Further search for virulence factors encoded by Salmonella serovar-specific plasmids.

The so called Salmonella virulence plasmids which are specifically prevalent among some of the S. enterica serovars were shown to contribute only marginally to the virulence make-up of salmonella, which is in contrast to Shigella and Yersinia spp. The experiments reported in this paper failed to find encoded plasmid factors which contribute to serum resistance, surface antigens, immunoinsufficiency or to up-regulation of chromosomally encoded factors such as toxins, surface antigens etc. Taking into consideration the rare prevalence of these plasmids among S. enterica but their common occurrence among a few of its serovars, their virulence implication remains an enigma.

-Cytokine level in malignant ascites and peripheral blood of patients with advanced ovarian carcinoma-. / Zytokinspiegel im malignen Aszites und peripheren Blut von Patientinnen mit fortgeschrittenem Ovarialkarzinom.

The concentrations of various cytokines were examined by ELISA in blood and in ascites from 14 patients with advanced ovarian cancer (stage IV). The control group consisted of 6 patients with benign gynaecological disorders. Compared with patients with benign gynaecological disorders, ascites and/or plasma of patients with ovarian cancer showed significantly higher levels of IL-6, IL-8, IL-10, TNF-alpha, and sIL-2R. There were no increases of IL-1 alpha, IL-1 beta, IL-2, IFN-gamma, and sCD14 levels. The possible pathogenetic significance of cytokines in ovarian cancer is discussed.

Recombinant Treponema pallidum antigens in syphilis serology.

Treponema pallidum, the etiological agent of syphilis, is characterized by a paucity of surface exposed outer membrane proteins and a high content of cytoplasma membrane associated lipoproteins. At all stages of infection intense antibody responses against lipoproteins are detectable. In order to provide antigens for syphilis diagnosis the highly immunogenic lipoproteins TpN17, TpN29-35 (TpD), TpN44.5 (TmpA), TpN47, and TpN35 (TmpC) and the membrane protein TpN39 (BMP) were cloned. Insertion of PCR amplified DNA into an E. coli expression vector resulted in high level expression of antigens. N-terminal hexahistidine sequence allowed efficient purification of fusion proteins by metal chelate affinity chromatography. The recombinant antigens were tested in enzyme-linked immunosorbent assays. TpN17, TpN47, and TpN44.5 antigens showed high antibody titers. Assays with the three antigens combined resulted in a further improvement of diagnostic sensitivity in comparison with single antigens. Antibodies were found in 17 of 18 patients in all stages of syphilis, whereas 42 normal human sera were nonreactive. No cross-reactivity was detected in 24 sera of patients with Lyme borreliosis.

[Lymphohistiocytic infiltration of endometrial adenocarcinoma]. / Lymphohistiozytäre Infiltration des endometrialen Adenokarzinoms.

Mononuclear cells adjacent to endometrial adenocarcinoma were evaluated in 8 biopsies by the immunoperoxidase technique. Between benign and malignant tissue a remarkable infiltration of CD45+, CD3+, CD4+, CD8+, and CD14+ cells could be observed. B cells (CD19) and NK cells (CD57) were rare. Tumor-infiltrating lymphocytes were only sparsely distributed. The findings suggest a cellular, yet ineffective, immune reaction of the host against the tumor at the borderline between benign and malignant tissue.

[Effect of granulocyte colony stimulating factor (G-CSF) on peripheral blood leukocytes and lymphocytes in patients with chemotherapy-induced leukopenia]. / Der Einfluss des Granulozyten-Kolonie-stimulierenden Faktors (G-CSF) auf die Leukozyten und Lymphozyten des peripheren Blutes von Patientinnen mit chemotherapie-induzierter Leukopenie.

In patients with extensive chemotherapy, G-CSF abrogated leukopenia following administration of cytotoxic agents. Six women with ovarian cancer and chemotherapy-induced leukopenia received 300 micrograms Filgrastrim (r-metHuG-CSF, Neupogen 30; AMGEN, Germany) daily for 10 days. Leukocytes and lymphocyte subsets of peripheral blood were determined before, throughout and after subcutaneous injections of G-CSF by flow cytometry using monoclonal antibodies to CD3, CD4, CD8, CD14, CD16/56, CD19 and CD45. It could be observed that not only neutrophils (23 fold) but also lymphocytes (6 fold) and monocytes (10 fold) showed a dramatic increase in cell counts throughout and after G-CSF administration. This is in contrast to previous reports, where only effects on neutrophils were described. In spite of the increase in lymphocytes the relative percentage of CD3+, CD19+, CD3-CD16/CD56+, CD3+, CD8+ and CD3+ CD4+ lymphocyte subsets did not change throughout and after therapy, except for an increased expression of HLA-DR on CD3+ lymphocytes.

[Leukocyte and lymphocyte populations in peripheral blood and malignant ascites in patients with ovarian carcinoma]. / Leukozyten- und Lymphozytenpopulationen im peripheren Blut und malignen Aszites von Patientinnen mit Ovarialkarzinom.

Ascites and peripheral blood of 12 patients with advanced ovarian cancer (stage IV) have been investigated by two-color flow cytometry for leukocytes and lymphocyte subsets with monoclonal antibodies, against CD3, CD4, CD8, CD14, CD16/56, CD19, CD25, CD45, CD57, and HLA-DR. Ascites compared with blood showed a significant raise of CD3-positive lymphocytes (80 +/- 14% vs. 69 +/- 8%) and a significant reduction of CD57-positive lymphocytes (13.6 +/- 13% vs 24 +/- 21%). There was an increased expression of HLA-DR on CD3-positive lymphocytes in malignant ascites. The results are discussed with regard to a supposedly defective local immune defense against the tumor.

[Determination of thyroglobulin autoantibodies using enzyme immunoassay, radioimmunoassay and passive hemagglutination]. / Untersuchungen zum Nachweis von Thyreoglobulin-autoantikörpern mit Enzymimmuntest, Radioimmuntest und passiver Hämagglutination.

A solid-phase enzyme immunoassay, a radioimmunoassay and the passive hemagglutination were tested for the determination of antithyroglobulin autoantibodies. The antibody-titres detected with these methods showed a good correlation. The sensitivities of the solid-phase immunoassays are four times higher than the sensitivity of the passive hemagglutination. For the diagnosis of thyroid autoimmune diseases the enzyme immunoassay.

[Axial filament antigen enzyme immunoassay (AF-ELISA) in syphilis serology]. / Der Axialfilamentantigen-Enzymimmunassay (AF-ELISA) in der Syphilisserologie.

179 sera from patients with various stages of syphilis were investigated in the axial filament enzyme linked immunosorbent assay (AF-ELISA), the Treponema pallidum haemagglutination assay (TPHA), the fluorescence treponemal antibody absorption (FTA-ABS) test and the cardiolipin micro precipitation test (CMT). For the control of specificity 30 problem sera from non Treponema pallidum infected patients with a high quantity of Treponema genus specific antibodies and 40 sera from people without syphilis were also investigated. The sensitivity of the AF-ELISA (98.3%) was comparable with the TPHA (99.4%) and the FTA-ABS-test (98.9%). It was a high unspecificity in the group of problem sera.

[Characterization of treponemal axial filament antigens using the Western blot]. / Charakterisierung der Treponemen-Axialfilamentantigene im Westernblot.

Analysis by SDS-PAGE of axial filaments of cultivable treponemes showed 3 major bands: a 33/34 kD doublet and a 37 kD polypeptide. In sera from patients with various stages of syphilis, from biological false positive reactors, and from negative controls the most consistent reaction was detected against the doublet, but only sera from patients with primary or secondary syphilis reacted with the 37 kD axial filament polypeptides. About 6 weeks after antibiotic treatment sera had lost their reactivity with 37 kD antigens in agreement with decreasing antibodies in VDRL-test. Because of their recognition early in infection 37 kD-proteins are postulated to use in serodiagnostic assays, but our data emphasize also the need for use further antigens in tests for diagnosis in later stages of syphilis.

[Incidence and significance of humoral antibodies in females with recurrent spontaneous abortions]. / Auftreten und Bedeutung humoraler Antikörper bei Frauen mit wiederholten Spontanaborten.

Serum samples from 70 patients with two or more early spontaneous abortions and, for comparison, sera from 20 healthy and fertile women were assayed by indirect immunofluorescence for the presence of antibodies to sperms and genital organ antigens and for non-organ specific and organ specific antibodies. All sera were tested for thyreoglobulin antibodies by hemagglutination. The frequency of sperm antibodies and autoantibodies was significantly higher in patients with recurrent abortions than in the control group. There was no relation between the type or titer of antibodies and the clinical data of the patients.

[Clinical, immunologic and therapeutic aspects of endangiitis obliterans]. / Klinische, immunologische und therapeutische Aspekte der Endangiitis obliterans.

In the period of 1975-1983 twenty-three patients with thromboangiitis obliterans were examined at the Surgical Clinic of the Medical Academy of Magdeburg. The diagnosis was established clinically, angiographically histologically and immunologically. In 7 out of 12 patients histology revealed inflammatory vascular alterations including lymphocyte infiltrates throughout the entire vessel wall and also in the perivascular area. Immunohistology showed in 10 out of 15 patients segmental granular fluorescence identifiable as deposits of IgM and IgG. In 6 cases a complement formation was found. The endangiitis group revealed more frequent and increased immune complexes concentrations. Circulating immune complexes were established by phase-locked radio-immunoassay. The therapy is depended on the localization of the obliteration and can be a lumbar sympathectomy or a vascular reconstruction.

[Determination of immune complexes in the serum of pregnant patients and puerperal females with EPH gestosis]. / Bestimmung von Immunkomplexen im Serum von Schwangeren und Wöchnerinnen mit EPH-Gestose.

We analyzed the blood sera from 100 patients with EPH gestosis and from 50 women with normal pregnancy to explore the immunological reactions during gestosis. The sera of cord blood were included in the investigations. We determined the circulating immune complexes by conglutinin and C1q solid phase radioimmunoassay and by polyethyleneglycol precipitation. It has been established by conglutinin RIA that during pregnancy the women with gestosis have significant higher concentrations of immune complexes than the control group. During puerperium and in the cord blood the values are significantly lower than during pregnancy in both groups. The estimations of immune complexes by C1q RIA have shown that the differences between the two groups are insignificant and the lowest values are in the cord blood. The concentrations of immune complexes estimated by PEG precipitation were not significantly different between the two groups. It is concluded that immune complexes are in connection with the pathogenesis of preeclampsia.

[Detection of circulating immune complexes in patients with EPH gestosis or intrauterine fetal growth retardation]. / Nachweis von zirkulierenden Immunkomplexen bei Patientinnen mit EPH-Gestose oder intrauteriner fetaler Retardierung.

Sera from 19 patients with EPH gestosis, 22 patients with hypotrophic newborns, and a control group of ten women with normal pregnancy and fetuses were studied for the presence of circulating immune complexes. Both the C1q solid-phase radioimmunoassay and the polyethylene glycol precipitation test revealed significantly higher levels of circulating immune complexes in both groups of patients.

[Detection and differentiation of autoantibodies to extractable nuclear antigens with immunoenzyme tests]. / Untersuchungen zum Nachweis und zur Differenzierung von Autoantikörpern gegen extrahierbare Kernantigene mit Enzymimmuntesten.

Nuclear antigens were extracted from calf thymus with phosphate-buffered saline The 60-80% ammonium sulfate fraction (ASF) of thymus extract contained SS-B-, the 30-60% ASF Sm- and U1-RNP-, and the chromatographically purified 30-60% ASF only Sm antigen. With these fractions enzyme immunoassays were developed for quantitative determination of Sm-, U1-RNP-, and SS-B autoantibodies. Out of 144 sera with antinuclear antibodies 85% were positive in the enzyme immunoassays, 41% in immunofluorescence tests on liver sections (12% speckled, 29% homogeneous immunofluorescence pattern), and 10% in Ouchterlony tests. 26% of the sera were positive in enzyme immunoassays and immunofluorescence tests. All antibody specificities detected by immunodiffusion could be confirmed by enzyme immunoassay. The enzyme immunoassay is far more sensitive then immunodiffusion and in contrast to immunofluorescence allows antibody specificities to be determined. Enzyme immunoassays are recommended for the diagnosis of rheumatic diseases.

[Effect of drugs on superoxide formation by neutrophilic granulocytes]. / Einfluss von Arzneimitteln auf die Superoxidbildung der neutrophilen Granulozyten.

The in-vitro influence of drugs on the superoxide production of human neutrophils was investigated. The superoxide production is inhibited by colchicine, g-strophanthin, propranolol, fluphenazine and stimulated by levamisole and lithium. The superoxide production is inhibited or stimulated by chloramphenicol, hydrocortisone and amphotericin B in concentration dependent manner.

[Pathogenetic mechanisms in autoimmune diseases]. / Pathogenetische Mechanismen bei Autoimmunkrankheiten.

In addition to an earlier review [Z. ges. inn. Med. 35 (1980) 58] newer concepts in the pathogenesis of autoimmune diseases are presented. Acantholysis in pemphigus is caused by proteases released from cells of stratified squamous epithelium stimulated by antibodies to cell membrane antigens. The dermoepidermal cleavage in pemphigoid is presumably due to proteases from granulocytes accumulating at the dermoepidermal junction by antibody-mediated complement activation. Receptor autoantibodies can directly stimulate or block receptor function, increase cell membrane receptor degradation, or activate complement with lysis of receptor-carrying membrane structures. Tissue injury in immune complex diseases is caused by reactive oxygen metabolites and proteases released from stimulated phagocytes.

[Animal experiment studies of the modification of pregnancy by heterologous antibodies to placental antigens]. / Tierexperimentelle Untersuchungen zur Beeinflussung der Schwangerschaft durch heterologe Antikörper gegen Plazentaantigene.

Pregnancy impairment in mice with abortion and fetal resorption was caused by rabbit antibodies to total mitochondrial, mitochondrial membrane and microsomal fractions of mouse placentas injected intraperitoneally at 12-14 days of gestation. Antibodies to cytosol and soluble mitochondrial constituents were ineffective. According to the marker distribution in the biologically active and inactive fractions membrane-bound antigens are thought to be responsible for the abortigenic effect.