Kazue Ohishi

Pathological and serological evidence of Brucella-infection in baleen whales (Mysticeti) in the western North Pacific

Abnormal testes and uterus were observed in 13 males (33%) and one female (3%) out of 40 common minke whales (Balaenoptera acutorostrata) in the western North Pacific. Similar lesions were found in testis and ovary, respectively, in one male (2%) and female (2%) out of 43 Brydes whales (Balaenoptera edeni) in the western North Pacific. Grossly, granular lesions with caseation and calcification were main pathological signs, and they were restricted to reproductive organs of mature whales. Chronic purulent or granulomatous orchitis was observed by microscopic analysis. Antibodies to Brucella species were detected in the serum samples of 15/40 (38%) of common minke whales and 4/43 (9%) of Brydes whales. Neither pathological nor serological change was found in the examined sperm whales (Physeter macrocephalus) in the western North Pacific and Antarctic minke whales (Balaenoptera bonaerensis). These results strongly suggest that Brucella infection was involved in two species of baleen whales (Mysticeti) in the North Pacific.

Protective Immunity Against Bovine Leukaemia Virus (BLV) Induced In Carrier Sheep By Inoculation With A Vaccinia Virus-BLV ENV Recombinant: Association With Cell-Mediated Immunity

The effects of vaccination of sheep with a recombinant vaccinia virus (rVV) expressing the bovine leukaemia virus (BLV) envelope glycoprotein (gp60) were studied by determining BLV titres in peripheral blood leukocytes after vaccination and challenge. The proliferation of BLV was suppressed markedly, not only when rVV was inoculated prior to challenge with BLV, but also when it was inoculated after challenge. These results indicate that vaccination with rVV induces protective immunity that can suppress the growth of BLV in carrier animals. Since rVV induced a strong anti-BLV delayed-type hypersensitivity response without producing detectable levels of binding or neutralizing antibodies, and there was no apparent correlation between the humoral immune response and BLV proliferation, a cell-mediated immune response was assumed to play a major role in protective immunity.

Serological evidence of transmission of human influenza A and B viruses to Caspian seals (Phoca caspica)

Seroepidemiological surveillance of influenza in Caspian seals (Phoca caspica) was conducted. Antibodies to influenza A virus were detected in 54% (7/13), 57% (4/7), 40% (6/15) and 26% (11/42) of the serum samples collected in 1993, 1997, 1998 and 2000 by enzyme‐linked immunosorbent assay (ELISA). In an hemagglutination‐inhibition (HI) test using H1‐H15 reference influenza A viruses as antigens, more than half of the examined ELISA‐positive sera reacted with an H3N2 prototype strain A/Aichi/2/68. These sera were then examined by HI test with a series of naturally occurring antigenic variants of human H3N2 virus, and H3 viruses of swine, duck, and equine origin. The sera reacted strongly with the A/Bangkok/1/79 (H3N2) strain, which was prevalent in humans in 1979–1981. The present results indicate that human A/Bangkok/1/79‐like virus was transmitted to Caspian seals probably in the early 1980s, and was circulated in the population. Antibodies to influenza B virus were detected by ELISA in 14% (1/7) and 10% (4/42) serum samples collected from Caspian seals in 1997 and 2000, respectively. Our findings indicate that seal might be a reservoir of both influenza A and B viruses originated from humans.

Long-term protective immunity to rinderpest in cattle following a single vaccination with a recombinant vaccinia virus expressing the virus haemagglutinin protein

A recombinant vaccine, produced by using a highly attenuated smallpox vaccine (LC16mO) as a vector and which expresses the rinderpest virus (RPV) haemagglutinin protein, has been developed. The properties of this vaccine, including its heat stability, efficacy in short-term trials, safety and genetic stability, have been confirmed in an earlier report. In the present study, the duration of the protective immunity generated by the vaccine in cattle was examined for up to 3 years following the administration of a single vaccination dose of 10(8) p.f.u. The vaccinated cattle were kept for 2 (group I) or 3 years (group II) and then challenged with a highly virulent strain of RPV. Four of five vaccinated cattle in group I and all six cattle in group II survived the challenge, some showing solid immunity without any clinical signs of rinderpest. Neutralizing antibodies were maintained at a significant level for up to 3 years and they increased rapidly following challenge. Lymphocyte proliferative responses to RPV were examined in group II cattle and were observed in four of the six vaccinated cattle in this group. The long-lasting protective immunity, in addition to the other properties confirmed previously, indicate the practical usefulness of this vaccine for field use.

Host-virus specificity of morbilliviruses predicted by structural modeling of the marine mammal SLAM, a receptor.

Signaling lymphocyte activation molecule (SLAM) is thought to be a major cellular receptor for high-host specificity morbilliviruses, which cause devastating and highly infectious diseases in mammals. We determined the sequences of SLAM cDNA from five species of marine mammal, including two cetaceans, two pinnipeds and one sirenian, and generated three-dimensional models to understand the receptor-virus interaction. Twenty-one amino acid residues in the immunoglobulin-like V domains of the SLAMs were shown to bind the viral protein. Notably, the sequences from pinnipeds and dogs were highly homologous, which is consistent with the fact that canine distemper virus was previously shown to cause a mass die-off of seals. Among these twenty-one residues, eight (63, 66, 68, 72, 84, 119, 121 and 130) were shared by animal groups susceptible to a particular morbillivirus species. This set of residues appears to determine host-virus specificity and may be useful for risk estimation for morbilliviruses.

An effective peptide vaccine to eliminate bovine leukaemia virus (BLV) infected cells in carrier sheep

Protective effects of the gp51 of bovine leukaemia virus (BLV) expressed by a recombinant baculovirus (rgp51) and synthetic multiple antigenic peptides (MAP) of T-helper, T-cytotoxic, and B-cell epitopes of gp51 were investigated against BLV challenge. Two and three sheep were immunized with rgp51 and a mixture of peptides with Freunds complete adjuvant, respectively. BLV was detected from all the immunized sheep at 2 weeks and showed peak levels at 4 weeks after the challenge. However, in two sheep immunized with the mixed peptides, the titer of BLV gradually decreased and one sheep eliminated BLV completely at 28 weeks after the challenge. These two sheep showed higher lymphocyte proliferative responses against the immunized peptides than the other sheep. One of the sheep also showed the specific cytotoxic lymphocyte activity against the BLV gp51-expressing target in vitro. These results suggest the possibility of the peptide vaccine for elimination of BLV in carrier animals in vivo.

CELL-MEDIATED IMMUNE RESPONSES IN CATTLE VACCINATED WITH A VACCINIA VIRUS RECOMBINANT EXPRESSING THE NUCLEOCAPSID PROTEIN OF RINDERPEST VIRUS

Rinderpest virus (RPV) is a member of the genus Morbillivirus in the family Paramyxoviridae which causes an acute and often fatal disease in large ruminants. To examine the immune response to the virus nucleocapsid (N) protein, a recombinant vaccinia virus expressing RPV nucleocapsid protein (rVV-RPV-N) was used to vaccinate cattle. The recombinant vaccine induced low levels of non-neutralizing anti-N antibodies. RPV-specific cell-mediated immunity induced by the recombinant was assessed by measuring both the lymphocyte proliferation and cytotoxic T-lymphocyte responses. The protective immune response was examined by challenging the vaccinated cattle with either a highly virulent (Saudi 1/81) or a mild (Kenya/eland/96) strain of the virus. The vaccinated cattle were not protected against challenge with the virulent RPV strain, except they showed a slight delay in the onset of disease when compared with the unvaccinated controls. In cattle challenged with the mild strain, apart from a transient fever, no clinical signs of rinderpest infection were seen in the vaccinated cattle. One out of two control cattle showed a similar response but the other died from classic rinderpest disease. Virus-neutralizing antibodies were induced more quickly following challenge with the mild strain in vaccinated cattle compared to the control animals. These data suggested that the cell-mediated immunity induced by rVV-RPV-N could stimulate the rapid production of neutralizing antibodies following RPV challenge but this response was not sufficient to protect against challenge with a virulent strain of the virus. Protection was seen in one of three animals challenged with a mild strain of the virus; however, a greater number of animals would need to be tested to estimate the significance of the protection afforded by the N protein.

Antibodies to Human‐Related H3 Influenza A Virus in Baikal Seals (Phoca sibirica) and Ringed Seals (Phoca hispida) in Russia

Antibodies to influenza A virus were detected using enzyme‐linked immunosorbent assay (ELISA) in the sera from two of seven Baikal seals (Phoca sibrica) and from five of six ringed seals (Phoca hispida) in Russia. In a hemagglutination‐inhibition test using H1–H15 reference influenza A viruses, ELISA‐positive sera from one Baikal seal and four ringed seals reacted to A/Aichi/2/68 (H3N2) and A/Bangkok/1/79 (H3N2) strains. One ringed seal serum sample reacted to A/seal/Massachusetts/1/80 (H7N7). The present results suggested that human‐related H3 viruses were prevalent in Baikal seals and ringed seals inhabiting the central Russian Arctic.

Augmentation of Bovine Leukemia Virus (BLV)‐Specific Lymphocyte Proliferation Responses in Ruminants by Inoculation with BLV env‐Recombinant Vaccinia Virus: Their Role in the Suppression of BLV Replication

Lymphocyte proliferation responses were investigated in sheep and cattle, in which the replication of bovine leukemia virus (BLV) had been known to be suppressed by inoculation with recombinant vaccinia virus (rVV) expressing BLV envelope glycoprotein (gp60). Enhanced lymphocyte proliferation responses were observed in animals inoculated with rVV, regardless of whether they were naive or BLV carriers. These responses were roughly inversely correlated to the growth of BLV in the peripheral blood leukocytes. In contrast, there was no apparent correlation between humoral immune response and BLV growth. Based on these results, it was suggested that rVV rendered its suppressive effect of BLV replication primarily via augmentation of cell‐mediated immunity.

RECENT HOST RANGE EXPANSION OF CANINE DISTEMPER VIRUS AND VARIATION IN ITS RECEPTOR, THE SIGNALING LYMPHOCYTE ACTIVATION MOLECULE, IN CARNIVORES

Abstract The signaling lymphocyte activation molecule (SLAM) is a receptor for morbilliviruses. To understand the recent host range expansion of canine distemper virus (CDV) in carnivores, we determined the nucleotide sequences of SLAMs of various carnivores and generated three-dimensional homology SLAM models. Thirty-four amino acid residues were found for the candidates binding to CDV on the interface of the carnivore SLAMs. SLAM of the domestic dog (Canis lupus familiaris) were similar to those of other members of the suborder Caniformia, indicating that the animals in this group have similar sensitivity to dog CDV. However, they were different at nine positions from those of felids. Among the nine residues, four of domestic cat (Felis catus) SLAM (72, 76, 82, and 129) and three of lion (Panthera leo persica) SLAM (72, 82, and 129) were associated with charge alterations, suggesting that the felid interfaces have lower affinities to dog CDV. Only the residue at 76 was different between domestic cat and lion SLAM interfaces. The domestic cat SLAM had threonine at 76, whereas the lion SLAM had arginine, a positively charged residue like that of the dog SLAM. The cat SLAM with threonine is likely to have lower affinity to CDV-H and to confer higher resistance against dog CDV. Thus, the four residues (72, 76, 82, and 129) on carnivore SLAMs are important for the determination of affinity and sensitivity with CDV. Additionally, the CDV-H protein of felid strains had a substitution of histidine for tyrosine at 549 of dog CDV-H and may have higher affinity to lion SLAM. Three-dimensional model construction is a new risk assessment method of morbillivirus infectivity. Because the method is applicable to animals that have no information about virus infection, it is especially useful for morbillivirus risk assessment and wildlife conservation.