IMMUNOLOGY OF DNA. III. <i>CRITHIDIA LUCILIAE</i>, A SIMPLE SUBSTRATE FOR THE DETERMINATION OF ANTI‐dsDNA WITH THE IMMUNOFLUORESCENCE TECHNIQUE*

La, Aarden; Groot, Els R. de; Feltkamp, T. E. W.

doi:10.1111/j.1749-6632.1975.tb29197.x

Cited by 534 publications

(168 citation statements)

References 19 publications

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“…Relevant control antibodies were included in each ELISA for intraassay validation of results (22,29). The Crithidia luciliae immunofluorescence test (CLIFT) was performed to detect anti-dsDNA antibodies, as previously described (31).…”

Section: Methodsmentioning

confidence: 99%

Critical comparative analyses of anti–α‐actinin and glomerulus‐bound antibodies in human and murine lupus nephritis

Kalaaji¹,

Sturfelt²,

Mjelle³

et al. 2006

Arthritis & Rheumatism

102

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Objective. Although anti-double-stranded DNA (anti-dsDNA) antibodies are important in lupus nephritis, the question regarding which glomerular structures (␣-actinin, nucleosomes, or others) are recognized by nephritogenic anti-dsDNA antibodies is still controversial. In this study, we determined which glomerular structures are recognized by monoclonal and in vivobound nephritogenic antibodies.Methods.Western blotting was used to analyze the ability of nephritogenic anti-dsDNA antibodies to recognize glomerular and nucleosomal structures. Sera from patients with lupus nephritis, sera from random antinuclear antibody-positive patients, and paired antibodies from sera and kidney eluates from nephritic (NZB ؋ NZW)F 1 mice were analyzed for activity against proteins identified by monoclonal nephritogenic antibodies, and against ␣-actinin, dsDNA, nucleosomes, histone H1, heparan sulfate, DNase I, and type IV collagen. Immunoelectron microscopy was used to determine the glomerular localization of ␣-actinin and in vivo-bound autoantibodies in nephritic (NZB ؋ NZW)F 1 mouse kidneys.Results. Anti-␣-actinin antibodies were observed in human and murine lupus nephritis sera and in sera from patients without systemic lupus erythematosus and were not detected in kidney eluates from nephritic mice. Antibodies to dsDNA and histone H1 were detected in all eluates. Western blot analyses revealed that nephritogenic anti-dsDNA antibodies recognized a 32-kd band, identified as histone H1. Competitive enzyme-linked immunosorbent assay demonstrated that nephritogenic monoclonal antibodies, and dominant antibodies eluted from nephritic kidneys, cross-reacted with dsDNA and H1. This cross-reactive anti-H1 specificity was largely absent in sera from those mice. Immunoelectron microscopic analysis of nephritic (NZB ؋ NZW)F 1 mouse kidneys revealed that antibodies eluted from kidneys, but not anti-␣-actinin antibodies, bound to distinct nephritis-associated electrondense structures linked to glomerular basement membranes. Conclusion.Cross-reactive anti-dsDNA/antihistone H1 antibodies, but not anti-␣-actinin antibodies, are central among those deposited in nephritic glomeruli.

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Section: Methodsmentioning

confidence: 99%

Critical comparative analyses of anti–α‐actinin and glomerulus‐bound antibodies in human and murine lupus nephritis

Kalaaji¹,

Sturfelt²,

Mjelle³

et al. 2006

Arthritis & Rheumatism

102

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“…C luciliae assay. This method used C luciliae as a substrate and fluorescein isothiocyanate-labeled anti-Ig (Central Laboratory of the Red Cross Blood Transfusion Service, Amsterdam, The Netherlands) as a conjugated antibody (24). Values were expressed in titers of 2-fold dilutions, starting at a dilution of 1: 10.…”

Section: Patients An11 Methodsmentioning

confidence: 99%

Measurement of increases in anti‐double‐stranded dna antibody levels as a predictor of disease exacerbation in systemic lupus erythematosus

Borg

Horst

Hummel

et al. 1990

Arthritis & Rheumatism

507

227

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To evaluate the predictive power of changes in levels of antibodies to double-stranded DNA (antidsDNA) as a predictor of disease exacerbations in systemic lupus erythematosus (SLE), we performed a prospective study on 72 unselected patients with SLE (mean duration of study 18.5 months, range 6-35 months). Patients were seen at least once every 3 months, and disease activity was scored according to a specific protocol. Plasma samples were obtained at least once every month and were assessed for anti-dsDNA antibody (by the Crifhidiu luciliue assay, an enzymelinked immunosorbent assay [ELISA], and the Farr assay) and for complement components C3 and C4. Twenty-seven of 33 disease exacerbations observed during the study period were accompanied by a positive test result for anti-dsDNA antibody (27 by the Farr assay, 19 by the C luciliae assay, and 23 by the ELISA). Twentyfour of these exacerbations were preceded by a significant increase in anti-dsDNA antibody levels (23 by the Farr assay, 12 by the C luciliae assay, and 17 by the ELISA). The first observance of a significant increase in anti-dsDNA antibody levels preceded the exacerbation by 8-10 weeks. Significant increases in anti-dsDNA antibody levels not followed by an exacerbation were observed in 5 cases by the Farr assay, in 7 cases by the C luciliue assay, and in 3 cases by the ELISA; however, in 3 cases, 2 cases, and 1 case, respectively, these increases were followed by an increase in disease activity that did not fulfill the criteria for an exacerbation. Serial measurement of anti-dsDNA antibody levels was more sensitive for predicting exacerbations than was measurement of C3 andor C4 levels (P < 0.03). Serial assessment of anti-dsDNA antibody levels, especially by the Farr assay, is a sensitive and reasonably specific method for predicting disease exacerbations in SLE.Systemic lupus erythematosus (SLE) is characterized by protean clinical manifestations and the occurrence of a variety of autoantibodies. Among these, anti-double-stranded DNA (anti-dsDNA) antibodies are highly specific for SLE and are detected at a high frequency (75-

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“…21 Blood was obtained for biochemical (renal and liver function, C-Reactive Protein), hematological (hemoglobin level, hematocrit, leucocytes and differentiation, platelets), auto-immunity (anti-DNA antibodies and immune-complexes (ICX)) and CEA level assays, before each vaccination and during follow-up visits. Anti-DNA antibodies, 22 ICX levels 23 and immune responses against ALVAC and p53 were measured as previously described. 24 IgG antibody titers against ALVAC are presented as mean OD measured at 415 nm (dilution 1:800) minus the background.…”

Section: Treatment and Follow-up Schedulementioning

confidence: 99%

Safety of intravenous administration of a canarypox virus encoding the human wild-type p53 gene in colorectal cancer patients

et al. 2003

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Overexpression of p53 occurs in more than 50% of colorectal cancers. Therefore, p53 represents an attractive target antigen for immunotherapy. We assessed the safety of a canarypox virus encoding the human wild-type p53 gene given intravenously to endstage colorectal cancer patients in a three-step dose escalation study aimed at inducing p53 immune responses. Patients with metastatic disease of p53-overexpressing colorectal cancers were vaccinated three times at 3-week intervals, each time with 10 6.5 CCID 50 (CCID 50 ¼ cell culture infectious dose 50%; group 1, n ¼ 5), 10 7.0 CCID 50 (group 2, n ¼ 5) or 10 7.5 CCID 50 (group 3, n ¼ 6). Vital signs and the occurrence of adverse events were monitored and blood was analyzed for biochemical and hematological parameters as well as signs of auto-immune safety. In all, 16 patients were enrolled and 15 patients completed three vaccinations. No anaphylactic reaction or unwanted auto-immune reactions were observed. A total of 16 serious adverse events (SAEs) occurred: 10 in group 1, three in group 2 and three in group 3. All SAEs were tumor-related complications. There was no difference in the frequency of adverse events between the three groups, except for fever. Fever was the only vaccination-related adverse event consistently observed and was most frequent and outspoken in the group 3 patients. The majority was a grade 1 or 2 fever (93%) and grade 3 fever (7%) was observed in three patients of group 3. Some patients showed humoral and cellular responses against p53, following vaccinations. After having completed his initial treatment cycle, one patient (group 2) received a second treatment cycle of three doses of 10 7.5 CCID 50 and subsequently showed stable disease. All other patients showed progressive disease. We conclude that ALVAC-p53 can be administered intravenously to colorectal cancer patients without serious toxicity or pathological autoimmunity and can induce immune responses against p53.

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IMMUNOLOGY OF DNA. III. CRITHIDIA LUCILIAE, A SIMPLE SUBSTRATE FOR THE DETERMINATION OF ANTI‐dsDNA WITH THE IMMUNOFLUORESCENCE TECHNIQUE*

Cited by 534 publications

References 19 publications

Critical comparative analyses of anti–α‐actinin and glomerulus‐bound antibodies in human and murine lupus nephritis

Critical comparative analyses of anti–α‐actinin and glomerulus‐bound antibodies in human and murine lupus nephritis

Measurement of increases in anti‐double‐stranded dna antibody levels as a predictor of disease exacerbation in systemic lupus erythematosus

Safety of intravenous administration of a canarypox virus encoding the human wild-type p53 gene in colorectal cancer patients

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