Yaba monkey tumor virus encodes a functional inhibitor of interleukin-18.

Interleukin-18 (IL-18) is a critical proinflammatory cytokine whose extracellular bioactivity is regulated by a cellular IL-18 binding protein (IL-18BP). Many poxviruses have acquired variants of this IL-18BP gene, some of which have been shown to act as viral virulence factors. Yaba monkey tumor virus (YMTV) encodes a related family member, 14L, which is similar to the orthopoxvirus IL-18BPs. YMTV 14L was expressed from a baculovirus system and tested for its ability to bind and inhibit IL-18. We found that YMTV 14L bound both human IL-18 (hIL-18) and murine IL-18 with high affinity, at 4.1 nM and 6.5 nM, respectively. YMTV 14L was able to fully sequester hIL-18 but could only partially inhibit the biological activity of hIL-18 as measured by gamma interferon secretion from KG-1 cells. Additionally, 17 hIL-18 point mutants were tested by surface plasmon resonance for their ability to bind to YMTV 14L. Two clusters of hIL-18 surface residues were found to be important for the hIL-18-YMTV 14L interaction, in contrast to results for the Variola virus IL-18BP, which has been shown to primarily interact with a single cluster of three amino acids. The altered binding specificity of YMTV 14L most likely represents an adaptation resulting in increased fitness of the virus and affirms the plasticity of poxviral inhibitor domains that target cytokines like IL-18.

Serological investigations into the presence of antibodies to Yaba 1--Lednice 110 virus in Romania.

Sera collected from several migratory birds of the Danube Delta, from humans and domestic birds and animals of three Romanian counties were tested by hemagglutination-inhibition reaction for the presence of antibodies to Yaba 1--Lednice 110 virus. Antibodies could be detected in only 8 of the 14 species of migratory birds included in the study, especially in wild geese and ducks.

Surface membrane redistribution and stabilization of concanavalin A-specific receptors following Yaba tumor poxvirus infection.

Monkey kidney cells productively infected with Yaba tumor poxvirus clearly exhibit plasma membrane alterations when treated with both fluorescein-labeled and unlabeled concanavalin A. The convanavalin A-mediated cytoagglutination reaction for Yaba-infected Jinet and CV-1 cells increased linearly from 12 to 16 h post-infection, reaching a maximum by 24-28 h. Treatment of either Yaba-infected CVC-1 or Jinet cells with methyl-D-glucopyranoside before or after addition of concanavalin A completely blocked or reversed the cytoaglutination response. Trypsin treatment of uninfected CV-1 or Jinet cells enhanced concanavalin A-mediated cytoagglutination properties. Conversely, trypsin treatment of Yaba-infected Jinet cells resulted in a reduced cytoagglutination response. Increasing temperature and lectin concentration enhance concanavalin A-mediated cytoagglutination for uninfected, trypsin-treated and Yaba-infected CV-1 cells. Cytosine arabinoside has little or no effect on the Yaba-induced cell cytoagglutination reaction while cycloheximide blocks the cytoagglutinatin response if added prior to 12 h post-infection. Fluorescein-labeled concanavalin A binding studies have revealed that at 4 degrees C, Yaba-infected CV-1 cells display a predominantly 'patchy' pattern of topological fluorescence, while trypsin-treated and uninfected CV-1 cells at 4 degrees C display a uniform pattern of fluorescence binding. Patchy fluorescence, indicative of concanavalin A-suspeptible, receptor-site clustering on the surface membrane, was reduced 50% if Yaba-infected CV-1 cells were treated with glutaraldehyde (2.5%) before addition of fluorescein-labeled concanavalin A at 4 degrees C. Similar pre-fixatin of trypsin-treated CV-1 cells resulted in uniform, fluorescent labelling patterns at all assay temperatures.

Cultivation of Lednice (Yaba 1) virus in goose, duck, and chick embryo cells.

Lednice (Yaba 1) virus was found to multiply in primary cultures of goose, duck and chick embryo cells. The size of inoculum and age of embryos used for preparation of primary cultures was decisive for the amount of multiplied virus. One strain (110) of Lednice virus showed reduced ability to induce a cytopathic effect in the first passage in chick embryo cells. Virus multiplication in all three kinds of cells was confirmed by immunofluorescence.

Experimental infection of Macaca mulatta monkeys with Lednice (Yaba 1) virus.

Macaca mulatta monkeys were inoculated with Lednice (Yaba 1) virus strain 6118. The animals developed no clinical signs and the body temperature and values of complete haematological examination remained within normal limits. In spite of positive immunofluorescence in regional lymph nodes, no virus could be recovered from them by passages in mice. Viraemia was not demonstrated. The antibody response was directly proportional to the amount of virus inoculated, i.e. it was significant after a higher virus dose or after repeated inoculation. The results obtained are discussed in respect of and in comparison with serological findings in man.

Studies on viremia and antibody formation in ducklings and goslings after experimental infection with Lednice (Yaba 1) virus.

A very low virus level was found for a short time in the blood of ducklings and goslings after experimental Lednice (Yaba 1) virus infection by subcutaneous (scut) and intranasal (inas) route and in blood of ducklings also after infection by intracerebral (icer) route. Subsequently virus neutralizing antibody formation was examined. Antibody formation was found in most of animals. In goslings onset of antibody formation seems to be slower than in ducklings after the same virus infection dose; vice versa, in ducklings antibody titer decreased after 6 weeks. Using low infection virus dose, neutralizing antibody level in ducklings was lower than in the former case. On examining the blood and the organs, the virus was detected in a very low amount and for a very short time in blood and spleen of goslings and only in blood of ducklings.

Serologic examination of birds from the area of Southern Moravia for the presence of antibodies against arboviruses of the groups alfa, flavo, Bunyamwera supergroup, and the virus Yaba 1-Lednice 110. I. Domestic fowls.

196 geese, 141 ducks, and 100 hens from 9 localities of the South Moravian region (District of Breclav) were examined by means of the haemagglutination-inhibition test (HIT) for the presence of antibodies against 12 arbovirus antigens of the groups Alfavirus (Western Equine Encephalitis (WEE), Eastern Equine Encephalitis (EEE), Semliki, Sindbis, Chikungunya, O'nyong-nyong), Flavovirus (Tick-borne Encephalitis (TBE), Dengue and West Nile Virus (WN), Bunyamwera Supergroup (Tahna and Beta Calovo), and the Yaba 1-Lednice 110 Virus. In tested fowls antibodies were mostly found against Yaba 1-Lednice 110 Virus, namely 3.6% in geese and 17.7% in ducks. Antibodies against Calovo Virus were found in only 1 of the tested ducks. All sera of hens were negative. Antibodies against arboviruses of the Alfa-and Flavovirus groups were not detected.

Distribution of Lednice (Yaba 1) virus in the chick embryo.

Distribution of Lednice (Yaba 1) virus antigen (LVA) was followed by immunofluorescene (IF) in chick embryos inoculated into the yolk sacs. Positive fluorescence of LVA was observed in neurons and neuroblasts of the developing brain, spinal cord and spinal ganglia as well as in skeletal muscles, heart muscle, vascular endothelium and lung mesenchyma. In the yolk sac, foci of specific fluorescence were occasionally seen in endothelium cells of vessels and in islands of extraembryonic haematopoesis. At sites corresponding to the occurrence of LVA, extensive oedema was accompanied by extravazation of erythrocytes and accumulation of white blood cells. The nature of tissues in which the virus replicates was discussed from the point of view of LVA distribution and the morphological lesions observed.

Serological evidence of infection with Tana and Yaba pox viruses among several species of monkey.

Sera from cynomolgus monkeys from Malaysia, from Indian rhesus monkeys, from various species of monkeys from Africa and from South America have been examined for neutralizing antibody to Tanapox and Yaba viruses. No antibody was found to either virus in the sera of rhesus monkeys or South American monkeys. A certain proportion of sera from cynomolgus monkeys and various species of African monkey showed antibody to one or other of the viruses, but few of the positive sera showed antibody to both. The results would seem to suggest that infection with the two viruses is endemic in African and Malaysian monkeys but does not occur or is very rare in Indian rhesus and New World monkeys.

Microcomplement-fixation inhibition: a rapid and economical test to detect non-complement-fixing antibodies.

Complement-fixation inhibition test was adapted to the microtiter system. When cat and monkey non-complement-fixing antibody were used, the microtest was shown to be as sensitive as the conventional tube complement-fixation inhibition test.

Differences in clinical and convalescent-phase antibodies of Rhesus monkeys infected with monkey pox, tanapox, and Yaba poxviruses.

Complement-fixing and complement-fixing inhibiting (CFI) antibodies were demonstrated in the clinical and convalescent stages, respectively, of rhesus monkeys infected with either monkey poxvirus, Tanapoxvirus, or Yaba poxvirus. Specificity of the CFI antibody was confirmed by its failure to cross-react with heterologous poxvirus antigens and by experiments demonstrating the CFI test as being antigen dependent. Serum containing CFI antibody neutralized homologous poxvirus but failed to agglutinate antigen-coated, tanned red blood cells. The application of CFI test as a seroepidemiologic tool for studies of poxvirus infection of man and simian monkeys and the biologic role of CFI antibody in pathogenesis were discussed.

Sero-epidemiological survey on Yaba and 1211 virus infections among several species of monkeys.

The distribution of neutralizing antibody to Yaba virus and 1211 agent in the sera of three Asian and one African monkey species was examined.Cynomolgus (Macaca irus), bonnet (M. radiata) and rhesus (M. mulatta) monkeys possessed antibody to Yaba virus at incidences of 19.9, 8.4 and 0%, respectively. In African green monkeys (Cercopithecus aethiops) the incidence was as high as 76.4%.As for 1211 agent, no Asian monkeys had neutralizing antibody and 5.5% of African green monkey sera neutralized the virus.