Immunotherapy for malignant tumors using combination of allogeneic intra-bone marrow-bone marrow transplantation, donor lymphocyte infusion and dendritic cells

We have previously shown that the combination of allogeneic intra-bone marrow-bone marrow transplantation (IBM-BMT) and donor lymphocyte infusion (DLI) using CD4+ cell-depleted spleen cells is effective in suppressing tumor growth, but that this does not induce graft-versus-host disease (GVHD) in mice. In this report, we show that formalin-fixed tumor cell-pulsed dendritic cells (FFTCP DCs) have an additive effect with IBM-BMT plus DLI on the suppression of tumor growth, but that the DCs do not augment GVHD. BALB/c mice, which had been subcutaneously inoculated with Meth A (BALB/c-derived fibrosarcoma), were irradiated at a low dose (5 Gy) and were transplanted with bone marrow cells (BMCs) from C57BL/6 (B6) mice into the bone marrow cavity (IBM-BMT). Simultaneously, the mice were intravenously injected with spleen cells from B6 mice, and subcutaneously injected with FFTCP DCs derived from the bone marrow (BM) of B6 mice. At the point of the induction of DCs from BMCs, formalin-fixed Meth A cells were added into the culture. The mice treated with the combination of FFTCP DCs, IBM-BMT and DLI using CD4+ cell-depleted spleen cells showed smaller tumor sizes and longer survival than the mice treated with IBM-BMT plus FFTCP DCs or IBM-BMT plus DLI using CD4+ cell-depleted spleen cells. These results suggest that the combination of FFTCP DCs, IBM-BMT plus DLI using CD4+ cell-depleted spleen cells has potent anti-tumor effects without showing GVHD.     

Results of survey conducted on perioperative chemotherapy and supportive care in primary breast cancer (JBCRG01)

We carried out a survey of supportive care at institutions that participated in the JBCRG01 study (FEC followed by docetaxel) as neoadjuvant therapy for operable breast cancer. The purpose was to share the information of supportive care for the treatment effect of perioperative intensive chemotherapy among institutions. Appropriate supportive care for nausea, vomiting, edema and febrile neutropenia (FN) is important with respect to the safety of chemotherapy. According to the results of the questionnaire, support from the family and the relationships with doctors, nurses and pharmacists familiar with the chemotherapy were important. The equipment and service for outpatients' cancer chemotherapy center are also important. This multicenter study enhances the exchange of information among institutes. The results of this survey suggest that adequate supportive care makes anthracycline and taxane chemotherapy manageable in the outpatient setting.     

Investigation of the clinical effect of large volume leukocytapheresis on methotrexate-resistant rheumatoid arthritis

Leukocytapheresis (LCAP) is already being used in a clinical setting for the treatment of autoimmune diseases such as inflammatory bowel disease and rheumatoid arthritis, and it has been reported to be effective. However, it is totally or partially ineffective in some patients, which has forced clinicians to rethink therapeutic strategies and concurrent treatment. With the aim of enhancing the therapeutic effect, we carried out large volume leukocytapheresis, with a throughput of 5000 mL instead of the 3000-mL throughput of conventional leukocytapheresis in nine patients with rheumatoid arthritis resistant to methotrexate treatment. Using Cellsorba, the column filled with the unwoven fabric made of the polyethylene phthalate, a leukocyte removal filter, large volume leukocytapheresis was carried out once a week for a total of five sessions. The observation period was the 12-week period following completion of treatment. The American College of Rheumatology (ACR) core set was used for assessment of efficacy. Eight weeks after completion of treatment, a 20% improvement in ACR was observed in 77.8% (7/9) of subjects, a 50% improvement in ACR was seen in 55.6% (5/9) of subjects, and a 70% improvement in ACR was observed in 22.2% (2/9) of subjects. C-reactive protein decreased gradually as treatment progressed, and a significant decrease was observed 4 weeks after completion of treatment. The fact that some subjects had an ACR70 response, few reports of which are observed in the case of conventional leukocytapheresis, and the fact that the effect continued up to 12 weeks after completion of treatment suggests that the degree and duration of the effect of large volume leukocytapheresis might be longer than those of conventional leukocytapheresis.     

Effects of extracorporeal treatment with Lixelle on the mortality and inflammatory responses to endotoxin-induced shock in rats

Endotoxemia and endotoxic shock are common problems in intensive care units and are associated with a very high mortality. Several previous studies have shown that Lixelle, which absorbs beta2-microglobulin for the treatment of dialysis-related amyloidosis, is also useful for the adsorption of inflammatory cytokines and endotoxins. The current study examined the use of Lixelle and its effects on the mortality and inflammatory responses to endotoxin-induced shock in rats. Male Sprague-Dawley rats were anesthetized and assigned to one of four groups (N = 13 per group): Escherichia coli endotoxin (15 mg/kg, i.v.) alone (endotoxemic); direct hemoperfusion apheresis without Lixelle for 120 min (direct hemoperfusion (DHP) alone); Lixelle treatment with Lixelle for 120 min immediately after endotoxin injection (Lixelle treatment); or Lixelle treatment with Lixelle for 120 min 2 h after endotoxin injection (Lixelle post-treatment). Hemodynamics and plasma lactate and cytokine concentrations were measured during observation. Mortality was assessed up to 8 h after the endotoxin injection. The mortality rates at 8 h after endotoxin injection were 92%, 85%, 23% and 46% for the endotoxemic, DHP-alone, Lixelle treatment, and Lixelle post-treatment groups, respectively. Elevated plasma cytokine concentrations were less conspicuous in the Lixelle treatment group than in the other three groups. Thus, Lixelle treatment drastically reduced the high mortality and the inflammatory responses in endotoxin-exposed rats. Moreover, Lixelle post-treatment also suppressed hypotension and a high mortality, although the inflammatory responses were the same as for endotoxin alone. These findings indicate that Lixelle treatment might be an effective therapy for endotoxemia and endotoxic shock.     

Phase I/II study of tandem high-dose chemotherapy with autologous peripheral blood stem cell transplantation for advanced multiple myeloma

The efficacy and safety of high-dose chemotherapy with tandem autologous peripheral blood stem cell transplantation (auto-PBSCT) were evaluated in a multicenter clinical study of patients with advanced multiple myeloma. Eligible patients (n = 40) were consecutively enrolled in the phase I/II study and received 2–4 cycles of vincristine–adriamycin–dexamethasone regimen. The responding patients underwent PBSC harvesting following high-dose cyclophosphamide and filgrastim administration. The first auto-PBSCT (n = 32) following high-dose melphalan (200 mg/m²) was performed within 2 months of PBSC harvesting; the second auto-PBSCT (n = 28) was scheduled 3–6 months later. Treatment-related mortality was 2.5% (n = 1) throughout the protocol. Grade 4 nonhematologic toxicity occurred in 12.5 and 14.3% of the first and second auto-PBSCT patients, respectively. All but one patient (who died) achieved hematopoietic recovery. For the 28 patients completing the second auto-PBSCT, the results were favorable with a response rate of 65% (complete response rate = 27.5%, n = 11); the five-year progression-free survival and overall survival were 20.3 and 66.5%, respectively. In conclusion, high-dose chemotherapy with tandem auto-PBSCT is feasible and safe with a favorable response rate in treating advanced multiple myeloma in Japan.     

Interobserver concordance of Ki67 labeling index in breast cancer: Japan Breast Cancer Research Group Ki67 Ring Study

The standardized assessment of Ki67 labeling index (LI) is of clinical importance to identify patients with primary breast cancer who could benefit from chemotherapy. In this study, we evaluated the interobserver concordance of Ki67 LI assessment. Six surgical pathologists participated and all the slides were prepared from archival breast cancer tissues fixed in 10% buffered formalin for 24 h and stained with MIB‐1. Three independent studies were conducted. In the first study, 30 stained slides were assessed using two different methods: the scoring system, with a positive rate scored from 1 (0–9%) to 10 (90–100%) by visual estimate; and the counting method, with approximately 1000 cells counted in hot spots. In the second study, 20 tumors with Ki67 LI 5–25% were assessed, and in the third study, 15 printed photographs of stained slides were assessed to avoid variations by selecting different fields. In study 1, the counting system (intraclass correlation coefficient [ICC], 0.66 [95% confidence interval 0.52–0.78]) demonstrated a better correlation than the scoring system (ICC, 0.57 [0.42–0.72]). In study 2, the assessment for Ki67 LI of 5–25% demonstrated a correlation (ICC, 0.68 [0.50–0.81]) similar to that of study 1 (unrestricted range of Ki67 LI). In study 3, the assessment of Ki67 LI by counting yielded a good concordance (ICC, 0.94 [0.88–0.97]). In conclusion, there was better concordance with the counting system, and concordance was high when the assessed field was predetermined, indicating that the selection of the evaluation area is critical for obtaining reproducible Ki67 LI in breast cancer.