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M Onuma

Publications and source records attributed to M Onuma.

At least 19 recordsLinked to original sources
PubMed1992

CD3+4-8- alpha beta T cell population with biased T cell receptor V gene usage. Presence in bone marrow and possible involvement of IL-3 for their extrathymic development.

Analysis of TCR of a series of CD4-8- (double negative; DN) alpha beta T cell lines induced with IL-3 revealed that their V gene usage was biased for V alpha 4 and V beta 2. This has been confirmed in the primary short-term cultures. Thus, IL-3 induced the generation of DN alpha beta T cells with predominant V beta 2 gene expression from the CD4+/CD8+ T cell-depleted spleen or bone marrow (BM) cells of both normal and nude BALB/c mice within 10 days. It was further indicated that the V beta 2+ beta-chain genes contained few junctional N regions in both IL-3-induced primary DN alpha beta T cells and continuous lines. Search for the in vivo counterpart of in vitro IL-3-induced DN alpha beta T cells revealed that BM, but not spleens, of normal BALB/c and B6 mice did contain a significant proportion of DN alpha beta T cells, and that the majority of them expressed V beta 2+ beta-chain genes with few junctional N regions. The presence of V beta 2+ DN alpha beta T cells was similarly observed in the BM of BALB/c nude mice, but their proportion varied markedly among various strains of mice, which was not linked to H-2 haplotypes. The results indicated that V beta 2+ DN alpha beta T cells in the BM represented one of the thymus-independent T cell populations, whose development was under the major histocompatibility Ag complex-unlinked genetic control. TCR of these T cells were shown to be functional as judged by the proliferative response to anti-V beta 2 antibody. Taken together, present results suggested that IL-3 could induce differentiation and/or proliferation of DN alpha beta T cells with uniquely limited repertoire, which existed preferentially in BM in vivo, and implied the possible involvement of extrathymic endogenous ligands as a positive selection force.

Animals↗
PubMed1992

Phenotyping of lymphocyte subsets in the vascular and epithelial lesions of a cow with malignant catarrhal fever.

Surface marker analysis of lymphoid cells infiltrating the vascular and epithelial lesions of a cow with malignant catarrhal fever (MCF) was conducted by immunohistochemistry using ten monoclonal antibodies. The majority of lymphoid cells in these lesions had BoCD8, BoCD6 or BoCD2, but they rarely possessed N-cell (BoCD5+/BoCD4-/BoCD8-, non-T non-B) markers. Similar reactivity was seen in lymphoid cells of perivascular infiltrates in the liver, heart and brain, and in T-dependent areas of lymph nodes. These results suggest involvement of cytotoxic T-lymphocytes in the pathogenesis of MCF.

Animals↗
PubMed1992

Genomic analysis of Theileria sergenti stocks in Japan with DNA probes.

Restriction fragment length polymorphisms of Theileria sergenti DNA from 18 different infections of cattle in 14 locations in Japan were analyzed by Southern blotting using T. sergenti genomic DNA fragments as probes. Probe pTs 2 hybridized with four fragments in BamHI digested piroplasm DNA, at 8.0, 7.3, 6.0 and 3.4 kb. Probe pTs 11-D1 hybridized with multiple fragments. With each probe, polymorphisms were observed among stocks from different locations. However, there was no correlation between the patterns of hybridization bands and the locations where parasites were collected. Analysis of the hybridization patterns of stocks obtained from individual cattle in the same grazing areas showed an almost identical pattern.

Animals↗
PubMed1992

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.

Animals↗
PubMed1992

Expression of the Marek's disease virus (MDV) homolog of glycoprotein B of herpes simplex virus by a recombinant baculovirus and its identification as the B antigen (gp100, gp60, gp49) of MDV.

A gene encoding a homolog of glycoprotein B of herpes simplex virus (gB homolog) has been identified on the Marek's disease virus (MDV) genome (L. J. N. Ross, M. Sanderson, S. D. Scott, M. M. Binns, T. Doel, and B. Milne, J. Gen. Virol. 70:1789-1804, 1989); however, the molecular and immunological characteristics of the gene product(s) are still not clear. In the present study, the gB homolog of MDV was expressed in insect cells by a recombinant baculovirus, and it was characterized to determine its molecular and antigenic properties. The expressed recombinant protein had three molecular sizes (88 to 110, 58, and 49 kDa) and was recognized by antisera from chickens inoculated with each of the three serotypes of MDV. By immunofluorescence analysis, it was shown that the protein was expressed in the cytoplasm and on the surface of the recombinant baculovirus-infected cells. The gB homolog of MDV was processed similarly to pseudorabies virus and varicella-zoster virus with respect to cleavage and the intramolecular disulfide bond between the cleaved products. Interestingly, the expressed protein reacted with monoclonal antibody M51, specific to the B antigen (gp100, gp60, gp49) of MDV, although the locations of the gene encoding the B antigen and of the gene encoding the gB homolog were reported to be different. Moreover, competitive experiments revealed that anti-gB homolog serum and monoclonal antibody M51 recognized the same molecules. From these results, the gB homolog and the B antigen of MDV seem to be the same glycoprotein.

Animals↗
PubMed1992

Activation of bovine peripheral blood monocyte and its suppressive effect on parasitemia in Theileria sergenti infected calves.

Activation of bovine peripheral blood monocytes and its suppressive effect on parasite growth was examined in Theileria sergenti-infected calves by using a rosette assay that detects changes in Fc receptor expression and by luminol-dependent chemiluminescence response. Monocyte activation preceded the peak of parasitemia but was depressed parallelly with the growth of the parasites. When four calves were treated with prednisolone, three showed a good correlation between the suppression of monocyte activity and an increase of parasitemia.

Animals↗
PubMed1992

Association of tumor-associated antigen on the proliferation of bovine leukemia virus-infected lymphoblastoid B-cell lines.

The association of tumor-associated antigen (TAA) on the proliferation of BLV-infected lymphoblastoid B-cell lines (BL2M3 and BL312) was investigated. Flow cytometric analysis of the expression of TAA with monoclonal antibody (mAb) c143 showed high expression of TAA on the surfaces of BL2M3 and BL312 cells. A large amount of TAA was found in the culture supernatant of BL2M3 and BL312 cells as well as in the lysates of BL2M3 and BL312 cells. Culture supernatant but not lysates of BL2M3 and BL312 cells promoted the growth of either BL2M3 cells or BL312 cells. Furthermore, this growth promoting activity in culture supernatants of BL2M3 and BL312 cells was inhibited in a dose-dependent manner when cultured with mAb c143. These results suggested that TAA may be involved in the growth factor-mediated cell growth of bovine B-lymphoblastoid cell lines expressing TAA on their cell surface.

Animals↗
PubMed1992

Detection and characterization of Theileria sergenti proteinases.

The lysate of Theileria sergenti piroplasms was tested for proteinases using sodium dodecyl sulfate-polyacrylamide gel electrophoresis in which substrate was included in gel matrix. Six proteinases of molecular weight 330, 125, 98, 94, 67 and 58 kilodalton (kDa) were detected. From the results of the Triton X-114 phase partition, 330, 125 and 58 kDa proteinases were partitioned into aqueous phase, which indicated that they were not associated with parasite membranes. All these three enzymes were classified into metalloproteinase family because of their sensitivities to metal-ion chelating compounds, ethylenediaminetetraacetic acid (EDTA) and 1,10-phenanthroline. On the other hand, 98 and 94 kDa proteinases were membrane-associated metalloproteinases which were preferentially inhibited by 1,10-phenanthroline. Another metalloproteinase of 67 kDa which was inhibited by EDTA and 1,10-phenanthroline was not associated with parasite membranes. Proteinases of 98 and 94 kDa degraded heat-denatured hemoglobin.

Animals↗
PubMed1992

Infection and dysfunction of monocytes induced by experimental inoculation of calves with bovine immunodeficiency-like virus.

Three calves were experimentally inoculated with bovine immunodeficiency-like virus (BIV) to examine BIV pathogenesis. Inoculated calves produced specific antibody that could be detected from 3 to 5 weeks up to 1 year postinoculation (pi). Virus was isolated from peripheral blood mononuclear cells (PBMC) 3-4 weeks pi by syncytia assay. Thereafter, the virus could be continually isolated. BIV could be isolated from monocytes but not from T cells. Likewise, monocytes could be infected with BIV in vitro. Various monocyte functions of these BIV-infected calves and age-matched uninfected calves were tested; superoxide anion release, phagocytic activity, and chemotactic responsiveness of monocytes were depressed in BIV-infected calves compared with control calves. A slight delay in the humoral immune response against mouse serum protein was also evident. During the observation period of approximately 1 year, no significant clinical symptoms could be observed. One calf, however, was killed at 15 months pi. At the time of necropsy, BIV could be isolated from PBMC as well as from cells of the spleen, liver, and lymph nodes.

Animals↗
PubMed1991

Activation of bovine peripheral blood macrophages in Theileria sergenti-infected calves.

Macrophage activation in Theileria sergenti-infected calves was studied by testing the production of oxygen metabolites in macrophages following specific and non-specific stimulation with T sergenti merozoites or zymosan, respectively. Six calves were inoculated with merozoites and three calves with sporozoites. All showed significant macrophage activation within one month after inoculation (P less than 0.05). Activation of macrophages appeared earlier than parasitaemia or the peak of antibody titre against T sergenti. The highest chemiluminescence response, indicative of macrophage activation, was observed when the merozoites were opsonised with immune sera.

Animals↗
PubMed1991

Characterization of haemagglutinin-neuraminidase glycoprotein of Newcastle disease virus expressed by a recombinant baculovirus.

A recombinant baculovirus containing a cDNA which encodes haemagglutinin-neuraminidase (HN) of Newcastle disease virus (NDV) was constructed. Spodoptera frugiperda cells infected with this recombinant virus produced a large amount of HN glycoprotein similar to the authentic HN in size. The recombinant HN glycoprotein was localized on the surface of the infected cells and conserved its haemadsorption and neuraminidase activities. The antigenic properties of the recombinant HN glycoprotein seemed to be slightly different from the authentic one, as judging by the reactivity with a panel of monoclonal antibodies specific to the antigenic sites responsible for neutralization of viral infectivity. Chickens inoculated with the cells infected with the recombinant virus developed haemagglutination-inhibition and virus neutralization antibodies, and were completely protected from the NDV challenge.

Animals↗
PubMed1991

Responses of peripheral blood mononuclear cells in calves infected with Theileria sergenti.

The role of peripheral blood mononuclear cell (PBMC) in Theileria sergenti-infected calves was studied by various in vitro assay systems. Proliferation of T cells in mixed lymphocyte protozoa culture (MLPC) increased with parasitemia, and the addition of monoclonal antibodies against T. sergenti merozoites in this MLPC enhanced the response. However, the addition of antibody-positive autologous serum resulted in the suppression of the response. Cell-mediated cytotoxicity of PBMC increased after peak parasitemia. This cytotoxicity increased on co-cultivation of PBMC with T. sergenti merozoites, but the addition of autologous serum suppressed the response.

Animals↗
PubMed1991

Production of monoclonal antibodies specific for ganglioside GD3.

Four kinds of anti-GD3 monoclonal antibodies, DSG-1, -2, -3, and -4, of the IgM class were obtained by the immunization of BALB/c mice with enzootic bovine leukosis tumor tissue-derived ganglioside GD3 inserted into liposomes with Salmonella minnesota R595 lipopolysaccharides. The specificities of the monoclonal antibodies obtained were defined by complement-dependent liposome immune lysis assay and by enzyme immunostaining on thin-layer chromatography. The reactivities of the monoclonal antibodies obtained to four ganglioside GD3 variants [GD3(NeuAc-NeuAc), GD3(NeuAc-NeuGc), GD3(NeuGc-NeuAc), and GD3(NeuGc-NeuGc)] were tested. All of the monoclonal antibodies were found to react with GD3(NeuAc-NeuAc) and GD3(NeuAc-NeuGc) but not with GD3(NeuGc-NeuAc) or GD3(NeuGc-NeuGc). Furthermore, various purified glycosphingolipids were used to determine the specificity of these monoclonal antibodies. All 4 antibodies reacted only with ganglioside GD3 [GD3(NeuAc-NeuAc) and GD3(NeuAc-NeuGc)], but not with several gangliosides linking the GalNAc, Gal beta 1-3GalNAc, NeuAc alpha 2-3Gal beta 1-3GalNAc, or NeuAc alpha 2-8NeuAc alpha 2-3Gal beta 1-3GalNAc residue to the Gal moiety of ganglioside GD3 (GD2, GD1b, GT1b, or GQ1b, respectively), ganglioside GT1a having the same terminal NeuAc alpha 2-8NeuAc alpha 2-3Gal residue as ganglioside GD3, other gangliosides, and neutral glycosphingolipids. These findings suggest that the 4 monoclonal antibodies obtained may be specific for the epitope of NeuAc-alpha 2-8Sia alpha 2-3Gal beta 1-4Glc residue of ganglioside GD3.

Animals↗
PubMed1991

Expression of the A antigen (gp57-65) of Marek's disease virus by a recombinant baculovirus.

A recombinant baculovirus expressing the A antigen (A Ag) of Marek's disease virus (MDV) was constructed. In Spodoptera fruiperda (Sf) cells infected with the recombinant virus, A Ag expression was localized to the cell surface. Only a small amount of recombinant A Ag was detected in the culture supernatant of infected Sf cells, but authentic A Ag is mainly secreted into the culture supernatant of MDV-infected chicken embryo fibroblasts. Cell surface-associated recombinant A Ag seemed to be slightly larger than authentic A Ag, whereas the secreted recombinant A Ag seemed to be smaller. The recombinant A Ag was shown to be reactive with the sera of MDV-infected chickens by immunodiffusion studies and ELISA. Sera of chickens immunized with recombinant A Ag formed a precipitin line with the culture supernatant of MDV-infected chicken embryo fibroblasts in an immunodiffusion test. These results indicate that the recombinant A Ag expressed by the recombinant baculovirus retains the antigenic and immunogenic properties of the authentic A Ag.

Animals↗
PubMed1991

Analysis of the protective effect of the haemagglutinin-neuraminidase protein in Newcastle disease virus infection.

The role of immune responses to haemagglutinin-neuraminidase (HN) protein in protection against a Newcastle disease virus (NDV) infection was investigated using a recombinant vaccinia virus expressing HN (HN-RVV). Live HN-RVV replicated in chickens and completely protected them from lethal infection with virulent NDV. Inactivated HN-RVV also protected chickens when administered with adjuvant but not when administered without adjuvant. However, large amounts of the inactivated HN-RVV (100-fold excess) without adjuvant provided protection. Specific antibodies against the HN protein of NDV were detected in sera from survivors but not from dying birds. However, the kinetics of antibody responses in chickens inoculated with live HN-RVV and inactivated HN-RVV were considerably different. These results clearly confirm that immune response(s) solely to the HN protein of NDV can provide chickens with protection against NDV challenge, and show that the presence of antibodies to the HN protein correlates significantly with the protection from NDV infection at least in HN-immunized chickens.

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