ABSTRACT
Context: The diagnosis of giant cell tumor of bone (GCTB) is difficult in small biopsies with unusual age of presentation, location, and extensive secondary changes. Most of the GCTBs harbor H3F3A G34W mutations with a subset of cases showing alternate G34V, G34R, and G34L mutations. Objectives: To analyze the expression of anti-histone H3.3G34W antibody in different cellular components of GCTB across different locations and presentations (including the unusual ones) and validate the utility of this antibody in the diagnosis of GCTB and differentiate it from the other osteoclast-like giant-cell-rich lesions. Design: Immunohistochemistry was performed using anti-histone H3.3G34W antibody in the diagnosed cases of GCTB (136 cases of GCTB from 133 patients, including two malignant GCTBs) and other giant cell-containing lesions (62 cases). The presence of unequivocal crisp nuclear staining was considered positive. Results: Immunohistochemistry revealed unequivocal nuclear positivity in the mononuclear cells in 87.3% of the cases of GCTB. Of these, most showed diffuse expression with moderate to strong intensity staining. The positive staining was restricted to the nuclei of mononuclear cells with the nuclei of osteoclastic giant cells being distinctly negative. In addition to conventional GCTBs, two cases each of multicentric and malignant GCTB showed positive staining. The other giant-cell containing lesions were distinctly negative. The present study showed a sensitivity of 87.3% with specificity and positive predictive value of 100%. Conclusion: The anti-histone G34W antibody is a highly sensitive and specific marker for the diagnosis of GCTB and differentiating it from its mimics. The positive staining is restricted to the mononuclear cell component of GCTB with sparing the osteoclastic giant cells further reiterating the fact that the mononuclear stromal cells are the true neoplastic component of GCTB.
ABSTRACT
Introducción: La prueba de anticuerpos antinucleares es una poderosa herramienta en el diagnóstico de las enfermedades reumáticas. Los anticuerpos antinucleares se determinan en el laboratorio por un algoritmo o secuencia que se inicia con prueba de cribado y sigue con la identificación de las especificidades antinucleares más comunes. Pero, ¿cómo interpretar los resultados discordantes entre los dos niveles de estudio de anticuerpos antinucleares? Objetivo: Determinar las especificidades antinucleares menos frecuentes en pacientes positivos de cribado de ANA y negativos de las especificidades más comunes. Métodos: Estudio prospectivo de 88 pacientes consecutivos remitidos para la detección rutinaria de ANA con resultado positivo de cribado por ensayo inmuno-adsorbente ligado a enzima (ELISA) pero negativo de anticuerpos anti-ADN de doble cadena (dc, IgG) y anti-antígenos nucleares extraíbles comunes (ENAc). Las muestras séricas correspondientes fueron evaluadas por inmunofluorescencia indirecta sobre células de carcinoma epidermoide laríngeo humano (IFI-HEp-2) y por ELISA para la detección individual de ANA específicos. Resultados: La prueba de ANA por IFI/HEp-2 resultó positiva en 56/88 (63,6 por ciento) y las especificidades antinucleares se detectaron en 57/88 (64,8 por ciento) muestras, en el orden decreciente de Anti-Nucs: 16/88 (18,2 por ciento); anti-centrómero (CENP-B): 15/88 (17,0 por ciento); -histona: 15/88 (17 por ciento); -PM/Scl: 13/88 (14,8 por ciento); -ADNsc: 11/88 (12,5 por ciento) y -ENAc individuales: 8/88 (9,1 por ciento). La sensibilidad de la IFI-HEp-2 para las especificidades antinucleares fue de 0,83 (IC95 por ciento: 0,72-0,93). De los pacientes negativos de subserología (26/31), 83,9 por ciento no tenían antecedentes de enfermedad reumática asociada a ANA. Conclusiones: La mayoría de los pacientes con resultados discordantes entre el primer y segundo nivel de ANA fueron positivos de especificidades antinucleares menos comunes, pero de reconocido valor diagnóstico(AU)
Introduction: The antinuclear antibody test is a powerful tool for diagnosing rheumatic diseases. Antinuclear antibodies are determined in the laboratory by an algorithm or sequence that starts with a screening test and continues with the identification of the most common antinuclear specificities. But how to interpret the discordant results between the two levels of study of antinuclear antibodies? Objective: To determine the less frequent antinuclear specificities in positive patients of ANA screening and negative of the most common specificities. Methods: A prospective study was done on 88 consecutive patients referred for the routine ANA screening with a positive result of screening by enzyme-linked immunosorbent assay (ELISA) but negative for anti-double-stranded DNA (dc, IgG) and common extractable anti-nuclear antigens (ENAc). The corresponding serum samples were evaluated by indirect immunofluorescence on human laryngeal epidermoid carcinoma cells (IFI-HEp-2) and by ELISA for the individual detection of specific ANA. Results: The ANA test by IFI / HEp-2 was positive in 56/88 (63.6 percent) and the antinuclear specificities were detected in 57/88 (64.8 percent) samples, in decreasing Anti-Nucs order: 16/88 (18.2 percent); anti-centromere (CENP-B): 15/88 (17.0 percent); -histona: 15/88 (17 percent); -PM / Scl: 13/88 (14.8 percent); -ADNsc: 11/88 (12.5 percent) and -ENAc individual: 8/88 (9.1 percent). The sensitivity of IFI-HEp-2 for antinuclear specificities was 0.83 (95 percent CI: 0.72-0.93). No history of rheumatic disease associated with ANA was read in (26/31) 83.9 percent patients with negative subserology. Conclusions: The majority of patients with discordant results between the first and second level of ANA were positive of less common antinuclear specificities, but of recognized diagnostic value(AU)
Subject(s)
Humans , Algorithms , Mass Screening , Antibodies, Antinuclear , Rheumatic Diseases/diagnosis , Prospective StudiesABSTRACT
Objective To investigate the injury effects and mechanisms of circulating histones on the hepatocytes in patients with hepatitis B virus (HBV )-related liver failure (HBV-LF) .Methods Serum samples from patients with HBV-LF were collected . The levels of serum histone H3 , histone H4 , prothrombin activity (PTA ) ,total bilirubin (TBil) ,creatinine (Cr) and international normalized ratio (INR) of the patients were measured .Model for end-stage liver disease (MELD) score was calculated in the patients .The serum levels of histones were compared between patients and the healthy volunteers . The correlation of histones with the MELD score and PTA was analyzed .The human liver cell line L-02 cells were cultured and treated with the serum of patients or L-02 cellular lysate supernatant preincubated with or without anti-histone H3 and H4 antibodies .The cellular morphology and rate of apoptosis were observed .Intracellular calcium ion concentration and Caspase-3 activity were detected in the cultured L-02 cells treated with histones .Mean of two independent samples was compared using t tests .Relationship between histones and the MELD score or PTA was conducted using Spearman correlation analysis .Results The levels of serum histones in the patients with HBV-LF were much higher than those in the healthy volunteers (H3 :[5 390 .3 ± 1 032 .0] μg/mL vs [42 .7 ± 12 .8] μg/mL , t = 32 .76 , P < 0 .01 ; H4 :[4 205 .1 ± 662 .3] μg/mL vs [40 .3 ± 14 .6] μg/mL ,t = 39 .74 , P< 0 .01) .In addition ,serum histones (H3/H4) levels in patients were negatively correlated with serum PTA (r= - 0 .325 ,P= 0 .038 and r =- 0 .572 ,P= 0 .028 ,respectively) ,but positively correlated with the MELD score (r= 0 .359 ,P= 0 .021 and r = 0 .568 , P = 0 .007 , respectively ) . Both serum of patients with HBV-LF and L-02 lysate supernatant were toxic to cultured L-02 cells .The injury effect was inhibited by anti-histone antibodies ([9 .3 ± 1 .5]% vs [14 .3 ± 0 .6]% , t = 4 .259 , P= 0 .02) .L-02 cells treated with calf thymus histone were cultured for 4 h . Cellular toxicity of histones resulted in Caspase-3 activation . The effect was inhibited by anti-histone antibodies ([3 .5 ± 0 .5]% vs [5 .2 ± 0 .6]% ,t= 4 .243 ,P= 0 .02) .Conclusion The elevated circulating histones in the patients with HBV-LF may aggravate the liver damage .
ABSTRACT
Background: Detection of anti-nucleosome antibodies (anti-nuc) in patients with systemic lupus erythematosus (SLE) has been well established and it is claimed that their presence is associated with disease activity. Aims: The aim of this study is to evaluate the incidence of anti-nuc antibodies and to correlate them with disease activity and its association with other autoantibodies like anti-nuclear antibodies (ANA), anti-double stranded DNA (anti-dsDNA), anti-histone antibodies (AHA), as well as autoantibodies to histone subfractions like H1, (H2A-H4) complex, H2B, and H3. Methods: This cross-sectional study included 100 SLE patients referred from the Rheumatology, Dermatology, and Nephrology Departments. SLE disease activity was evaluated by using SLE-Disease Activity Index (SLEDAI) score. A patient was defined as having active SLE when the SLEDAI score was more than 5.0. Fifty normal controls were also tested as a healthy control group. Anti-nuc antibodies, anti-dsDNA, and AHA were tested by Enzyme-Linked Immunosorbent Assay (ELISA) and ANA was detected by an indirect immunofluorescence test. Results: All patients studied were in an active stage of disease and were untreated, of which 44 patients had renal biopsy-proven kidney involvement, which was categorized as lupus nephritis (LN) and 56 patients did not show any renal manifestations (SLE without LN). Anti-nuc antibodies were positive in 88%, anti-dsDNA in 80%, and AHA in 38% of the cases. ANA was positive in all SLE patients studied. None of the normal controls was found to be positive for these antibodies. Although a slightly higher incidence of autoantibodies were noted in LN, there was no statistical difference noted between LN and SLE without LN groups for anti-nuc and anti-dsDNA antibodies (p > 0.05). A higher incidence of autoantibodies to ANA specificities were noted in anti-nuc positive cases, but there was no statistical difference between anti-nuc positive and anti-nuc negative cases for ANA specificities among LN and SLE without nephritis groups (p > 0.05). Conclusions: Anti-nuc antibody detection could be a better tool for the diagnosis of SLE. Although there was no significant difference in LN and SLE without LN groups, this study suggests that anti-nuc detection can be useful as an additional disease activity marker to other laboratory tests.
ABSTRACT
Objective: To explore the influence of estradiol benzoate in induction of BALB/c mouse systemic lupus erythematosus model. Methods: Gonadectomied BALB/c mice were immunized with homologous spleen cell activated by concanavalin A three times,and at the same time injected with different doses estradial benzoate. Serum antinuclear antibody, antihistone antibody and estradiol in serum and kidney tissue homogenate were detected by enzyme linked immunosorbent assay. Kidney lesions were detected by hematoxylin-eosine(HE) and periodic acid-Schiff(PAS) staining, and by transmission electron microscope for detecting electron-dense agent as well as by direct fluorometric method for detecting immunoglobulin G antigen-antibody complex. Results: BALB/c mice immunized with homologous spleen cells activated by concanavalin A all developed systemic lupus erythematosus, but a statistically significant variation was observed in both autoantibody production and kidney damage were observed with a statistically significant variatron in the groups receiving different dosage of estradiol benzoate group(P
ABSTRACT
Objective:To study the relationship between the dose of active DNA and the induction of SLE-like syndrome.Methods: DNA from extracted ConA-activated spleen lymphocytes and immunized syngenic mice with different quantities of active DNA, the anti-dsDNA antibodies and anti-histone antibodies as well as the antibody subclass were detected by ELISA.The patterns of antinuclear antibodies and immune complexes in glomeruli were observed by immunofluorescent-stain. Results: 10 pig active DNA could induce all the animal to produce anti-dsDNA and anti-histone antibodies,and the induced autoann'bodies were mainly IgGl type.Only 25%animals produced autoantibodies immunized by 5 fjig active DNA. Conclusion:The minimum dose of active DNA to induce SLE-like syndrome was 10