This Article Abstract Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Email this article to a colleague Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Save to my personal folders Download to citation manager Rights & Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Raez, L. E. Articles by Podack, E. R. Search for Related Content PubMed PubMed Citation Articles by Raez, L. E. Articles by Podack, E. R. Social Bookmarking                            What's this?

Allogeneic Vaccination With a B7.1 HLA-A Gene-Modified Adenocarcinoma Cell Line in Patients With Advanced Non–Small-Cell Lung Cancer

From the Division of Hematology/Oncology, Department of Medicine, Department of Microbiology and Immunology, and Department of Epidemiology and Public Health, Sylvester Comprehensive Cancer Center, University of Miami School of Medicine, Miami, FL

Address reprint requests to Luis E. Raez, MD, FACP, Division of Hematology/Oncology, Sylvester Comprehensive Cancer Center, University of Miami School of Medicine, 1475 NW 12 Ave # 3510, Miami, FL 33136; e-mail: lraez{at}med.miami.edu

	ABSTRACT

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION Authors' Disclosures of... REFERENCES

 
PURPOSE: To determine the safety, immunogenicity, and clinical response to an allogeneic tumor vaccine for non–small-cell lung cancer, we conducted a phase I trial in patients with advanced metastatic disease.

PATIENTS AND METHODS: We treated 19 patients with a vaccine based on an adenocarcinoma line (AD100) transfected with B7.1 (CD80) and HLA A1 or A2. Patients were vaccinated intradermally with 5 x 10⁷ cells once every 2 weeks. Three vaccinations represented one course of treatment. If patients had complete response, partial response, or stable disease, they continued with the vaccinations for up to three courses (nine vaccinations). Immune response was assessed by a change between pre-study and postvaccination enzyme-linked immunospot frequency of purified CD8 T-cells secreting interferon-gamma in response to in vitro challenge with AD100.

RESULTS: Four patients experienced serious adverse events that were unrelated to vaccine. Another four patients experienced only minimal skin erythema. All but one patient had a measurable CD8 response after three immunizations. The immune response of six surviving, clinically responding patients shows that CD8 titers continue to be elevated up to 150 weeks, even after cessation of vaccination. Overall, one patient had a partial response, and five had stable disease. Median survival for all patients is 18 months (90% CI, 7 to 23 months), with corresponding estimates of 1-year, 2-year, and 3-year survival of 52%, 30%, and 30%, respectively. HLA matching of vaccine, age, sex, race, and pathology did not bear a significant relation to response.

CONCLUSION: Minimal toxicity and good survival in this small population suggest clinical benefit from vaccination.

	INTRODUCTION

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION Authors' Disclosures of... REFERENCES

 
Carcinoma of the lung is the leading cause of cancer death and the second most commonly occurring cancer in both men and women in the United States.¹ Non–small-cell lung cancers (NSCLC) are considered to be minimally or nonimmunogenic, and may contain CD4 regulatory cells that suppress generation of cytotoxic lymphocytes (CTL).² Although NSCLC has not been considered a good candidate for immunotherapy, we hypothesized that NSCLC is indeed suitable for successful vaccine therapy because the tumor cells have not been exposed to immune attack and have not developed resistance mechanisms. Immunotherapy trials for lung cancer have yielded no consistent benefit to date in humans.^3-5 Vaccine trials with B7.1 (CD80) transfected allogeneic or autologous cells have not been reported in patients with NSCLC, although similar vaccines have shown good activity in other human studies.^6-9 The objectives of this phase I study were to assess the safety, immunogenicity, and clinical response to an allogeneic whole cell tumor vaccine transfected with CD80 and HLA A1 or A2 administered to patients with advanced metastatic NSCLC. Here we report on vaccine safety, clinical response, and overall survival.

	PATIENTS AND METHODS

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION Authors' Disclosures of... REFERENCES

 
Patients
In this University of Miami (Miami, FL) institutional review board-approved phase I clinical trial, which was also approved by the National Institutes of Health Recombinant DNA Advisory Committee and by the US Food and Drug Administration, we treated 19 patients who had relapsed metastatic NSCLC. Most had already been unsuccessfully treated with chemotherapy, radiotherapy, surgery, or a combination of all three. The eligibility criteria were as follows: age older than 18 years, histologically confirmed NSCLC (stage IIIB with malignant pleural effusion, stage IV, or recurrent), measurable disease, Eastern Cooperative Oncology Group performance status 0 to 2, and signed informed consent. Patients with brain metastases were included if these had been successfully treated. Patients were not eligible for study if they had received chemotherapy, radiation therapy or a biologic modifying agent during the preceding 4 weeks. Patients were also not eligible if they had symptomatic cardiac disease, materially reduced lung function, known HIV infection, or were on corticosteroids or other immunosuppressive therapy. All patients were treated at the University of Miami/Sylvester Comprehensive Cancer Center and gave signed informed consent. A complete history and physical exam were performed and the following tests made before enrollment: complete blood count; platelet count; chemistries (calcium, lacate dehydrogenase, blood urea nitrogen, creatinine, electrolytes, liver function tests); EKG; and HLA typing. Patients were followed twice monthly while being vaccinated, with tumor response assessed by computed tomography scans.

Vaccine Cell Line
A human lung adenocarcinoma cell line (designated AD100) was established in 1994 at the University of Miami, derived from a patient with NSCLC. This cell line has been kept in culture in standard medium and is free of contamination by Mycoplasma, virus, or other adventitious agents. It is homogeneous, adherent to plastic, and grows at a rate of division of approximately 26 hours.

Genetic Modification
AD100 was transfected with plasmid cDNA, pBMG-Neo-B7.1, and with either pBMG-His-HLA A2 or with B45-Neo-CM-A1-B7.1.¹⁰ Transfected cells were selected with G418 and histidinol. Verification of correct sequences was based on restriction analysis and the expression of the relevant gene products, namely G418 or histidinol resistance for the vector sequence, HLA A1, A2, and B7.1 expression for the transfected cDNA. The cells were irradiated with 120 Gy in a Co irradiator and stored frozen in 10% dimethyl sulfoxide in aliquots of 5 x 10⁷ cells until use. In tissue culture, the cells were viable for approximately 14 days but were unable to form colonies, indicating their inability to replicate. The cells were thus considered safe for use as vaccine cells. The minimum requirement for such use was coexpression of HLA A1 or A2 plus B7.1 on at least 70% of the transfected cells. The untransfected AD100 line was negative by flow cytometry for staining with anti HLA A1 or A2 or B7.1. Figure 1 shows the quality control by flow cytometric analysis of CD80 and HLA A1 or A2 transfected AD100 vaccine cells used for immunization.

View larger version (29K): [in this window] [in a new window]   Fig 1. Coexpression of transfected HLA A1 (A) or A2 (B) and CD80 on vaccine cells used for the treatment of non–small-cell lung cancer; two-color flow cytometry with anti-CD80-phycoerythrin and anti-HLA A1 or A2-fluorescein-isothiocyanate.

Immunologic Testing
Immunologic tests were done by enzyme-linked immunospot (ELISPOT) assays for interferon-gamma (IFN-). Purified CD8 cells were obtained from patients before and after each course of three vaccinations. CD8 cells were enriched by negative depletion with anti-CD56, anti-CD4, and other antibodies using the Spin-sep prep (Stem Cell Technologies, Vancouver, Canada). Purity was better than 80%, the primary contaminating cells being B-cells. CD8 cells were frozen in 10% dimethyl sulfoxide and 20% fetal cult serum-containing medium for analysis until all vaccinations of a study patient were completed. Analysis for pre- and postvaccination ELISPOT frequency was carried out on the same day in the same microtiter plate. Assays were done in quadruplicate, stimulating 2 x 10⁴ purified patient CD8 cells with 10³ A1 or A2 transfected or untransfected AD100, respectively, with K562 or with media only for 3 days, and determining the frequency of IFN- producing cells by ELISPOT. Immune assays were performed before vaccination and after three, six, and nine vaccinations.

Statistical Analysis
Patient characteristics are presented as counts with percentages, or as mean values and range. Overall survival, estimated by the Kaplan-Meier product-limit method,¹¹ is defined as time from enrollment onto study until death from any cause. In the absence of death, follow-up was censored at the date of last patient contact. Univariate and multivariate proportional hazards regression¹¹ were used to determine whether patients' survival time was related to age (continuous), sex, race (other v white non-Hispanic), tumor pathology (adenocarcinoma v other), and HLA-matching of vaccine. Logistic regression was used for the corresponding analyses of clinical response. For hazard ratios and the percentage of patients surviving, we report 90% CIs L₉₀ -U₉₀. These can be interpreted as providing 95% confidence that the parameter being estimated, such as the hazard ratio, exceeds L₉₀.

	RESULTS

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION Authors' Disclosures of... REFERENCES

 
Patient Characteristics
The characteristics of the 19 study patients are outlined in Table 1. Eastern Cooperative Oncology Group performance status was 0 to 1 in 18 patients (95%). Thirteen patients received vaccine matched for HLA, either A1 (three patients) or A2 (10 patients), whereas the six patients who were non-A1 and non-A2 received unmatched vaccine (ie, HLA A1 vaccine). While HLA A matched patients may be able to mediate CD8 responses by direct antigen presentation by the vaccine cells, we reasoned that unmatched patients may, nonetheless, mount a CD8 response via indirect antigen presentation after vaccine cell death and antigen uptake by antigen presenting cells. Before being enrolled on study, all patients had been previously treated: nine (47%) with surgery, six (32%) with radiation therapy, and 17 (89%) with chemotherapy. Among the chemotherapy-treated patients, 10 (53%) had been unsuccessfully treated with more than one chemotherapy regimen.

Regarding other side effects, one patient complained of transient pain at the injection site. Four patients developed some erythema at the vaccination site that resolved within a week. One patient experienced moderate arthritic pain in several joints after the first course. We did not find any patients with significant alteration of their laboratory parameters, including: complete blood and platelet counts, creatinine/BUN, calcium, and liver function tests.

One patient had a partial response lasting 13 months, and five showed stable disease ranging from 1.6 to 39+ months (Table 3). The clinical response rate was 32% (six of 19 patients). As of February 2004, these patients had survival times ranging from 23 to 40+ months, and five patients were still alive.

Among the other 13 patients who did not respond to therapy, only two were alive as of February 2004. One of these patients experienced disease stabilization with gefitinib, and the other is undergoing palliative chemotherapy.

Logistic regression analyses of age, sex, race, pathology, and HLA-matching of vaccine showed that none of these factors were statistically significantly related (P .10 in all instances) to clinical response (ie, to partial response or stable disease).

Figure 2 shows the Kaplan-Meier estimate of overall survival for the 19 study patients (vertical tick marks indicate censored follow-up). The estimated median survival time is 18 months (90% CI, 7 to 23 months). Estimates of 1-year, 2-year, and 3-year overall survival are 52% (90% CI, 32% to 71%), 30% (90% CI, 11% to 49%), and 30% (90% CI, 11% to 49%), respectively. As of February 2004, death had occurred in 12 patients from 1 to 23 months after entry on study (Table 2). For the seven patients who are still alive, follow-up from study entry currently ranges from 10 to 40 months, with a median follow-up time of 36 months.

View larger version (11K): [in this window] [in a new window]   Fig 2. Overall survival for the 19 B7 vaccine-treated non–small-cell lung cancer study patients.

Immune Response to Vaccination
This cohort of patients had been heavily pretreated and carried large tumor burdens that are believed to be immunosuppressive. It was important, therefore, to establish whether the tumor vaccination protocol was able to induce a specific immune response in these patients. Since the CD8 CTL response is thought to be critical for tumor rejection, we focused on this arm of the immune system. To distinguish between nonspecific natural killer (NK) activity and CD8 CTL activity, we employed a two-fold strategy. First, CD8 cells were purified to eliminate NK cells by including anti-CD56 in the negative selection cocktail of antibodies. Second, the CD8 cells were challenged with K562, an NK target. NK contamination would result in high titers of cells responding to K562 challenge.

All but one patient had a measurable CD8 response after 6 weeks (three vaccinations) that tended to increase after 12 weeks and stabilize by 18 weeks (Table 4). In vitro challenge of patient CD8 cells with wild type A1 or A2 transfected AD100 did not reveal significant differences. Two patients (patient Nos. 1012 and 1019) could not be evaluated immunologically because there was no follow-up sample available for analysis due to early disease progression or adverse events. One patient had only a very modest response, while most other patients showed a strong, highly statistically significant response to vaccination (see pre- and postimmunization titers on challenge with vaccine cells, and lack of response to K562 control; Fig 3, top panels).

View larger version (25K): [in this window] [in a new window]   Fig 3. CD8 immune response. Top two panels (A): CD8 prior to immunization or at 6, 12, and 18 weeks after challenged with untransfected (AD wt) vaccine cells or K562 control. Lower six panels (B): CD8 response after termination of vaccination (arrow) in patients with clinical response. Inf-, interferon gamma.

The immune response of the six clinically-responding patients (Fig 3, lower panels) shows that CD8 titers to AD100 stimulation continue to be elevated up to 150 weeks after cessation of vaccination. Sometimes CD8 cells, after prolonged immunization, secrete IFN- in response to K562, suggesting the acquisition of promiscuous CTL activity.

	DISCUSSION

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION Authors' Disclosures of... REFERENCES

 
Current recommendations for NSCLC patients with locally-advanced inoperable disease (stage IIIB) include platinum-based chemotherapy plus radiation therapy, and chemotherapy alone for patients with metastases (stage IV).¹² Results of these approaches are nevertheless poor, and the increase in survival is limited. The largest meta-analysis published to date concluded that chemotherapy increases the chance of 1-year survival by 10% and median survival by 6 weeks.¹³ A recent report from the Big Lung Trial group (BLT) reported similar results.¹⁴ In phase III clinical trials, patients with metastatic disease have a median survival of less than 1 year.¹⁵ Two-phase III trials showed that after failure of first-line chemotherapy, only 6% of patients receiving standard second-line chemotherapy could expect to respond, with median survival being approximately 6 months.^16,17 Our group of patients had a very poor prognosis as a result of their relapsed or metastatic disease status, and most patients had been unsuccessfully treated with surgery, radiation, and/or palliative chemotherapy, resulting in a projected survival of less than 6 months.

A vaccination approach such as used here may be an effective means of inducing immune response in patients with nonimmunogenic tumors. There is evidence that NSCLC tumors contain tumor antigens;^10,18-20 however, it is thought that lung tumors are poor candidates for immunotherapy because they are poorly immunogenic and potentially immunosuppressive,^2,21-24 thereby anergizing or tolerizing T-cells.^25-26 Lung tumors, therefore, have not been subjected to immune attack, and hence have not been able to evolve evasive mechanisms to resist immune effector cells. Lung tumors, unlike immunogenic tumors that harbor tumor-infiltrating lymphocytes, thus may succumb to killer CTLs, especially in light of the involvement of CD8 CTLs in tumor rejection in a number of model systems.²⁷ This led to considering that a potentially immunogenic tumor vaccine like ours could generate an appropriate immune response.

We chose an allogeneic whole cell vaccine, because whole cell vaccines have given the best clinical results so far. For example, statistically significant survival benefit occurred when a whole cell melanoma vaccine was administered.²⁸ In contrast, vaccine directed at a single epitope may have limited utility due to tumor escape mutants.²⁹ The additional advantage of a whole cell vaccine approach is that it does not require a priori delineation of specific lung tumor antigens. If vaccination is successful and CTLs are generated, as seems likely from our experience, the responsible antigenic sites can be identified later. Allogeneic cell-based vaccines offer a good alternative to autologous vaccines under the assumption that lung tumor antigens are shared in lung tumors of different patients, and the antigens can be cross-presented by patients' antigen-presenting cells. Although there is only limited evidence for shared antigens in lung tumors,^10,20 this has been shown in other tumors.^30,31

To obtain direct evidence that the CD8 cells generated in response to allogeneic vaccination recognize autologous tumor cells, it is necessary to obtain tumor specimens at the time of surgery. Tumor specimens were not available in this trial of patients with advanced disease. However, the prolonged maintenance of a high frequency of patient CD8 cells reacting to AD100 in vitro, and their increase in some patients (No. 1004 and No. 1007; Fig 3) even after cessation of external vaccination, is consistent with the hypothesis of immune stimulation of patient CD8 cells by the autologous tumor and their cross-reaction with the allogeneic vaccine.

Although only one patient had a partial response, five other patients had stable disease. Enhanced immune reactivity was demonstrated by a CD8-mediated tumor-specific immune response. The fact that six (32%) of 19 patients with very poor prognosis exhibited disease stabilization of a rapidly lethal condition, with median survival of the whole cohort reaching 18 months despite far-advanced disease, is encouraging. Our study suggests that tumor progression is slowed by vaccination, and that this effect occurs regardless of whether or not patients are allogeneic to the HLA A1 or A2 locus of the vaccine. The findings also suggest that indirect antigen presentation may be effective in promoting antitumor activity and that allogeneic major histocompatibility complex molecules enhance the effect. However, since this is a phase I study and the number of patients we treated is small, it is premature to draw firm conclusions from our data.

The vaccine was well tolerated and the patients' quality of life was very good. Because this is an immunologic product, we assumed that some immune-mediated side effects would be anticipated. Probable examples of such phenomena were the local erythema at the vaccination site in five patients, and the episode of arthritic pain experienced by one patient.

Given the advanced stage of disease in patients enrolled in this pilot phase I trial, we were surprised to find suggestive evidence of some clinical benefit. Moreover, since the B7-vaccine tested here induced CD8 CTL responses, it may be that the CD8 response is causally related to the clinical outcome seen here. We believe that the data shown here justify additional, larger studies to corroborate our results. We therefore intend to mount a phase II clinical trial in the setting of minimal disease. Patients with early stage NSCLC (stage I/II) will be vaccinated after surgery to decrease the chance of relapse and potentially prolong survival.

	Authors' Disclosures of Potential Conflicts of Interest

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION Authors' Disclosures of... REFERENCES

 
The authors indicated no potential conflicts of interest.

	NOTES

 
Supported by the Research Career Development Award given by the American Society of Clinical Oncology, University of Miami/WCA, RO1-CA39201-14, and the family of Iris Zeitler.

Preliminary results were presented at the 92nd Annual Meeting of the American Association of Cancer Research (AACR), San Francisco, CA, April 6–10, 2002, and the 10th World Conference on Lung Cancer, Vancouver, Canada, August 10–14, 2003.

In memory of Dr Kasi Sridhar (1953–2001).

Authors' disclosures of potential conflicts of interest are found at the end of this article.

	REFERENCES

TOP ABSTRACT INTRODUCTION PATIENTS AND METHODS RESULTS DISCUSSION Authors' Disclosures of... REFERENCES

 
1. Jemal A, Murray T, Samuels A, et al: Cancer statistics, 2003. CA Cancer J Clin 53:5–43, 2003[Abstract/Free Full Text]

2. 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[Abstract/Free Full Text]

3. 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]

4. 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]

5. Ratto GB, Cafferata MA, Scolaro T, et al: Phase 1I study of combined immunotherapy, chemotherapy, and radiotherapy in the postoperative treatment of advanced non-small-cell lung cancer. J Immunother 23:161–167, 2000

6. Antonia SJ, Seigne J, Diaz J, et al: Phase 1 trial of a B7-1 (CD80) gene modified autologous tumor cell vaccine in combination with systemic interleukin-2 in patients with metastatic renal cell carcinoma. J Urol 167:1995–2000, 2002[CrossRef][Medline]

7. Horig H, Lee DS, Conkright W, et al: Phase 1 clinical trial of a recombinant canarypoxvirus (ALVAC) vaccine expressing human carcinoembryonic antigen and the B7.1 co-stimulatory molecule. Cancer Immunol Immunother 49:504–514, 2000[CrossRef][Medline]

8. Hull GW, McCurdy MA, Nasu Y, et al: Prostate cancer gene therapy: Comparison of adenovirus-mediated expression of interleukin 12 with interleukin 12 plus B7-1 for in situ gene therapy and gene-modified, cell-based vaccines. Clin Cancer Res 6:4101–4109, 2000[Abstract/Free Full Text]

9. von Mehren M, Arlen P, Tsang KY, et al: Pilot study of a dual gene recombinant avipox vaccine containing both carcinoembryonic antigen (CEA) and B7.1 transgenes in patients with recurrent CEA-expressing adenocarcinomas. Clin Cancer Res 6:2219–2228, 2000[Abstract/Free Full Text]

10. 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[Abstract/Free Full Text]

11. Parmar MKB, Machin D: Survival Analysis: A Practical Approach. New York, Wiley, 1995

12. Clinical practice guidelines for the treatment of unresectable non-small-cell lung cancer. Adopted on May 16, 1997 by the American Society of Clinical Oncology. J Clin Oncol 15:2996–3018, 1997[Abstract]

13. Chemotherapy in non-small cell lung cancer: A meta-analysis using updated data on individual patients from 52 randomised clinical trials. Non-Small Cell Lung Cancer Collaborative Group. BMJ 311:899, 1995[Abstract/Free Full Text]

14. Stephens RJ, Fairlamb D, Gower N, et al: The big lung trial (BLT): determining the value of cisplatin-based chemotherapy for all patients with non-small cell lung cancer (NSCLC). Preliminary results in the supportive care setting. Proc Am Soc Clin Oncol 21:291a, 2002 (abstr 1161)

15. 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[Abstract/Free Full Text]

16. 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[Abstract/Free Full Text]

17. 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[Abstract/Free Full Text]

18. 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[Abstract/Free Full Text]

19. 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]

20. 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[Abstract/Free Full Text]

21. 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[Abstract/Free Full Text]

22. 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]

23. Neuner A, Schindel M, Wildenberg U, et al: Cytokine secretion: Clinical relevance of immunosuppression in non-small cell lung cancer. Lung Cancer 34:S79–82, 2001 (suppl 2)

24. 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[Abstract/Free Full Text]

25. Schwartz RH: Models of T cell anergy: Is there a common molecular mechanism? J Exp Med 184:1–8, 1996[Free Full Text]

26. Lombardi G, Sidhu S, Batchelor R, et al: Anergic T cells as suppressor cells in vitro. Science 264:1587–1589, 1994[Abstract/Free Full Text]

27. Podack ER: Functional significance of two cytolytic pathways of cytotoxic T lymphocytes. J Leukoc Biol 57:548–552, 1995[Abstract]

28. Morton DL, Hsueh EC, Essner R, et al: Prolonged survival of patients receiving active immunotherapy with Canvaxin therapeutic polyvalent vaccine after complete resection of melanoma metastatic to regional lymph nodes. Ann Surg 236:438–449, 2002[CrossRef][Medline]

29. Velders MP, Schreiber H, Kast WM: Active immunization against cancer cells: Impediments and advances. Semin Oncol 25:697–706, 1998[Medline]

30. Fong L, Engleman EG: Dendritic cells in cancer immunotherapy. Annu Rev Immunol 18:245–273, 2000[CrossRef][Medline]

31. Boon T, Cerottini JC, Van den Eynde B, et al: Tumor antigens recognized by T lymphocytes. Annu Rev Immunol 12:337–365, 1994[CrossRef][Medline]

Submitted October 31, 2003; accepted April 28, 2004.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

				X. Zang and J. P. Allison The B7 Family and Cancer Therapy: Costimulation and Coinhibition Clin. Cancer Res., September 15, 2007; 13(18): 5271 - 5279. [Abstract] [Full Text] [PDF]

				I. Bolonaki, A. Kotsakis, E. Papadimitraki, D. Aggouraki, G. Konsolakis, A. Vagia, C. Christophylakis, I. Nikoloudi, E. Magganas, A. Galanis, et al. Vaccination of Patients With Advanced Non-Small-Cell Lung Cancer With an Optimized Cryptic Human Telomerase Reverse Transcriptase Peptide J. Clin. Oncol., July 1, 2007; 25(19): 2727 - 2734. [Abstract] [Full Text] [PDF]

				J. Nemunaitis, R. O. Dillman, P. O. Schwarzenberger, N. Senzer, C. Cunningham, J. Cutler, A. Tong, P. Kumar, B. Pappen, C. Hamilton, et al. 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 J. Clin. Oncol., October 10, 2006; 24(29): 4721 - 4730. [Abstract] [Full Text] [PDF]

				D. O'Mahony, S. Kummar, and M. E. Gutierrez Non-Small-Cell Lung Cancer Vaccine Therapy: A Concise Review J. Clin. Oncol., December 10, 2005; 23(35): 9022 - 9028. [Abstract] [Full Text] [PDF]

				L. E. Raez, S. Fein, and E. R. Podack Lung Cancer Immunotherapy Clin. Med. Res., November 1, 2005; 3(4): 221 - 228. [Abstract] [Full Text] [PDF]

				S. Gnjatic Immunogenic Targets in Non-Small Cell Lung Cancer: More Is More Clin. Cancer Res., August 1, 2005; 11(15): 5331 - 5332. [Full Text] [PDF]

This Article Abstract Full Text (PDF) Purchase Article View Shopping Cart Alert me when this article is cited Alert me if a correction is posted Services Email this article to a colleague Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Save to my personal folders Download to citation manager Rights & Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Raez, L. E. Articles by Podack, E. R. Search for Related Content PubMed PubMed Citation Articles by Raez, L. E. Articles by Podack, E. R. Social Bookmarking                            What's this?