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PubMed2000

Comparison of different diagnostic methods for lupus pleuritis and pericarditis: a prospective three-year study.

BACKGROUND AND PURPOSE: Pleural or pericardial effusions, or both, are commonly encountered, but the differential diagnosis is sometimes difficult. We evaluated the diagnostic value of effusion immunofluorescent antinuclear antibody (ANA) titer, systemic lupus erythematosus (SLE) latex agglutination slide test, and cytologic LE cell examination in patients with pleural and/or pericardial effusions of various etiologies. METHODS: A total of 153 pleural and/or pericardial effusion specimens were collected by aspiration from 152 patients (14 SLE and 138 non-SLE patients). All specimens were sent for routine biochemistry testing, determination of ANA titer, SLE latex agglutination slide test, and LE cell examination. RESULTS: Ten of the 14 SLE patients had lupus serositis and all of them had high ANA titers (> or = 1:160) in their effusions. SLE latex and LE cell tests were positive in seven and eight patients with lupus serositis, respectively. The remaining four SLE patients with effusion of etiologies other than lupus serositis had low or negative effusion ANA titers. Among the non-SLE patients, 29 of 112 patients (26%) with pleural effusion and six of 26 patients (23%) with pericardial effusion had positive ANA tests (> or = 1:40). None of them had a positive SLE latex or LE cell test result. Thirteen of the 138 non-SLE patients (11%) had high effusion ANA titers (> or = 1:160). Effusion in 11 of 13 non-SLE patients (85%) was due to malignancy. CONCLUSIONS: Effusion ANA titer detection is a very sensitive but nonspecific test for the diagnosis of lupus serositis. SLE latex and cytologic LE cell tests can aid in the differential diagnosis as complementary tools. The specificity, positive and negative predictive values of these two tests are excellent for the diagnosis of lupus serositis.

Adolescent↗
PubMed1996

Retinoic acid: identification of specific receptors through which it may mediate transcriptional regulation of fibronectin gene in bovine lens epithelial cells.

To elucidate in detail the regulatory mechanisms by which the fibronectin mRNA and protein are altered by all-trans retinoic acid (RA) in bovine lens epithelial (LE) cells, we have investigated the transcription mechanism and the occurrence of nuclear receptors for RA in the nuclei of cultured LE cells. The rate of transcription of fibronectin (FN) mRNA increased by 130% in response to RA. The RA receptors in nuclear extracts prepared from LE cells show specific all-trans [3H]-RA binding sites. Saturation binding of all-trans RA to these receptors and the Scatchard plot analyses of the binding data yielded an apparent dissociation constant (Kd) of about 12 nM and Bmax of about 48 pmoles/mg protein. Bovine retinoic acid receptors (RARs) have not been reported previously. Our results demonstrate for the first time that RA regulates FN expression and that regulation is at the transcriptional level in LE cells. Thus, retinoids may have an important role in the functional adhesion of epithelium to the lens capsule.

Animals↗
PubMed1993

[Antinuclear antibodies detected in autoimmune diseases].

In 1948, Hargraves discovered LE cells in the bone marrow and peripheral blood of a patient with systemic lupus erythematodes (SLE). Since then, the usefulness of the cell in the diagnosis of SLE has not changed. LE factor, a component of LE cell, is an anti-nuclear antibody. To date, many kinds of anti-nuclear antibodies have been discovered. Among others anti-dsDNA, anti-Sm and anti-ribosome antibodies were detected at a high rate and/or specifically in SLE. Therefore, they are used as markers for SLE. Recently, the antigen corresponding to each auto-antibody and the gene coding for it were clarified. In this paper, we describe methods for detecting anti-nuclear antibodies and LE cells, the significance of each auto-antibody in the diagnosis of auto-immune diseases, especially SLE and the relation between autoantibodies and symptoms in patients with the antibodies.

Antibodies, Antinuclear↗
PubMed2000

The lupus erythematosus cell phenomenon: comparative analysis of antichromatin antibody specificity in lupus erythematosus cell-positive and -negative sera.

OBJECTIVE: To compare and investigate antihistone and antichromatin antibody responses as well as clinical variables in patients with systemic lupus erythematosus (SLE) who were either positive (LEC+) or negative (LEC-) for the lupus erythematosus (LE) cell phenomenon. METHODS: The binding properties of LEC+ and LEC- SLE sera to chromatin-associated nuclear antigens (histones H1, H2A, H2B, H3, H4; complexes of H2A-H2B, [H2A-H2B]-DNA, H1-DNA; total and H1-stripped chromatin; native and denatured DNA) were investigated. In addition, sera from patients with drug-induced lupus (by procainamide, hydralazine, or quinidine), as well as from patients with rheumatoid arthritis and osteoarthritis, were assessed. Enzyme-linked immunosorbent assay was used to detect specific antibody binding. RESULTS: Mirroring the important role of histone H1 in the formation of LE cells, anti-histone H1 reactivity was 8-fold higher in LEC+ sera than in LEC- sera. In addition, reactivities to most of the other antigens tested, i.e., other histones and histone-DNA complexes as well as chromatin and DNA, were significantly higher in LEC+ sera than in LEC- sera. All but 1 serum sample from the patients with drug-induced lupus were negative for LE cell formation as well as for anti-histone H1 reactivity, but displayed high antibody reactivities to histone-DNA complexes, including chromatin. Sera from patients with rheumatoid arthritis and osteoarthritis did not show significant binding to these antigens. When comparing the clinical features of LEC+ and LEC- SLE patients, severe organ involvement, including nephritis and central nervous system involvement, was common in the LEC+ group, but rare in the LEC- group. CONCLUSION: A positive LE cell phenomenon not only correlated with the presence of high anti-histone H1 antibody levels in SLE, but also indicated serologically and clinically active disease with major organ involvement.

Antibody Specificity↗
PubMed1971

Canine systemic lupus erythematosus. Genetic analysis of an established breeding colony.

Three breeding lines, originating from dogs with SLE, have been established. Two lines were initiated by mating a female with SLE with a normal male. The third line resulted from a mating of two affected dogs. Brother-to-sister matings have reached the third generation in each line. In addition, backcross and outcross matings were carried out. More than one-third of the autopsied dogs had thymic abnormalities. The commonest lesion was a lymphoid follicle; the thymus of one dog contained multiple granulomas, and in one animal a reticulum cell sarcoma of the thymus was found. Multiple serological abnormalities, including positive LE cell tests, anti-nuclear antibodies (ANA), and rheumatoid factor, were found in the progeny. The development of ANA appeared unrelated to the incidence of positive LE cell tests. About 10% of the animals had rheumatoid factor in their serum. Control populations of dogs; including house pets; two other, unrelated lines of inbred dogs; and normal dogs housed in the same facility as the SLE colony did not have these abnormalities. The incidence of positive LE cell tests in the inbred, backcross, and outcross matings was not consistent with any conventional genetic mechanism of inheritance. It is conceivable that the results can be explained by vertical transmission of an infectious agent in a genetically susceptible individual.

Animals↗
PubMed1994

Practice parameter--the lupus erythematosus cell test. An obsolete test now superseded by definitive immunologic tests.

The lupus erythematosus (LE) cell test (synonyms: LE prep, LE phenomenon; CPT No. 85544, LE Cell Prep) is a diagnostic test for systemic lupus erythematosus (SLE) that is based on an in vitro immunologic reaction between the patient's autoantibodies to nuclear antigens and damaged nuclei in the testing medium. It is subject to numerous experimental variables and dependent on subjective interpretation. It should be abandoned in favor of more definitive, quantitative immunologic tests for this condition.

Cytodiagnosis↗
PubMed1975

DNA-antibodies in sera from patients with systemic lupus erythematosus.

DNA-antibody determinations using the Farr technique and LE cell tests were performed on single serum samples from 55 patients with SLE according to well defined criteria. It was found that the DNA-antibody test was just as sensitive as the LE cell test, and studies of sera from patients with questionable SLE, discoid LE, other connective-tissue diseases, psoriasis, and from healthy persons, indicated that the DNA-antibody test is highly specific for SLE. Neither the DNA-antibody test nor the LE cell test could discriminate between SLE patients with active and inactive disease as estimated by clinical criteria or by the serum concentrations of complement C3 and C4.

Antibodies, Antinuclear↗
PubMed1997

Detection of polyanion-restricted anti-histone antibodies in patients with systemic lupus erythematosus.

The nature of the antibodies responsible for lupus erythematosus (LE) cells in systemic lupus erythematosus (SLE) remains obscure. We examined whether polyanion-restricted anti-histone antibodies were present in serum of patients with SLE using Western blotting analysis. Dextran sulfate or alginate was used as a polyanion compound in place of DNA. Antibodies which recognized dextran sulfate-histone complexes were present in serum of patients with SLE (17/34, 50%). These antibodies were detected in most SLE patients positive for LE cells (17/18, 94%) but not in those negative for LE cells or in patients with other collagen diseases. Similar results were obtained using alginate-histone complexes as antigens for Western blotting analysis. The antibodies to dextran sulfate-histone or alginate-histone complexes in serum of SLE patients were completely absorbed by treatment of serum with DNA-histone complexes, while they were unaffected by treatment with DNA only. The presence of antibodies to free histones and dextran sulfate-histone complexes did not seem to be related to the titer of anti-single stranded DNA antibody and anti-double stranded DNA antibody. We demonstrated the presence of polyanion-restricted antibodies in SLE, which may be responsible for the LE factor.

Adolescent↗
PubMedDate not supplied

[Studies on the genesis of so-called Sjögren cells in supravital preparations].

1. The effect of the LE-cell factor lies in nuclear blockade; the normal degenerative nuclear swelling and lysis is inhibited. 2. The time of the nuclear blockade and thus the appearance of the LE-bodies depend on the activity of the LE-serum. 3. High-titred sera lead to an early nuclear blockade. Small, rigid, dark, clump-like LE-bodies appear rapidly and are fully phagocytosed. 4. Our experiments have shown that with decreasing activity of the LE-sera the nuclear blockade appears later and more slowly. Such LE-bodies are relatively large and homogenous and have lost some rigidity. They are correspondingly late and only partly phagocytosed. So-called granulocytic and monocytic Sjögren-cells appear. 5. We found that quantitative differences of the causal factors lead on the one hand to LE-cell formation, on the other to the partial phagocytosis phenomenon. Therefore, we prefer not to label the Sjögren-cells as pseudo-LE-cells. 6. It has again been shown that the standardized supravital preparation is not only a simple method for demonstrating LE-cells, but is also superior to other methods used because of its greater specifity and sensitivity. Besides the judgement of cell vitality, many hours after making the preparation phagocytosis can be observed.

Antibodies, Antinuclear↗
PubMed2007

Curcumin inhibits the formation of capillary-like tubes by rat lymphatic endothelial cells.

The natural pigments curcumin and berberine have been shown to exhibit a variety of pharmacologic effects including anti-inflammatory, anti-cancer, and anti-metastatic properties. Here, we investigated the anti-lymphangiogenic effect with an in vitro tube-forming model using conditionally immortalized lymphatic endothelial TR-LE cells, a newly established cell line originating from the thoracic duct of a transgenic rat expressing the temperature-sensitive SV40 large T-antigen. Curcumin, but not berberine, exhibited a dose-dependent inhibition of the formation of capillary-like tubes by TR-LE cells without affecting cell viability and adhesion to Matrigel. To address the molecular mechanisms involved, we performed experiments with specific inhibitors against putative targets of curcumin, including IkappaB kinase (IKK), epidermal growth factor receptor (EGFR), phosphatidylinositol-3 kinase (PI3K)/Akt, and matrix metalloproteinases (MMPs). While the IKK-2 inhibitor VI and EGFR tyrosine kinase inhibitors gefitinib and PD153035 had no effect, both the PI3K inhibitor LY294002 and the MMP inhibitor GM6001 shortened the tubes by approximately 50%. Western blot analysis and gelatin zymography revealed that curcumin, but not berberine, has an inhibitory effect on the phosphorylation of Akt and enzymatic activity of MMP-2 in TR-LE cells. These results suggest that curcumin exerts its inhibitory effect on lymphangiogenesis partly through Akt and MMP-2.

Animals↗
PubMed1984

Uterine growth responses of the mature castrate rat to estradiol-17B.

To examine estrogen-stimulated uterine growth we have monitored changes in uterine DNA synthesis, ornithine decarboxylase (ODC) activity and protein content as well as luminal epithelial (LE) cell mitotic index and ultrastructural changes. We have utilized this model to examine castrate mature rat uterine growth as a function of time between 18 and 40 hours following a single injection of 25.0 ug of estradiol-17B. LE cell mitotic index and protein content increases were maximally elevated as early as 18 hours postinjection while uterine ODC activity was maximal at 28 hours; uterine DNA synthesis increases continued throughout the experiment. In addition, the infusion of either 1 or 2 ug E2 plus progesterone over a 24 hour period, stimulated elevated ODC activity under both treatment regimens and LE cell mitotic index which was inversely related to E2 dose.

Animals↗
PubMed1987

Lupus erythematosus cells in serum and pleural fluid of a patient with negative fluorescent antinuclear antibody test.

The authors report here a 61-year-old male with anemia, arthritis, and pleural effusions who had lupus erythematosus (LE) cells in the pleural fluid and a strongly positive LE cell test in the blood. However, the fluorescent antinuclear antibody (FANA) assay performed repeatedly with the serum always had negative results, despite the use of various technics. The patient was considered to have systemic LE and responded well to treatment with corticosteroids. Although the authors could not explain the LE cell-FANA incongruity, they suggest that the LE test not be deleted from the clinical laboratories' tests.

Antibodies, Antinuclear↗
PubMed1976

Unusual cells in lupus erythematosus bullae.

We saw a case of bullous lupus erythematosus (LE) in which there were pre-LE cells and hematoxylin bodies in clear bullous fluid. Pre-LE cells are degenerated leukocytes with pyknotic nuclei. The diagnosis of lupus erythematosus was substantiated by immunofluorescence studies.

Adolescent↗
PubMed1975

[Immunodiagnostic methods in lupus erythematosus disseminatus].

In 27 patients with lupus erythematodes diseminatus the determinations of the LE-cells according to the macromethod (Zimmer and Hargraves) and the micromethod (Mudrik and co-workers) were compared with the demonstration of antinuclear factors according to the indirect immunofluorescence and immune enzyme technique. The sensitiveness of the two last-mentioned immunomorphological methods is somewhat larger. In these cases the size of the titre of the antinuclear factor almost always correlates positively with the number of the LE-cells. For the purpose of the initial diagnostics and the judgment of the course a morphological method cannot be renounced, since in the acute episode a high consumption of the antinuclear factor the immunological methods negatively correlate with the number of the LE-cells. The immune enzyme technique is to be recommended on account of the smaller expenditure, permanence of the preparations and high sensitiveness as alternative method of the immunofluorescence technique. In the micromethod the large variation is opposite to the advantage of the slight quantity of blood and to an always existing evaluability. Investigations of the lymphocytes of patients with lupus erythematodes disseminatus by means of the lymphocyte transformation test and the determination of the B-cells with the help of the direct immune peroxidase technique refer to the close pathogenetic connections of cellular and humoral immune reactions in this disease.

Antibodies, Antinuclear↗
PubMed2003

Embryonic enhancers in the dpp disk region regulate a second round of Dpp signaling from the dorsal ectoderm to the mesoderm that represses Zfh-1 expression in a subset of pericardial cells.

During germ band elongation, widespread decapentaplegic (dpp) expression in the dorsal ectoderm patterns the underlying mesoderm. These Dpp signals specify cardial and pericardial cell fates in the developing heart. At maximum germ band extension, dpp dorsal ectoderm expression becomes restricted to the dorsal-most or leading edge cells (LE). A second round of Dpp signaling then specifies cell shape changes in ectodermal cells leading to dorsal closure. Here we show that a third round of dpp dorsal ectoderm expression initiates during germ band retraction. This round of dpp expression is also restricted to LE cells but Dpp signaling specifies the repression of the transcription factor Zfh-1 in a subset of pericardial cells in the underlying mesoderm. Surprisingly, we found that cis-regulatory sequences that activate the third round of dpp dorsal ectoderm expression are found in the dpp disk region. We also show that the activation of this round of dpp expression is dependent upon prior Dpp signals, the signal transducer Medea, and possibly release from dTCF-mediated repression. Our results demonstrate that a second round of Dpp signaling from the dorsal ectoderm to the mesoderm is required to pattern the developing heart and that this round of dpp expression may be activated by combinatorial interactions between Dpp and Wingless.

Animals↗
PubMed1989

Oxytocin production and oxytocin messenger ribonucleic acid levels in bovine granulosa cells are regulated by insulin and insulin-like growth factor-I: dependence on developmental status of the ovarian follicle.

Oxytocin is a major peptide product of the ruminant corpus luteum, and the release of oxytocin from serum-free cultures of bovine granulosa cells is stimulated by insulin and insulin-like growth factor-I (IGF-I). Here we have assessed the effects of insulin and IGF-I on oxytocin gene expression in bovine granulosa cells and the dependence of these effects on the developmental status of the cells. When cells from individual follicles were cultured, the estradiol concentration of the follicular fluid was highly correlated with insulin-stimulated oxytocin release. Subsequently, cells were pooled from follicles selected on the basis of estradiol content, and two subsets of cells were distinguished. The first contained highly differentiated cells, as judged by the high estradiol (HE-cells) concentration of the follicular fluid (greater than 40 ng/ml), high levels of LH receptors, and high hCG-stimulated cAMP accumulation. The second subset contained cells from follicles with low estradiol (less than 1 ng/ml; LE-cells) which have fewer LH receptors and low hCG-stimulated cAMP accumulation. Oxytocin production was increased more than 50-fold by insulin (EC50, 230 +/- 57 ng/ml) and IGF-I (EC50, greater than 10 ng/ml), but only in the HE-cells. Oxytocin mRNA was also greatly increased by insulin and IGF-I in the HE-cells only. In contrast, insulin and IGF-I stimulated progesterone release from both HE- and LE-cells. Since oxytocin production is a characteristic of bovine luteal cells, our results support possible roles for IGF-I and insulin in regulation of luteinization or luteal activity. The data suggest that effects of insulin and IGF-I on oxytocin production reflect their effects on oxytocin gene transcription, and that granulosa cells must be appropriately primed (presumably by the in vivo hormonal environment) before they are able to produce oxytocin in response to these polypeptides.

Animals↗
PubMed1996

F protein induced fusion of Sendai viral envelopes with mouse teratocarcinoma cells through Le(x)-Le(x) interaction.

The efficiency of membrane fusion between reconstituted Sendai viral envelopes containing only the fusion protein (F-virosomes) and the plasma membrane of mouse teratocarcinoma cells (F9) in culture was assessed using an assay based on the relief of self-quenching of a lipid probe incorporated in the F-virosomes. The potential of F-virosomes was also evaluated for a targeted cytosolic delivery of lysozyme to F9 cells. [125I]Lysozyme entrapped into F-virosomes was taken to examine its fusion-mediated transfer to the F9 cells. Target specificity of the F-virosomes was confirmed by the interaction between the terminal Le(x) moiety (Gal beta 1-->4(Fuc alpha 1-->3)GlcNAc) of F protein and the Le(x) determinant on the membrane of F9 cells. Incubation of the loaded F-virosomes with cells led to fusion-mediated delivery, as inferred from the ability of cells to internalize lysozyme in the presence of azide (a potent inhibitor of endocytosis). These results suggest that carbohydrate-carbohydrate interaction is strong enough for target cell recognition followed by phospholipid bilayer melding induced by fusion glycoprotein of Sendai virus.

Animals↗
PubMed1998

Loss of imprinting of the insulin-like growth factor II gene in mouse hepatocellular carcinoma cell lines.

We investigated expression of insulin-like growth factor II (Igf2) in primary cultured hepatocytes, liver epithelial (LE) cell lines derived from normal hepatocytes, and hepatocellular carcinoma (HCC) cell lines from crosses between C3H/HeJ (C3H) and Mus musculus molossinus mice (MSM). Igf2 mRNA was detected by reverse transcriptase-polymerase chain reaction in primary cultured hepatocytes from 5 d after the start of cultivation and in all 12 LE and 16 HCC cell lines. Analysis of the untranslated region of Igf2 exon 6, which contains polymorphic CA repeats, revealed that 13 of the 16 HCC cell lines had biallelic expression, whereas monoallelic expression was retained in the primary cultured hepatocytes and all 12 LE cell lines. The Igf2 transcripts contained exons 1-3 in all the HCC cell lines but only exons 2 and 3 in cultures of hepatocytes and LE cell lines, indicating difference in promoter use. However, the biallelic HCC cell lines did not have larger amounts of Igf2 mRNA and protein than did the monoallelic lines, suggesting that loss of imprinting may not be directly related to the level of Igf2 expression.

Alleles↗