Kiyoaki Tanimoto

Family analysis of Werner's syndrome: A survey of 42 Japanese families with a review of the literature

Forty‐two Japanese families, including 80 individuals with Werners syndrome were studied, confirming that this syndrome is inherited as an autosomal recessive trait. The incidence of malignancy was relatively high in these families and individuals with Werners syndrome, although the incidence was not so high as was reported previously. HLA typing revealed no significant linkage with Werners syndrome. The frequency of Werners syndrome in Japan was estimated using two methods which indicated approximately 300 cases among 100 million people.

Association between HLA and Japanese patients with rheumatoid arthritis.

Japanese patients with rheumatoid arthritis (RA) were observed to have a statistical association with HLA-DR4, MT3. Strong association between the clinical severity of RA and HLA was also observed. Male patients had a stronger association with HLA than female patients. Males are more resistant to RA than females. This suggested that the threshold of liability for RA is higher in males than in females. Japanese patients with RA with systemic vasculitis were negative for HLA-Bw44 and had antilymphocytotoxic autoantibody, indicating that RA with systemic vasculitis is different in etiology from RA without systemic vasculitis.

Werner's Syndrome: Analysis of 15 Cases with a Review of the Japanese Literature

ABSTRACT: Fifteen patients with Werners syndrome were studied 12 males and 3 females, whose ages ranged from 17 to 59 years. Most of the previously reported clinical characteristics were confirmed, and an additional finding was the frequent occurrence of flat feet and hyperreflexia of the patellar and Achilles tendons. Also noted were an excessive urinary excretion of hyaluronic acid and a decrease in the T‐cell subpopulation. The significance of these findings is discussed.

Spontaneously enhanced in vitro immunoglobulin synthesis by B cells in systemic lupus erythematosus

Abstract In vitro immunoglobulin (Ig) synthesis was studied in peripheral blood lymphocytes (PBL) from patients with systemic lupus erythematosus (SLE) (SLE-PBL) in the presence or absence of pokeweed mitogen (PWM). In the presence of PWM, IgM and IgG syntheses were reduced in SLE-PBL, while in the absence of PWM, IgG synthesis was enhanced when compared with normal PBL although IgM synthesis was not significantly augmented. Preincubation and time course studies suggested that the enhancement of IgG synthesis in SLE is not due to the serum factors. The coculture experiments using separated T- and B-rich fractions indicated that reduced Ig synthesis in the presence of PWM may be the result of impaired T- and B-cell functions in SLE, and that raised Ig synthesis in the absence of PWM will be the result of primary B-cell activation. These results suggested that B cells in SLE-PBL are primarily activated and produce Ig without the aid of T cells.

Detection of Antilymphocyte Antibody with Two-Color Method in Systemic Lupus Erythematosus and Its Heterogeneous Specificities against Human T-Cell Subsets

The two-color method originally described by Van Rood et al. (Van Rood, J. J., A. Van Leeuwen, and J. S. Ploen. 1976. Simultaneous detection of two cell populations by two-color fluorescence and application to the recognition of B-cell determinants. Nature (Lond.). 262: 795-797) for the typing of homologous leukocytic antibodies, D-region was used for the detection of antilymphocyte antibody (ALA) in systemic lupus erythematosus. In this method, surface immunoglobulin-bearing cells were identified with fluorescein isothiocyanate-labeled anti-immunoglobulin and nuclei of killed cells were stained with ethidium bromide. Therefore, cell type (T or B) of the target cells can be identified without fractionating them. ALA was detected in 87% of lupus sera and had a preferential reactivity with T cells. Its major immunoglobulin class was shown to be immunoglobulin (Ig)M. The subspecificity of ALA was further analyzed using fractionated T-cell subsets as target cells. When T lymphocytes were separated into Fc receptor-bearing (Tgamma) and lacking (Tgamma[-]) cells, 64% of ALA showed preferential reactivity with Tgamma cells and 14% with Tgamma(-) cells. The remainder had no selective reactivity against Tgamma or Tgamma(-) cells. Tgamma cells were shown to have suppressor activity, whereas Tgamma(-) cells were indicated to contain helper cells. The above finding was in agreement with the observation that treatment of T cells with ALA that preferentially react with Tgamma cells considerably enhanced immunoglobulin synthesis in vitro, whereas treatment of T cells with ALA reactive with Tgamma(-) cells clearly suppressed the formation of immunoglobulins. Treatment of ALA with no selective reactivity showed variable effects on in vitro immunoglobulin synthesis. These results indicate that ALA in lupus have heterogeneous specificities against human T-cell subsets.

T lymphocyte interaction with immunoglobulin G antibody in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease with multiple immune disturbances whose mechanisms remain unclear. We examined the interaction of antilymphocyte antibodies with cultured normal T lymphocytes. T cells were prepared by E-rosetting after petri-dish removal of adherent cells and cultured for 2-7 d in the presence of SLE sera or normal human sera. Cultured T cells were washed and sonicated, and the amount of cell-associated IgG was quantitated by radioimmunoassay or enzyme-linked immunoassay (ELISA) methods. T cells cultured with 27 of 39 SLE sera showed marked increments of associated immunoglobulin G (IgG) although this was not observed with sera from mixed connective tissue disease patients containing high titers of ribonucleoprotein antibody or normal donors. The effective factors for IgG association in SLE sera were absorbed with normal peripheral blood lymphocytes or T cells. Anti-T cell IgG cytotoxic activity strongly correlated with T cell IgG association (P less than 0.01). T cell-associated IgG was not removed by stripping of cell membrane IgG from living cells by acid buffer treatment; indirect immunofluorescence of cells fixed after 2-4 d of culture revealed cytoplasmic IgG staining. IgG anti-T cell antibodies appeared to associate inside the cell membrane or to penetrate into the cytoplasm of cells. T cell Fc receptor blocking by heat-aggregated IgG or anti-beta 2-microglobulin antibody did not alter IgG cell association. Since pepsin-digested SLE sera showed no T cell association activity, whole IgG antibody molecules appeared to be necessary for interaction with cultured T cells. In addition, reduction and alkylation of active SLE sera completely nullified T cell reactivity. When normal T cells were cultured with SLE sera showing marked IgG T cell association, viability of cultured T cells decreased rapidly after 4 d, which suggests that IgG anti-T cell antibodies were associated with cell destruction. IgG cell-associating antilymphocyte antibodies present in SLE sera may cause T cell disturbances in vivo and may be related to the lymphocytopenia present in SLE patients.

Human anticentriole autoantibody in patients with scleroderma and Raynaud's phenomenon.

Unique autoantibody was found in sera from two different patients that reacted with centrioles of both cultured mammalian cells and human peripheral leukocytes as detected by the indirect immunofluorescent method. Sera from the same individuals, one with scleroderma and the other suffering from Raynauds phenomenon, also stained the basal bodies of rat tracheal ciliated cells by the identical technique. Data from subsequent investigations have suggested that the antigen(s) involved in the reaction are water-insoluble protein(s) or polypeptide(s) associated with centrioles and are distinct from microtubular proteins or purine nucleoside phosphorylase.

Genetic contribution of the tumour necrosis factor (TNF) B + 252*2/2 genotype, but not the TNFa,b microsatellite alleles, to systemic lupus erythematosus in Japanese patients

The contribution of the tumour necrosis factor (TNF) B + 252 (TNFB) dimorphism and microsatellite polymorphisms of TNFa and TNFb to the pathogenesis of systemic lupus erythematosus (SLE) was studied in Japanese patients. The TNFB dimorphism was determined using the restriction fragment length polymorphism (RFLP) method with NcoI digestion followed by specific polymerase chain reaction (PCR) amplification. TNFa and TNFb microsatellite polymorphisms were determined using the DNA sequencer and GeneScan program (Applera Corporation, Foster City, CA) followed by specific PCR amplification. HLA‐DRB1*15 typing was carried out by the PCR‐sequence specific conformational polymorphism (SSCP) method. In SLE, the allele frequency of TNFB*2 significantly increased (68.9%, P < 0.05) and the genotype frequency of TNFB*2/2 also increased (52.8%, P < 0.05). TNFB*2 showed no significant linkage disequilibrium with HLA‐DRB1*1501. The prevalence of TNFa13 and TNFb4 showed very slight increases, but these increases were not significant. An association analysis indicated that TNFB*2/2 conferred greater, or at least equal, susceptibility to SLE in Japanese patients in comparison with HLA‐DRB1*1501. The TNFB*2/2 genotype may contribute additively with DRB1*1501 to SLE in Japanese patients. No association was observed between auto‐antibodies and TNF. TNFB*2 is a genetic marker for SLE in Japanese patients, while TNFa and TNFb microsatellites are not associated with SLE.

Fc-Rosette Inhibition by Pregnant Women’s Sera and by Rabbit Anti-β2-Microglobulin

Some anti-HLA sera showed Fc-rosette inhibitory activity, which had little correlation with type-specific anti-HLA activity. Cytotoxic titers did not correlate with Fc-rosette inhibition rate and these antisera showed considerable inhibition of rosette formation even after absorption of anti-HLA activity with T lymphocytes. These results suggest that some anti-HLA sera contain B-cell-specific inhibitory activity against Fc-receptor like anti-Ia antibody in mice. Rabbit antiserum against human beta2-microglobulin showed specific inhibition of Fc-rosette formation. It was suggested that Fc-receptor or Ia-like antigen in human has a close relation with beta2-microglobulin.

Effect of free radicals on lymphocyte response to mitogens and rosette formation

Abstract When normal human lymphocytes were exposed to hypoxanthine plus xanthine oxidase system in vitro , the blastogenic response of lymphocytes to mitogens was markedly suppressed. The lymphocytes which respond to Con A were most sensitive to toxic effects of free radicals. In contrast, lymphocyte blastogenesis induced by PWM was less affected by free radicals. Superoxide dismutase was relatively ineffective in preventing the toxicity of free radicals on lymphocyte blastogenesis induced by PHA. In contrast, catalase showed a preventive effect on the oxidase-induced suppression of lymphocyte response to PHA. Uric acid plus uricase also suppressed blastogenic response of the lymphocytes to PHA. The suppressive effects of uric acid plus uricase on lymphocyte proliferation were reversed by catalase. Furthermore, hypoxanthine plus xanthine oxidase were demonstrated to have effects on the lymphocyte membrane receptors, by markedly inhibiting E rosettes; EAC rosettes were only slightly inhibited. These results suggest that H 2 O 2 suppresses lymphocyte function through damage to the cell membrane, and that T cells are more sensitive than B cells to the toxic effects of free radicals.