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1.
J Lab Clin Med ; 137(1): 28-37, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11150021

ABSTRACT

Immune function is markedly attenuated in endotoxemia. Zinc is involved in the regulation of cellular functions and maintenance of immune function, and its level in the serum is low in endotoxemia. We mainly investigated the effects of zinc acetate (ZA) on splenocytes in mice with endotoxemia. After we confirmed increased plasma zinc level by ZA treatment, C57BL/6 mice were randomly divided into four groups: 10 control mice received 500 microL saline solution as vehicle; 10 control mice received ZA at 3 mg/kg body weight; 20 endotoxemic mice received a 40 mg/kg lethal dose of lipopolysaccharide (LPS); 20 mice received ZA followed by LPS as the above dose. In vivo, we confirmed that ZA pretreatment did not significantly affect the plasma cytokine level in endotoxemic mice. In vitro, splenocytes from ZA-plus-LPS mice showed drastic effects, in that ZA abrogated LPS-induced suppression of cellular proliferation and production of interleukin-2 and interferon-gamma. The percentage of apoptotic splenocytes was significantly reduced in ZA-plus-LPS mice (23.4%) as compared with LPS mice (41.6%). Furthermore, the expression of HSP-70 mRNA in splenocytes was strongly enhanced in both ZA and ZA-plus-LPS mice, especially in the latter group. Finally, studies monitoring survival rates for 6 days showed that LPS caused 100% mortality while ZA-plus-LPS mice showed 75% survival. Our results suggest that zinc normalized the immune response and reduced apoptosis of splenocytes. These changes were probably caused by increased synthesis of HSP-70 by splenocytes, which might enhance survival of mice with LPS-induced endotoxemia.


Subject(s)
Apoptosis/drug effects , Endotoxemia/drug therapy , Endotoxemia/immunology , HSP70 Heat-Shock Proteins/genetics , Spleen/cytology , Zinc Acetate/pharmacology , Animals , Apoptosis/immunology , Blotting, Northern , Cell Division/drug effects , Cell Division/immunology , Cells, Cultured , Concanavalin A/pharmacology , Cytokines/blood , Endotoxemia/mortality , Female , Flow Cytometry , Gene Expression/drug effects , Gene Expression/immunology , Lipopolysaccharides , Mice , Mice, Inbred C57BL , RNA, Messenger/analysis , Specific Pathogen-Free Organisms , Spleen/immunology , Survival Rate , Zinc/blood
2.
J Virol ; 72(8): 6907-10, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9658144

ABSTRACT

Subcutaneous administration in mice of recombinant Sindbis viruses expressing a class I major histocompatibility complex-restricted 9-mer epitope of the Plasmodium yoelii circumsporozoite protein or the nucleoprotein of influenza virus induces a large epitope-specific CD8(+) T-cell response. This immunization also elicits a high degree of protection against infection with malaria or influenza A virus.


Subject(s)
Epitopes, T-Lymphocyte/immunology , Genetic Vectors , Influenza A virus/immunology , Influenza, Human/prevention & control , Malaria/prevention & control , Plasmodium yoelii/immunology , RNA-Binding Proteins , Sindbis Virus , T-Lymphocytes, Cytotoxic/immunology , Animals , Drug Administration Routes , Genetic Vectors/genetics , Humans , Influenza A virus/growth & development , Influenza Vaccines/immunology , Influenza, Human/immunology , Mice , Mice, Inbred BALB C , Nucleocapsid Proteins , Nucleoproteins/genetics , Nucleoproteins/immunology , Plasmodium yoelii/genetics , Plasmodium yoelii/growth & development , Protozoan Proteins/genetics , Protozoan Proteins/immunology , Protozoan Vaccines/immunology , RNA, Protozoan , Sindbis Virus/genetics , Vaccination , Vaccines, Synthetic/immunology , Viral Core Proteins/genetics , Viral Core Proteins/immunology
3.
J Med Virol ; 53(3): 266-72, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9365894

ABSTRACT

The nucleotide sequences of the core upstream and precore regions (371 nucleotide length, nt. 1604-1974) of hepatitis B virus (HBV) were analysed sequentially in three subjects who were positive serologically for anti-HBe and had acute clinical exacerbation after immunosuppressive treatment. These patients were asymptomatic HBV carriers before therapy. The results revealed that the mutant with an 8-bp deletion (nt. 1768-1775) located in the basic core promoter region was dominant in the asymptomatic HBV carrier phase in two of three subjects. After exacerbation, however, such mutant clones possessing 8-bp deletion disappeared or decreased in number and were replaced by the clones possessing a precore stop codon mutation G to A (nt. 1896) or by the clones possessing additional contiguous point mutations A to T (nt. 1762) and G to A (nt. 1764) and a new point mutation C to T (nt. 1653). Possible relationships between acute exacerbation of liver function accompanied by mutation and the transition of the dominant clones were discussed.


Subject(s)
Carrier State , Genome, Viral , Hepatitis B Antibodies/blood , Hepatitis B e Antigens/blood , Hepatitis B virus/genetics , Hepatitis B/virology , Mutation , Acute Disease , Base Sequence , Brain Neoplasms/complications , DNA, Viral , Dermatomyositis/complications , Female , Glioblastoma/complications , Hepatitis B/complications , Hepatitis B/immunology , Hepatitis B/physiopathology , Hepatitis B Antibodies/immunology , Hepatitis B e Antigens/immunology , Hepatitis B virus/immunology , Hodgkin Disease/complications , Humans , Male , Middle Aged , Molecular Sequence Data , Protein Precursors/genetics , Sequence Analysis, DNA , Sequence Homology, Nucleic Acid
4.
Exp Parasitol ; 84(2): 223-30, 1996 Nov.
Article in English | MEDLINE | ID: mdl-8932772

ABSTRACT

In this study we characterized the CD4+ T cell response directed against two distinct epitopes located in the circumsporozoite (CS) protein of Plasmodium yoelii. The immunization of mice with P. yoelii sporozoites induced CD4+ T cells which were mostly directed against one of these peptides, Py-1, previously reported to contain a CD4+ epitope. The CD4+ T cells directed against this immunodominant epitope were mostly of the Th-1 type. Another newly identified peptide, AS44, induced a specific CD4+ T cell response, which was mainly detectable after immunization with the corresponding peptide. Several CD4+ T cell clones, recognizing this epitope, were generated and their lymphokine expression was characterized, as well as their surface markers and their anti-parasite activity in vivo. It was noteworthy that some of these CD4+ T cell clones, which recognize this cryptic epitope and were of different Th subtypes, were shown to have a strong inhibitory effect on the development of liver stages of malaria parasites.


Subject(s)
Antigens, Protozoan/immunology , CD4-Positive T-Lymphocytes/immunology , Malaria/immunology , Peptides/immunology , Plasmodium yoelii/immunology , Protozoan Proteins/immunology , Adoptive Transfer , Amino Acid Sequence , Animals , Female , Immunization , Immunodominant Epitopes , Interferon-gamma/metabolism , Interleukin-5/metabolism , Malaria/parasitology , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Plasmodium yoelii/growth & development , Th1 Cells/immunology
5.
Leuk Lymphoma ; 15(3-4): 303-10, 1994 Oct.
Article in English | MEDLINE | ID: mdl-7866279

ABSTRACT

Recently, several cases of adult T-cell leukemia (ATL) with CD30 antigen have been reported, but its clinical significance remains unknown. Accordingly, we studied CD30 antigen expression in ATL cases and documented the clinicopathological characteristics of these cases. Immunohistochemical and clinical characteristics were studied in 46 patients with malignant lymphoma or benign lesions of lymphoid tissue, who had antibodies against human T-cell leukemia virus type I (HTLV-I). Monoclonal integration of HTLV-I provirus was demonstrated in the tumor cells in 36 (ATL) of the 46 cases. CD30 antigen expression was evident in seven of these 36 cases (19.4%), however it was not seen in any of the ten cases lacking the integration of HTLV-I provirus. A comparison of ATL cases with and without CD30 antigen expression revealed significantly larger numbers of abnormal lymphocytes in the peripheral blood and lower serum calcium levels in ATL expressing CD30 antigen.


Subject(s)
Ki-1 Antigen/physiology , Leukemia, T-Cell/immunology , Adult , Aged , Aged, 80 and over , DNA, Neoplasm/genetics , DNA, Viral/genetics , Female , HTLV-I Infections/immunology , Human T-lymphotropic virus 1/genetics , Humans , Immunohistochemistry , Leukemia, T-Cell/pathology , Leukemia, T-Cell/virology , Male , Middle Aged , Virus Integration
6.
J Gen Virol ; 75 ( Pt 6): 1339-44, 1994 Jun.
Article in English | MEDLINE | ID: mdl-8207400

ABSTRACT

Mice vaccinated intraperitoneally (i.p.) with 10(7) p.f.u. of a vaccinia virus recombinant expressing either the glycoprotein (rVac-G) or nucleoprotein (rVac-N) of rabies virus 3 weeks before challenge were protected against peripheral lethal infection. Similarly, by post-exposure vaccination in which mice were first infected with rabies virus and subsequently vaccinated i.p. with the recombinant, rVac-G conferred protection when given immediately following infection and up to 24 h after infection. Prior treatment of those mice with anti-CD8 monoclonal antibodies (MAb) did not significantly affect the outcome of the infection. In contrast, rVac-N failed to confer protection even with higher doses (10(8) p.f.u.) of the virus or even when administered by the intradermal route. Anti-nucleoprotein antibody production by these mice was not suppressed by prior rabies virus infection and the levels and the time of antibody production were similar to those of anti-glycoprotein antibody production in mice vaccinated with rVac-G after rabies virus infection. The cytotoxic T lymphocyte response was also not down-regulated by rabies virus in the mice that were given rVac-N. Possible mechanism(s) for the ineffectiveness of rVac-N by post-exposure vaccination in contrast to pre-exposure vaccination was discussed.


Subject(s)
Antigens, Viral , Capsid/immunology , Glycoproteins/immunology , Rabies Vaccines/immunology , Rabies virus/immunology , Rabies/prevention & control , Viral Core Proteins/immunology , Viral Envelope Proteins/immunology , Animals , Antibodies, Viral/biosynthesis , Antibody Formation , CD8 Antigens/immunology , Cytotoxicity, Immunologic , Immunity, Cellular , Mice , Mice, Inbred A , Rabies/immunology , Recombinant Proteins/immunology , T-Lymphocyte Subsets/immunology , Time Factors , Vaccines, Synthetic/immunology , Vaccinia virus
7.
Microbiol Immunol ; 38(9): 721-6, 1994.
Article in English | MEDLINE | ID: mdl-7854213

ABSTRACT

Target cells of cytotoxic T lymphocytes (CTL) directed to the individual structural proteins (except for the large polymerase (L) protein) of rabies virus were established by expressing only the respective protein in murine neuroblastoma (NA) and murine macrophage (J774-1) cell lines. Mice infected with the ERA strain of rabies virus developed CTL responses to all of these rabies virus proteins. The cytotoxic activity was abrogated by pretreatment of the effector cells with anti-CD8 monoclonal antibody (MAb) and complement but not with anti-CD4 MAb. Cell lysis by CTL was blocked in the presence of anti-major histocompatibility complex (MHC) class 1 antibodies in J774-1 cell lines. Rabies virus-infected cells express these proteins at the surface, which can be recognized and lysed by the respective CTL. Mice immunized with beta-propiolactone-inactivated virus induced a CTL response against glycoprotein but not against internal viral components. This assay system might be useful for further analysis of the possible contribution of these proteins in the cell-mediated immune protection against rabies.


Subject(s)
Rabies virus/immunology , T-Lymphocytes, Cytotoxic/immunology , Viral Structural Proteins/immunology , Animals , Antibodies, Viral/immunology , Cell Line , Cytotoxicity Tests, Immunologic , Cytotoxicity, Immunologic/immunology , Gene Expression Regulation, Viral , Macrophages/virology , Male , Mice , Mice, Inbred A , Mice, Inbred BALB C , Mice, Inbred C57BL , Neuroblastoma/virology , Rabies/immunology , Rabies/microbiology , Rabies virus/genetics , Rabies virus/growth & development , Tumor Cells, Cultured , Viral Structural Proteins/analysis
8.
Virus Genes ; 7(1): 83-8, 1993 Feb.
Article in English | MEDLINE | ID: mdl-8470369

ABSTRACT

Nucleotide sequences and the deduced amino acid sequences of the gene encoding the matrix (M) protein of the Nishigahara and the CVS strains of rabies virus have been determined. The M gene is 609 nucleotides long and is capable of coding for a peptide composed of 202 amino acids. Sequence comparison of these M genes with those of other stains [Pasteur (PV), ERA, Avol] revealed that there is 89.7-91.5% homology at the nucleotide level, and 90.1-92.1% homology at amino acid level, between almost all combinations of these strains. However, in the combinations of the PV and ERA strains, and the virulent CVS and the avirulent CVS-derived Avol strains, much higher homology was observed both at the nucleotide and amino acid levels. The predicted secondary structure and hydropathy profiles also exhibited similar features. Recombinant vaccinia virus containing the M gene was constructed. Sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel electrophoresis of the precipitates obtained by immune reaction of the recombinant virus-infected cell lysate with a monoclonal antibody against the M protein revealed that electrophoretic mobility of the expressed protein is indistinguishable from that of the authentic M protein from rabies virions.


Subject(s)
Genes, Viral , Rabies virus/genetics , Vaccinia virus/genetics , Viral Matrix Proteins/genetics , Amino Acid Sequence , Base Sequence , DNA, Viral/genetics , Gene Expression , Molecular Sequence Data , Recombination, Genetic , Sequence Analysis, DNA , Species Specificity
9.
Arch Virol ; 132(1-2): 51-65, 1993.
Article in English | MEDLINE | ID: mdl-8352659

ABSTRACT

Mice were vaccinated with recombinant vaccinia virus (rVac) expressing the glycoprotein (G), nucleoprotein (N), phosphoprotein (NS) or matrix protein (M) of rabies virus and their resistance to peripheral lethal infection with street rabies virus was examined. Mice vaccinated with rVac-G or rVac-N developed strong antibody responses to the corresponding proteins and essentially all mice survived challenge infection. Mice vaccinated with rVac-NS or rVac-M developed only a slight antibody response, however, a significant protection (59%) was observed in the rVac-NS-vaccinated mice, whereas rVac-M-vaccinated mice were not protected. No anti-G antibodies were detected in the sera of mice which has been vaccinated with rVac-N or rVac-NS and survived challenge infection. Passive transfer of anti-N monoclonal antibodies (MAbs) recognizing an epitope located on amino acids 1-224 of the protein prior to challenge resulted in significant protection, although the protection was not complete even with a high amount of antibodies. In contrast, none of the mice given MAbs recognizing an epitope of amino acids 247-415 or F(ab')2 fragments from a protective MAb IgG were protected. Administration of anti-CD 8 MAb to rVac-N-vaccinated mice showed no significant effect on protection. Our observations suggest that a considerable part of the protection achieved by the vaccination with rVac-N can be ascribed to the intact anti-N antibodies recognizing an epitope located on amino acids 1-224 of the protein.


Subject(s)
Rabies Vaccines/administration & dosage , Rabies/prevention & control , Viral Structural Proteins/immunology , Animals , Antibodies, Monoclonal/administration & dosage , Antibodies, Monoclonal/immunology , Antibodies, Viral/administration & dosage , Antibodies, Viral/biosynthesis , Base Sequence , CD8 Antigens/immunology , Capsid/administration & dosage , Capsid/immunology , Cells, Cultured , Cricetinae , DNA, Viral , Immunity, Innate , Immunization, Passive , Male , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Rabbits , Rabies Vaccines/immunology , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/immunology , Vaccinia virus/genetics , Viral Core Proteins/administration & dosage , Viral Core Proteins/immunology , Viral Matrix Proteins/administration & dosage , Viral Matrix Proteins/immunology , Viral Nonstructural Proteins/administration & dosage , Viral Nonstructural Proteins/immunology , Viral Structural Proteins/administration & dosage
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