The rinderpest virus non-structural C protein blocks the induction of type 1 interferon.

The innate immune response, in particular the production of type 1 interferons, is an essential part of the mammalian host response to viral infection. We have previously shown that rinderpest virus, a morbillivirus closely related to the human pathogen measles virus, blocks the actions of type 1 and type 2 interferons. We show here that this virus can also block the induction of type 1 interferon. The viral non-structural C protein appears to be the active agent, since expressing this protein in cells makes them resistant to activation of the interferon-beta promoter while recombinant virus that does not express the C protein activates this promoter much more than virus expressing the C protein. In addition, differences in activation of the interferon-beta promoter by different strains of rinderpest virus are reflected in differing abilities of their respective C proteins to block activation of the promoter by dsRNA. The C protein blocks the activation of this promoter induced by either cytoplasmic dsRNA or by Newcastle disease virus (NDV) infection, as well as activation induced by overexpression of several elements of the signalling pathway, including mda-5, RIG-I and IRF-3. The RPV C protein also blocks transcription from promoters responsive individually to the three transcription factors that make up the interferon-beta promoter enhanceosome, although it does not appear to block the activation of IRF-3.

Inhibition of host peripheral blood mononuclear cell proliferation ex vivo by Rinderpest virus.

Rinderpest, or cattle plague, is caused by Rinderpest virus (RPV), which is related most closely to human Measles virus (MV), both being members of the genus Morbillivirus, a group of viruses known to have strong immunosuppressive effects in vitro and in vivo. Here, it was shown that peripheral blood mononuclear cells (PBMCs) isolated from cattle experimentally infected with either wild-type or vaccine strains of RPV impaired the proliferation of PBMCs derived from uninfected animals; however, in contrast to either mild or virulent strains of wild-type virus, the inhibition induced by the vaccine was both weak and transient. Flow-cytometric analysis of PBMCs obtained from cattle infected with different strains of RPV showed that the proportion of infected cells was virus dose-dependent and correlated with lymphoproliferative suppression.

Mapping of T-helper epitopes of Rinderpest virus hemagglutinin protein.

Rinderpest virus (RPV) is a highly contagious and often fatal disease of domestic and wild ruminants, caused by rinderpest virus of the genus Morbillivirus under the family Paramyxoviridae. Hemagglutinin (H) and fusion (F) proteins of this enveloped virus confer protective immunity against experimental challenge with virulent rinderpest virus. We have earlier demonstrated that immunization with a single dose of recombinant extracellular baculovirus expressing H protein elicits H-specific humoral and lymphoproliferative responses in cattle. The lymphoproliferative responses are predominantly BoLA class II restricted. In this work, we have analyzed lymphoproliferative responses of peripheral lymphocytes from immunized cattle to truncated H protein fragments expressed in E. coli for locating domains harboring Th epitopes. One region (aa 113-182) recognized by immune T cells is conserved in the H protein of measles virus, which was earlier shown to contain a dominant Th epitope in mouse. Synthetic peptides within this region of measles virus H protein were used to identify a Th epitope conserved in the H protein of RPV virus (aa 123-137) in cattle. A second Th epitope located at the C-terminus of RPV-H was mapped to the region corresponding to aa 512-609 using truncated protein fragments expressed in E. coli. The C-terminal epitope (aa 575-583) was mapped using synthetic peptides corresponding to measles virus H as well as RPV-H protein.

Protection of rabbits against lapinized rinderpest virus with purified envelope glycoproteins of peste-des-petits-ruminants and rinderpest viruses.

Haemagglutinin (HA) and fusion (F) proteins of peste-des-petits-ruminants virus (PPRV) and rinderpest virus (RPV) were purified by immunoaffinity chromatography. The purified proteins were characterized by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Rabbit hyperimmune sera were raised against the purified HA and F proteins and assayed by enzyme-linked immunosorbent assay (ELISA), haemagglutination-inhibition (HAI) and virus neutralization (VN) tests. The immunized animals were challenged with a virulent lapinized (rabbit-adapted) strain of RPV. Both HA and F proteins of PPRV protected rabbits against a lethal challenge with lapinized RPV. As expected, RPV HA and F proteins also conferred a similar protection against the homologous challenge. The postchallenge antibody responses were of a true anamnestic type.

Recombinant capripoxvirus expressing the hemagglutinin protein gene of rinderpest virus: protection of cattle against rinderpest and lumpy skin disease viruses.

A cDNA clone containing the complete coding sequence of the hemagglutinin (H) protein gene of the RBOK vaccine strain of rinderpest virus, under the control of the vaccinia late promoter p11, was inserted by homologous recombination into the thymidine kinase gene of the KS-1 strain of capripoxvirus. The recombinant virus produced authentic H protein as judged by its electrophoretic mobility, transport to the cell surface of infected lamb testis cells, and reactivity with monoclonal antibodies specific for the H protein of rinderpest virus. The recombinant virus induced significant levels of rinderpest virus neutralizing antibodies in vaccinated cattle and protected them from clinical rinderpest after challenge with a lethal dose of a highly virulent heterologous strain of the virus. Protection was achieved using vaccine doses lower than those used with a similar recombinant expressing the fusion protein gene of rinderpest. The parental KS-1 virus is widely used as a vaccine against capripox viruses and so the rinderpest recombinant acts as a dual vaccine to protect cattle against both rinderpest and lumpy skin disease.

Immunological responses of mice and cattle to baculovirus-expressed F and H proteins of rinderpest virus: lack of protection in the presence of neutralizing antibody.

Rinderpest is a highly contagious viral disease of ruminants and has greater than 95% morbidity and mortality. The etiological agent, rinderpest virus (RPV), is a member of the family Paramyxoviridae and the genus Morbillivirus. Immune responses to both the hemagglutinin (H) and the fusion (F) antigens of morbilliviruses play an important role in the prevention of infection, and only attenuated live vaccines have been shown to provide protective immunity against the group. The lack of protection with inactivated vaccines has been attributed to the denaturation of the F glycoprotein of the virus. Our previous study, however, demonstrated complete protection of cattle vaccinated with infectious vaccinia virus recombinants expressing the H (vRVH) or F (vRVF) protein alone, even in the presence of only 4 U of serum-neutralizing (SN) antibody to RPV (T. Yilma, D. Hsu, L. Jones, S. Owens, M. Grubman, C. Mebus, M. Yamanaka, and B. Dale, Science 242:1058-1061, 1988). We have constructed recombinant baculoviruses that express the F (Fb) and H (Hb) glycoproteins of RPV. Furthermore, we have analyzed the immune responses of mice and cattle to these antigens. Cattle vaccinated with Fb or Hb or a mixture of both antigens were not protected from challenge inoculation with RPV, even when the SN titer was greater than in cattle vaccinated with vRVF alone. This lack of protection, in the presence of SN antibody, would indicate that live attenuated and recombinant vaccines induce immune responses necessary for protection (e.g., cell-mediated immunity) that are not generated by subunit or inactivated whole-virus vaccines.

[An immunoenzyme test used for the detection of antibodies to rinderpest: the advantage of using a purified virus cultivated in a cell line]. / Un test immunoenzymatique appliqué à la détection des anticorps dirigés contre la peste bovine: l'interêt d'utiliser un virus purifié cultivé sur cellules de lignée.

Two types of in vitro assays (enzyme immunoassay and sero-neutralization test) were compared for their ability to detect antibodies to rinderpest virus in field sera from West African bovines. Purified virus grown on Vero cells (Ag/Vero) or bovine kidney cells (Ag/BK), were tested as antigen in enzyme-linked immunoassays (EIA). The results of the comparative evaluation of the two antigens by EIA, prove that Ag/Vero did enhance the sensitivity and the specificity of the test in comparison to Ag/BK and offers a 94% correspondence with seroneutralization.

Simple and rapid dot-enzyme immunoassay for visual detection of rinderpest antibodies in bovine and caprine sera.

A modified solid phase enzyme immunoassay (EIA) is described for visual detection of anti-rinderpest virus (RPV) antibodies in cattle and goat sera. Dots of RPV antigens were adsorbed to nitrocellulose (NC) paper (hence Dot-EIA) and the adsorptive reactive sites were blocked with skim milk powder. After immersion in bovine or caprine test serum bound antibodies were reacted with a peroxidase-conjugated anti-bovine or anti-caprine IgG (H & L), respectively. Positive reactions were easily visualized as red-brown dots after enzyme degradation of a substrate containing hydrogen peroxide and amino-ethylcarbazole (AEC). The Dot-EIA was comparable to the serum neutralisation (SN) test in its ability to detect antibody in bovine sera seven or ten days after experimental infection (DPI) with live attenuated Kabete "O" (RBOK) strain of RPV (grown in Vero cells) by a combination of subcutaneous (s/c), intravenous (i/v) or intranasal (i/n) routes. Early (seven DPI) RPV antibodies were detected in a serum sample from one goat experimentally infected with RPV by combined s/c-i/v routes but not in another goat only infected intranasally. The specificity of the Dot-EIA was equal to that of the SN test, as serum samples, collected from these experimental animals and those inoculated with non infected Vero cell culture fluid, with SN titres of 0.3 or lower were all negative by Dot-EIA. The Dot-EIA may have potential application as a rapid, simple and economical field test in diagnosis of rinderpest, vaccination surveillance and other seroepidemiological studies.

Use of an enzyme-linked immunosorbent assay for the detection of IgG antibodies to rinderpest virus in epidemiological surveys.

A comparison was made between an indirect enzyme-linked immunosorbent assay (ELISA) and the virus neutralisation test (VNT) for the detection of antibodies to rinderpest virus in field sera. The results did not agree for 6 per cent of the sera tested where 3 per cent of the samples gave ELISA positive/VNT negative and 3 per cent gave ELISA negative/VNT positive. The latter sera all had high levels of IgM antibody, which may indicate animals being at an early stage of infection or detection of a non-specific reaction. The ELISA results give a representative picture of the immune status for field surveys and a greater number of sera can be assayed with relative ease, compared to the traditional serum neutralisation test.

[Effect of estrogens on infection of rabbits with cattle plague virus previously administered to rabbits]. / Influence des aestrogènes sur l'infection du Lapin par le virus bovipestique lapinisé.

The Rabbit bovipest virus selectively destroys the lymphoid B line in the Rabbit. Estrogens inhibit this destruction when they are administered before the infection which leads are to think that they act on membrane sites of the lymphocytes.