|
Advertisement |
|
|
||
 |
|
|||||
|
|||||
|
 |
||||||
|
||||||
|
||||||
|
Originally published as JCO Early Release 10.1200/JCO.2005.05.5335 on September 11 2006 © 2006 American Society of Clinical Oncology. Phase II Study of Belagenpumatucel-L, a Transforming Growth Factor Beta-2 Antisense Gene-Modified Allogeneic Tumor Cell Vaccine in Non–Small-Cell Lung Cancer
From the Mary Crowley Medical Research Center/Texas Oncology Professional Association; Baylor Sammons Cancer Center, Baylor University Medical Center; Murex Pharmaceutical Inc; Institute for Health Care Research and Improvement, Baylor Health Care System, Dallas, TX; Hoag Cancer Center, Newport Beach; NovaRx Corporation, San Diego, CA; and Louisiana State University, New Orleans, LA Address reprint requests to John Nemunaitis, MD, 1717 Main St, 60th Floor, Dallas, TX 75201; e-mail: jnemunaitis{at}mcmrc.com
PURPOSE: Belagenpumatucel-L is a nonviral gene-based allogeneic tumor cell vaccine that demonstrates enhancement of tumor antigen recognition as a result of transforming growth factor beta-2 inhibition. PATIENTS AND METHODS: We performed a randomized, dose-variable, phase II trial involving stages II, IIIA, IIIB, and IV non–small-cell lung cancer patients. Each patient received one of three doses (1.25, 2.5, or 5.0 x 107 cells/injection) of belagenpumatucel-L on a monthly or every other month schedule to a maximum of 16 injections. Immune function, safety, and anticancer activity were monitored.
RESULTS: Seventy-five patients (two stage II, 12 stage IIIA, 15 stage IIIB, and 46 stage IV patients) received a total of 550 vaccinations. No significant adverse events were observed. A dose-related survival difference was demonstrated in patients who received CONCLUSION: Belagenpumatucel-L is well tolerated, and the survival advantage justifies further phase III evaluation.
Non–small-cell lung cancer (NSCLC) is the most common and one of the most debilitating cancers affecting society.1 The majority of patients present at an advanced stage2 for which chemotherapy, despite its limited efficacy, is usually the only therapeutic option.3-5 Expected survival of patients who experience progression after initial chemotherapy is 8 months or less, and generally, less than 30% of patients survive 1 year.6-11 Recently, we demonstrated evidence of antitumor activity in advanced-stage NSCLC patients with an adenoviral gene-based autologous tumor vaccine (GVAX; Cell Genesys, San Francisco, CA)12,13 designed to enhance tumor antigen recognition by immune effector cells. Although encouraging, the requirement for harvest of autologous tumor necessarily imposed limits on this approach. Therefore, to further explore anticancer immune therapy through enhancement of effective immunogenic tumor antigen recognition, we studied the use of a nonviral gene-based allogeneic vaccine belagenpumatucel-L (Lucanix; NovaRx Corporation, San Diego, CA) incorporating transforming growth factor beta-2 (TGF-ß2) antisense gene modification of tumor cells. Transforming growth factor beta (TGF-ß) is one of a family of multifunctional proteins that regulate growth and function of normal and neoplastic cells.14-17 Elevated levels of TGF-ß2, in particular, are linked to immunosuppression in cancer patients,15-20 and the level of TGF-ß2 is inversely correlated with prognosis in patients with NSCLC.21 TGF-ß2 has antagonistic effects on natural killer cells, lymphokine activated killer cells, and dendritic cells.22-27 Using an antisense gene to inhibit TGF-ß2, we and others have demonstrated inhibition of cellular TGF-ß2 expression with resultant increased immunogenicity of gene-modified cancer cells.28-36 The results of this work led us to conduct a phase II trial of vaccination with belagenpumatucel-L in NSCLC patients.
Study Design This was an open-label, three-arm, phase II study designed to evaluate the safety and efficacy of intradermal immunization with three dose levels of an allogeneic tumor cell vaccine administered once a month or once every other month. Patients were randomly assigned to one of the three dose cohorts (1.25, 2.5, or 5 x 107 cells/injection) and were treated after a minimum workout of 30 days from prior cytotoxic therapy. Tumor staging was performed at baseline, at weeks 8 and 16, and quarterly thereafter. Patients were serially monitored for immune response every 4 weeks up to week 28 and quarterly thereafter. Toxicity was monitored throughout the study. Patients demonstrating benefit from treatment at 16 weeks could be given up to 12 additional vaccinations. All patients signed local institutional review board–approved informed consent. Two patients received concurrent radiation therapy.
Study Population
Humoral Immune Response Assessment Multiplex flow cytometric analysis was used to determine both the presence and HLA specificity of lymphocytotoxic antibodies in sera. Reactivity was determined with microbeads coated with purified class I or class II HLA antigens (LABScreen; One Lambda Inc, Canoga Park, CA). Antibody (Ab) reactivity in each test serum was determined with the use of the Lambda Array Beads Multianalyte System and LABScan 100 flow analyzer (One Lambda Inc) for data acquisition and analysis.
Enzyme-Linked Immunospot Assay for Interferon Gamma Release
Detection of Intracellular Cytokines in Pre- and Post-Treatment PBMCs Cytokine levels in pre- and post-treatment PBMCs were analyzed by immunofluorescent staining of intracellular cytokines by flow cytometric analysis. Fresh, unstimulated PBMCs in whole blood were incubated with protein transport blocking reagent (Brefeldin A; eBiosciences, San Diego, CA) for 4 hours, followed by RBC lysis using PhamLyse solution (BD Biosciences). The PBMCs were fixed and permiabilized using the Cytofix/Cytofirm solution (BD Biosciences), followed by incubation with fluorochrome-conjugated Ab to the cytokine. The stained cells were analyzed by flow cytometry (BD FACScan, BD Biosciences) with the CELLQUEST software (BD Biosciences).
Vaccine Production
Antisense Plasmid The plasmid used for transfection was pCHEK/HBA-2. It contains an EBV origin of replication and EBNA-1 genes, a hygromycin resistance gene under the control of SV-40 promoter, and a 929–base pair fragment of the human TGF-ß2 molecule in antisense orientation under the control of the CMV immediate-early promoter and enhancer. The SV-40 promoter/intron unit was incorporated to increase expression of the hygromycin resistance gene used to facilitate selection of gene-modified cells in culture. The pCHEK/HBA-2 vector was electroporated into each cell line ex vivo.
Investigational Product
Statistical Methods
In analyzing adverse events, a severity analysis approach was pursued. Homogeneity in the rate of serious adverse events across dose cohorts was tested using a Pearson
A Pearson
Statistical analysis of the effect of tumor vaccination on intracellular cytokine expression was performed to compare pretreatment value with post-treatment samples for advanced-stage patients or by cohort or clinical response. The nonparametric Kruskal-Wallis test was used for cohort analyses. The Fisher’s exact test was used in the contingency table analysis of HLA-Ab expression or ELISPOT response with clinical responsiveness. A Pearson
Seventy-five patients were entered onto the trial between May 30, 2002 and March 26, 2004. Demographics of advanced-stage (20 in cohort 1, 20 in cohort 2, and 21 in cohort 3) and early-stage patients are listed separately in Table 1. Toxicity and survival were assessable in all patients. One patient was not assessable for time to progression. Immune function samples were not assessable from site 004 as a result of handling complications. Five hundred fifty vaccinations were administered. Demographic comparison of sex, age, stage (IIIB and IV), tumor volume (< 12 or  12 cm3), prior chemotherapy, and date of diagnosis found no statistical difference among cohorts.
Safety Analysis of severity of adverse events demonstrated no detectable difference in the rate of serious adverse events across dose cohorts (P = .5698). All toxic events occurring at a 5% frequency are listed in Table 2. Separation of toxic events between late-stage and early-stage patients and between cohorts suggests no difference. All grade 3 and 4 events were attributed to disease progression except two events (Table 2).
Response The analysis of patient response in the advanced-stage patients (using Response Evaluation Criteria in Solid Tumors criteria) revealed a partial response rate of 15% at week 16. Cohort-specific response of patients with measurable disease (n = 40) was one of 16, three of 11, and two of 13 patients in cohorts 1, 2, and 3, respectively (Fig 1). The median tumor shrinkage at 16 weeks was 63% (maximum shrinkage, 80%). Fifty-nine percent of all patients showed no evidence of progression at week 16. Progression-free survival across dose cohorts was not different (P = .3816). The median age of the five responding patients was 70 years (range, 54 to 81 years); all five were females; two had stage IIIB disease, and three had stage IV disease; and two patients had adenocarcinoma, two had squamous cell carcinoma, and one had large-cell carcinoma. Two of the responding patients had received combination chemotherapy and radiation therapy, and all two of these patients had achieved an initial response to the combination. A fourth patient received radiation therapy without chemotherapy and demonstrated progressive disease, along with two other patients who demonstrated progressive disease after attempt at surgical resection without follow-up adjuvant therapy. The median number of measurable lesions per patient was two (range, one to five lesions). Disease was limited to multiple lung sites in two of five patients. Prior treatment was discontinued a median of 2 months (range, 2 to 4 months) before random assignment of vaccination in all five responding patients. Median volume of measurable disease was 5.63 cm3 (range, 0.27 to 51 cm3).
Survival Range of follow-up time for surviving patients was 351 to 967 days. Median overall survival time of all three cohorts was 441 days (Fig 2). Dose-related survival differences were found between cohorts (P = .0155; Fig 3). The analysis suggested that, at doses 2.5 x 107 cells/injection (cohorts 2 and 3), there was a significant difference in survival compared with cohort 1 (P = .0069). This difference is less than the required P value .05/2 = .0250 cutoff specified by the Bonferroni adjustment. The estimated 1- and 2-year survival probabilities were 68% (95% CI, 55% to 80%) and 52% (95% CI, 35% to 68%), respectively, for the two higher dose groups combined and 39% (95% CI, 22% to 56%) and 20% (95% CI, 4% to 36%), respectively, for the low-dose group. There was no significant difference in survival between cohorts 1 and 2 versus 3 (P = .9198).
An estimated 64% (95% CI, 50% to 78%) and 47% (95% CI, 30% to 65%) of the advanced-stage patients in combined cohorts 2 and 3 survived 1 and 2 years, respectively, compared with 37% (95% CI, 18% to 55%) and 18% (95% CI, 2% to 35%) of advanced-stage patients in cohort 1. The overall difference in survival across dose cohorts for advanced-stage patients was only marginally significant (P = .0605). The estimated median survival time of 581 days in dose cohorts 2 and 3 was significantly higher than the median survival time for dose cohort 1 (252 days; P = .0186; Fig 2). Median survival time of all stage IIIB and IV patients was 441 days; 1- and 2-year survival rates were 54% (95% CI, 42% to 67%) and 35% (95% CI, 20% to 51%), respectively. The difference in survival between dose cohorts 1 and 2 versus 3 in advanced-stage patients was not significant (P = .5148).
Activation of Immune Responses
Before vaccination, nine of 36 patients lacked a significant ELISPOT response when stimulated with PMA+I (Table 3). However, seven of these nine patients generated a positive ELISPOT polyclonal response (PMA+I) at week 12 after belagenpumatucel-L vaccination. Furthermore, a majority of advanced-stage patients with stable disease or better produced a markedly elevated IFN-  response (ELISPOT activity) when challenged with belagenpumatucel-L (Table 3). Twelve of 20 patients with clinical response (partial response or stable disease) displayed a two-fold response. By comparison, five of 16 patients with progressive disease (P = .086, Pearson’s  2; n = 36) produced an elevated ELISPOT response (Table 3).
Ab-Mediated Immune Response
Abs reactive with belagenpumatucel-L were determined. Seven of 57 patients showed an increase in an IgM titer
Historically, NSCLC has generally been regarded as a nonimmunogenic cancer.37 Immunotherapy for lung cancer has yielded little benefit in humans,38-40 although preliminary results of more recent vaccine studies designed to enhance tumor antigen recognition have demonstrated beneficial outcome in subsets of patients.12,13,41-43 In the current study, we defined a clear dose-related effect of belagenpumatucel-L involving both early- and late-stage patients. We also found an impressive survival advantage at dose levels 2.5 x 107 cells/injection, with an estimated 2-year survival rate of 47% in response to belagenpumatucel-L in advanced-stage patients. This compares favorably to the historical 2-year survival rate of less than 20% in stage IIIB and IV NSCLC patients. Furthermore, a correlation of positive clinical outcome with induction of immune enhancement of tumor antigen was observed. Hypotheses as to why previous immune approaches to NSCLC have yielded disappointing results include ineffective priming of tumor-specific T cells, lack of high-avidity tumor-specific T cells, and physical or functional disabling of primed tumor-specific T cells by primary host and/or tumor-related mechanisms. For example, a high proportion of the tumor-infiltrating lymphocytes in NSCLC tissue are immunosuppressive regulatory T cells (CD4+CD25+) with spontaneous TGF-ß secretion and constitutive high-level expression of cytotoxic T-cell lymphocyte-4.44,45 These have been shown to specifically prevent effective immune activation44,46-52 and, thus, could lead to tolerance (or cross tolerance) of T cells rather than cross priming.44,45,53-57 In support of this hypothesis, elevated levels of IL-10 and TGF-ß are found in patients with NSCLC.
Indeed, evidence suggests that TGF-ß is a key immunosuppressive factor in NSCLC.15-21 TGF-ß is able to convert CD4+CD25––naïve T cells to CD4+CD25+ regulatory T cells by induction of transcription factor Foxp3.58,59 The process of cancer recruitment and subsequent recognition by immune-processing cells (eg, natural killer T cells, There is evidence for shared antigens in lung cancers46-52 (Nemunaitis et al, submitted for publication), as is seen in other tumor types.79,80 Dendritic cells, as opposed to tumor cells, may be responsible for the induction of antitumor immunity in tumor-bearing hosts by a process of antigenic cross presentation.81,82 One of the critical factors in dendritic cell–based immunization is the activation status of dendritic cells. Immature dendritic cells are able to process antigen but are not able to stimulate potent immune responses. The secretion of TGF-ß2 by cancer cells may contribute to immunosuppression by blocking maturation of dendritic cells83 and reversibly regulating dendritic cell chemotaxis by modifying chemokine receptor expression.84 TGF-ß2 has been shown to immobilize dendritic cells within the tumor and reduce the number of dendritic cells migrating to drainage lymph nodes.85 Those dendritic cells reaching the drainage nodes are less likely to be completely activated, thereby favoring the induction of cross tolerance rather than cross priming.86 Our findings suggest that overall immune activation may contribute to a favorable clinical outcome. The current vaccination protocol used whole tumor cells, and the immunodominant antigens have not been characterized. Hence, immune response evaluations have been limited to the use of unfractionated belagenpumatucel-L preparations to encompass allogeneic as well as potential tumor-specific activational events. The selected immune assays represented attempts to document collateral immune activation events in the cellular (ELISPOT analyses) and humoral components (overall Ab production and HLA-directed Ab responses) that accompany belagenpumatucel-L vaccination. Vaccination-induced alloreactivity indicates that belagenpumatucel-L is effective across allogeneic barriers and is likely to translate into clinical effectiveness among immunocompetent patients who have the capacity to mount antigen-presentation/activation events against novel allo- and/or tumor-associated antigens. Our findings of upregulated immune responses that correlated with clinical responsiveness support the possibility that enhanced tumor antigen sensitization may be achieved through multiple treatments with the TGF-ß2 antisense-modified cancer vaccine. In conclusion, in view of the acceptable safety profile and suggested survival advantage, further investigation is recommended. Data presented support the hypothesis that inhibition of TGF-ß2 in an allogeneic cell vaccine results in sufficient repression of inhibiting factors to allow for immune recognition and effector cell activation in patients with advanced NSCLC.
Although all authors completed the disclosure declaration, the following authors or their immediate family members indicated a financial interest. No conflict exists for drugs or devices used in a study if they are not being evaluated as part of the investigation. For a detailed description of the disclosure categories, or for more information about ASCO’s conflict of interest policy, please refer to the Author Disclosure Declaration and the Disclosures of Potential Conflicts of Interest section in Information for Contributors.
Dollar Amount Codes (A) < $10,000 (B) $10,000–$99,999 (C)
We thank Brenda Marr and Jennifer Hartley for manuscript preparation; Linda Jennings, PhD, for statistical analysis; Douglas Smith, MD, for HLA-related analyses; and Steven Dubinett, MD, for the cell line RH-2 analysis.
published online ahead of print at www.jco.org on September 11, 2006. Supported in part by Small Business Innovative Research Grant No. 1 R44 CA96025 (H.F.). Terms in blue are defined in the glossary, found at the end of this article and online at www.jco.org. Authors’ disclosures of potential conflicts of interest and author contributions are found at the end of this article.
1. Parkin DM, Bray FI, Devesa SS: Cancer burden in the year 2000: The global picture. Eur J Cancer 37:S4-S66, 2001 (suppl 8)[Medline] 2. Jemal A, Murray T, Samuels A, et al: Cancer statistics, 2003. CA Cancer J Clin 53:5-26, 2003 3. Non-Small Cell Lung Cancer Collaborative Group: Chemotherapy in non-small cell lung cancer: A meta-analysis using updated data on individual patients from 52 randomised clinical trials. BMJ 311:899-890, 1995 4. Grilli R, Oxman AD, Julian JA: Chemotherapy for advanced non-small-cell lung cancer: How much benefit is enough? J Clin Oncol 11:1866-1872, 1993 5. Schiller JH, Harrington D, Belani CP, et al: Comparison of four chemotherapy regimens for advanced non-small-cell lung cancer. N Engl J Med 346:92-98, 2002 6. Shepherd FA, Dancey J, Ramlau R, et al: Prospective randomized trial of docetaxel versus best supportive care in patients with non-small-cell lung cancer previously treated with platinum-based chemotherapy. J Clin Oncol 18:2095-2103, 2000 7. Fossella FV, DeVore R, Kerr RN, et al: Randomized phase III trial of docetaxel versus vinorelbine or ifosfamide in patients with advanced non-small-cell lung cancer previously treated with platinum-containing chemotherapy regimens: The TAX 320 Non-Small Cell Lung Cancer Study Group. J Clin Oncol 18:2354-2362, 2000 8. Hanna N, Shepherd FA, Fossella FV, et al: Randomized phase III trial of pemetrexed versus docetaxel in patients with non-small-cell lung cancer previously treated with chemotherapy. J Clin Oncol 22:1589-1597, 2004 9. Tsao MS, Sakurada A, Cutz JC, et al: Erlotinib in lung cancer: Molecular and clinical predictors of outcome. N Engl J Med 353:133-144, 2005 10. Shepherd FA, Rodrigues Pereira J, Ciuleanu T, et al: Erlotinib in previously treated non-small-cell lung cancer. N Engl J Med 353:123-132, 2005 11. Kris MG, Natale RB, Herbst RS, et al: Efficacy of gefitinib, an inhibitor of the epidermal growth factor receptor tyrosine kinase, in symptomatic patients with non-small cell lung cancer: A randomized trial. JAMA 290:2149-2158, 2003 12. Nemunaitis J: Vaccines in cancer: GVAX, a GM-CSF gene vaccine. Expert Rev Vaccines 4:259-274, 2005[CrossRef][Medline] 13. Nemunaitis J, Sterman D, Jablons D, et al: Granulocyte-macrophage colony-stimulating factor gene-modified autologous tumor vaccines in non-small-cell lung cancer. J Natl Cancer Inst 96:326-331, 2004 14. Fakhrai H, Gramatikova S, Safaei R: Down-regulation of TGF-b2 as a therapeutic approach, in Brain Tumor Immunotherapy. Totowa, NJ, Humana Press, 2001, pp 289-305 15. Sporn MB, Roberts AB, Wakefield LM, et al: Transforming growth factor-beta: Biological function and chemical structure. Science 233:532-534, 1986 16. Massague J: The TGF-beta family of growth and differentiation factors. Cell 49:437-438, 1987[CrossRef][Medline] 17. Border WA, Ruoslahti E: Transforming growth factor-beta in disease: The dark side of tissue repair. J Clin Invest 90:1-7, 1992[Medline] 18. Bodmer S, Strommer K, Frei K, et al: Immunosuppression and transforming growth factor-beta in glioblastoma: Preferential production of transforming growth factor-beta 2. J Immunol 143:3222-3229, 1989[Abstract] 19. Jakowlew SB, Mathias A, Chung P, et al: Expression of transforming growth factor beta ligand and receptor messenger RNAs in lung cancer cell lines. Cell Growth Differ 6:465-476, 1995[Abstract] 20. Constam DB, Philipp J, Malipiero UV, et al: Differential expression of transforming growth factor-beta 1, -beta 2, and -beta 3 by glioblastoma cells, astrocytes, and microglia. J Immunol 148:1404-1410, 1992[Abstract] 21. Kong F, Jirtle RL, Huang DH, et al: Plasma transforming growth factor-beta1 level before radiotherapy correlates with long term outcome of patients with lung carcinoma. Cancer 86:1712-1719, 1999[CrossRef][Medline] 22. Rook AH, Kehrl JH, Wakefield LM, et al: Effects of transforming growth factor beta on the functions of natural killer cells: Depressed cytolytic activity and blunting of interferon responsiveness. J Immunol 136:3916-3920, 1986[Abstract] 23. Kasid A, Bell GI, Director EP: Effects of transforming growth factor-beta on human lymphokine-activated killer cell precursors: Autocrine inhibition of cellular proliferation and differentiation to immune killer cells. J Immunol 141:690-698, 1988[Abstract] 24. Tsunawaki S, Sporn M, Ding A, et al: Deactivation of macrophages by transforming growth factor-beta. Nature 334:260-262, 1988[CrossRef][Medline] 25. Hirte H, Clark DA: Generation of lymphokine-activated killer cells in human ovarian carcinoma ascitic fluid: Identification of transforming growth factor-beta as a suppressive factor. Cancer Immunol Immunother 32:296-302, 1991[CrossRef][Medline] 26. Naganuma H, Sasaki A, Satoh E, et al: Transforming growth factor-beta inhibits interferon-gamma secretion by lymphokine-activated killer cells stimulated with tumor cells. Neurol Med Chir (Tokyo) 36:789-795, 1996[Medline] 27. Ruffini PA, Rivoltini L, Silvani A, et al: Factors, including transforming growth factor beta, released in the glioblastoma residual cavity, impair activity of adherent lymphokine-activated killer cells. Cancer Immunol Immunother 36:409-416, 1993[CrossRef][Medline] 28. Fakhrai H, Mantil JC, Liu L, et al: Phase I clinical trial of TGF-beta antisense-modified tumor cell vaccine in patients with advanced glioma. Cancer Gene Therapy (in press) 29. Liau LM, Fakhrai H, Black KL: Prolonged survival of rats with intracranial C6 gliomas by treatment with TGF-beta antisense gene. Neurol Res 20:742-747, 1998[Medline] 30. Dorigo O, Shawler DL, Royston I, et al: Combination of transforming growth factor beta antisense and interleukin-2 gene therapy in the murine ovarian teratoma model. Gynecol Oncol 71:204-210, 1998[CrossRef][Medline] 31. Tzai TS, Shiau AL, Liu LL, et al: Immunization with TGF-beta antisense oligonucleotide-modified autologous tumor vaccine enhances the antitumor immunity of MBT-2 tumor-bearing mice through upregulation of MHC class I and Fas expressions. Anticancer Res 20:1557-1562, 2000[Medline] 32. Tzai TS, Lin CI, Shiau AL, et al: Antisense oligonucleotide specific for transforming growth factor-beta 1 inhibit both in vitro and in vivo growth of MBT-2 murine bladder cancer. Anticancer Res 18:1585-1589, 1998[Medline] 33. Marzo AL, Fitzpatrick DR, Robinson BW, et al: Antisense oligonucleotides specific for transforming growth factor beta2 inhibit the growth of malignant mesothelioma both in vitro and in vivo. Cancer Res 57:3200-3207, 1997 34. Park JA, Wang E, Kurt RA, et al: Expression of an antisense transforming growth factor-beta1 transgene reduces tumorigenicity of EMT6 mammary tumor cells. Cancer Gene Ther 4:42-50, 1997[Medline] 35. Kettering JD, Mohamedali AM, Green LM, et al: IL-2 gene and antisense TGF-beta1 strategies counteract HSV-2 transformed tumor progression. Technol Cancer Res Treat 2:211-221, 2003[Medline] 36. Fakhrai H, Dorigo O, Shawler DL, et al: Eradication of established intracranial rat gliomas by transforming growth factor beta antisense gene therapy. Proc Natl Acad Sci U S A 93:2909-2914, 1996 37. Salgaller ML: The development of immunotherapies for non-small cell lung cancer. Expert Opin Biol Ther 2:265-278, 2002[CrossRef][Medline] 38. Ratto GB, Zino P, Mirabelli S, et al: A randomized trial of adoptive immunotherapy with tumor-infiltrating lymphocytes and interleukin-2 versus standard therapy in the postoperative treatment of resected nonsmall cell lung carcinoma. Cancer 78:244-251, 1996[CrossRef][Medline] 39. Lissoni P, Meregalli S, Fossati V, et al: A randomized study of immunotherapy with low-dose subcutaneous interleukin-2 plus melatonin vs chemotherapy with cisplatin and etoposide as first-line therapy for advanced non-small cell lung cancer. Tumori 80:464-467, 1994[Medline] 40. Ratto GB, Cafferata MA, Scolaro T, et al: Phase II study of combined immunotherapy, chemotherapy, and radiotherapy in the postoperative treatment of advanced non-small-cell lung cancer. J Immunother 23:161-167, 2000[CrossRef][Medline] 41. Salgia R, Lynch T, Skarin A, et al: Vaccination with irradiated autologous tumor cells engineered to secrete granulocyte-macrophage colony-stimulating factor augments antitumor immunity in some patients with metastatic non–small-cell lung carcinoma. J Clin Oncol 21:624-630, 2003 42. Soiffer R, Hodi FS, Haluska F, et al: Vaccination with irradiated, autologous melanoma cells engineered to secrete granulocyte-macrophage colony-stimulating factor by adenoviral-mediated gene transfer augments antitumor immunity in patients with metastatic melanoma. J Clin Oncol 21:3343-3350, 2003 43. Morse MA: Technology evaluation: BLP-25, Biomira Inc. Curr Opin Mol Ther 3:102-105, 2001[Medline] 44. Woo EY, Chu CS, Goletz TJ, et al: Regulatory CD4(+)CD25(+) T cells in tumors from patients with early-stage non-small cell lung cancer and late-stage ovarian cancer. Cancer Res 61:4766-4772, 2001 45. Woo EY, Yeh H, Chu CS, et al: Cutting edge: Regulatory T cells from lung cancer patients directly inhibit autologous T cell proliferation. J Immunol 168:4272-4276, 2002 46. Sienel W, Varwerk C, Linder A, et al: Melanoma associated antigen (MAGE)-A3 expression in stages I and II non-small cell lung cancer: Results of a multi-center study. Eur J Cardiothorac Surg 25:131-134, 2004 47. Yamazaki K, Spruill G, Rhoderick J, et al: Small cell lung carcinomas express shared and private tumor antigens presented by HLA-A1 or HLA-A2. Cancer Res 59:4642-4650, 1999 48. Weynants P, Thonnard J, Marchand M, et al: Derivation of tumor-specific cytolytic T-cell clones from two lung cancer patients with long survival. Am J Respir Crit Care Med 159:55-62, 1999 49. Bixby DL, Yannelli JR: CD80 expression in an HLA-A2-positive human non-small cell lung cancer cell line enhances tumor-specific cytotoxicity of HLA-A2-positive T cells derived from a normal donor and a patient with non-small cell lung cancer. Int J Cancer 78:685-694, 1998[CrossRef][Medline] 50. Yamada A, Kawano K, Koga M, et al: Gene and peptide analyses of newly defined lung cancer antigens recognized by HLA-A2402-restricted tumor-specific cytotoxic T lymphocytes. Cancer Res 63:2829-2835, 2003 51. Jiang F, Yin Z, Caraway NP, et al: Genomic profiles in stage I primary non small cell lung cancer using comparative genomic hybridization analysis of cDNA microarrays. Neoplasia 6:623-635, 2004[CrossRef][Medline] 52. Melloni G, Ferreri AJ, Russo V, et al: Prognostic significance of cancer-testis gene expression in resected non-small cell lung cancer patients. Oncol Rep 12:145-151, 2004[Medline] 53. Neuner A, Schindel M, Wildenberg U, et al: Prognostic significance of cytokine modulation in non-small cell lung cancer. Int J Cancer 101:287-292, 2002[CrossRef][Medline] 54. Neuner A, Schindel M, Wildenberg U, et al: Cytokine secretion: Clinical relevance of immunosuppression in non-small cell lung cancer. Lung Cancer 34:S79-S82, 2001 (suppl 2)[Medline] 55. Dohadwala M, Luo J, Zhu L, et al: Non-small cell lung cancer cyclooxygenase-2-dependent invasion is mediated by CD44. J Biol Chem 276:20809-20812, 2001 56. Schwartz RH: Models of T cell anergy: Is there a common molecular mechanism? J Exp Med 184:1-8, 1996 57. Lombardi G, Sidhu S, Batchelor R, et al: Anergic T cells as suppressor cells in vitro. Science 264:1587-1589, 1994 58. Chen W, Jin W, Hardegen N, et al: Conversion of peripheral CD4+CD25- naive T cells to CD4+CD25+ regulatory T cells by TGF-beta induction of transcription factor Foxp3. J Exp Med 198:1875-1886, 2003 59. Fantini MC, Becker C, Monteleone G, et al: Cutting edge: TGF-beta induces a regulatory phenotype in CD4+CD25- T cells through Foxp3 induction and down-regulation of Smad7. J Immunol 172:5149-5153, 2004 60. Groh V, Rhinehart R, Secrist H, et al: Broad tumor-associated expression and recognition by tumor-derived gamma delta T cells of MICA and MICB. Proc Natl Acad Sci U S A 96:6879-6884, 1999 61. Bauer S, Groh V, Wu J, et al: Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science 285:727-729, 1999 62. Friese MA, Wischhusen J, Wick W, et al: RNA interference targeting transforming growth factor-beta enhances NKG2D-mediated antiglioma immune response, inhibits glioma cell migration and invasiveness, and abrogates tumorigenicity in vivo. Cancer Res 64:7596-7603, 2004 63. Nakamura K, Kitani A, Strober W: Cell contact-dependent immunosuppression by CD4(+)CD25(+) regulatory T cells is mediated by cell surface-bound transforming growth factor beta. J Exp Med 194:629-644, 2001 64. Annunziato F, Cosmi L, Liotta F, et al: Phenotype, localization, and mechanism of suppression of CD4(+)CD25(+) human thymocytes. J Exp Med 196:379-387, 2002 65. Parekh VV, Prasad DV, Banerjee PP, et al: B cells activated by lipopolysaccharide, but not by anti-Ig and anti-CD40 antibody, induce anergy in CD8+ T cells: Role of TGF-beta 1. J Immunol 170:5897-5911, 2003 66. Castelli C, Rivoltini L, Andreola G, et al: T-cell recognition of melanoma-associated antigens. J Cell Physiol 182:323-331, 2000[CrossRef][Medline] 67. Roszman T, Elliott L, Brooks W: Modulation of T-cell function by gliomas. Immunol Today 12:370-374, 1991[CrossRef][Medline] 68. Smith KA: Interleukin-2: Inception, impact, and implications. Science 240:1169-1176, 1988 69. Smith KA: Lowest dose interleukin-2 immunotherapy. Blood 81:1414-1423, 1993 70. Tigges MA, Casey LS, Koshland ME: Mechanism of interleukin-2 signaling: Mediation of different outcomes by a single receptor and transduction pathway. Science 243:781-786, 1989 71. Fontana A, Frei K, Bodmer S, et al: Transforming growth factor-beta inhibits the generation of cytotoxic T cells in virus-infected mice. J Immunol 143:3230-3234, 1989[Abstract] 72. Hirte HW, Clark DA, O’Connell G, et al: Reversal of suppression of lymphokine-activated killer cells by transforming growth factor-beta in ovarian carcinoma ascitic fluid requires interleukin-2 combined with anti-CD3 antibody. Cell Immunol 142:207-216, 1992[CrossRef][Medline] 73. Ranges GE, Figari IS, Espevik T, et al: Inhibition of cytotoxic T cell development by transforming growth factor beta and reversal by recombinant tumor necrosis factor alpha. J Exp Med 166:991-998, 1987 74. Ahmadzadeh M, Rosenberg SA: TGF-beta 1 attenuates the acquisition and expression of effector function by tumor antigen-specific human memory CD8 T cells. J Immunol 174:5215-5223, 2005 75. Gorelik L, Constant S, Flavell RA: Mechanism of transforming growth factor beta-induced inhibition of T helper type 1 differentiation. J Exp Med 195:1499-1505, 2002 76. Maggard M, Meng L, Ke B, et al: Antisense TGF-beta2 immunotherapy for hepatocellular carcinoma: Treatment in a rat tumor model. Ann Surg Oncol 8:32-37, 2001 77. Hirschowitz EA, Foody T, Kryscio R, et al: Autologous dendritic cell vaccines for non-small-cell lung cancer. J Clin Oncol 22:2808-2815, 2004 78. Raez LE, Cassileth PA, Schlesselman JJ, et al: Allogeneic vaccination with a B7.1 HLA-A gene-modified adenocarcinoma cell line in patients with advanced non-small-cell lung cancer. J Clin Oncol 22:2800-2807, 2004 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
TOP
ABSTRACT
INTRODUCTION
125 mL (excluding nodal or bone disease). Patients were required to have completed or refused conventional therapy and have an Eastern Cooperative Oncology Group performance status of 
, tumor necrosis factor alpha; GM-CSF, granulocyte-macrophage colony-stimulating factor.
T cells, macrophages, and CD8+