Tumor necrosis factor-beta in the serum of adult T-cell leukemia with hypercalcemia.

Serum tumor necrosis factor-beta (TNF-beta) from patients with adult T-cell leukemia (ATL) was studied by a sandwich enzyme-linked immunosorbent assay (ELISA) developed in our laboratory using biotinylated monoclonal anti-TNF-beta and recombinant TNF-beta. Seven of eight patients with hypercalcemia showed elevation of serum TNF-beta. On the other hand, TNF-beta could not be detected by the ELISA in 28 patients without hypercalcemia. The lower detection limit in this assay was 100 pg/mL, corresponding to 500 pg/mL by the conventional method. In two patients serum TNF-beta level decreased after treatment in association with the level of serum calcium. Furthermore, immuno-staining using anti-TNF-beta and avidin-biotin complex showed the presence of cytoplasmic TNF-beta in not only human T-cell leukemia virus type I infected cell lines, but also freshly isolated cells from ATL patients with hypercalcemia. The actual biologic activity of TNF-beta in serum was confirmed by a conventional bioassay in a patient with hypercalcemia, and its cytotoxic activity was inhibited by the addition of anti-TNF-beta antibody in the assay. These results suggested that serum TNF-beta might be one of the factors contributing to the hypercalcemia, at least in patients with ATL.

An enzyme-linked immunosorbent assay for immune complex of HTLV-I.

A sandwich enzyme-linked immunosorbent assay (ELISA) for immune complexes of human T cell leukemia virus type I (HTLV-I) was developed using monoclonal antibody (MoAb) 3G1 which recognizes a different epitope on HTLV-I to that with which natural human anti-HTLV-I antibody binds. The assay was capable of titrating artificial immune complexes not only at antigen-antibody equivalence but also at antibody excess. Although the antigen-antibody ratios could not be determined in the individual sera from patients with overt ATL, the level of immune complexes in three out of four sera was estimated to be 250 +/- 36 ng/ml. Immune complexes of HTLV-I could not be identified in sera obtained from one patient with overt ATL, three healthy HTLV-I carriers and three normal human controls.

[Classification of asymptomatic autoimmune thyroiditis by thyrotropin-releasing hormone loading].

Thyrotropin-releasing hormone loading was performed on 91 patients with asymptomatic autoimmune thyroiditis. Four women had no response to this loading test and had high levels in serum total and free thyroxine (TT4, FT4) and in serum total and free triiodothyronine (TT3, FT3). These patients might be classified as subclinical hyperthyroidism (Group G). Twenty-four patients had normal levels of both basal and peak thyrotropin after loading and were classified as Group I. There were no significant differences between 45 controls (Group C) and Group I patients in serum thyroid hormone levels. Patients with normal basal and high peak levels of thyrotropin were included in Group II. The number of patients in this group was 53. The mean levels of basal and peak thyrotropin were 4.8 microU/ml and 39.6 microU/ml, respectively, and were significantly higher than in Group C and Group I (P less than 0.005). In 10 patients classified as Group III with high levels of both basal and peak thyrotropin, serum concentrations of TT4, FT4 and FT3 were significantly lower than in the other groups (P less than 0.025); however, significant differences in TT3 could not be seen among them. Serum cholesterol levels gradually increased from Group C to Group III. There were significant differences between Group C and Group II (P less than 0.05).

Islet cell antibodies in the Japanese population and subjects with type 1 (insulin-dependent) diabetes.

Islet cell antibodies were studied in 1,112 non-diabetic adults, 473 normal school children and 162 Type 1 (insulin-dependent) diabetic patients in a Japanese population. The prevalence of islet cell antibodies was 0.5%, 0.4% and 32%, respectively. Most islet cell antibodies positive subjects with Type 1 diabetes had short duration of the disease. No patients who had over 10 years from the onset had islet cell antibodies. Six non-diabetic adults with islet cell antibodies were followed for 4 years. Only one with Hashimoto's thyroiditis showed a diabetic pattern in her oral glucose tolerance test. However, none developed overt insulin-dependent diabetes until 1984. Two out of these six subjects continued to be positive for both islet cell antibodies and antithyroid antibodies or antinuclear antibodies. Islet cell antibodies in the remaining four patients disappeared during the second year. It is difficult to predict the onset of Type 1 diabetes by islet cell antibodies in non-diabetic individuals because they may be transient.

[Immunohistochemical localization of cytoskeletal proteins and thyroglobulin in the follicular cells of Hashimoto's thyroiditis].

Using antisera against three kinds of cytoskeletal proteins (keratin proteins, actin protein and myosin protein) and thyroglobulin, immunoperoxidase staining was performed on the follicular cells of 30 patients with Hashimoto's thyroiditis. These patients were subdivided into three types by Woolner's classification: 9 patients of lymphoid type (L-type), 12 patients of oxiphilic cell type (O-type), and 9 patients of pronounced epithelial destruction type (P-type). The results obtained were as follows: (1) In three-ninths to seven-twelfths of the patients of O-type and P-type, the cytoskeletal proteins were identified in the epithelial cells forming degenerating or atrophic thyroid follicles. In the patients of L-type, however, the cytoskeletal proteins which form large follicles containing much colloid were not found in the epithelium. (2) In some patients of O-type, keratin proteins were abundantly present in the epithelial cells with squamous cell metaplasia. (3) In the patients of L-type, thyroglobulin was found in most of the epithelium forming large follicles, but it was not found in those forming degenerating or atrophic follicles in the patients of O-type and P-type.

A population survey of pancreatic islet cell antibodies and antithyroid antibodies.

In a survey of one Japanese population, we detected pancreatic islet cell antibodies (ICA), antithyroid antibodies (ATA) and antinuclear antibodies (ANA). The prevalence of ICA was 6 out of 1125 cases, or 0.5%. ATA and ANA were detected in 8.9% and 1%, respectively. There were no cases of either type I or type II diabetes in subjects with ICA. But there was one case who had ATA and another with ANA. Serum samples from this population had been obtained once a year from 1979 and one case with neither ATA nor ANA was found positive for ICA in 1980. Identical tests for ICA were performed on 80 childhood diabetics as were carried out on type I diabetics. Pancreatic isles cell surface antibodies (ICsA), ATA and ANA were studied simultaneously. The prevalence of ICA in 80 cases was 36.3% and that of ICsA was 13.8%. 4 cases had both ICA and ICsA. The prevalence of ATA was 11.2% and that of ANA was 16.3%.

Antinuclear antibodies in childhood diabetics.

Antinuclear antibodies (ANA) were detected both in type 1 diabetic children and in control subjects. The incidence of ANA in eighty of these diabetics was 16.3%, as determined using two different substrates, human pancreas and human peripheral leucocytes. The incidence and the patterns in the detection of ANA were the same. Four hundred and seventy-three children and one thousand one hundred and twenty-five adults served as the controls. The incidence of ANA in non-diabetic children was 0.8% and that in one adult population was 1.1%. Therefore, the incidence of ANA in childhood diabetics was significantly higher. We studied autoantibodies in childhood diabetics, in normal children and in one adult population. Pancreatic islet cell antibodies (ICA) were detected in 29 out of 80 type 1 diabetics (36.3%) in two out of 473 normal children (0.4%) and in six cases in one population (0.5%). thyroid microsomal antibodies (MCHA) were detected in 9 out of 80 childhood diabetics and the incidence of MCHA in type 1 diabetics was significantly higher than in the controls.

Relationship between reduced B cell susceptibility to IgM antibodies and reduced IgD-bearing B cells in patients with systemic lupus erythematosus.

Peripheral blood lymphocytes from 14 patients with systemic lupus erythematosus, 5 patients with rheumatoid arthritis, and 10 normal subjects were cultured for 7 days with or without anti-IgM or anti-IgD antibodies, and IgG- and IgM-secreting cells were assayed by reverse hemolytic plaque assay. Surface immunoglobulin (Ig) isotypes on peripheral blood B cells were also examined by a direct anti-Ig rosetting reaction. In normal subjects and rheumatoid arthritis patients, the spontaneous development of IgG- and IgM-secreting cells was markedly suppressed by anti-IgM or anti-IgD antibodies. Over 50% of peripheral blood B cells were IgD- and IgM-bearing cells in normal subjects and in most patients with rheumatoid arthritis. In lupus patients, however, the suppression of IgG and IgM production by anti-IgM or anti-IgD antibodies was remarkably reduced, especially in the active stage. Furthermore, the percentage of IgD-bearing cells in peripheral blood B cells was remarkably reduced, especially in patients with active disease. There was a good correlation between reduced susceptibility of B cells to anti-IgM antibody-mediated suppression and reduced percentage of IgD-bearing cells in lupus patients.

HLA-DR antigens in Japanese with Hashimoto's thyroiditis and Graves' disease.

HLA-A, -B, -C and -DR specificities were determined to study the association of HLA in 49 unrelated patients with Hashimoto's thyroiditis and 51 patients with Graves' disease. The results were compared with 144 control subjects from the Japanese population at large. Much less HLA-DR2 was found in the patients with Hashimoto's thyroiditis than in the normal controls. It was present in 16% of the thyroiditis patients compared to 39% in the controls. Also the frequency of HLA-DRl was less in the patients with Graves' disease (0%) than in the controls (15%). No positive association was found for any HLA-antigens in the two diseases.

HLA-DR specificities among Japanese with several autoimmune diseases.

HLA-A, -B, -C, and -DR antigens were determined in order to study the association of HLA in Japanese patients with several autoimmune diseases, hypertrophic cardiomyopathy, and Hodgkin's disease. The frequency of HLA-DR4 was significantly increased in the patients with rheumatoid arthritis, juvenile-onset insulin-dependent diabetes mellitus (IDDM) and hypertrophic obstructive cardiomyopathy. In this study, no significant associations with A, B, or C specificities were observed except BW22 in IDDM. In contrast, the negative association with HLA-DR2 was observed in Hashimoto's thyroiditis, pemphigus vulgaris and hypertrophic non-obstructive cardiomyopathy.

HLA-DR specifications in Japanese with juvenile-onset insulin-dependent diabetes mellitus.

Specific allelic associations vary among ethnic groups. We studied the distribution of HLA-A,-B,-C, and -DR antigens in 34 Japanese juvenile-onset diabetic patients. The focus of our current work was HLS-DR antigens because there have been few studies of Japanese with this disease. A significant increase in the frequency of HLA-DR4 was found in patients but not in unaffected persons: DR4 was found in 56.3% of the patients versus 32.6% of the unaffected persons. However, the negative correlation between DR2 and patients was not statistically significant.

Reduced susceptibility to anti-Fab antibodies of B cells in patients with systemic lupus erythematosus.

We have observed the effects of rabbit F(ab')2 anti-Fab antibodies (a-Fab Ab) on the differentiation of B cells in normal subjects and in patients with systemic lupus erythematosus (SLE). Peripheral blood lymphocytes were obtained and cultured with or without the addition of a-Fab Ab, and IgG- and IgM-secreting cells were assayed by reverse hemolytic plaque assay after 7 days of culture. In normal subjects, the spontaneous development of IgG- and IgM-secreting cells in the cultures of lymphocytes without pokeweed mitogen (PWM) was markedly suppressed by a-Fab Ab. In patients with SLE, however, the spontaneous development of IgG- and IgM-secreting cells was almost unchanged in the presence of a-Fab Ab. The effects of a-Fab Ab on immunoglobulin production induced by PWM were different in active and inactive SLE. The inhibitory effect of a-Fab Ab on PWM-induced development of IgG- and IgM-secreting cells was observed in normal subjects and inactive SLE patients. In active SLE patients, however, the production of IgG and IgM was not significantly increased by PWM and almost unchanged with the addition of a-Fab Ab. These findings show that B cells in SLE are much less susceptible to a-Fab Ab than normal B cells.