anti-NCS Antibodies Research Articles

Antinucleosomal Antibodies and Its Correlation to SLE Manifestations

Subject: SLE is an autoimmune disease with skin, joint, renal, cardiovascular, and nervous manifestations. The disease is classified as an immune complex-mediated disease and is characterized by the production of various autoantibodies. Until now, more than 100 autoantibodies have been identified. Patients and methods: This study was performed on 200 SLE patients. All of them were females and their ages ranged from 20 to 49 years. All participants in this study were subjected to physical examination, thorough history taking including age, sex, age of onset, duration of SLE disease, family history, and SLEDAI. Laboratory investigations included: CBC, ESR, liver function tests, renal function tests, ANA, Anti-dsDNA, CRP, Complements C3, C4, and Anti-NCS. Results: There was a highly significant inverse correlation between anti-NCS antibodies and Hb level, a significant direct correlation between anti-NCS antibodies and 24hr proteinuria, a significant inverse correlation between anti-NCS antibodies and complements (C3 and C4), a significant correlation between anti-NCS antibodies and anti dsDNA antibodies, a significant correlation between anti-NCS and disease activity, SLEDAI, and renal affection. Conclusion: Anti-NCS antibody can be a useful tool in the diagnosis of SLE especially in patients who are negative for anti-ds DNA antibodies.

Antinucleosome antibodies as new marker in early diagnosis of systemic lupus erythematosus (SLE) in comparison with other autoantibodies

Systemic lupus erythematosus (SLE) is an autoimmune disorder of the multisystem, a multifactorial disease with generalized persistent inflammation that can involve multiple tissues and organs. Nucleosomes are essential chromatin molecules, and are thought to be the main antigens in SLE pathogenesis. A case-control study was performed to show the relationship between antinucleosome antibodies and systemic lupus erythematosus in Najaf province. This study was carried out in the Department of Microbiology in the Faculty of Medicine- University of Kufa, for the period of (January 2018 to December 2018), subjects were included in present study, divided into three groups: SLE patient (30), RA patient (30), and Healthy controls(20). Anti-NCS antibodies, ANA and anti-dsDNA antibodies were done for all individuals those included in this study. The findings of the current study showed that, in sera of SLE patients the mean level of antibodies against the following parameters NCS, ANA and ds-DNA were high in comparison with the mean levels of these parameters in sera of the normal control group (p<0.001). As well as, the levels of antibodies against NCS significantly higher in sera of the SLE group than in the rheumatoid arthritis RA (diseased control group) (p<0.001). On the other hand, the mean levels of antibodies against NCS were statically insignificant (p value 0.356) among the healthy individuals and the diseased control group. Anti-NCS antibodies have an important role in SLE diagnosis in comparison to other autoantibodies, and may have a role to predict the disease outcome.

Antinucleosome antibodies in systemic lupus erythematosus patients: Relation to disease activity and lupus nephritis

Aim of the workTo evaluate the level of anti-nucleosome (anti-NCS) antibodies in systemic lupus erythematosus (SLE) patients and study their association with disease activity and lupus nephritis. Patients and methodsThe study was carried out on 50 SLE patients; 47 females and 3 males and 30 matched controls. The SLE disease activity index (SLEDAI) was assessed. Plasma levels of anti-NCS antibodies were measured. Renal biopsy was done in those with lupus nephritis. ResultsThe mean age of patients was 28.3 ± 12.9 years; disease duration was 5.5 ± 6.02 years and the SLEDAI.5 ± 7.8. LN was present in 60% of the patients. Anti-nuclear antibodies (ANA) were positive in 48 (96%) and the anti-double stranded deoxyribonucleic acid positive in 38 (76% of the patients. The anti-NCS antibodies level was significantly increased in the patients (470.1 ± 369.4 U/ml) compared to the control (36.43 ± 14.58 U/ml) (p = 0.0001). The anti-NCS was detected in 48/50(96%) patients at a cut-off of 65 U/ml. There was no significant difference in the anti-NCS antibodies levels between those with nephritis (456.53 ± 362.59 U/ml) and those without (490.4 ± 387.96 U/ml) (p = 0.84). A significant correlation was present between the anti-NCS antibodies and photosensitivity (p = 0.032), ESR (p = 0.03), complement (C3) consumption (p = 0.01) and urinary casts (p = 0.04). A non-significant correlation was detected between level of anti-NCS and grades of nephritis (p = 0.49) or SLEDAI (p = 0.09). ConclusionAnti-NSC antibodies could be a useful addition to the laboratory tests that can help in the diagnosis of SLE. Also, it has a significant association to photosensitivity but no relation to with disease activity or renal involvement.

Anti-nucleosome antibodies in systemic lupus erythematosus patients: Relation to anti-double stranded deoxyribonucleic acid and disease activity

Aim of the workTo measure the level of anti-nucleosome (anti-NCS) antibodies in systemic lupus erythematosus (SLE) patients and to evaluate their relation with anti-double stranded deoxyribonucleic acid (anti-dsDNA) antibodies and SLE disease activity. Patients and methods66 Egyptian SLE patients were investigated for the detection of anti-NCS antibodies and anti-dsDNA antibodies. Disease activity was assessed using the SLE disease activity index (SLEDAI) and the European consensus lupus activity measurement (ECLAM). ResultsThe median age of the patients was 25.5years (12–48years) and disease duration 3years (1month to 26years). anti-NCS antibody was found in 48 (72.7%) patients. Non-significant difference was found between both those positive or negative anti-NCS antibodies regarding the clinical features apart from fever (p=0.019). Lupus nephritis was present in 35/48(72.9%) of those with positive and in 11/18 (61.1%) of those with negative anti-NCS (p=0.35) A significant correlation was found between anti-NCS antibodies with SLEDAI (r=0.36, p=0.003) and ECLAM (r=0.29, p=0.019). No significant relation was found between anti-NCS antibodies and clinical features of SLE, apart from fatigue (r=0.3, p=0.015). A significant correlation with hypocomplementemia (C3 r=−0.37, p=0.002 and C4 r=−0.32, p=0.018) and anaemia (r=−0.32, p=0.009). anti-dsDNA antibodies were detected in 35(53%) SLE patients; 70.8% of those with positive and 5.6% of those with negative anti-NCS antibodies. ConclusionAnti-NCS antibodies could play a role in the pathogenesis of SLE and is related to disease activity. Its association with anti-dsDNA antibodies and its presence in those with negative anti-ds DNA may aid in the diagnosis of SLE.

Desempenho diagnóstico e associações clínicas dos anticorpos contra componentes da cromatina no lúpus eritematoso sistêmico juvenil

To determine the frequency of antibodies to chromatin components in juvenile systemic lupus erythematosus (JSLE), and to correlate the presence of these autoantibodies with clinical manifestations and disease activity. Anti-chromatin (anti-CHR), anti-nucleosome core particle (anti-NCS) and anti-dsDNA antibodies were measured in 175 individuals, including 37 patients with active JSLE and 41 with inactive disease, 47 non-lupus autoimmune disease patients (non-lupus AD), and 50 healthy children. An in-house ELISA was developed with purified nucleosome core particles from calf thymus to determine IgG and IgG3 anti-NCS antibodies. Anti-CHR and anti-dsDNA antibodies were detected by commercial ELISA kits (INOVA). Anti-NCS and anti-CHR antibodies exhibited high specificity for JSLE and similar frequency in active and inactive JSLE. Anti-CHR and IgG/IgG3 anti-NCS serum levels did not differ between active and inactive JSLE. SLEDAI correlated with anti-dsDNA antibodies but not with antibodies to other chromatin components. There was association of anti-dsDNA, anti-CHR and IgG/IgG3 anti-NCS antibodies with proteinuria and low C4 serum levels. Anti-NCS antibodies in the absence of anti-dsDNA were observed in 14% of the JSLE patients. Our data indicate that anti-NCS and anti-CHR antibodies are relevant diagnostic markers for JSLE and appear to be correlated with JSLE lupus nephritis activity. IgG3 isotype anti-NCS antibodies do not seem to be more relevant than IgG anti-NCS antibodies as markers of disease activity or active nephritis in JSLE.

Primary antiphospholipid syndrome evolving into systemic lupus erythematosus: may antinucleosome antibodies be predictive?

It was reported by several groups that patients diagnosed as primary antiphospholipid syndrome (PAPS) had developed a full-blown systemic lupus erythematosus (SLE) even after many years of follow-up. Little is known about clinical and/or serological factors that may help predict such evolution. Antinucleosome antibodies (anti-NCS) were described to appear in early stages of SLE, in particular before anti-dsDNA antibodies. The aim of the study is to evaluate the prevalence of anti-NCS in a large cohort of PAPS patients. IgG and IgM anti-NCS antibodies were detected using a home made assay with H1-stripped chromatin as antigen. Sera from 106 PAPS patients were tested; 52 of them were also tested during the follow-up, at least 2 years apart form the basal sample. Medium-high titre anti-NCS were found in nearly half of the patients (49/106, 46%), more frequently in those presenting features of "lupus like disease". Most of patients displayed an unchanged pattern of anti-NCS over time. We describe three cases of PAPS patients that developed SLE after many years of follow-up; high titre and low titre anti-NCS were present in two and one of them respectively several years before evolving into SLE. A significant proportion of PAPS patients displayed medium-high titre anti-NCS, suggesting that the autoimmune response against chromatin may be a relevant event not only in patients with SLE. Further studies are warranted to explore the predictive value of anti-NCS with respect to the evolution from PAPS to SLE.

Anti-nucleosome and anti-chromatin antibodies are present in active systemic lupus erythematosus but not in the cutaneous form of the disease

The objective of this study is to investigate the presence of anti-nucleosome (anti-NCS) and anti-chromatin (anti-CRT) antibodies in patients with cutaneous lupus erythematosus (CLE) compared with active and inactive systemic lupus erythematosus (SLE). A total of 154 subjects were evaluated: 54 patients presenting CLE, 66 patients with active SLE and 34 with inactive SLE. Lupus activity was assessed using the disease activity index (SLEDAI). Anti-NCS and anti-CRT antibodies were detected by enzyme-linked immunosorbent assay (ELISA). Only one of 54 patients with CLE tested positive for both anti-NCS and anti-CRT antibodies. The prevalence of anti-CRT antibodies was significantly higher in active SLE (84.8%) when compared with inactive SLE (26.4%) and CLE (1.8%) (P < 0.001). Anti-NCS antibodies were also more prevalent in active SLE patients (74.2%) than inactive SLE (11.7%) and CLE patients (1.8%) (P < 0.001). The presence of anti-CRT and anti-NCS antibodies was correlated to disease activity in patients with SLE (r = 0.4937, r = 0.5621, respectively). Furthermore, the detection of both antibodies was correlated with disease activity in patients with SLE who tested negative for anti-dsDNA antibodies (r = 0.4754 for anti-NCS and r = 0.4281 for anti-CRT). The presence of these two auto-antibodies was strongly associated with renal damage in patients with SLE (OR = 13.1, for anti-CRT antibodies and OR = 25.83, for anti-NCS antibodies). The anti-NCS and anti-CRT antibodies were not found in CLE. In patients with SLE, there is a correlation of these antibodies with disease activity and active nephritis. When compared with anti-dsDNA antibodies, anti-NCS and anti-CRT antibodies were more sensitive in detecting disease activity and kidney damage in lupus patients.

Association of antinucleosome antibodies with disease flare in serologically active clinically quiescent patients with systemic lupus erythematosus

To identify the prevalence of serologically active clinically quiescent (SACQ) patients in a cohort of 290 patients with systemic lupus erythematosus (SLE). We investigated if the presence of anti-double-stranded DNA (anti-dsDNA) or antinucleosome (anti-NCS) antibodies during the SACQ period was associated with future flares. SACQ patients defined as clinically inactive for 6 months (global British Isles Lupus Activity Group index [BILAG] scores <6) and serologically active (anti-dsDNA antibodies >50 units/ml on at least 2 occasions by enzyme-linked immunosorbent assay [ELISA]) were identified. Patient sera collected during the defined SACQ period were also tested for anti-NCS antibodies (ELISA). We retrospectively reviewed patient clinical details and episodes of flare using the BILAG activity index. Twenty-seven (9%) patients were SACQ. Seventeen (81%) patients experienced a flare (total of 91 flares, up to 12 flares per person) in the next 5 years. Median duration to first flare was 15 months (range 2-46). Time to first flare after SACQ period was significantly correlated with the presence of anti-NCS (P = 0.0012), high anti-NCS antibody titers (P = 0.0006), and anti-dsDNA titers 5 times above the normal limit (P = 0.02). Patients with higher absolute anti-NCS antibody titers showed a significant correlation with the number of flares (r = 0.57, P = 0.007). A minority of patients with SLE are SACQ. The majority of these patients experience a flare in the next 5 years and close followup is recommended. Anti-NCS antibodies may be a better predictor than anti-dsDNA antibodies for future flares.

Anti-nucleosome antibodies in patients with systemic lupus erythematosus of recent onset. Potential utility as a diagnostic tool and disease activity marker.

To compare the utility of anti-chromatin antibodies for the diagnosis of systemic lupus erythematosus (SLE) and as markers of disease activity. We included 73 consecutive patients (62 female) with SLE (four or more ACR criteria) of recent onset (<1 yr since diagnosis). As control groups we included 130 healthy blood donors and 261 patients with 11 systemic autoimmune diseases (SAD). Disease activity was assessed using the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). A venous blood sample was drawn to measure three anti-chromatin antibodies [anti-nucleosome (anti-NCS), anti-double-stranded DNA (anti-dsDNA) and anti-histones (anti-HST)] by enzyme-linked immunosorbent assay. The prevalence of anti-chromatin antibodies in SLE patients and healthy controls was 100 and 3% respectively for anti-NCS, 63 and 5% for anti-dsDNA, and 15 and 3% for anti-HST. Anti-NCS had a sensitivity of 100% and specificity of 97% for SLE diagnosis. When SLE and SAD patients were compared [excluding mixed connective tissue disease (MCTD)], the sensitivity of anti-NCS, anti-dsDNA and anti-HST antibodies for SLE diagnosis was 93, 71 and 40% respectively and the specificity was 97, 98 and 98%. Anti-chromatin antibodies were not useful in differentiating between SLE and MCTD patients. Anti-NCS antibodies showed the highest correlation with disease activity (r = 0.45, P < 0.0001), especially in patients negative for anti-dsDNA antibodies (r = 0.58, P = 0.001). Anti-NCS antibodies also showed strong association with renal damage (odds ratio 4.1, 95% confidence interval 1.2-13.6, P = 0.01). Anti-NCS antibodies could be a useful tool in the diagnosis and assessment of disease activity in SLE patients, especially in patients who are negative for anti-dsDNA antibodies.

Uptake and Localization of Neocarzinostatin in Malignant Glioma

The authors studied the localization of neocarzinostatin (NCS) in cultured cells and in tumor-bearing rats by means of immunofluorescent staining. Anti-NCS antibody was obtained through immunization of rabbits with NCS. Cellular uptake of NCS was dose-dependent (1.0 to 1000 micrograms/ml) in 9L rat gliosarcoma cells in monolayer. In monolayer cells of 9L, KMG-4 (derived from human glioblastoma), and KMS II (human ependymoma) treated with 1 mg/ml of NCS, drug uptake occurred within a few seconds. Accumulation was much higher in the cytoplasm than in the nucleus and, although nuclear uptake increased slightly over time, there appeared to be no increase in total cellular uptake. Mitotic cells, which were spherical in culture, showed greater intracellular accumulation than other cells. There was no significant difference in uptake among non-mitotic cells. Cells surviving 20 hours of treatment retained accumulation as high as that in killed cells. In KMG-4 monolayers, cytoplasmic and nuclear NCS distribution still differed, whereas 9L monolayers exhibited more even intracellular distribution. In 9L spheroid models treated with 1 mg/ml of NCS, the drug permeated almost all layers within 10 minutes, and at 120 minutes had heavily accumulated in the central necrotic areas. In rats with transplanted brain tumors, NCS selectively accumulated in neoplastic tissues following intra-arterial administration. However, NCS uptake by arterial endothelium was also seen, which suggests the potential for vascular toxicity. The therapeutic value of NCS is discussed in terms of its pharmacokinetic characteristics.

Pharmacokinetics of Neocarzinostatin in Patients with Malignant Glioma

Because of technical difficulties, the pharmacokinetics of neocarzinostatin (NCS) have not been thoroughly evaluated in patients with malignant glioma. The authors produced anti-NCS antibody by immunizing rabbits with NCS and established a means of quantifying tissue levels of NCS with enzyme-linked immunosorbent assay. In one patient given a bolus injection of 1 mg of NCS intra-arterially, the concentration of drug in neoplastic tissue at 25 minutes (0.1136 micrograms/g) was higher than that in blood at 20 minutes and was retained for a longer period. Rapid entry of NCS into the tumor cavity was observed at 5 minutes. In two postoperative cases, NCS applied topically to the tumor site (50 and 100 micrograms) was retained at high levels (0.2941 and 3.33 micrograms/ml) even after 48 hours, although no NCS was detected in blood after 60 minutes. NCS concentrations as low as 1 microgram/ml demonstrated cytocidal effects, and a delay in tumor growth was observed even at an NCS level of 0.1 microgram/ml, despite the fact that the half-life of NCS is extremely short (3 seconds in serum). Because its cytotoxic effect seems to be very rapid, it appears more important to obtain a high initial NCS concentration than to maintain a constant blood level.

Enzyme immunoassay with high sensitivity and accuracy for specific antibody to neocarzinostatin

A quantitative enzyme immunoassay (EIA) for specific antibody to neocarzinostatin (NCS) is described which uses enzyme-labeled anti-rabbit IgG antibody, solid-phase NCS and standard purified specific antibody to NCS. The dose of the standard was determined by sandwich EIA for rabbit IgG. The lower detection limit was 3 ng of the specific antibody per tube. The accuracy of the assay was excellent and a comparative study with the sandwich EIA for rabbit IgG showed good correlation. The antiserum to NCS of the highest titer was found to contain 0.6 mg and 40 mg per ml of specific antibody to NCS and of normal IgG, respectively. The accuracy of the assay results and the purity of the standard was established by 2 recovery tests for anti-NCS antibody.

Production of tumor antibody-neocarzinostatin(NCS) conjugate and its biological activities

Rabbit xenoantiserum was produced against a human leukemia cell line (NALL-1) derived from a patient with acute lymphoblastic leukemia, and IgG was purified. Anti-NALL-1 rabbit IgG was reacted with NCS, an unique membrane-reactive anticancer antibiotic, in the presence of water-soluble carbodiimide. The resulting mixture was concentrated and chromatographed on a Sephadex G-200 column. The first and second fractions were shown by immunoelectrophoresis and the Ouchterlony double-diffusion method to contain NCS-IgG but not free NCS. The conjugates inhibited the growth of Sarcina lutea, and the growth and 3H-TdR incorporation of NALL-1 cells. A membrane immunofluorescent test with FITC-labeled rabbit anti-NCS and goat anti-rabbit IgG antibodies demonstrated specific localization of NCS-IgG on NALL-1 cell surfaces. These results indicate that IgG-bound NCS retained both NCS and antibody activities, and thus should be useful for cancer therapy.