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In 1994, we reported on a series of patients treated with T-cell therapy (Study #1). This paper (Study #2) is an update of our experience through 1999 in the production of tumor-derived activated cells (TDAC), also called tumor-infiltrating lymphocytes (TIL), from tumor biopsies. TDAC were successfully grown in medium containing Interleukin-2 from 75% of the 366 tumor biopsies tested. There was no significant difference in success (growth to 1 x 10(9) cells) comparing primary and metastatic tumors. Many of the tumors were shipped to the laboratory by overnight delivery from distant sites. Success rate did decrease with the length of time for tumor transport in excess of 24 hours. Certain additional cytokines were tested when cultures did not grow. Interleukin-4 was beneficial in the development of 1 of 4 TDAC cultures which did not grow with IL-2 alone. In order to produce TDAC to treat patients, cells were grown in gas permeable plastic bags or in artificial capillary bioreactor cultures. Approximately 1 x 10(9) were seeded from an initially successful "feasibility study" to bulk produce cells for treatment. Harvest was carried out after about 3 weeks. Sixty-three patients were treated at least once with a minimum of 1 x 10(10) TDAC given by intravenous infusion. On the average, the number of cells per treatment was 3 x 10(10) with a viability of 87%. TDAC cultures contained T cells with variable ratios of CD4 to CD8 cells. Secreted granulocyte-monocyte colony stimulating factor, interferon gamma and tumor necrosis factor alpha were measured in TDAC conditioned medium. Only 34 patients received the full course of 4 TDAC treatments. The cells were well tolerated with mild fever and dyspnea. Partial responses were observed in 8 patients, including the dramatic regression of scalp nodules in a patient with renal cancer. These results showed that therapeutic amounts of TDAC can be produced in cell culture in a reasonable and cost-effective manner. The cells were well tolerated and responses were seen in renal and melanoma patients resistant to IL-2 with bulky, advanced cancer.

Background: Biochemical modulation of 5-Fluorouracil activity with Leucovorin has been well documented in colorectal cancer. Several studies have shown increased efficacy of 5-fluorouracil in combination with alpha interferon. We therefore initiated a phase II trial of dual modulation of 5-fluorouracil with leucovorin and alpha interferon to evaluate outcomes in patients with metastatic carcinoma of the colon., Methods: Patients with metastatic colon carcinoma with expected survival > 4 months and performance status of ECOG < or = 2 were treated weekly with Leucovorin 400 mg i.v. followed by 5-FU 600 mg/m2 i.v. bolus. Alpha interferon 3-9 million units was administered subcutaneously every Monday, Wednesday and Friday. Patients were analyzed for toxicity, tumor response and survival., Results: Sixteen patients with a median age of 66 years were treated. Three patients were not evaluable for response but were evaluable for toxicity. Grades 3 and 4 toxicities were neutropenia, diarrhea, mucositis, nausea and vomiting, fatigue, fever, asthenia and elevated hepatic enzymes. One patient died from complications associated with diverticulitis and neutropenia. Objective response rate was 23% (95% confidence interval 4-46%) and median survival was 11.5 months (95% confidence interval 6.3-19 months). Thirty-eight percent of the patients were alive at one year and 19% at two years., Conclusion: The combination of 5-fluorouracil, leucovorin and alpha interferon as administered in this phase II study did not result in enhanced response rate or survival. However this regimen was associated with considerable toxicity.

CBT-1, a natural product, was studied as an MDR modulator with Taxol (135 mg/m2) in an escalating dose Phase I clinical trial. CBT-1 was administered orally at doses from 300 mg/m2 to 500 mg/m2 daily x 7. The MTD was determined to be 500 mg/m2 with moderate nausea and occasional emesis. Side effects were mainly attributable to Taxol rather than the study drug. A total of 18 patients were registered on study with only one patient determined to be intolerant of CBT-1 due to nausea and emesis. In this Phase I study four patients (3 breast, 1 NSCLC) remained stable for greater than two cycles of treatment. No complete or partial responses were seen in this Taxol resistant population of patients with advanced cancer.

The use of monoclonal antibodies (MoAbs) for immunotherapy and radioimmunotherapy has ushered in a new era in the treatment of non-Hodgkin's lymphoma.

CBT-1, a natural product, was studied in an escalating dose Phase I clinical trial with doxorubicin at 60 mg/m2. CBT-1 was administered by mouth at doses from 200 mg/m2 to 600 mg/m2. The drug was given for 7 days and doxorubicin administered intravenously on day 6. The MTD was determined to be 500 mg/m2 although some patients did tolerate 600 mg/m2 with moderate nausea and occasional vomiting. Side effects were otherwise mild in the 23 patients treated. Pharmacokinetic determinations in an additional 11 patients demonstrated that CBT-1 did not significantly alter the pharmacokinetics of doxorubicin. In this Phase I study, 25 of 34 patients were evaluable for response and 5 patients demonstrated tumor shrinkage.

The Cancer Biotherapy Research Group [CBRG], formerly known as the National Biotherapy Study Group [NBSG], celebrated its 10th anniversary in 1997. CBRG is a not-for-profit cancer clinical trials group of community oncologists whose activities for the most part have been funded by their associated community hospitals. From its inception the Group has focused on clinical investigations of biological agents in the treatment of patients with advanced cancer. This article reviews the history of the Group, its structure, accomplishments, and its current objectives and challenges.

Purpose: Between 1987-1990, 612 patients received high-dose continuous intravenous interleukin-2 (IL-2) in phase II clinical trials of the National Biotherapy Study Group (NBSG). The purpose of this analysis was to determine the long-term survival rates associated with such therapy and the correlation, if any, between objective tumor response, and survival., Methods: Patients who are known to have survived at least 3 years or more were identified. Actual and actuarial survival rates were determined for various malignancies and by tumor response., Results: At least 37 (6.0%) survived > or = 3 years from the initiation of IL-2 therapy, and it is possible the 3-year survival rate is as high as 20%. This included 14/168 (8%) of patients with renal cell carcinoma, 10/175 (6%) melanoma, 2/51 (4%) lung cancer, 0/61 colorectal, 0/36 breast, 1/17 sarcoma, 1/14 pancreas, 1/19 ovary, 3/11 lymphoma, 2 adenocystic carcinomas, 2 carcinoid, and one Hodgkin's disease. Four hundred ninety-one of 612 (80%) are known to have died, 121 were still alive at the time of the last follow-up. Median survival was 7.9 months. Among 547 evaluable patients, there were eight complete responses (CR) and 42 partial responses (PR), for an objective response rate of 9%. An additional 32 patients had mixed or minimal responses (MR) for a total response rate of 15%. The 3-year survival rates were 25% for PR, 17% for PR, 16% for MR, 8% for stable disease (SD), and 2% for patients with progressive disease (PD). Responders made up a higher proportion of patients who survived > or = 3 years than of patients who survived < or = 3 years (14/37 = 38% vs 68/500 = 14%; p < .0001, X2). A higher proportion of responders survived > or = 3 years than non-responders (14/82 = 17% vs 23/230 = 4%, p < .0001, X2). Patients with CR, PR, or MR had a > or = 3-year survival rate (14/82 = 17%) than patients who had SD (20/249 = 8%; p = .02, x2, who in turn had a > or = 3-year survival rate that was greater than patients who had PD (3/206 = 2%; p = .001, X2). For individual trials in which 10 or more patients were enrolled, the percentage of patients surviving > or = 3 years ranged between 4% and 8%., Conclusion: We conclude that in the setting of IL-2 therapy for metastatic cancer, the probability of surviving > or = 3 years was approximately 12 times greater for responders, and five times greater for patients with SD; as compared to patients who had PD. Furthermore, despite diagnoses of metastatic cancer, often in settings in which disease had been refractory to standard therapy there was an actual 3-year survival rate of at least 6% to 8% for patients with metastatic melanoma and renal cell carcinoma.

The National Biotherapy Study Group conducted a phase I/II trial of alpha-interferon (IFN) plus radiation therapy (RT) in glioma patients to confirm the feasibility of combining these two modalities. Patients newly diagnosed gliomasreceived external beam RT as 180 cGy in 33 fractions over six to seven weeks, five days a week, and IFN at a dose of 3 MIU SC Monday, Wednesday and Friday of each week. IFN was increased to 5 MIU after two weeks and was given for up to 16 weeks. Patients were monitored for toxicity and failure-free and overall survival. There were 12 men and seven women with an age range of 24-77, and a median age of 64 years. There were 12 glioblastomas and seven advanced astrocytomas. Complete surgical resection was carried out in two patients, nine had a partial resection, and eight had a biopsy only. Two patients in the latter group deteriorated rapidly and received < 2 weeks of RT/IFN. One patient stopped IFN because of a skin rash, another stopped because of concurrent pneumonia, and one patient was noncompliant. RT and IFN were well-tolerated; 14 of the 19 patients completed the eight weeks of IFN/RT. However, only three patients took IFN for the maximum of 16 weeks. The only grade 4 toxicities noted were increases SGOT in three, increases alk phos in two, and severe fatigue in four patients. The median failure-free survival was two months, median survival was 7.5 months, and four patients survived beyond one year. The longest survivor was 29.1 months, and one patient is still alive after 20.7 months. IFN/RT can be safely co-administered in patients with gliomas. A randomized trial would be needed to establish clinical benefit.

Interleukin-15 was tested to determine whether this recently discovered cytokine was capable of stimulating the growth of tumor derived activated T cells in culture (TDAC, also referred to as tumor infiltrating lymphocytes). When established cultures of IL-2 induced, IL-2 dependent TDAC were tested, IL-15 stimulated growth in a dose dependent manner, alone or in the presence of IL-2. One established TDAC was cultured with IL-15 alone for 18 passages over a 10 week period. Comparing IL-2 and IL-15 treated cultures, growth rate with IL-15 was slower. IL-15 doubled the secreted interferon alpha and granulocyte-macrophage colony stimulating factor. IL-15 and IL-2 were compared in primary TDAC cultures. IL-15 induced TDAC outgrowth in 3 of 6 cultures. IL-2 induced outgrowth in all 6. Tumor cells were eliminated as TDAC grew out in both IL-2 and IL-15 treated cultures. These results suggested that IL-15 like IL-2, is capable of stimulating the growth of TDAC with antitumor activity, but with certain distinct effects which may be of interest therapeutically.

This report describes the production of Tumor Derived Activated Cell cultures (TDAC, also called tumor infiltrating lymphocytes) from patient tumor biopsies and our preliminary experience growing these cells to therapeutic levels using artificial capillary bioreactor cultures. TDAC were successfully grown in medium containing Interleukin 2 from 80% of the 113 tumor biopsies tested. There was no significant difference in success (growth to 1 x 10(9) cells) comparing primary and metastatic tumors. Many of the tumors were shipped to the laboratory from distant sites. Success rate did decrease with the length of time for tumor transport. Interleukin 4 was beneficial in the development of 1 of 4 TDAC cultures which did not grow with IL-2 only. Seventy-seven bioreactor cultures were initiated for 31 patients. On the average, 1.9 x 10(9) TDAC were inoculated per bioreactor; 3.3 x 10(10) were harvested in 22 days. Twelve liters of medium were required per 1 x 10(10) TDAC produced. TDAC cultures contained T cells with variable ratios of CD4 to CD8 cells. Secreted granulocyte monocyte colony stimulating factor, interferon gamma and tumor necrosis factor were measured in the bioreactor cartridge conditioned medium. Twenty three patients were evaluated. Partial responses were observed in 4 patients including a dramatic remission of scalp nodules in a patient with renal cancer. Results showed that therapeutic amounts of TDAC cells may be produced in a reasonable and cost effective manner using artificial capillary bioreactor cultures.