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Kazuaki Mannen

Publications and source records attributed to Kazuaki Mannen.

10 recordsLinked to original sources
PubMed2005

Construction of human Fab library and isolation of monoclonal Fabs with rabies virus-neutralizing ability.

A combinatorial human Fab library was constructed using RNAs from peripheral blood lymphocytes of 6 rabies vaccine-boosted volunteers using pComb3X phagemid vector. The size of the constructed library was approximately 7.0 x 10(7) Escherichia coli transformants. The library was selected against purified rabies virus (RV) virion or purified RV glycoprotein for isolation of phages displaying RVneutralizing human Fab antibody. Among 132 selected clones, two Fab preparations revealed neutralizing activities against RV strain CVS when assayed in the rapid fluorescent focus inhibition test (RFFIT). The Fab preparation EP5G3 exhibited neutralizing activity with an infected cell count reduction of 76% at a dilution of 1: 2, and of 20% at a dilution of 1: 4. The Fab preparation GD2D12 also exhibited neutralizing activity with a 57% reduction at 1: 2 and 41% reduction at 1: 4. In the co-immunoprecipitation using strain CVS, the RV glycoprotein was precipitated in reactions with both Fab preparations. The RV neutralizing ability of the Fab preparations described in the study were not directly correlated with their binding specificity for RV antigens detected by ELISA.

Amino Acid Sequence↗
PubMed2004

Transgenic mouse expressing human mutant alpha-galactosidase A in an endogenous enzyme deficient background: a biochemical animal model for studying active-site specific chaperone therapy for Fabry disease.

Fabry disease is an inborn error of glycosphingolipid metabolism caused by the deficiency of lysosomal alpha-galactosidase A (alpha-Gal A). We have established transgenic mice that exclusively express human mutant alpha-Gal A (R301Q) in an alpha-Gal A knock-out background (TgM/KO mice). This serves as a biochemical model to study and evaluate active-site specific chaperone (ASSC) therapy for Fabry disease, which is specific for those missense mutations that cause misfolding of alpha-Gal A. The alpha-Gal A activities in the heart, kidney, spleen, and liver of homozygous TgM/KO mice were 52.6, 9.9, 29.6 and 44.4 unit/mg protein, respectively, corresponding to 16.4-, 0.8-, 0.6- and 1.4-fold of the endogenous enzyme activities in the same tissues of non-transgenic mice with a similar genetic background. Oral administration of 1-deoxygalactonojirimycin (DGJ), a competitive inhibitor of alpha-Gal A and an effective ASSC for Fabry disease, at 0.05 mM in the drinking water of the mice for 2 weeks resulted in 13.8-, 3.3-, 3.9-, and 2.6-fold increases in enzyme activities in the heart, kidney, spleen and liver, respectively. No accumulation of globotriaosylceramide, a natural substrate of alpha-Gal A, could be detected in the heart of TgM/KO mice after DGJ treatment, indicating that degradation of the glycolipid in the heart was not inhibited by DGJ at that dosage. The alpha-Gal A activity in homozygous or heterozygous fibroblasts established from TgM/KO mice (TMK cells) was approximately 39 and 20 unit/mg protein, respectively. These TgM/KO mice and TMK cells are useful tools for studying the mechanism of ASSC therapy, and for screening ASSCs for Fabry disease.

1-Deoxynojirimycin↗
PubMed2004

Use of herpesvirus papio 2 as an alternative antigen in immunoblotting assay for B virus diagnosis.

Herpesvirus papio 2 (HVP2), which infects baboons, is much more closely related genetically and antigenically to monkey B virus (BV) than to human herpes simplex virus 1(HSV1) and other related herpes viruses. The usefulness of HVP2 as an alternative test antigen in immunoblotting assays to detect BV-antibody in macaque monkey sera was assessed. Six HVP2 proteins reacted with BV-positive sera in immunoblotting. No specific bands could be detected with BV-negative sera. These results show the usefulness of HVP2 antigen as an alternative and safer antigen than authentic BV antigen in detecting BV antibody in immunoblotting.

Animals↗
PubMed2004

Role of IL-6 and IL-1beta in reactivation by acetylcholine of latently infecting pseudorabies virus.

We previously reported that the latently infecting Pseudorabies virus (PrV) could be reactivated by injection of swine or mice with acetylcholine. However, the mechanism of the reactivation was not clear yet. In this study, we analyzed the kinetics of cytokines related to stress to clarify the relationship between virus reactivation by acetylcholine and the immune system. IL-6 and IL-1beta were detected in mice after stimulation with acetylcholine. This shows that acetylcholine induced physiological stress conditions. However, there seemed to be no relationship between the kinetics of the cytokine levels and PrV excretion. Moreover, neither IL-6 nor IL-1beta alone could reactivate latently infecting PrV. Thus, acetylcholine causes the reactivation of latent PrV via a mechanism not involving these immunological factors.

Acetylcholine↗
PubMed2003

Mapping of the low pH-sensitive conformational epitope of rabies virus glycoprotein recognized by a monoclonal antibody #1-30-44.

Monoclonal antibody (mAb) #1-30-44 recognized an acid-sensitive conformational epitope of rabies virus glycoprotein (G). The antigenicity of G protein exposed on the cell surface was lost when the infected cells were exposed to pH 5.8. By comparing the deduced amino acid sequence of G protein between the HEP-Flury strain and the epitope-negative CVS strain as well as the mAb-resistant escape mutants, two distant sites that contained Lys-202 and Asn-336 were shown to be involved in the epitope formation. Lys-202 is located in the so-called neurotoxin-like sequence, while Asn-336 is included in antigenic site III and is very near the amino acid at position 333, which is known to affect greatly the neuropathogenicity of rabies virus when changed. Consistent with this finding, antigenicity of a neurovirulent revertant of the HEP-Flury strain, in which Gln-333 of G protein was replaced by Arg, was also affected as shown by its greatly decreased reactivity with mAb #1-30-44 compared to that of the original avirulent HEP virus. Based on these results, we hypothesize that the neurotoxin-like domain and some amino acids in antigenic site III come into contact with each other to form a conformational epitope for mAb #1-30-44, and such a configuration would be lost when exposed to acidic conditions to perform a certain low pH-dependent function of G protein.

Amino Acid Sequence↗
PubMed2003

Effect of mild stress in mice latently infected pseudorabies virus.

Stress is one of the important factors that induces reactivation of pseudorabies virus (PrV) in latently infected pigs. We established a murine model of latent PrV infection and examined the effects of mild stress treatment in order to demonstrate that this model simulates natural infection in the pig. Latently infected mice excreted PrV from the nasal cavity under stress treatments consisting of restraint, exposure to cold or transport. Similar reactions have been observed upon treatment with acetylcholine and dexamethasone. The present findings demonstrate that these kinds of mild stress reactivate the virus in murine latent infection models in a manner similar to the induction of latent infection in pigs in the field.

Acetylcholine↗
PubMed2003

[Rabies virus].

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Animals↗
PubMed2002

Genetic analysis of rabies virus isolates in the Philippines.

To determine the genetic characteristics of the rabies virus in the Philippines, 59 rabies virus isolates were obtained from domestic rabid dogs and their partial nucleotide sequences of nucleoprotein (N) gene were compared. Based on comparison with reported sequences, phylogenetic analysis revealed that all isolates from the Philippines had close genetic relations and formed two subgroups. The Philippines isolates belonged to a different lineage from other Asian isolates but were closer to them than to terrestrial isolates and laboratory strains. Several specific nucleotide and amino acid substitutions were observed among the Philippines isolates. Our results suggest that rabies viruses in the Philippines might have a characteristic evolution.

Amino Acid Sequence↗