Tetsuya Wada

Methotrexate for steroid-resistant systemic lupus erythematosus.

SummaryWe here report two patients with steroid-resistant systemic lupus erythematosus (SLE) who were successfully treated with methotrexate (MTX). In both cases, a steroid resistant high fever, associated with mild myositis and pancytopenia were the main common findings, and all these symptoms were alleviated within a few days either by 7.5 mg or 5 mg MTX per week. The number of CD4+ cells increased along with the clinical improvement, whereas the number of CD20+ cells and HLA-DR expressing cells also decreased. Taking into account the side effects of high dose corticosteroids and cyclophosphamides, treatment with a weekly low dose of MTX is known to contribute to an improvement in the long-term prognosis for patients with refractory SLE.

Multicentric reticulohistiocytosis associated with subclinical Sjögren's syndrome

SummaryA case of multicentric reticulohistiocytosis in a 60-year-old Japanese woman associated with subclinical Sjögrens syndrome is presented. The clinical features along with the light microscopic studies are commented. Partial improvement of the skin lesions and tendon sheath swelling was achieved after treatment with cyclophosphamide but the patients general condition remained unchanged. Taking the rarity of multicentric reticulohistiocytosis into account, coexistence of these two conditions in the present and previously reported cases suggests that an autoimmune mechanism may play a part in the pathogenesis of multicentric reticulohistiocytosis.

Coexistence of ochronosis and rheumatoid arthritis

SummaryWe describe a 64-year-old female patient with ochronosis and rheumatoid arthritis. Magnetic resonance imaging of the spinal column disclosed the destruction of vertebral disks, and a bony bridging in Th12 to L2. In addition, we observed joint space narrowing in the wrists as well as among the carpal bones, positive rheumatoid factor and the presence of rheumatoid nodules, in which the histological findings were compatible with those of rheumatoid arthritis. The coexistence of these two diseases has not yet been previously reported. Pre-existing ochronotic arthropathy might have masked the manifestation of rheumatoid arthritis and made the diagnosis of rheumatoid arthritis rather difficult.

Enhanced chemosensitivity of cells from malignant effusions under condition of exposure to high temperature

Utilizing a clonogenic assay, the effects of hyperthermia and selected chemotherapeutic drugs on growth of cells from malignant effusions were studied. Fourteen of 25 samples obtained from 25 patients with various carcinomas formed at least 30 colonies per plate. Exposure of the cells to heat at 42°C for 1 hr before the plating slightly inhibited the colony growth. The drugs, adriamycin (AM) and mitomycin C (MMC), were tested at 3 different concentrations. When the cells were treated with these two drugs for 1 hr at 42°C, the percent of surviving colonies was significantly decreased, as compared to findings at 37°C, in both groups, at 3 different concentrations. The combination of drugs and hyperthermia appeared to function synergistically in one-third of such cases. These results suggest that cells from malignant effusions in patients with various carcinomas were more sensitive to AM or MMC, under condition of a higher temperature (42°C).

Additive two DMARD therapy of the patients with rheumatoid arthritis.

SummaryFrom the beginning of 1987 to the end of 1989, 72 rheumatoid arthritis patients (RA) whose disease could not be controlled by a single disease modifying antirheumatic drug (DMARD) were selected for the trial treatment. They continued the DMARD treatment used initially at its regular dose, and then started another DMARD regimen at 1/3 to 1/2 of the regular dose as an additive DMARD treatment, which we have designated as Additive Two DMARD Therapy (ATDT). The patients were followed until the end of 1992. In the 3 months of ATDT, the effectiveness of ATDT was obtained in 42 (58.3%) patients who showed more than a 30% decrease in the initial Lansburys activity index (AI). The rate of side effects at 3 months were 5.6%. Tiopronin, bucillamine or salazopirine added to gold sodium thiomalate or tiopronin were suggested as the recommended DMARD combinations for ATDT. The suppressive effects on AI, ESR, CRP and rheumatoid factor continued for as long as 18 to 24 months. The mean period of ATDT was 21.7 months and that at which ATDT proved useful was 31.9 months. A discontinuation of the first DMARD treatment without any following disease aggravation was obtained in 10 of 15 patients whose disease activity had been sufficiently suppressed for longer than a year. In conclusion, ATDT was suggested to be a useful way of treating RA patients whose disease activity could not be controlled by a single DMARD treatment, as well as a way of evaluating the next DMARD while the ongoing DMARD was observed to gradually lose its initial drug effect.

Combination chemoimmunotherapy for advanced gastric carcinoma

Eighty-nine patients with advanced gastric carcinoma were treated with a combination chemo-immunotherapy regimen that consisted of active immunotherapy with Vibrio cholerae neuraminidase (VCN) treated autologous tumor cells admixed with BCG and drugs including cyclophosphamide, mitomycin C (MMC) and 5-fluorouracil, followed by long term tegafur (FT) and immunomodulators. This treatment significantly improved survival rate of patients in Stages III, IV and unresectable or recurrent carcinoma, compared to that of historical controls. As compared to controls treated with MMC followed by long term FT and immunodulators concurrently, survival rate of those in Stage III tended to improve (p<0.1) and survival rate at 4.5 years in Stage III was significantly higher (p<0.01), although it was not improved in Stage IV. In patients with unresectable or recurrent tumor, survival time was not significantly lengthened with this therapy when compared with that in patients given BCG alone in the same treatment schedule (CCI-BCG group). However, none of 19 patients in CCI-BCG group survived more than 15 months, although 4 of 28 patients receiving this therapy survived. These results suggest that this combination chemo-immunotherapy is effective for a selected group of patients with advanced gastric carcinoma.

Cell-mediated cytotoxic activity of spleen cells from patients with gastric carcinoma

The cell-mediated cytotoxic activities of cells from the spleens (SP cells) of patients with gastric carcinoma were assayed in comparison with the activities of peripheral blood mononuclear cells (PBM cells) from the same patients, and from patients with benign lesions. The natural killer cell (NK) activity of the SP cells and their capacity to generate allogeneic cytotoxicity in mixed lymphocyte culture (MLC) were very similar to those of the PBM cells. The cytotoxic activity of SP cells induced by alloactivation in MLC, however, was significantly higher than that of the PBM cells from the same patient as well as from patients with benign lesions. The production of interleukin 2 (IL 2) and the ability to induce cytotoxic cells after activation with IL 2 (LAK) were therefore examined. Both the ability to produce IL 2 and to generate LAK cells were shown to be significantly increased in SP cells when compared to PBM cells. These results indicate that the spleen may be a potential reservoir for the precursors of these activated killer cells in patients with gastric carcinoma. Furthermore, it may play an important role in the defence against tumors in these patients.

A simplified tritiated thymidine incorporation assay for chemosensitivity testing of human tumors

Chemosensitivity testing was performed using a simplified rapid assay based on tritiated thymidine incorporation. In this assay, 1.5 X 10(5) tumor cells were cultured for 72 h in double-layer agarose in 25 mm plastic dishes, which had a Millipore membrane sealed to the underside. After 24 h labeling of the cells with tritiated thymidine, cells were collected on the membrane by evacuating the solubilized medium through the filter sealed to the underside of the dish, which was placed in a boiling water bath. The radioactivity of the collected cells was counted by liquid scintillation. We found that 65% of 68 malignant tumors gave evaluable chemosensitivity results. Overall, chemotherapeutic drugs caused a 38% response rate in vitro. Twenty instances were evaluable for in vitro-in vivo correlations. The predictive accuracies were 93% for resistance and 67% for sensitivity. The results indicate that, with technical simplicity and shorter time course (4 days), this assay may be useful for chemosensitivity testing in human tumors.

Cell-mediated cytotoxic activity of regional lymph node cells from patients with gastric carcinoma

The cell-mediated cytotoxic activities of cells from the perigastric lymph nodes (LNC) were assayed in patients with gastric carcinoma. These activities were compared with those of the peripheral blood mononuclear cells (PBM), and also with the LNC of patients with benign lesions. The capacity of LNC to be converted to cytotoxic cells in mixed cell culture was significantly impaired in the cancer patients as compared to that of either the PBM from the same patients, or the LNC from patients with benign lesions. The natural killer cell (NK) activity of LNC was significantly lower than that of the PBM in both groups of patients. The cytotoxic activity induced by phytohemagglutinin activation (PAK) in the LNC from patients with carcinoma, as well as from those with benign lesions was also significantly decreased, when compared to that of the PBM, although the ability of LNC to produce interleukin 2 (IL 2) was significantly increased. The ability of these cells to generate cytotoxicity after activation with IL 2 (LAK) was therefore examined, and a decreased capacity in LNC was observed. These results indicated that the ability of T cells in LNC to develop into cytotoxic cells in patients with gastric carcinoma was impaired, and that the nonspecific cytotoxicity, including NK or PAK, as well as LAK activity, was essentially low in the perigastric lymph nodes.

A simplified method for determination of tritiated thymidine incorporation into cells from tissue in soft agar culture

We designed a simple method forin vitro chemosensitivity testing of tritiated thymidine incorporation by cells from tumor tissue plated in soft agar. Cells from tumor tissue were cultured in a plastic dish, of which a Millipore membrane had been sealed to the underside, containing two layers of agar. After labeling the cells with tritiated thymidine, the dishes were placed in a boiling water bath to solubilize the agar. The labeled cells could be readily collected on the membrane by evacuating the solubilized medium through membrane filters sealed underside of the dish. Using this system, 11 (55 per cent) of 20 tumor specimens from 20 patients with various carcinomas showed an incorporation of over 200 dpm above background and 80 per cent or greater inhibition of thymidine uptake by sodium azide treatment. This method offered several advantages over the thymidine incorporation assay, including technical simplicity and a shorter time course.