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At least 19 recordsLinked to original sources
PubMed1985

[The role of methods using immunofluorescence (platelet suspension immunofluorescence test and granulocyte immunofluorescence test) in the diagnosis of autoimmune thrombocytopenia and neutropenia].

The authors examined 46 patients with autoimmune thrombocytopenic Purpura and 17 patients with autoimmune neutropenia, with the direct and indirect immunofluorescence method. They intended to verify the sensibility of such methods and to determine the class of immunoglobulins. In the PTA the direct PSIFT was positive in 26 of the 46 examined patients, while the indirect in 15 cases. In the GPA the direct GIFT was positive in 9 of the 17 patients and the indirect in 6 cases. By using FITC-labelled monospecific anti-immunoglobulin reagents, in the PTA we have found IgG in the 57.7% of the patients, IgM in the 11.5% and IgG + IgM in 30.8%; in the GPA we have found IgG in the 66% of the cases, IgM in 11% and IgG + IgM in 23%. These methods have shown a good reliability and sensibility, but also delicacy.

Agranulocytosis↗
PubMed1982

Double immunofluorescence microscopy: a method for localizing immune deposits in skin diseases associated with linear basement membrane zone immunofluorescence.

Direct immunofluorescence microscopy has shown that a linear pattern of immunoglobulin and/or complement deposition at the cutaneous basement membrane zone is a characteristic feature in a number of acquired bullous diseases and is occasionally observed in systemic lupus erythematosus. Immunoelectron microscopy has shown the linear pattern of immunofluorescence may be produced by immune deposits located either above the basal lamina (in the lamina lucida) or below the basal lamina (in the upper dermis). Distinguishing between these sites of immune reactant deposition may be of value in differential diagnosis. In this study we report a double immunofluorescent method by which skin biopsies with linear IgG immunofluorescence due to deposits above the basal lamina (bullous pemphigoid) could be distinguished from biopsies with deposits beneath the basal lamina (bullous systemic lupus erythematosus and epidermolysis bullosa acquisita). When skin sections were treated sequentially with rhodamine-labeled anti-human IgG followed by fluorescein-labeled antilamina lucida (pemphigoid) antibody and examined by fluorescence microscopy, the following results were obtained. In biopsies with IgG deposits in the lamina lucida, a single green fluorescent band was observed. In tissues with subbasal lamina deposits, either parallel and contiguous bands of green and yellow-orange fluorescence or a single band of yellow-orange fluorescence was observed. The method is simpler, quicker, and less expensive than immunoelectron microscopy and should be a useful technique for evaluating skin diseases with linear immunofluorescence at the basement membrane zone.

Basement Membrane↗
PubMed1983

Pitfalls in immunofluorescence testing in dermatology. II. Pemphigus-like antibodies in the cat, and direct immunofluorescence testing of normal dog nose and lip.

Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 75 cats: 25 cats with various nonpemphigus dermatologic diseases, 25 cats with various nondermatologic diseases, and 25 normal cats. Pemphigus-like antibodies were not detected. It was concluded that neither true pemphigus antibodies, nor pemphigus-like antibodies are commonly detected by indirect immunofluorescence testing in the cat. Direct immunofluorescence testing for IgG, IgA, IgM, and C3 was performed on the nose and lip of 15 normal dogs. Granular deposition of IgM at the basement membrane zone of the nose was demonstrated in 11 of the 15 dogs. Lip was consistently negative. It was concluded that direct immunofluorescence testing of canine nose by only polyvalent immunoglobulin antisera or anti-IgM antisera may lead to misinterpretation and misdiagnosis in up to 73% of all dogs tested.

Animals↗
PubMed1984

Pitfalls in immunofluorescence testing in dermatology. III. Pemphigus-like antibodies in the horse and direct immunofluorescence testing in equine dermatophilosis.

Indirect immunofluorescence testing for pemphigus-like antibodies was performed on 79 horses: 28 horses with various nonpemphigus dermatologic diseases, 21 horses with various nondermatologic diseases, and 30 normal horses. Pemphigus-like antibodies were detected in 6 horses: 3 normal horses with titers of 1:40, 2 horses with dermatophilosis at titers of 1:10 and 1:80, and 1 horse with lymphosarcoma at a titer of 1:320. It was concluded that equine pemphigus-like antibodies are a potential source of misinterpretation and misdiagnosis in indirect immunofluorescence testing. Direct immunofluorescence testing for whole immunoglobulin, IgG, IgM, and IgA was performed on skin lesions from 2 horses with dermatophilosis. Diffuse intercellular deposition of whole immunoglobulin and IgG was found in both horses. It was concluded that equine dermatophilosis is a potential source of misinterpretation and misdiagnosis in direct immunofluorescence testing.

Actinomycetales Infections↗
PubMed1997

Circulating pemphigus IgG in families of patients with pemphigus: comparison of indirect immunofluorescence, direct immunofluorescence, and immunoblotting.

BACKGROUND: Patients with pemphigus vulgaris (PV) are genetically linked to two alleles of the HLA subgroup, and circulating antibodies were found in first-degree relatives of these patients, thus showing genetic predisposition. OBJECTIVE: Our purpose was to determine the occurrence of circulating true PV-IgG in patients' relatives. METHODS: Circulating PV-IgG was determined in 21 first-degree relatives of 12 patients with PV by indirect immunofluorescence on monkey esophagus, carcinoma A431 cultures, and Western immunoblotting. Direct immunofluorescence was performed on skin biopsy specimens of 20 relatives. RESULTS: Circulating PV-IgG was detected in 15 relatives (71%) by all methods tested. Good correlation was found between immunoblot reactivity and immunofluorescence. Of the 15 "positive" relatives, only five showed fixation of IgG to epidermal cells in vivo. CONCLUSION: The permeability of the epidermis or epidermal cell reactivity in vivo probably controls the expression of disease in patients' relatives.

Adult↗
PubMed1986

Rapid diagnosis of respiratory syncytial virus infection by antigen immunofluorescence detection with monoclonal antibodies and immunoglobulin M immunofluorescence test.

During a respiratory syncytial virus (RSV) infection outbreak in a pediatric hospital, diagnosis was made by immunofluorescence on smears by using an anti-RSV monoclonal antibody (IFm). Immunoglobulins M and G were titrated by indirect immunofluorescence on HEp-2 cells infected with an RSV strain. The IFm was sensitive (89%) and specific (75%) when compared with the cell culture method. We showed that the specimens which were found positive by IFm and negative by cell culture were truly positive. Under these conditions, the IFm test appears more sensitive and more specific than cell culture, particularly when no care is taken to maintain the specimens in the cold during transport. In this study the immunoglobulin M immunofluorescence test had a low sensitivity (34%), especially on serum samples taken on days 0 to 4 after the onset of illness.

Antibodies, Monoclonal↗
PubMed1978

A comparison of complement fixation, indirect immunofluorescence for viral late antigens, and anti-complement immunofluorescence tests for the detection of cytomegalovirus specific serum antibodies.

It was shown that, on average, the titres of the sera obtained by both the anti-complement immunofluorescence (ACIF) and indirect immunofluorescence for viral late antigens (IFA-LA) procedures were approximately eight times higher than those obtained by complement fixation. There was no significant difference between the mean ACIF and IFA-LA titres, but the former method was preferred because it was not affected by non-specific staining. When 406 sera were screened for the presence or absence of antibodies, only five sera (1.2%) gave discordant results with the three tests. None of the tests could detect all of the sera that contained specific antibodies while each test detected over 98% of these sera. Thus, the increased sensitivity of the immunofluorescence procedures did not result in an appreciably increased rate of detection of cytomegalovirus seropositive sera.

Antibodies, Viral↗
PubMed1994

Quantitative immunofluorescence data suggest a permanently enhanced GAD67/GAD65 ratio in nerve endings in rat cerebral cortex damaged by early postnatal hypoxia-ischemia: a comparison between two computer-assisted procedures for quantification of confocal laser scanning microscopic immunofluorescence images.

The aim of the present study was 2-fold: (1) to determine the ratio between the amount of GAD67 and GAD65 (two isoforms of the GABA synthetizing enzyme glutamic acid decarboxylase) in nerve endings in the mature rat cerebral cortex damaged by hypoxia-ischemia during early postnatal life; and (2) to compare two different computer-assisted procedures developed for quantitative analysis of immunofluorescence images obtained with a confocal laser scanning microscope (CLSM). One procedure was based on a program present in the standard Leica CLSM software packet for full-field analysis, the other on a specially written program for object-oriented analysis run on a Kontron IBAS-KAT image analysis system. To this end, rat pups were unilaterally exposed to hypoxic-ischemic conditions and, after a survival period of 6.5 months, sacrificed by perfusion fixation. After dissection of the brain and vibratome sectioning, three animals with substantial damage on one cortical side were selected. Sections of these animals were double-stained with primary antibodies against GAD67 and GAD65 and fluorophore-conjugated secondary antibodies and subsequently sampled with a CLSM. Analysis of the CLSM images with both computer-assisted procedures showed for all three animals a clear tendency to higher GAD67/GAD65 ratios in cortical GABAergic nerve endings on the hypoxia-damaged side than in matched areas on the contralateral side. This outcome led to the following conclusions. (1) The correspondence between the outcome of both analysis procedures indicates that both procedures are valid for quantification of immunofluorescence images of nerve endings obtained with a CLSM. (2) The outcome lends further support to our view that hypoxic-ischemic encephalopathy, sustained during early postnatal life, may result in an unstable cortical network generating abnormal synchronizations and oscillations which can be amplified and propagated as true epileptic discharges. In such a network both excitatory and inhibitory processes are tonically enhanced, the latter probably as a homeostatic reaction tending to keep abnormal excitation within physiological limits.

Animals↗
PubMed1986

Analysis of anti-HTLV-I antibody by strip radioimmunoassay--comparison with indirect immunofluorescence assay, enzyme-linked immunosorbent assay and membrane immunofluorescence assay.

Antibodies against human T-cell lymphotropic virus type I (HTLV-I) in the sera from 60 patients with adult T-cell leukemia and 21 carriers who were suspected of having HTLV-I infection were investigated by indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), membrane immunofluorescence assay (MIA) and strip radio immunoassay based on the western blotting technique (SRIA). The sera of 2 of the carriers who were seropositive in IFA and ELISA were negative in MIA and did not react with virus-specific proteins by SRIA. Two sera were negative in IFA and ELISA. These sera were positive in MIA and reacted with only the envelope-related glycoprotein (gp46) and not with gag-related proteins (p28, p24, p19) by SRIA. These findings suggest that the main antigens defined by IFA and ELISA are gag-related proteins and some sera which do not contain anti-HTLV-I antibodies give false-positive results because of the reaction to unknown cellular components. Also some sera may have antibodies against only envelope glycoproteins, and these sera may give false-negative results in IFA and ELISA.

Antibodies, Viral↗
PubMed1991

Rapid diagnosis of viral respiratory infections. Comparison between immunofluorescence on clinical samples and immunofluorescence on centrifuged cell cultures.

We compared the technique of immunofluorescence on clinical samples (IFCS) and immunofluorescence on centrifuged cell cultures (IFCC) for the detection of respiratory viruses, using monoclonal antibodies. Four hundred fourty five nasopharyngeal aspirates were tested prospectively for RSV, influenza- (A and B), parainfluenza viruses (1, 2, 3) and adenovirus. IFCS and IFCC detected 94% and 51% of RSV respectively, 78.5% and 82% of influenza viruses, 8.3% and 100% of parainfluenzaviruses and 0 and 100% of adenoviruses. We conclude that for routine rapid virus diagnosis the IFCS procedure is acceptable for RSV, that for influenza virus the IFCC procedure can be omitted for samples positive by IFCS, and that for the detection of parainfluenza and adenoviruses the IFCS procedure is too insensitive.

Cells, Cultured↗
PubMed1978

Tissue localization of zinc glycinate marker and carcinoembryonic antigen by immunofluorescence. II. Immunofluorescence microscopy.

Preliminary indirect immunofluorescence studies on the zinc glycinate marker (ZGM) were compared with carcinoembryonic antigen (CEA) immunofluorescence, ZGM, detected in 26 of 29 human colon adenocarcinomas, was associated with the epithelial component of the malignant glands. Fluorescence was generally less strong and more granular for ZGM than for CEA and was found in intraglandular spaces, luminal border areas, and cytoplasm. ZGM concentration and tissue localization appeared to be related to tumor differentiation. ZGM was also detected in benign colon mucosae (adjacent to and distant from the carcinomas) from patients with colon carcinoma, but differed from CEA in that it was present in the deep crypt portion only. Gastric, pancreatic, esophageal, and anal adenocarcinomas, as well as benign gastric pyloric and small bowel mucosae had detectable ZGM. CEA, but not ZGM, was observed in 20 nongastrointestinal carcinomas to date. Studies are under way to determine whether ZGM is a marker associated with colon and gastrointestinal adenocarcinoma specifically or undifferentiated crypt cells of the colon and digestive tract in general.

Adenocarcinoma↗
PubMed1980

Studies in cutaneous immunofluorescence. I. The effect of storage time on direct immunofluorescence of skin biopsies from bullous disease and lupus erythematosus.

Forty-eight biopsy samples submitted to a diagnostic immunofluorescence service have been stored at -20 degrees C and sequentially re-tested at monthly intervals to ascertain the duration for which immunoglobulins and complement can be identified in these samples in typical and diagnostic patterns of distribution. Four biopsies from pemphigus vulgaris, 14 biopsies from bullous pemphigoid, nine biopsies from dermatitis herpetiformis and 21 from chronic discoid lupus erythematosus have been studied in this manner. In all cases diagnostic immunofluorescence patterns were present up to 4 months after biopsy and in several biopsies a diagnostic pattern was observed at 9 months. It is concluded that adequate storage facilities for a diagnostic immunfluorescence service can be achieved with a -20 degrees C storage system and that known positive material can be retrieved from this system for use as controls or in research studies for a minimum of 4 months.

Biopsy↗
PubMed1999

Comparison of sensitivity of immunofluorescent microscopy to that of a combination of immunofluorescent microscopy and immunomagnetic separation for detection of Cryptosporidium parvum oocysts in adult bovine feces.

A direct immunofluorescence assay (DFA) (Merifluor; Meridian Diagnostics, Inc., Cincinnati, Ohio) was compared to an immunomagnetic separation (IMS) assay (Dynabeads; Dynal, Inc., Lake Success, N.Y.) coupled with immunofluorescent microscopy (Waterborne, Inc., New Orleans, La.) for their ability to detect low concentrations of Cryptosporidium parvum oocysts in adult bovine fecal material. IMS-DFA resulted in a 2-log-unit increase in sensitivity (10 oocysts/g) compared to DFA alone (1,000 oocysts/g). The higher sensitivity obtained with IMS-DFA resulted from testing 2 g of fecal material instead of the 13 to 19 mg of fecal material tested in the DFA; the increased sensitivity was not attributable to a higher percent recovery.

Animals↗
PubMed1988

Detection of benzo[a]pyrene-DNA adducts in cultured cells treated with benzo[a]pyrene diol-epoxide by quantitative immunofluorescence microscopy and 32P-postlabelling; immunofluorescence analysis of benzo[a]pyrene-DNA adducts in bronchial cells from smoking individuals.

Monoclonal antibodies were raised against the reaction product of benzo[a]pyrene diol-epoxide (BPDE) and deoxyguanosine-5'-monophosphate. The antibodies were used for detection of DNA adducts in situ in BPDE-treated cultured human fibroblasts by immunofluorescence microscopy. Analogue-digital conversion of the fluorescence signal and further image processing allowed measurement of the immunospecific fluorescence in the nuclei of these cells. The results are compared with the adduct levels measured in isolated DNA by 32P-postlabelling. Preliminary results are shown of the application of the immunofluorescence method to the analysis of DNA adducts in bronchial cells obtained from smoking individuals.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1977

Antitumor antibodies in human breast cancer sera as detected by fixed cell immunofluorescence and living cell membrane immunofluorescence assays.

The reactions of patients' sera with cultured human breast cancer cells were studied by fixed cell immunofluorescence (FCF) and living cell membrane immunofluorescence (MF) tests. The results suggested that the FCF reaction detected antibodies that were cell or organ specific, whereas the MF test was more indicative of an antineoplastic immune response.

Adenofibroma↗
PubMed1975

Uses for immunofluorescence tests of skin and sera. Utilization of immunofluorescence in the diagnosis of bullous diseases, lupus erythematosus, and certain other dermatoses.

Sera and frozen biopsy specimens taken for immunofluorescence can be of diagnostic value in studies of vesiculobullous eruptions and connective tissue diseases. Appropriate handling of specimens is important. In cases of pemphigus and pemphigoid, findings of typical antibodies in sera and immunoglobulin deposits in skin sections serve to establish the diagnoses. Fluctuations in titers of pemphigus antibodies afford a prognistic guide. IgA deposits at the dermoepidermal junction in patients with vesiculobullous eruptions are diagnostic of dermatitis herpetiformis. Elevated titers of antinuclear antibodies, frequently with a peripheral pattern, and DNA antibodies as well as junctional deposits of immunoglobulins or complement or both in the apparently uninvolved skin are highly characteristic of systemic lupus erythematosus, more or less typical patterns of immunoglobulin and complement deposits in the lesion appear in discoid lupus erythematosus, porphyrias, and some other dermatoses.

Acantholysis↗
PubMed1975

Immunofluorescence and anti-complement immunofluorescence absorption tests for quantitation of Epstein-Barr virus-associated antigens.

Immunofluorescence absorption methods are described which permit quantitative estimation and differentiation of Epstein-Barr virus (EBV)-associated antigens (virus capsid antigen, VCA, early antigen, EA and EBV-determined nuclear antigen, EBNA) in cell extracts. EBNA was present in all cell lines (producer and non-producer) which carried the EBV-genome, while VCA and EA were present in producer lines only. All the antigens were absent from a lymphoid cell line (MOLT-4) which lacked the EBV-genome, as well as from leukemia cells from peripheral blood. The techniques demonstrated antigenic identity of the various antigens when prepared from different cell lines.

Absorption↗
PubMed1981

The platelet adhesion immunofluorescence test: a modification of the platelet suspension immunofluorescence test.

Platelets can adhere to surfaces by forming pseudopods. This ability is made use of here to simplify conventional platelet suspension immunofluorescence tests, since steps like serum and FITC incubations, washing and microscopic evaluation are done on platelets adhered to glass surfaces. By eliminating time-consuming manipulations like washing in individual test tubes and mounting platelets to microscopic slides, large scale studies can now be performed (up to 480 tests per day and technician). Antibodies can be detected with fresh as well as with frozen platelets from a well defined cell panel. The PAIFT is 2 to 3 times more sensitive than the NIH-standard-micro-lymphocytotoxicity-test (NIH-LCT), thus allowing earlier recognition of alloimmunization of patients.

Female↗