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PubMed1976

Symptomatology and diagnosis in connective tissue disease. Antibodies to extractable ribonucleoprotein in 123 patients reacting with cell nuclei in the immunofluorescence test.

Sera with an antinuclear immunofluorescence titre of 1/000 were taken consecutively from the diagnostic routine flow and examined for agglutinating antibodies against desoxyribonucleic acid (DNA) and extractable nuclear antigens (ENA). Passive haemagglutination tests with antigen-coated tanned erythrocytes were used and the specificity of the reactions was corroborated by testing against enzyme-treated cells. After the exclusion of the DNA-reacting 15%, three major groups and one minor could be distinguished on a serological basis. The largest group (41%) contained cases with a speckled immunofluorescence pattern and a RNase-trypsin sensitive agglutination reaction with ENA coated cells (sRNP). Nearly all cases of mixed connective tissue disease and scleroderma fell into this group which also contained 44% of the SLE cases. Symptomatically the group was characterized by remarkably high incidences of Raynaud's syndrome and myositis. The major group next in size comprised cases with a homogeneous immunofluorescence pattern but no reaction against DNA or ENA. Half of the cases within this group had the diagnosis SLE; they also constituted 42% of all SLE cases. The only other diagnosis of significant frequency within the group was unspecified collagenosis (23%). The symptomatology of the group was rather uncharacteristic, with the exception of the low incidence of Raynaud's syndrome. The third major group comprised cases with a speckled immunofluorescence pattern but no agglutination reaction against ENA or DNA. This group had a very high incidence of rheumatoid factor and also the highest incidence of visceral lesions among the groups. Yet the group contained only a small proportion (14%) of the SLE cases and the rheumatoid arthritis cases were about equally shared between this and the first group. The most common diagnosis in the group was unspecified collagenosis (40%). A fourth, small but homogeneous group contained cases with a speckled immunofluorescence pattern and a reaction with Sm antigen, i.e. an enzyme-resistant agglutination reaction with ENA. Six cases in this group had the diagnosis SLE. No diagnosis was available in two cases.

Antibodies, Antinuclear↗
PubMed1976

Immunofluorescent detection of adenovirus antigen in epidemic keratoconjunctivitis.

An immunofluorescent technique was used to demonstrate soluble adenoviral antigens in epithelial cells on conjunctival scrapings of patients with epidemic keratoconjunctivitis (EKC). Conjunctival scrapings for immunofluorescence and viral isolation studies were performed on 79 patients suspected of having EKC or other acute follicular or papillary conjunctivitides. Of 41 patients with clinical findings consistent with a diagnosis of EKC and three patients with pharyngoconjunctional fever, 43 were positive by immunofluorescence. All 39 patients with adenovirus isolation had positive immunofluorescence studies. The five remaining cases had other documented evidence of EKC. No false-positive responses were encountered in relation to the clinical diagnoses. The fluorescent staining was predominantly found in the cytoplasm. Speckled nuclear fluorescence was also noted. The cytoplasmic fluorescence is compatible with the adenovirus cellular replication cycle. The immunofluorescent technique was found to be a reliable, sensitive, specific, and rapid diagnostic technique for detection of group-reaction adenoviral antigens in conjunctival scrapings.

Adenoviridae↗
PubMed1977

[Immunofluorescent method of studying autopsy material in acute pneumonia].

The immunofluorescent procedure in examinations of the autopsy material from 304 fatal cases hospitalized for acute pneumonia permitted to diagnose influenza A2 in 29.0%, influenza B in 18.1%, parainfluenza in 3.5%, adenovirus infection in 9.2% and respiratory syncytial virus infection in 3.5% of the cases. In the period of a high incidence of acute respiratory infection, influenza A2 was detected by this method in 40.9% and influenza B in 50% of the cases. Simultaneous examinations of the material in the influenza epidemic period by virological and immunofluorescent methods (63 cases) in 13 cases positive results were obtained with both methods, in 6 cases where influenza viruses were detected the immunofluorescent test was negative, and in 28 cases the positive diagnosis by the immunofluorescent test could not be confirmed virologically. Among the cases examined, 33 were found by the immunofluorescence test to have a mixed respiratory infection, including influenza A2 with other forms of respiratory infection in 18, and influenza B with other respiratory infections in 19 cases. Serological examinations by the complement fixation and hemagglutination inhibition tests on the blood from fatal cases irrespective of the time of examination, as a rule, revealed antibody in low titres which did not confirm the diagnosis.

Acute Disease↗
PubMed1977

Skin basement membrane immunofluorescence in rheumatoid arthritis: lack of diagnostic correlation.

Thirty-nine patients with rheumatoid arthritis were studied for the presence of skin basement membrane immunofluorescence. Punch biopsies from normal sun-exposed skin of the forearm were negative for basement membrane immunofluorescence in all cases, except one which was read as questionable. No correlation with serum antinuclear antibody or lupus erythematosus cells was observed. Skin immunofluorescence studies are helpful in differential diagnosis when patients with a clinical picture of rheumatoid arthritis present with serum antinuclear antibodies and lupus erythematosus cells. Positive basement membrane immunofluorescence is strong evidence of systemic lupus erythematosus.

Adolescent↗
PubMed1978

Identification of young megakaryocytes by immunofluorescence and cytophotometry.

The DNA-content of fluoresceine-labeled platelet antigen containing cells of mouse bone marrow was measured. For immunofluorescence highly specific anti-mouse-platelet-serum and fluoresceine-conjugated antigammaglobuline was used, applying the "sandwich" technique. Three hundred panoptically identifable megakaryocytes served as control group. The DNA-polyploidization pattern of megakaryocytes and immunofluorescence positive cells was almost identical. However, among the immunofluorescence positive cells a considerable amount of cells showed DNA-values lower than 4c, whereas the megakaryocytes of the Pappenheim stained smears revealed no DNA-values lower than 4c. The percentages of diploid and tetraploid cells, respectively, was 6 and 7% compared with 0 and 1% of panoptically identifiable megakaryoctyes. The results suggest that young megakaryocytic cells with diploid and tetraploid DNA-values can be detected by immunofluorescence technique, indicating that the flow from the uncommited to the committed megakaryocytic precursor cell appears at this early stage of megakaryocyte production.

Age Factors↗
PubMed1977

Effects of an immunofluorescent transport solution on the ultrastructure of human biopsies.

Although immunofluorescent tests have been demonstrated to be of value in the diagnosis of many oral and skin lesions, the use of this technique has until recently been cumbersome because of the requirement for frozen tissue specimens. This has been circumvented by the development of a solution in which tissue can be transported to the testing laboratory at ambient temperatures. To verify that the immunofluorescent findings in gingiva and skin kept in this solution are not artifactual, ultrastructural studies were performed on such tissue. These revealed that over-all tissue architecture, individual cells, and the epidermal-connective tissue interface remain intact. Since the majority of oral lesions submitted for immunofluorescent testing are suspected of being pemiphigus or pemphigoid, the finding that these tissue features remain intact indicates that the distribution of immunofluorescence is related to definite morphologic structures and provides additional evidence for the validity of using the transport solution.

Ammonium Sulfate↗
PubMed1977

Utility of protease-digested human peripheral blood lymphocytes for the detection of lymphocyte-reactive alloantibodies by indirect immunofluorescence.

Human peripheral blood lymphocytes were digested briefly with protease prior to application of indirect immunofluorescence techniques for detecting alloantibodies in sera of patients with chronic renal failure on maintenance hemodialysis. Background staining of intrinsic surface IgM and cytophilic IgG bound to Fc receptors was eliminated or greatly reduced, enabling detection of B cell specific antibodies, including cold-reactive types not demonstrable by conventional immunofluorescence or complement-dependent lymphocytotoxicity. The antigenicity of HLA and other surface membrane determinants was not decreased by protease, although reactivity with certain sera was enhanced. In experiments comparing indirect immunofluorescence using protease-treated cells with complement-dependent lymphocytotoxicity and antibody-dependent, lymphocyte-mediated cytotoxicity assays, indirect immunofluorescence was more sensitive and comprehensive, but not less specific, in defining alloantibodies of a variety of types.

Cytotoxicity Tests, Immunologic↗
PubMed1978

An immunofluorescence study of pityriasis lichenoides.

In a study of 27 patients with pityriasis lichenoides IgM and C3 have been observed on direct immunofluorescence of fresh lesions. Other immunoglobulins and complement components were not observed. Immunofluorescence was seen in 31 (72%) of 43 fresh lesions. It occurred in the walls of superficial dermal vessels and along the dermal-epidermal junction. This pattern of immunofluorescence appears to be characteristic of the disease. Uninvolved skin showed the immunofluorescence less frequently and old scaly lesions none. The concept that pityriasis lichenoides is an immune complex disorder is discussed.

Complement C3↗
PubMed1977

Immunofluorescence of the skin in gold rashes - with particular reference to IgE.

Immunofluorescence studies have been carried out on rashes from 36 patients with rheumatoid arthritis receiving gold therapy. 24 of the rashes were clinically attributed to gold and 12 were diagnosed as coincidental rashes. IgE was found in 6 of the gold rashes and in 4 of the coincidental rashes. Immunofluorescence changes of immune complex vasculitis, lichen planus, or pemphigoid were found in 9 gold rashes while 2 coincidental rashes showed vascular fluorescence for immunoglobulins but nor for complement. Two definite gold rashes showing no changes on immunofluorescence showed perivascular infiltration with lymphocytes on light microscopy. Thus, while immunofluorescence is only marginally helpful in the diagnoses of gold rashes, evidence of an immunological reaction tends to favour a diagnosis of a gold-induced rash.

Arthritis, Rheumatoid↗
PubMed1978

[Identification of various porcine mycoplasmas using epi-immunofluorescence].

The responses of the field strains and type strains of Mycoplasma hyorhinis are compared in epi-immunofluorescence, in growth-inhibition tests, and growth-precipitation tests. The type strains, together with the field strains, could be definitely identified by direct and indirect epi-immunofluorescence. In indirect epi-immunofluorescence, all rabbit antiserums against the strains of Mycoplasma hyorhinis tested by us always gave positive cross reactions with all the tested strains of Mycoplasma hyorhinis. On the other hand, in the growth-inhibition cross test and growth precipitation cross test, the type strains of Mycoplasma hyorhinis always reacted positively only with the homologous antiserums, whereas the heterologous reactions were mostly negative or dubious. The clearly positive epi-immunofluorescent reactions with serums against any strain of Mycoplasma hyorhinis suggest that this method is more suitable for the identification of Myocplasma species than the other two tests studied.

Animals↗
PubMed1975

Production of ferritin by rat hepatoma cells in vitro. Demonstration of protein subunits and ferritin by immunofluorescence.

Using precipitating antibodies to ACI rat liver ferritin and to sodium-dodecyl-sulfate-dissociated protein subunits of ACI rat liver ferritin, we have demonstrated the presence of ferritin-positive sites and subunit-positive sites in situ in several rat hepatoma cell lines by immunofluorescence. Hepatoma cells from three transplantable rat hepatomas (Reuber H-139, Reuber H-35, and Morris 5123) were explanted and propagated. Rabbit antibodies specific for either protein subunits of ferritin or ferritin were prepared by affinity chromatography or by dissociation of antibody-antigen complexes with 0.1 M acetic acid followed by differential ultracentrifugation. Explants of Reuber H-139, Reuber H-35, and Morris 5123 hepatoma cells, grown either in ordinary McCoy's 5a medium or in such medium enriched with iron (0.002% Fe), gave positive immunofluorescence for subunits as well as ferritin. Exposure of a clonal strain of Morris 5123 hepatoma cells to iron-enriched culture medium for varying lengths of time of up to 24 hours resulted in progressive increase in the quantity of ferritin-specific immunofluorescent cytoplasmic material, which was at first present diffusely, and later in clumps. By contrast, during the initial 24-hour period, subunit-specific immunofluorescence remained at relatively low intensity, with diffuse distribution through the cytoplasma. Our findings indicate a) the presence, in the cytoplasm, of the three kinds of hepatoma cells, of unassembled or only partly assembled subunits of fragments of subunits as well as of ferritin, and b) rapid assembly of the protein subunits into apoferritin and ferritin after administration of iron, so that the concentration of subunits in the cytoplasm was not significantly increased.

Animals↗
PubMedDate not supplied

[Recognition of influenza and other acute respiratory virus infections and their combinations by means of immunofluorescence].

The results of postmortem examinations performed in 237 fatal cases of influenza and other acute respiratory diseases are presented. In smears of organs and tissue culture cells infected with these materials and heart blood the immunofluorescent procedure detected influenza virus antigens in 67% of the cases, antigens of other respiratory viruses in 53% including 18-19% of mixed infections. Parallel immunofluorescent studies in living patients and isolation of influenza virus and adenoviruses increased per cent of influenza case confirmation to 82%, other respiratory infections to 63%. Mixed infection was recorded in 22-28%. Antigens of influenza and parainfluenza viruses were more frequently found in smears of bronchi and trachea, antigens of respiratory syncytial virus and adenovirus in lung smears. Adenovirus antigen was also found in lymphoid cells of smears of the spleen, bronchial lymph node, tonsils. In immunofluorescent examinations of tissue cultures infected with the heart blood, virus antigen was detected most frequently in influenza and adenovirus infection (24% and 26% of cases, respectively), less frequently in respiratory syncytial virus infection (11%) and parainfluenza (5%). Virus antigens were also demonstrated in tissue culture cells infected with specimens of the liver, spleen, bronchial lymph node, tonsils, brain. The immunofluorescent studies demonstrated long-term persistence of virus antigens in the body. In a portion of cases of laboratory confirmed mixed infection there were no clinical-morphological signs of two virus diseases. In these, one of the infections was latent or was due to a previously experienced disease.

Adenoviridae↗
PubMed1977

Diagnosis of encephalitozoonosis in experimentally infected rabbits by intradermal and immunofluorescence tests.

The indirect immunofluorescence antibody test was performed on serial blood samples from eight young New Zealand White rabbits with experimental encephalitozoonosis. The test showed seroconversion in six of the eight infected rabbits by the 8th day after inoculation and in all rabbits by the 15th day. Antibody titers reached a peak by about the 36th day after inoculation and remained significantly elevated until the termination of the experiment at 84 days after inoculation. None of four sham-inoculated rabbits showed an immunofluorescence response by the 60th day after inoculation. Immunofluorescence and intradermal test responses were compared before infection and at the 60th day after inoculation in a total of 32 experimentally infected rabbits. Both tests were equally effective (100%) in detecting infected animals. Six of eight (first group) and 22 of 24 (second group) experimentally infected rabbits were confirmed histologically to have lesions compatible with encephalitozoonosis. No cross reactions were observed between Encephalitozoon cuniculi and Toxoplasma gondii, Eimeria perforans, or Eimeria stiedai by intradermal test or immunofluorescence test.

Animals↗
PubMed1977

[Diagnostic value of synovial membrane immunofluorescence].

The immunofluorescent study of 93 synovial membranes of persons suffering from various types of arthropathy has shown that the only element sufficiently specific to have real diagnostic value is the presence of cells with a fluorescent cytoplasm. This appearance was in fact found in 63% of the cases of rheumatoid arthritis, irrespective of their being either seropositive or seronegative, in 69% of the cases of probable rheumatoid arthritis, in only 15% of the unclassified cases of arthritis, in 28% of the cases of various types of arthritis (20% with exclusion of a case of mixed connectivitis and of a case of Waldenström's macroglobulinemia with rheumatoid arthritis) and in 0% of the cases of mechanical arthropathy. The results of immunofluorescent examination compare favorably with those of standard histology: the classical histologic appearance of rheumatoid synovitis with a node-forming tendency of the infiltrate was observed in only 36% of the verified cases of rheumatoid arthritis, while immunofluorescence was positive in 63% of the cases in this group. In the category of unclassified arthritis, these percentages were comparable, viz. 19% and 20%, repectively. The presence of cells with fluorescent cytoplasm during immunofluorescent examination of the synovial membrane may be regarded as an additional criterion supporting the diagnosis of rheumatoid arthritis.

Adult↗
PubMed1979

Antibodies to native DNA in connective tissue disease. A comparison of radioimmunoassay, counterimmunoelectrophoresis and indirect immunofluorescence on Crithidia luciliae substrate.

Sera of patients with symptoms of connective tissue diseases were investigated for the presence of antinuclear antibodies by indirect immunofluorescence on rat liver substrate. In positive cases antibodies to native DNA were determined by radioimmunoassay and by counterimmunoelectrophoresis. Twenty seven selected sera were also tested by indirect immunofluorescence on Crithidia luciliae substrate to test the sensitivity and specificity of this method. Immunofluorescent antinuclear antibodies could be found in all groups of connective tissue diseases while anti-native DNA antibody was demonstrated in higher amount only in the cases of systemic lupus erythematosus and therefore determination of anti-native DNA antibody may be helpful in the diagnosis of oligosymptomatic SLE. Counterimmunoelectrophoresis and immunofluorescence on Crithidia smears may serve as screening methods for anti-native DNA while radioimmunoassay provides a quantitative determination, although there are minor differences in the sensitivity and the specificity of these three methods.

Adolescent↗
PubMed1979

Urticaria. An immunofluorescence and histopathology study.

Fourteen randomly chosen patients with "garden variety" urticaria were studied for the presence of vasculitis and immunoglobulins and complement. Results of direct immunofluorescence (DIF) of the involved skin were negative, although two patients had immunoglobulins and complement demonstrable in the cytoplasm of the epidermal cells. Results of DIF of uninvolved skin were also predominantly negative. Findings from serum samples tested by indirect immunofluorescence (IIF) were negative, except for one positive in low titer (1:10, the basement membrane zone). Serum C3 and C4 levels were normal in five patients, both levels were low in two, and the C4 level was low in one patient. No skin-reactive immunoglobulins were found in these three patients by DIF or IIF. The ESR was measured and found to be elevated in four patients. Results of immunofluorescence proved negative in these cases. Of the 12 patients studied by hematoxylineosin staining to determine histology, none exhibited vasculitis. We believe that vasculitis with antigen-antibody reactions is not the rule in "garden variety" urticaria.

Adult↗
PubMed1977

Immunofluorescent localization and ultrastructural characterization of gonadotrophe cells in the adenohypophysis of the barbary drake (Cairina moschata L.) using anti-chicken LH serum.

An indirect immunofluorescence technique and an anti-chicken LH serum were used to localize cells in the adenohypophyses of drakes at different stages of their breeding cycle, after castration, and after castration combined with thyroxine treatment. Immunofluorescent cells were distributed throughout both lobes of the adenohypophyses from control and experimental birds and were shown to be alcian blue positive. PAS negative, basophiles. Immunofluorescent cells were as numerous in castrated birds as in castrated birds treated with thyroxine. Adjacent thin and semi-thin sections were used to study the cells binding anti-LH serum at light microscope and ultrastructural levels. The cells contained spherical granules with variable densities and diameters ranging between 40 and 280 nm in the rostral (=cephalic) lobe, and between 60 and 260 nm in the caudal lobe. The light microscope and ultrastructural observations showed that the anti-LH serum binds to cells which have been classified by other authors in the Pekin duck, quail and pigeon as TSH producing delta cells. The experimental technique used did not permit a distinction to be made between cells producing FSH and LH.

Animals↗
PubMed1975

Immunofluorescent localization of sites binding anti-synthetic LHRH serum in the median eminence of the greenfinch (Chloris chloris L.).

With an indirect immunofluorescence technique and an antiserum raised against synthetic LHRH, immunofluorescent granules were observed in fine radially oriented fibres in the palisade layer of the zoma externa in both the anterior and the posterior divisions of the median eminence in the greenfinch. There was no specific immunofluorescence in the tractus supraoptico-hypophyseus. It is concluded that the greenfinch median eminence contains material which is similar to mammalian LHRH.

Animals↗