Effect of IFN-γ receptor gene deletion on vaccinia virus virulence

Summary.  Two vaccinia virus (VV) strains, WR and Praha, were selected for a study undertaken to determine whether the virus-encoded interferon-γ receptor (IFN-γR) plays any role in virus virulence. Both of the viruses expressed the B8R gene coding for IFN-γR in infected cell cultures. The nucleotide sequence of the Praha virus B8R gene was determined, and, when compared with the published sequence of the WR virus, it only displayed one silent nucleotide substitution. Mutants of the WR and Praha viruses with deleted B8R gene were constructed. In rabbits, skin lesions produced by the WR B8R-deleted mutants were smaller and tended to disappear earlier than those caused by wild-type WR virus. Similar results were obtained with both independently prepared WR B8R-deleted mutants. These data strongly suggested that the product of B8R gene did play a role in virus virulence. A similar comparison of the wild-type Praha virus and its mutant could not be done because of the very low virulence of the parental virus for rabbits. Received March 13, 2000 Accepted August 16, 2000     

Effects of melanotan II,a melanocortin agonist,on grooming and exploration in rats after repeated restraint/immobilization

2 mg/kg melanotan II (MTII, administered i.p.), a cyclic peptide analog of α-melanocyte stimulating hormone, at a single dose increased grooming in naive rats placed in an unfamiliar open-field device without changing locomotion or rearing. Male rats exposed to restraint/immobilization stress (IS) for 1 h on three consecutive days displayed increased grooming after the second stressor exposure, compared to pre-stress levels. MTII, administered to the rats after IS, enhanced the grooming response compared both to the pre- and post-stress values. The increase was greatest after the first dose and declined over the following two applications. As to the locomotion of rats in the entire experimental space, IS reduced the distance moved only after the first two stressor exposures; MTII did not influence these alterations. Locomotion in the central part of arena was not reduced by the stressor or by MTII, on the contrary, there was an increase in both groups after the third intervention. The only observed change in rearing was an increase in the MTII group after the third restraint exposure. Thus, MTII selectively increased grooming without markedly affecting the spatio-temporal structure of locomotor behavior in the open-field. The decline of MTII enhanced grooming over the three test days may be interpreted in terms of adaptation to the stressor and of the developing tolerance to the peptide.     

Enzymes in polymer chemistry, 5. Radical polymerization of different 11-methacryloylamino-undecanoic acid esters and oligoesters esterfied by lipases

11-Methacryloylaminoundecanoic acid ( 1 ) was esterified with the monoalcohols isobutyl alcohol, cyclohexanol, DL -menthol, cholesterol, testosterone and an excess of 12-hydroxylauric acid in the presence of lipase as catalyst. The kinetics of the esterification reactions were followed by ¹H NMR spectroscopy and the results were correlated with sterical effects. The monomers were polymerized radically. The monomeric 12-hydroxylauric acid oligoester 11 as well as its homopolymer 12 were characterized by DSC and viscosity measurements.     

Localization of the Antigen and Antibodies in the Placenta of Immunized Sheep

ABSTRACT: Placentomata of sheep immunized with human serum albumin (HSA) were examined. Both HSA and immunoglobulins were found in the maternal part and maternofetal border of the placenta using FITC labelled antisera on paraffin sections. Radiolabelled HSA was also detected in the fetal blood. The ultrastructure of placentomata revealed immunopathological process.     

Ein Vergleich der Glucose-, Ketonkörper- und Glycerinspiegel im Nabelschnurblut und im Placentarblut

Zusammenfassung 1. Die Bestimmung von Ketonkörpern, Glucose und Glycerin getrennt in Nabelartcrie, Nabelvene, Placentararterie und Placentarvene ergab für Glycerin und Ketonkörper im Placentarblut meist etwas höhere Werte als im Nabelschnurblut.2. Es folgt daraus, daß es nicht zulässig ist, aus experimentellen Gründen Placentarblut an Stelle von Nabelschnurblut zu verwenden.3. Durch die Analyse der möglichen Ursachen kamen wir zu dem Schluß, daß es sich bei diesen Veränderungen wahrscheinlich um eine Widerspiegelung des placentaren Stoffwechsels handelt. Hämodynamische Einflüsse jedoch sind nicht auszuschließen.4. Es ist möglich, daß der Vergleich von Placentarblut und Nabelschnurblut unter normalen und pathologischen Bedingungen zum Studium des Stoffwechsels der Placenta in situ einen Beitrag liefern kann.

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Summary 1. The separate determination of ketone bodies, glucose and glycerol in umbilical cord artery and vein, placental artery and vein resulted in elevated levels of glycerol and ketone bodies in placental blood compared to cord blood.2. It is concluded that for experimental purposes cord blood is not to be replaced by placental blood.3. In analysing the possible facts we drew the conclusion that these results indicate a representation of the placental metabolism. Hemodynamic factors were not to be excluded, however.4. The comparison of placental with cord blood under normal and pathological conditions will possibly add to the study of placental metabolism in situ.

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Herrn Prof. Dr.G. Joppich zum 65. Geburtstag gewidmet.

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Die Arbeit wurde von der Deutschen Forschungsgemeinschaft unterstützt (Wo 69/7).

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Stipendiat der Alexander von Humboldt-Stiftung, 1967/68.     

Reactivation of Epstein-Barr virus in Sj?gren''s syndrome

Summary Sjögren's syndrome (SS) is a chronic autoimmune disease characterized by severe dryness of the eyes and mouth, resulting from lymphocytic infiltration of the lacrimal and salivary glands. SS may exist as a primary condition (primary SS, 1° SS) or as a secondary condition (2° SS) in association with rheumatoid arthritis, systemic lupus erythematosus, or progressive systemic sclerosis. In some 1° SS patients, there may be involvement of the extraglandular organs, including skin, kidney, liver, lung and nervous system. Furthermore, these patients may develop a lymphoproliferative syndrome that includes lymphadenopathy and increased risk of lymphoma. In the pathogenesis of SS, a role for Epstein-Barr virus (EBV) has been suggested because: (a) EBV is present in salivary gland epithelial cells of normal individuals and exaggerated immune responses against EBV could play a role in the destruction of salivary glands in SS; (b) SS salivary gland biopsies contain increased levels of EBV DNA in comparison to normal salivary glands, indicating viral reactivation and inability of lymphoid infiltrates to control EBV replication in SS patients; and (c) salivary gland epithelial cells in SS patients express high levels of HLA-DR antigens and may present EBV-associated antigens to immune T cells in SS patients. Therefore, SS may represent a situation in which genetically predisposed individuals (i. e., HLA-DR3-DQA4-DQB2) have a persistent but ineffectual T cell immune response against EBV at its site of latency. Among 14 non-Hodgkin's lymphomas that developed in SS patients, EBV DNA was detected in increased amounts in the tumor tissue of one patient. Characterization of this tumor DNA revealed: (a) polyclonal immunoglobulin gene rearrangements; (b) EBV DNA with an unusual restriction fragment length polymorphism pattern involving the Bam M fragment; and (c) EBV terminal repeat sequences suggestive of viral replication, similar to those reported in EBV lymphomas occurring in other immunocompromised individuals. Early recognition of this clinical problem may allow beneficial use of antiviral agents.     

Monoclonal antibodies against transferrin. Precipitating mixtures and lack of inter-species cross-reactivity

Five stable hybridoma lines were prepared using the myeloma cell line P3-X63-Ag.653 and spleen cells of mice hyperimmunized by pig transferrin. All hybridomas grew well in mouse peritoneal cavity and produced antibodies of the IgG₁ subclass. Antibody preparations obtained from ascitic fluids tested for their capacity of antigen precipitation. No precipitation was obtained with single antibodies and with pairs of antibodies. Three out of 10 possible triads gave clear and sharp precipitation zones and rings in immunodiffusion tests performed in agar gel. All 5 antibodies were shown by quantitative enzyme-immunoassay to be specific for pig transferrin: no cross-reaction was obtained with mouse, human, horse and sheep transferrins.     

Autoimmunity to Polymorphonuclears: Functional Consequences of the Binding of Antibodies to Membrane and Cytoplasmic Target Antigens of Polymorphonuclear Leukocytes

Antineutrophil autoantibodies reacting with cytoplasmic antigens are associated with various types of vasculitides, whereas antibodies reacting with neutrophil membrane antigens are mostly related to autoimmune neutropenias. The aim of this study was the investigation of the effect of monoclonal antibodies (MoAbs) reacting with surface and cytoplasmic antigens of polymorphonuclear leukocytes (PMN) known to be targets for autoantibodies in human diseases. Blood of healthy volunteers was tested for several phagocytic functions in the presence of MoAbs against surface (CD16, GD11b, CD18, NB1) and cytoplasmic (proteinase 3; PR3) molecules. Candidacidal activity was significantly inhibited in the presence of all MoAbs but isotypic control. Phagocytic activity was inhibited by anti-CD11b and/or anti-CD18 MoAbs. Zymosan-induced chemiluminescence was reduced by MoAbs anti-CD16, CD18, and NB1, enhanced by anti-PR3 MoAb, and less enhanced by anti-CD11b. In conclusion, antimembrane antibodies diminished phagocytic functions at multiple steps; in contrast, anticytoplasmic MoAb promoted activation of oxidative burst in addition to impairment of microbicidal activity. This fact may be related to different pathogenic aspects of diseases associated with antimembrane and anticytoplasmic antibodies.