The establishment of a new glioma cell line, DBTRG-05MG, in a modified RPMI 1640 medium is described. The cells were derived from an adult female with glioblastoma multiforme who had been treated with local brain irradiation and multidrug chemotherapy; the tumor showed substantial change in histologic appearance compared to the original biopsy 13 mo. previously. The line has been successfully cryopreserved and passaged up to 20 times. The karyotype of the cells demonstrated it as a hypotetraploid line; the DNA index of 1.9 confirmed the karyotype analyses. By immunocytochemical analysis, the cell line reacted with polyclonal antibodies to vimentin, S100, and neuron specific enolase, reflecting its primitive neuroectodermal character. Positive immunostaining for epidermal growth factor receptor correlated with the excess of chromosome 7 seen in the karyotype. The cell line reacted negatively to antibodies against platelet-derived growth factor and its receptor, neuronal cell adhesion molecule, and glial fibrillary acidic protein. By flow cytometry, the cells were major histocompatibility class I antigen positive and class I antigen negative. Growth kinetic studies demonstrated an approximate population doubling time of 34 to 41 h and a colony forming efficiency of 71.4%. Western blot analysis showed the presence of low levels of normal-sized retinoblastoma protein. When compared to the patient's lymphocyte DNA, no loss of heterozygosity of the p53 tumor suppressor gene was observed in the DBTRG-05MG cell line DNA.

Between August 1986 and October 1987, the Denver Brain Tumor Research Group conducted a clinical trial using autologous human recombinant interleukin-2 (rIL-2)-activated lymphocytes to treat 20 patients with recurrent high-grade gliomas. The trial involved surgical resection and/or decompression followed by intracavitary implantation of lymphokine-activated killer (LAK) cells and autologous stimulated lymphocytes (ASL) along with rIL-2 in a plasma clot. One month later, stimulated lymphocytes and rIL-2 were infused through a Rickham reservoir attached to a catheter directed into the tumor bed. The LAK cells were rIL-2-activated peripheral blood lymphocytes cultured for 4 days; the ASL were lectin- and rIL-2-activated peripheral blood lymphocytes cultured for 10 days. Of the 20 patients treated, 11 were evaluated as a group (mean age, 44 years, range, 15-61 years; mean Karnofsky rating, 69, range, 50-100; mean Decadron dose at entry, 14 mg/d, range, 0-32). The average number of lymphocytes implanted was 7.6 x 10(9) (range, 1.9-27.5 x 10(9], together with 1 to 4 x 10(6) U of rIL-2. To date, 10 of the 11 patients died, all from recurrent tumor growth. The median overall survival time was 63 weeks (range, 36-201; mean, 86). The median survival time after immunotherapy was 18 weeks (range, 11-151; mean, 39). No significant difference in survival after immunotherapy was found between those patients who had received previous chemotherapy and those who had not. The use of steroids or prior chemotherapy did not influence the in vitro generation of ASL or LAK cells.(ABSTRACT TRUNCATED AT 250 WORDS)

A minimized cardiopulmonary bypass circuit provides less hemodilution, platelet consumption, chest tube output and lower post-operative blood loss than standard cardiopulmonary bypass. Red blood cell usage was also less. All differences are advantageous.

Recombinant interleukin 2 (rIL-2) and various effector cell populations were used for adoptive immunotherapy in the Fischer strain 9L rat gliosarcoma model. The in vivo cytotoxicities of nonspecifically activated lymphocytes and specifically activated cytotoxic T lymphocytes (CTLs) were assessed in a modified in vivo neutralization (Winn) assay.Effector cells (106) those who received syngeneic unstimulated T lymphocytes and rIL-2. For a set of animals given the same inoculum of 9L tumor, significantly improved survival was shown for groups treated with nonadherent or adherent LAK cells (P < 0.0003), syngeneic CTLs (P = 0.0327), or allogeneic CTLs (P = 0.0025) over untreated control animals by using Mantel-Haenzel nonparametric logrank equations. Only treatment with allogeneic CTLs prevented tumor take.The rat 9L gliosarcoma model provides an efficient and rapid means to explore the efficacy of lymphokine and cellular therapy for brain tumors. It is derived from an inbred strain of Fischer rats, with a major histocompatibility complex haplotype of RTII'i (1, 2). Intracranially implanted 9L glioma cells grow in a predictable fashion in the syngeneic Fischer 344 rat (3). Rats bearing 9L tumor and given systemic recombinant interleukin 2 (rIL-2) therapy show a small increase in survival time compared to untreated control animals (4).Adoptive immunotherapy has been investigated in other brain tumor models. Lymphokine-activated killer (LAK) cells and rIL-2 were combined with the F98 rat glioma tumor 18 hr before implantation into the rat cerebrum and the rats exhibited an increased survival (5). Similarly, a clone of syngeneic tumor-sensitized cytotoxic T lymphocytes (CTLs) was partially effective in the immunotherapy of 203 glioma in an animal model (6). The in vivo antitumor activity of the clone has been demonstrated both in a Winn neutralization assay against 203 glioma cells inoculated intracranially and when administered intravenously 7 days after intracranial inoculation of 203 glioma. In rats bearing T9 gliosarcoma, LAK cells administered intravenously and intratumorally increased survival, but immune spleen cells did not (7). Likewise, LAK cells administered to Wistar rats bearing C6 glioma showed antitumor activity in vitro and in vivo (8).The implication that the brain is immunologically privileged was based on demonstrations that allogeneic and xenogeneic glioma cells were maintained intracranially (9). More recent studies have revealed some immunological response to these tumors (10, 11); thus, the brain is now considered to be a semiprivileged immune site. On that premise and because it has been shown that CTLs from an allogeneic source are more effective against tumor than those from a syngeneic source (12), we have investigated whether brain tumors in rats could be cured by local adoptive transfer. The advantage of using allogeneic CTLs for therapy ofglioma patients is that the cells are from a healthy donor. This circumvents collecting the patients' own cells for therapy, which would ex...