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H Nomaguchi

Publications and source records attributed to H Nomaguchi.

At least 37 records · Page 2Linked to original sources
PubMed1994

Antibodies to 65Kd heat-shock protein were elevated in rheumatoid arthritis.

Antibodies to 65Kd heat-shock protein (hsp) of mycobacterium leprae were measured by enzyme-linked immunosorbent assay (ELISA) in the three immunoglobulin classes in paired sera and synovial fluids of patients with rheumatoid arthritis (RA). Titers of anti-hsp antibody were expressed by optical density (OD) values for sera or indexes (OD values divided by amounts of immunoglobulin in each class) for synovial fluids and for their paired sera. Indexes of anti-hsp antibody were higher in synovial fluids than those in sera at 15/18 for IgG, 17/18 for IgA and 16/18 for IgM class. These results suggest the participation of anti-hsp antibodies to synovitis in RA.

Antibodies↗
PubMed1994

Antibody to 65-kD stress protein (HSP65) of Mycobacterium leprae in various inflammatory skin diseases. A preliminary report.

Cells from prokaryotes and eukaryotes exposed to environmental changes produce a series of highly homologous proteins called stress proteins, or heat shock proteins (HSPs). Recent investigators suggested that the reaction to the shared antigenic epitope between HSPs may link infections with induction of autoimmune processes. In the present study, antibody level to HSP with 65 kDa (HSP65) of Mycobacterium leprae was investigated with enzyme-linked immunosorbent assay (ELISA) in various skin diseases. Comparing to normal group (n = 9) including patients with nevus cell nevus showing 0.097 +/- 0.039 (mean +/- SD) in anti-HSP65 IgG level (OD492), 20 patients with palmoplantar pustulosis (PPP) and 22 with psoriasis demonstrated elevated (0.170 +/- 0.079, 0.111 +/- 0.053, respectively) level. Among them patients judged as focal infection-related PPP or psoriasis showed significantly higher level of anti-HSP65 (p < 0.01) than those without focal infection. Anaphylactoid purpura (0.125 +/- 0.085, n = 5), Behcet disease (0.178 +/- 0.045), atopic dermatitis (0.218 +/- 0.096, n = 13), urticaria (0.185 +/- 0.079, n = 30), and herpes zoster (0.193 +/- 0.092, n = 13) showed likewise elevated anti-HSP65 antibody. Similar tendency was found in anti-HSP65 IgM level but not in anti-HSP65 IgA. Western blotting confirmed specific immunoreaction bands to HSP65 in blood samples with high titer. Immunomodulation by stress proteins of bacterial or host cells is assumed in pathophysiology of inflammatory skin disorders, especially in relation to focal bacterial infection as observed in cases with PPP and psoriasis.

Autoantibodies↗
PubMed1993

Presence in Kawasaki disease of antibodies to mycobacterial heat-shock protein HSP65 and autoantibodies to epitopes of human HSP65 cognate antigen.

The central features of Kawasaki disease are immune activation and cytokine-mediated generalized vasculitis. To identify the predisposing factors, we examined the antibody response to BCG antigens, since reactivation of a previous BCG inoculation site is an early, specific manifestation of this disease. BCG antigens were separated on SDS-PAGE, transferred to membrane, and incubated with acute- and convalescent-phase sera of 21 patients with Kawasaki disease. Sera were also examined for the presence of antibodies to mycobacterial 65-kDa heat-shock protein (HSP65), and to its human homolog P1 antigen using synthetic peptides of nonhomologous region. To demonstrate the HSP65-sensitized T cells, in vitro proliferation assay was performed. All convalescent, but not acute phase, sera showed a strong antibody reactivity against 65-kDa protein. The reactivity was directed to recombinant HSP65. Non-cross-reactive sequences between rHSP65 and human HSP65 cognate were synthesized. The sera recognized these peptides of rHSP65 and autologous P1 antigen. Peripheral lymphocytes proliferated following the addition of rHSP65 (stimulation indices, 2.16-7.82; mean, 4.54). These findings suggest that HSP65 may be the most potent factor predisposing to Kawasaki disease, and that an autoreactivity to the epitope of the human HSP65 homolog may be related to the susceptibility to the disease.

Amino Acid Sequence↗
PubMed1993

Detection of heat shock protein in patients with insulin-dependent diabetes mellitus.

We have investigated whether antibodies to heat shock protein (hsp) 65 are present in sera from patients with insulin-dependent diabetes mellitus by using Mycobacterium leprae hsp65. Fifty-two sera from patients with IDDM, 36 from patients with unclassified insulin-treated diabetes mellitus and 41 from normal healthy controls were examined by ELISA assay. Seventeen (32.7%) out of 52 IDDM sera and 10 (27.8%) out of unclassified insulin-treated diabetic sera were positive for anti-Mycobacterium (anti-M. leprae) hsp65 antibodies while none of the healthy control sera were positive. Based on western blot analysis, 12 of the 17 IDDM sera and 1 of 2 sera from the unclassified insulin-treated diabetics were positive for anti-M.leprae hsp65 antibodies while all normal control sera were negative. These results support the idea that hsp65 may play a role in the pathogenesis of IDDM. Future studies are necessary to elucidate the role of hsp65 in the pathogenesis of IDDM.

Antibodies, Bacterial↗
PubMed1993

Immunological properties of ribosomal proteins from Mycobacterium bovis BCG.

Two proteins with molecular mass 65 kDa, a heat shock protein, and an S1-like protein were found in a 30S ribosomal subunit from Mycobacterium bovis BCG. The 17-kDa protein in the 30S subunit was homologous to alpha-crystallin heat shock protein, and the 16-kDa protein in the 50S subunit was homologous to the L7/L12 protein. The latter provoked a strong delayed-type hypersensitivity reaction in the sensitized guinea pigs. The GroES-like protein (12 kDa) loosely associated with ribosomes.

Animals↗
PubMed1992

Constitutive expression of 65-kDa heat shock protein (HSP65)-like immunoreactivity in cultured mouse oligodendrocytes.

The expression of mycobacterial 65-kDa heat shock protein (HSP65)-like immunoreactivity in cultured mouse oligodendrocytes and astrocytes was investigated using three monoclonal antibodies (ML30, IA1, 3A) specific for the mycobacterial HSP65. In western blot analysis, these antibodies recognized the proteins with molecular weights approximately of 50-, 60-, and 70-kDa expressed in both heat-stressed and unstressed glial cells. When the cells were exposed to heat stress, the expression of both 50- and 70-kDa proteins was attenuated, whereas that of the 60-kDa protein was not affected. On immunocytochemical studies, an appreciable level of HSP65 immunolabelling was identified in most (> 90%) oligodendrocytes under both heat-stressed and unstressed conditions but only marginally detectable in most (> 95%) astrocytes. These results indicate that mouse oligodendrocytes in vitro express the mycobacterial HSP65-like immunoreactivity constitutively.

Animals↗
PubMed1989

Overproduction, affinity purification and characterization of 65-kDa protein of Mycobacterium leprae in Escherichia coli.

The 65-kDa protein of Mycobacterium leprae was produced in an Escherichia coli strain carrying a plasmid harboring the recloned gene coding for the protein. The protein was purified through affinity chromatography prepared with the IgG fraction of a monoclonal antibody which was prepared against the 65-kDa protein. The purified 65-kDa protein also reacted immunologically with the monoclonal antibody IIIE9, which recognizes the epitope for M. leprae, prepared by Buchanan, et al. BALB/c mice were inoculated with M. leprae and 4 months later were skin tested with the purified 65-kDa protein. Gross changes were observed at the skin-test site. The role of the protein in protective immunity against M. leprae foot pad infection in mice was also studied.

Animals↗
PubMed1987

Characterization of antibody-reactive epitopes on the 65-kilodalton protein of Mycobacterium leprae.

Twenty-three monoclonal antibodies (MAbs) prepared in seven different laboratories were studied, all of which recognized the 65-kilodalton (kDa) protein of Mycobacterium leprae as determined by Western blotting or gel radioimmunoassay or both. Fourteen of the MAbs recognized different epitopes, as evaluated by cross-competition studies using radiolabeled MAb and unlabeled inhibitors; the species specificity of these epitopes was defined by nitrocellulose dot blot immunoassays with bacterial sonic extract antigen preparations from 23 species of mycobacteria. Each of the 14 distinct MAbs recognized a 65-kDa protein produced by a lysogenized Escherichia coli Y1089 host containing cloned rDNA which included the gene for the M. leprae 65-kDa protein. Of the 14 distinct MAbs, 1 recognized an epitope found only on M. leprae, and the others recognized epitopes present on as few as 8 or as many as all 23 of the mycobacterial species studied. Identification of these distinct 65-kDa protein epitopes and use of the MAbs which recognize them should assist future structural studies of this protein and characterization of the T-cell reactive and serodiagnostically useful portions of the molecule.

Antibodies, Monoclonal↗
PubMed1986

Restored pathogenicity of attenuated Mycobacterium lepraemurium in mice.

The ability of Mycobacterium lepraemurium (Mlm) to adhere to A31 cells in culture decreased with the number of passages of the bacilli on Ogawa egg-yolk medium. Pathogenic Mlm consistently grew in tissue culture cells but growth was not seen with attenuated Mlm isolated from a smooth colony. After prolonged incubation, attenuated Mlm became adapted to tissue culture growth. The pathogenicity of the attenuated bacilli was restored partially by the adaptation to tissue culture cells and restored almost completely by passage in mice. After restoration of pathogenicity by these methods, the Mlm formed rough-type colonies on Ogawa egg-yolk medium although the colonies were not completely of the rough type. Attenuated Mlm did not interfere with the growth of in vivo-derived Mlm in tissue culture or in mice.

Adhesiveness↗
PubMed1980

Growth and drug sensitivity of M. lepraemurium by tissue culture applying monolayer and agar suspension technique.

M. lepraemurium grow well in a Balb/c 3T3 recloned cell line (A31). In monolayer culture, the average generation time of M. lepraemurium in A31 cells was 5.3 to 9.4 days at 37 degrees C. A31 cells are very sensitive to infection with M. lepraemurium. Bacterial increases were readily apparent 30 days after inoculating 2 X 10(5) A31 cells in monolayer culture with only six bacilli. The intracellular bacilli were well transferred without apparent losses by host cell transfer. The growth of intracellular bacilli was inhibited by streptomycin 100 micrograms/ml, clindamycin 25 micrograms/ml, INH 5 micrograms/ml, and rifampin 5 micrograms/ml. When streptomycin or clindamycin was removed from the culture medium after 41 days of treatment and the cultivation continued in drug-free medium, the intracellular bacilli began to multiply once more without a lag period. When the intracellular bacilli were treated with INH for 35 days or rifampin for ten days, growth resumed, but only after lag periods after removal of these drugs. We utilized agar suspension techniques for the cultivation of host cells M. lepraemurium because normal cells or transformed cells ceased undergoing cell division and remained healthy for long periods of time in agar medium. M. lepraemurium grew well in A31, A31 transformed by polyoma virus, nude mouse foot pad, chick embryo, and human neuroblastoma cells, utilizing the agar suspension technique. The agar suspension cell culture method should provide useful clues for the cultivation of M. leprae.

Agar↗