microgram quantities of chlorophyll accumulated after 1 hour of incubation. Part of the chlorophyll was not extractable in organic solvents; it is referred to as bound chlorophyll. The amount of bound chlorophyll depended on the degree of cotyledon greening at the time of plastid isolation. Etioplasts with or without a lag phase of chlorophyll biosynthesis synthesized nonphototransformable protochlorophyll and smaller amounts of extractable chlorophyll. As the greening of excised cotyledons progressed, more of the chlorophyll became bound before and after in vitro incubation. It is suggested that this increase in the fraction of bound chlorophyll reflects the biosynthesis of membrane-bound chlorophyll receptor sites. In the absence of cofactors, chlorophyll biosynthesis was blocked and porphyrins accumulated, indicating damage of the chlorophyll biosynthetic chain. It is concluded that chlorophyll accumulation constitutes a potentially convenient tool for the study of thylakoid membrane biogenesis in vitro.