Autologous cytotoxic T lymphocytes (CTL) against primary-cultured malignant gliomas were generated from peripheral blood mononuclear cells in vitro in 4 patients. Activities of the CTL were highly specific to the corresponding autologous glioma and were inhibited, in one patient, with antibodies against CD3, CD8 and MHC-class I molecules. When the CTL were injected 3 times into the primary-tumor-resected cavity via an Ommaya tube, reduction of the recurrent tumors with magnetic resonance imaging (MRI)-measured volumes exceeding 45 cm3 was observed in 3 patients. In a patient with glioblastoma multiforme (GBM), the tumor volume (estimated, 130 cm3) was rapidly reduced to 1/3, although re-recurrence of the tumor followed 40 days later. A slight but distinct rapid reduction of the tumor volume was observed in another GBM patient and in an anaplastic astrocytoma patient; essentially no change was observed in a further GBM patient. These results suggest that adoptive immunotherapy with autologous CTL will be clinically effective against end-stage malignant gliomas.
e13029 Background: We report a prospective phase I/IIa clinical trial of fractionated radiotherapy (FRT) concomitant with Temozolomide (TMZ) , followed by autologous formalin-fixed tumor vaccine (AFTV) and TMZ maintenance in patients with newly diagnosed glioblastoma multiforme (GBM). Methods: Twenty-four patients (aged 16-75 years; Karnofsky performance scale score 60% or more before initiation of FRT) with newly diagnosed GBM received a total dose of 60 Gy of FRT with daily concurrent TMZ. After a 4-week interval, the patients received three AFTV injections and the first course of TMZ maintenance chemotherapy for 5 days, followed by multiple courses of TMZ for 5 days in each 28-day cycle. Results: This treatment regimen was well toleratedby all patients. The percent of patients with progression-free survival (PFS) ≥24 months was 33%. The median PFS, median overall survival (OS), and the actuarial 2-year survival rate of the 24 patients were 8.2 months, 22.2 months, and 47%, respectively. The median PFS in patients with a delayed-type hypersensitivity (DTH) response after the third AFTV injection (DTH-2) of 10 mm or larger surpassed the follow-up period (29.5 months), which was significantly longer than in patients with a smaller DTH-2 response. Moreover, total resection of the tumor was associated with high DTH response. Conclusions: The treatment regimen resulted in favorable PFS and OS for newly diagnosed GBM patients. Total resection may enhance DTH response after vaccination. Clinical trial information: UMIN000001426.
Journal Article Inhibition of 6-Methylsalicylic Acid Synthesis by the Antibiotic Cerulenin Get access Hiroie OHNO, Hiroie OHNO The Kitasato Institute and Kitasato UniversityShirokane, Minato-ku, Tokyo 108 Search for other works by this author on: Oxford Academic PubMed Google Scholar Tadao OHNO, Tadao OHNO The Kitasato Institute and Kitasato UniversityShirokane, Minato-ku, Tokyo 108 Search for other works by this author on: Oxford Academic PubMed Google Scholar Juichi AWAYA, Juichi AWAYA The Kitasato Institute and Kitasato UniversityShirokane, Minato-ku, Tokyo 108 Search for other works by this author on: Oxford Academic PubMed Google Scholar Satoshi ÖMURA Satoshi ÖMURA 1 The Kitasato Institute and Kitasato UniversityShirokane, Minato-ku, Tokyo 108 1 To whom all of correspondence should be addressed. Search for other works by this author on: Oxford Academic PubMed Google Scholar The Journal of Biochemistry, Volume 78, Issue 6, December 1975, Pages 1149–1152, https://doi.org/10.1093/oxfordjournals.jbchem.a131010 Published: 01 December 1975 Article history Received: 28 May 1975 Published: 01 December 1975
Expression of N-ras gene was examined by northern blot analysis in two cases of astrocytoma (grade II), four cases of glioblastoma multiforme (six samples including two recurrent tumors), and normal brain tissues. Expression of N-ras gene was only found in the two recurrent glioblastoma multiforme samples. Southern blot analysis found no significant difference in copy numbers of N-ras gene in pre- and post-recurrence samples. The histological diagnosis of the two recurrent tumors was glioblastoma multiforme at both first recurrent presentation. However, the tumors had progressed during recurrence, because of worsened histological findings, such as increased cellularity, pleomorphism, and vascularity. These tumors recurred at very short intervals. Enhanced expression of N-ras gene by disorder of transcription may be a factor in the progression of glioblastoma multiforme.
The sensitivity of five kinds of cytotoxicity assays using ethanol on human hepatoblastoma cells (HUH-6 line), which were cultured as monolayers or spheroids, was compared. Ethanol was chosen as a test because it acts on cell membranes directly without being metabolized and exerts its cytotoxicity. The assay methods used were as follows: 3- (4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), lactate dehydrogenase (LDH), colony formation, cell growth and DNA assays. The sensitivity of the assays was: LDH < DNA < cell growth < MTT < colony formation. LDH assay had the advantage that the same culture could be used for multiple assays, but when a small number of cells were assayed, no significant increase in the release of LDH was detected in the assay cultures compared with the control cultures. Although the DNA and cell growth assays were more sensitive than the LDH assay, the extent of cell damage may be underestimated because the damaged cells and DNA present in the cultures are included in the assay samples. On the other hand, both MTT and colony formation assays showed a high sensitivity. The MTT assay was done within 24 h after ethanol was added to the cultures and was applicable to both monolayer and spheroid cultures, while the colony formation assay required 1-2 weeks and it was applicable only to monolayer cultures. Taken together, the MTT assay was the most suitable method to evaluate the cytotoxic effects of ethanol on HUH-6 cells cultured as either monolayers or spheroids.
Fibroblast-like cells from a BALB/c mouse embryo were propagated and transferred weekly in culture with a constant inoculation density of 5 × 103 cells/cm2. In this culture schedule a period of decreased proliferation (crisis) continued for 3 to 4 weeks before the initiation of continuous cell proliferation. This provided an experimental system suitable for studying proliferation conversion. The number and the proliferation rate of clonogenic cells increased early in crisis. Production of the colony-stimulating factor for mouse bone marrow cells by fibroblast-like cells was enhanced during the period of decreasing proliferation and crisis, then it decreased when continuous proliferation began. The results indicate that cells with infinite proliferation potential appear early in crisis, although the major population of cells terminates proliferation during crisis and enhances differentiated functions.
