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#17998268   2007/12/27 Save this To Up

Nanogold catalysis-based immunoresonance-scattering spectral assay for trace complement component 3.

Complement component 3 (C3) is an essential bridge linking innate immunity and adaptive immunity. We describe an immunonanogold catalytic resonance-scattering (RS) technique for assaying C3 in serum.

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#7507644   1994/03/01 Save this To Up

Hormonal regulation of complement components and receptors throughout the menstrual cycle.

Complement components have been recently demonstrated to be present in the reproductive tract. Among these components, C3 synthesis by glandular epithelium of the rat uterus has been shown to be regulated by estrogen; progesterone inhibits this synthesis. However, the hormonal regulation of C3 and the presence of other complement factors in the human has not been investigated to date. In this study we examined the presence and the hormonal regulation of different complement components and receptors in the human endometrium at various phases of the menstrual cycle of normally cycling women with no pelvic pathologic abnormalities.

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#2658844   1989/07/07 Save this To Up

Determination of IgG subclasses in patients with pemphigus with active disease and in remission.

IgG subclasses were determined in perilesional skin of 13 patients with pemphigus with active disease and of 14 patients in a state of clinical remission. Using indirect immunofluorescence technique, frozen sections were incubated with mouse monoclonal antihuman IgG1, IgG2, IgG3, and IgG4 followed by a second incubation with fluorescein isothiocyanate-conjugated goat antimouse IgG. The results showed that among patients with active disease, IgG1 was found in all of them and IgG4 in 85%, while IgG2 and IgG3 were found in 54% and 77%, respectively. For patients in remission the most common subclass was IgG4 in 79% of patients, and in a decreased order IgG1, 50%; IgG3, 29%; and IgG2, 14%. It appears that IgG1 and IgG4 are predominant among patients with active lesions. IgG1 seems to be the most sensitive indicator for activity of the disease. IgG4, normally found in the lowest concentration in human serum, is the most common subclass in patients who are in remission. IgG3 and C3 may have a predictive value for remission.

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#3258914   1988/06/03 Save this To Up

Complement fixing immune complexes containing antinuclear antibodies in patients with systemic lupus erythematosus.

Immune complexes (IC) in the sera of patients with systemic lupus erythematosus (SLE) were isolated using goat antihuman C3 and rabbit antihuman Clq immunoabsorbent columns. The sera from 12 patients with SLE were sequentially eluted from the columns with veronal buffer, 0.02 M EDTA, 0.5 M NaCl, and 1 M propionic acid. The isolated IC were analyzed for antinuclear antibodies (ANA) with an HEp-2 cell substrate. ANA were detected in 6 patients' eluates from the EDTA fractions and 2 patients' propionic acid fraction by the antihuman Clq column method. ANA were noted in 7 patients' isolates from antihuman C3 column, 6 EDTA fractions, and 2 patients' propionic acid fractions. The ANA showed speckled patterns in 6 patients and a diffuse pattern in 1. Our studies demonstrate complement fixing IC containing ANA can be detected in the sera of patients with SLE by isolation with the antihuman Clq and antihuman C3 columns.

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#3298445   1987/07/28 Save this To Up

IgA immune complexes in patients with dermatitis herpetiformis occur in the absence of IgA rheumatoid factor.

Thirty to forty percent of patients with dermatitis herpetiformis (DH) have IgA-containing circulating immune complexes (IgA-CIC); however, the antigenic composition of these complexes as well as the role they play in the pathogenesis of DH are unknown. The failure to detect wheat protein in these IgA-CIC, despite the association of DH with gluten-sensitive enteropathy, suggests that the IgA-CIC in DH may be similar to those seen in the IgA nephropathies and represent IgA rheumatoid factor (RF)-IgG complexes. We have examined the sera of 32 patients with DH, 16 non-DH patients positive for RF by latex fixation, and 15 normal subjects for IgA and IgM RF using enzyme-linked immunosorbent assays (ELISAs) and for IgA-CIC using an anti-C3 ELISA. Thirteen of 16 (81%) latex fixation test-positive patients had IgA RF by ELISA and 15/16 (94%) had IgM RF by ELISA. The total amount of RF detected by the ELISA (IgA + IgM RF) correlated with the latex fixation titer (r = 0.678, p = 0.004) in these latex fixation-positive patients. Six of the 16 (38%) latex fixation-positive patients also were found to have IgA-CIC. Solid phase absorption using goat antihuman C3 decreased the levels of immune complexes but not the level of IgA RF, suggesting the IgA-CIC detected do not represent uncomplexed IgA RF. In contrast, although 12 of 31 (39%) patients with DH had IgA-CIC ranging in amount from 0.331-26.0 micrograms IgA/ml (nl less than 0.150 microgram IgA/ml), only 1 of 32 (3%) DH patients had detectable levels of IgA RF (7.0 micrograms IgA/ml, nl less than 2.0 micrograms IgA/ml). Low levels of IgM RF were found in 8/32 (25%) of patients with DH (1.1-1.6 micrograms IgM/ml, nl less than 1.0 microgram IgM/ml). These data document that IgA RF is not present in the sera of patients with DH independent of the presence or absence of IgA-CIC and that it is unlikely that the IgA-CIC present are IgA RF complexed with autologous IgG.

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#3491333   1986/12/31 Save this To Up

Studies of circulating immune complexes and lymphocyte subpopulations in childhood IgM mesangial nephropathy.

T cell subsets, serum immunoglobulin (IgM) level, IgM-bearing lymphocytes, and circulating immune complexes (CIC) were studied in 12 children who suffered from IgM mesangial nephropathy (IgMN) during the acute nephrotic phase, in remission and relapse. Frequent relapses were observed in 11 cases, and 1 was partially responsive to steroid treatment. IgMN was diagnosed by the consistent pattern of IgM deposition by all four FITC-labelled antihuman IgM antibodies from rabbits and goats supplied by four different companies and by the 100% positivity of electron-dense mesangial deposits in an identical localization and distribution pattern of kidney biopsy specimen. CIC were detected by the 3.5% polyethylene glycol method. In sera from 12 patients IgM CIC were detected in 8 cases during the acute nephrotic phase. High levels of C3 CIC and C4 CIC were also found in these cases during the acute nephrotic phase. The CIC were undetectable in remission. Only 3 cases were detectable at low levels of IgM CIC during the second relapse. High serum IgM levels and IgM-bearing lymphocytes were noted in these patients. The patients also had a significant increase of OKT8 cells and a decrease in the OKT4/OKT8 ratio during the acute phase and in relapse. Taken together, the immunopathologic and clinical features suggest that IgMN is a disease entity with a chiefly classical pathway activation of complement components. The correlation between the changes of T cell subsets and the disease activity in IgMN suggests that this may serve as a therapeutic and prognostic guide.

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#4019777   1985/08/28 Save this To Up

Autoantibody to the C3b/C4b receptor and absence of this receptor from erythrocytes of a patient with systemic lupus erythematosus.

A 29-yr-old woman with systemic lupus erythematosus (SLE) was found to have no detectable C3b/C4b receptors (CR1) on her erythrocytes (E) when they were assayed by the binding of rabbit polyclonal and murine monoclonal (Yz-1) anti-CR1. Analysis by two-color fluorescent flow cytometry of CR1 expression on the patient's B lymphocytes that had been stained indirectly with monoclonal anti-B1 and rabbit F(ab')2 anti-CR1 also revealed a marked deficiency of CR1. Total cellular CR1 of neutrophils, assessed by a sandwich radioimmunoassay, was about half that of neutrophils from normal individuals. Because her E had expressed 173 sites/cell 2 yr before, the CR1 deficiency was considered to be acquired and a possible mechanism was sought. Autoantibody to CR1 was measured by a radioimmunoassay in which serum or its fractions were incubated in microtiter wells that had been coated with purified CR1, and binding of immunoglobulin to the wells was quantitated with 125I-labeled goat IgG antihuman F(ab')2. The CR1-specific binding of immunoglobulin from the patient's serum was 19.1 ng/well of the detecting antibody when her E had eight CR1 sites per cell; that of 28 healthy donors was 1.3 +/- 0.5 ng/well (mean +/- SEM), and that of 34 additional patients with SLE was 0.5 +/- 0.3 ng/well. The activity was present also in purified IgG and its F(ab')2 fragment, indicating that the binding of serum immunoglobulin to CR1 was not mediated by C3 fragments. The specificity of the patient's IgG for CR1 was confirmed when pretreatment of the CR1-coated wells with affinity-purified rabbit F(ab')2 anti-CR1 was shown to inhibit by 68% the binding of the IgG. The autoantibody also interacted with CR1 in cell membranes, as assessed by its capacity to inhibit the binding of indirectly fluoresceinated Yz-1 to neutrophils, and, when combined with goat IgG antihuman F(ab')2, to diminish the binding of dimeric C3b to normal E. During the period of the marked deficiency of CR1 the patient experienced an exacerbation of disease activity which was treated with prednisone. Clinical improvement was accompanied by a decrease in the serum concentration of anti-CR1 to levels present 2 yr earlier, and an increase of CR1 to 170 sites/E. The temporal association between high titers of an autoantibody to CR1, absence of CR1 from E, and heightened activity of SLE suggest that the former may have had a role in the other manifestations of the patient's disease.

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#6388973   1985/01/09 Save this To Up

An enzyme-linked immunoassay for detection of IgG- and C3-containing circulating immune complexes: comparison with a radioimmunoassay and quantitation in patients with rheumatic diseases.

We developed a simplified, relatively rapid, inexpensive, antigen-nonspecific, enzyme-linked immunosorbent assay (ELISA) for immunoglobulin G (IgG)- and C3-containing circulating immune complexes (CICs), adapted from a solid-phase anti-C3 radioimmunoassay (RIA). Standards (containing purified, heat-aggregated IgG and fresh human serum) or samples were allowed to react with goat F(ab')2 antihuman C3 bound to the matrix of microtiter plates. Then alkaline phosphatase conjugated to goat IgG fraction antihuman IgG was added, followed by p-nitrophenylphosphate, optical densities determined, and concentrations of CICs calculated. We found excellent correlations between serum and plasma CIC levels by either ELISA (r = 0.95, p less than 0.01) or RIA (r = 0.89, p less than 0.01). Furthermore, ELISA quantitation of CICs correlated well with RIA (serum, n = 75, r = 0.64, p less than 0.01; plasma, n = 101, r = 0.56, p less than 0.01). By ELISA we found 32 normal subjects had 38 +/- 12 micrograms CIC/ml in serum and 34 +/- 10 micrograms CIC/ml in plasma. Patients with systemic lupus erythematosus (39% of 27 patients, p less than 0.05) had significantly elevated CIC levels compared with normal (serum, 157 +/- 50 micrograms/ml, p less than 0.01; plasma, 89 +/- 23 micrograms/ml, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

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#7024428   1981/11/22 Save this To Up

C3b receptor in normal human skin.

Receptors for C3b in normal skin were studied. C3b was produced by treating normal human serum with cobra venom factor and by partial digestion of purified C3 with trypsin. Cryostat sections of normal human skin were incubated with C3b, followed by a direct immunofluorescent technique using monospecific goat antihuman C3. The histologic localization of C3b fluorescence was ascertained by fixing cryostat sections with glutaraldehyde and staining with hematoxylin and eosin. The following structures showed staining with anti-C3: (1) endothelial cells in capillaries, larger vessels, and arteries, (2) smooth muscle in arrector pilori muscles and artery walls, and (3) myoepithelial cells in the secretory portion of sweat glands. C3b did not bind to the intercellular substance nor to the basement membrane zone in normal human skin. Normal human sera treated with EDTA, EGTA, and heat (56 degrees C for 30 min) were negative, as was purified C3 by itself, thus indicating that native C3 did not bind to these receptors. Specificity for C3/C3b was shown by blocking with both unconjugated rabbit antihuman C3 and purified C3. The endothelial C3b receptor may have an important role in the localization of immune complexes in cutaneous vasculitis.

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#222928   1979/09/25 Save this To Up

Tumor-bound immunoglobulin in human gynecologic cancers.

By the use of immunofluorescence techniques, immunoglobulin of the IgG class was consistently found in touch preparations and in frozen sections of squamous cell carcinomas of the uterine cervix (both keratinizing and nonkeratinizing) and in an adenosquamous cell carcinoma of the cervix, an adenocarcinoma of the cervix, a mixed mesodermal-uterine tumor, and a uterine adenocarcinoma metastasized to the ovaries. Trace amountsof IgM were found in 1 squamous cell carcinoma and in 1 adenocarcinoma of the cervix. Except for 1 tumor specimen consisting primarily of infiltrating lymphocytes that stained positive for human IgG, IgM, IgA, and C3, the tumors were consistently negative for IgA and C3. Specimens made from normal cervical tissue were uniformly negative for all immunoglobulins and complement. Positive staining for human IgG could be eliminated by incubation of the tumor preparations with unconjugated goat antihuman IgG before the preparations were stained with fluorescein isothiocyanate-conjugated goat antihuman IgG. Attempts to elute the tumor-bound IgG with glycine-HCl buffer (pH 2.2) were most successful with the use of unfixed tissue, although the positive staining for IgG could not be entirely eliminated. The elution effects of the low-pH buffer on tissue fixed over 2 hours in 10% Formalin were minimal.

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