Chest ACCP Career Connection
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Guest Access | Sign In via User Name/Password
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF) Free
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Article Archive
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (15)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sugaya, M.
Right arrow Articles by Yasumoto, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sugaya, M.
Right arrow Articles by Yasumoto, K.
(Chest. 2002;122:282-288.)
© 2002 American College of Chest Physicians

Establishment of 15 Cancer Cell Lines From Patients With Lung Cancer and the Potential Tools for Immunotherapy*

Masakazu Sugaya, MD; Mitsuhiro Takenoyama, MD; Toshihiro Osaki, MD; Manabu Yasuda, MD; Akira Nagashima, MD; Kenji Sugio, MD and Kosei Yasumoto, MD

* From the Second Department of Surgery (Drs. Sugaya, Takenoyama, Osaki, Yasuda, Sugio, and Yasumoto), University of Occupational and Environmental Health; and Department of Chest Surgery (Dr. Nagashima), Kitakyushu Municipal Medical Center, Kitakyushu, Japan.

Correspondence to: Masakazu Sugaya, MD, the Second Department of Surgery, University of Occupational and Environmental Health, 1-1 Iseigaoka, Yahatanishi-ku, Kitakyushu 807-8555, Japan; e-mail: sugaya{at}med.uoeh-u.ac.jp


   Abstract
TOP
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
Background: Since lung cancer is the major cause of death not only in Japan but in many other industrialized countries, the development of new therapeutic modalities is quite important. In patients with melanoma, immunotherapy with some tumor antigens has been shown to result in tumor regression. However, little is known about specific immune responses and tumor antigens in lung cancer, due to difficulty in establishing appropriate lung cancer cell lines. In order to resolve these difficulties, we tried to establish and characterize lung cancer cell lines as useful tools for the analysis of tumor-specific immune responses in patients with lung cancer.

Materials and methods: We tried to establish lung cancer cell lines from 549 patients with resectable lung cancer and from 21 patients with pleural and pericardial effusions or lymph node metastasis. We characterized the established cell lines after the induction of tumor-specific cytotoxic T lymphocytes (CTLs), and analyzed both the major histocompatibility complex (MHC) class I and class II molecules on their surfaces.

Results: We succeeded in establishing 15 lung cancer cell lines from 570 specimens (2.6%). The success rate of the establishment of lung cancer cell lines was significantly higher in patients at such advanced stages as MHC III and IV than in those at MHC stages I and II (p = 0.004). MHC class I molecules were expressed in 12 of 15 cell lines (80%), while MHC class II molecules were found in 3 of 15 cell lines (20%) on their cell surfaces by flow cytometry. A haplotype loss of MHC class I antigens was found in 6 of 15 cell lines (40%). Although CTLs were induced in only two of eight cell lines tried by stimulation with nontransduced autologous tumor cell lines, CTLs were successfully induced in all of eight cell lines tested by stimulation with CD80-transfected autologous tumor cells.

Conclusions: These results suggested that the tumor antigens recognized by CTLs could thus exist in the tumor cells derived from many lung cancer patients. It is, therefore, possible that antigen-specific immunotherapies may be potentially effective for patients with lung cancer by adoptive transfer of CTLs, as well as by vaccine therapy using tumor-specific antigens.

Key Words: cytotoxic T lymphocyte • lung cancer • major histocompatibility complex class I and class II • tumor cell line


   Introduction
TOP
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Lung cancer is the leading cause of cancer death not only in Japan but in many other industrialized countries because of its increasing frequency of occurrence, as well as its resistance to currently available chemotherapy and radiation therapy.1 2 Therefore, the development of new modalities of treatment is important to provide improved cure rate for this disease.

Tumor regression antigens, which are recognized by cytotoxic T lymphocytes (CTLs), have been identified from melanoma complementary DNA,3 4 5 and have been shown to act as specific targets for immune recognition in patients with melanoma. Immunotherapy using some of these tumor-specific peptide antigens as vaccine therapy has been shown to result in tumor regression in some of melanoma patients.6 7 Up to now, little is still known about tumor antigens in lung cancer, because of the difficulty in establishing lung cancer cell lines from tumor samples. In the present study, we established 15 lung cancer cell lines from surgically resected specimens of lung cancer patients, and thus analyzed the expression of major histocompatibility complex (MHC) class I and class II molecules, and the ability of tumor cells to induce CTLs by transfecting with CD80 into the cell lines.


   Materials and Methods
TOP
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Specimens
Tumor samples were obtained from patients with lung cancer undergoing diagnostic and therapeutic procedures. All patients gave informed consent for the use of surgical samples for these investigational and analytical studies. Most of the tissue samples were obtained from surgically resected tumor tissue specimens (549 cases); but, when possible, pleural (10 cases) and pericardial effusions (5 cases) or metastasized lymph nodes (6 cases) were also obtained. All samples were immediately placed on ice following surgical resection, thoracentesis, or pericardiocentesis, and were transported to our laboratory. One half of the samples were used for these studies, and the rest of the samples were processed for pathologic or cytologic examination.

Culture Medium
The culture medium (CM) consisted of RPMI-1640 medium (Gibco; Grand Island, NY) supplemented with 10 mM hydroxyethyl piperazine-ethanesulfonic acid, 100 U/mL of penicillin, 100 mg/mL of streptomycin, and 10% heat-inactivated fetal calf serum (Gibco).

Preparation of Lung Cancer Cells From Resected Samples
Tumor cells were prepared as described previously.8 Briefly, fresh tumor tissues and lymph nodes with metastasis were excised from patients with lung cancer. Both necrotic tissue and apparently normal tissue were discarded, and the remainder of the tumor tissue was then minced into small pieces (approximately 1 mm3) with scissors. This minced tissue was shaken with a mixture of 0.1 mg/mL of deoxyribonuclease type I, 1 mg/mL collagenase type IV, and 0.5 mg/mL of hyaluronidase type V (all from Sigma Chemical; St. Louis, MO) in CM at 150 revolutions per minute in a water bath at 37°C for 1 h. This resultant cell suspension was washed in Hanks balanced salt solution, applied to a Ficoll-Hypaque gradient (LSM; Organon Teknika; Durham, NC) and then centrifuged (1,000g for 30 min). The pleural and pericardial effusions were also subject to Ficoll-Hypaque gradient separation. The interface was collected and washed three times with Hank’s balanced salt solution and suspended in CM in order to use as tumor cells. When a growth of fibroblasts was noted in the cultures, they were removed using a cell scraper. Fifteen tumor cell lines were generated from the above preparations and were maintained for > 12 months as a monolayer culture by serial passages in CM. The human leukocyte antigen (HLA) genotype of established lung cancer cell lines was identified by using polymerase chain reaction (Shionogi; Tokyo, Japan).

Transfection of CD80 Into Lung Cancer Cell Lines
Lung cancer cell lines were transfected with 2 µg of the pBj/hCD80 plasmid containing human CD80 gene (kindly provided by Dr. M. Azuma; National Children’s Medical Research Center)9 using a lipofectamin reagent (Gibco) according to an instruction manual. These cell lines were selected with G418, 1 mg/mL, for > 1 month.

Preparation of Regional Lymph Node Lymphocytes and Peripheral Blood Lymphocytes
Regional lymph nodes lymphocytes (RLNLs) and peripheral blood lymphocytes (PBLs) from patients with lung cancer were obtained at the time of surgery. Each lymphocyte was prepared using the method described in our previous report,8 and PBLs were subject to MHC class I serotyping (SRL; Tokyo, Japan).

Generation of Epstein-Barr Virus-Transformed B Cells
Autologous B cells immortalized by the Epstein-Barr virus (EBV-B) were produced from patients with lung cancer by infection of peripheral blood mononuclear cells with supernatant form the Epstein-Barr virus producer line B95.8. The HLA genotype of autologous EBV-B was identified by Shionogi Company (Tokyo, Japan).

Induction of CTLs
RLNLs were stimulated with solid-phase anti-CD3 monoclonal antibody (mAb) [Ortho Pharmaceutical Corporation; Raritan NJ] for 48 h, and then were expanded in CM containing 50 U/mL recombinant interleukin-2 (kindly donated by Takeda Chemical Industries; Osaka, Japan) in 24-well plates (Iwaki Glass; Tokyo, Japan) for 14 days at 37°C in a 5% CO2 atmosphere as previously reported.8 Subsequently, the lymphocytes were stimulated with irradiated (100 Gy) CD80-transfected autologous tumor cells weekly at a tumor-to-lymphocyte ratio of 1:10 in CM with 50 U/mL recombinant interleukin-2 for 3 weeks. The CTL activity was assessed at the end of the culture.

mAb
The culture supernatants of American Type Cell Culture HB-145 (IVA12; anti-HLA-DR, -DP, -DQ mAb), ATCC HB-95 (W6/32; anti-HLA-A, -B, -C), CRL-8002 (OKT 4; anti-CD4) and CRL-8014 (OKT 8; anti-CD8) were purchased (American Type Cell Culture; Rockville, MD) and used as sources for blocking mAb and for analyzing the expression of MHC class I and II molecules on the cell surfaces.

Flow Cytometry
For indirect staining, the cells (2 x 105) were incubated with either the first mAb or murine IgG (control) for 30 min at 4°C. They were washed and stained with fluorescein isothiocyanate-conjugated anti-mouse IgG. A fluorescence analysis was performed using a flow cytometer (EPICS XL; Coulter International; Fullerton, CA).10

Cytotoxicity Assay
The cytotoxicity of CTLs against the tumor cells was examined by a standard 51Cr-release cytotoxicity assay as previously described.8 Briefly, the 51Cr-labeled target cells were incubated with effector cells at different effector-to-target ratios in 96-well round-bottomed microplates (Nunc; Roskilde, Denmark). The supernatants were collected after 4 h of incubation at 37°C in 5% CO2 for the measurement of the chromium released from lysed cells.

Detection of Interferon-{gamma} by Enzyme-Linked Immunosorbent Assay
The CTLs (106/mL) were co-cultured with autologous tumor cells (105/mL) for 24 h, and the amount of interferon (IFN)-{gamma} in the supernatant was measured (Human Interferon Gamma ELISA Test Kit; Amersham International; Little Chalfont, Buckinghamshire, England). The sensitivity limit of the IFN-{gamma} (enzyme-linked immunosorbent assay) is 2 pg/mL

Statistical Test
Data were analyzed using Student t test for comparison of means, and {chi}2 analysis for comparison of proportions.


   Results
TOP
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Establishment of Lung Cancer Cell Lines and Clinical Features
From January 1996 to July 2000, tumor cells that were obtained mainly from digestion of 570 lung cancer tissues into single cells with a mixture of enzymes were put into a culture in an attempt to directly establish permanent lung cancer cell lines. Fifteen of the 570 specimens (2.6%) could thus be established as stable cell lines (14 cell lines from resected tumor tissues and 1 cell line from pericardial effusion). The 15 cell lines comprised eight adenocarcinomas, two squamous cell carcinomas, three large cell carcinomas, one adenosquamous carcinoma, and one small cell carcinoma. These cell lines can be maintained as adherent cells in continuous culture for > 12 months. Next, we investigated the clinical status of the patients whose tumor cells were established as in vitro culture cell line. Table 1 shows the characteristics of 570 patients, such as stage, sex, age, and histology. The success rate of the establishment of lung cancer cell lines was significantly higher in patients at advanced stages III and IV than in patients at stages I and II (p = 0.004). As shown in Table 2 , 10 of 15 patients (67%) have already died, and 11 of 15 patients (73%) were at extended stage (stage III or stage IV) at diagnosis.


View this table:
[in this window]
[in a new window]

  		Table 1.. Patient Characteristics*

 

View this table:
[in this window]
[in a new window]

  		Table 2.. In Vitro Cell Growth and Survival of Lung Cancer Patients

 
Expression of MHC Class I and Class II Molecules in Established Cancer Cell Lines
We examined the expression of MHC class I and class II molecules in the 15 established cell lines. As shown in Table 3 , MHC class I molecules were expressed in 12 of 15 cell lines (80%) on their cell surfaces by flow cytometry. In contrast, MHC class II molecules were expressed in 3 of 15 cell lines (20%). These results suggest that most of lung cancers express MHC class I molecules on their cell surfaces for the presentation of certain antigens recognized by CD8-positive T cells.


View this table:
[in this window]
[in a new window]

  		Table 3.. Expression of MHC Class I and MHC Class II Molecules in Lung Cancer Cell Lines

 
MHC Class I Genotypes of Lung Cancer Cell Lines
Since the loss of MHC class I molecules was found in 3 of 15 cell lines by flow cytometry, we next examined the MHC class I genotypes of established cell lines to determine whether MHC class I gene was lost or not, and whether or not a loss of MHC class I haplotype occurred. As shown in Table 4 , all of the lung cancer cell lines had MHC class I genes, and 6 of 15 cell lines (40%) had a haplotype loss of MHC class I antigens in comparison with those of either EBV-B or the serotyping of autologous PBLs. There was no relationship between the expression of MHC class I on cell surfaces by flow cytometry and the haplotype loss of MHC class I antigens. The expression of MHC class I molecules on the cell surfaces needed for the presentation of peptide antigens to CTLs of the immune system.11 The loss of MHC class I molecules on the cell surfaces and the haplotype loss of MHC class I antigens may be one of the mechanisms by which tumor cells escape from immune detection.


View this table:
[in this window]
[in a new window]

  		Table 4.. MHC Class I Haplotype Analysis of Lung Cancer Cell Lines*

 
Induction of CTLs Specific for Autologous Lung Cancer Cells
Since 12 of the established cell lines proved to express MHC class I molecules on their surfaces, we attempted to generate autologous tumor-specific CTLs by stimulation with either autologous tumor cells or CD80-transfected autologous tumor cells. We previously reported the increased success rate of induction of tumor-specific CTLs in patients with lung cancer using CD80-transfected autologous tumor cells.9 In this study, as shown in Table 5 , we successfully obtained tumor-specific CTLs from all eight patients tested by in vitro stimulation with CD80-transfected autologous tumor cells, although CTLs were induced in two of eight patients by stimulation with autologous tumor cells without CD80 transfection. These results suggest that costimulatory molecules expressed on tumor cells are needed to induce CTLs effectively, and that the tumor-specific antigens recognized by CTLs are frequently expressed in lung cancer cell lines. The characterization of CTLs from A925 patients is representatively shown in Figures 1 , 2 . Cytotoxic activity against autologous tumor cells of CTLs by stimulation with autologous tumor cells without CD80 transfection was apparent in comparison to those stimulated with autologous tumor cells or those without autologous tumor stimulation (Fig 1) . The CTLs produced IFN-{gamma} in response to autologous tumor cells. To further analyze the mechanism of recognition by the CTLs, blocking assays with anti-MHC class I, MHC class II, CD4, and CD8 mAbs were performed (Fig 2) . Anti-MHC class I mAb and CD8 mAb were thus found to significantly inhibit the IFN-{gamma} production from the CTLs, but not anti-class II or CD4 mAb. The CTLs from the other seven patients showed the similar activities. These results indicated that the recognition by the CTLs was restricted by MHC class I molecules.


View this table:
[in this window]
[in a new window]

  		Table 5.. Induction of CTLs Specific for Autologous Lung Cancer Cells

 


View larger version (14K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 1.. Cytotoxic activity of lymphocytes stimulated with CD80-transfected autologous tumor cells. Cytotoxic activities against parental autologous tumor cells stimulated either with autologous tumor cells (AT), CD80-transfected autologous tumor cells (CD80-AT), or without any tumors were assessed at 7 days after the last tumor stimulation (day 35) by a standard 4-h 51Cr release assay. The representative data (A925) from eight cases are shown. Open circles indicate CD80-transfected autologous tumor cells; open squares indicate no tumor; open triangles indicate autologous tumor cells. Effector-to-target ratio: effectors indicate lymphocytes stimulated either with autologous tumor cells or CD80-transfected autologous tumor cells, or without any tumors. Target indicates autologous tumor cells.

 


View larger version (32K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 2.. The blocking effects of mAbs on CTL activity. The CTLs generated from RLNLs were co-cultured with irradiated autologous tumor cells in the presence of the indicated mAbs for 24 h, and then the supernatant of the mixed culture was collected and assayed for IFN-{gamma}. Results are the mean ± SE of three experiments.

 

   Discussion
TOP
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Generally, it is more difficult to establish lung cancer cell lines compared with melanoma cell lines that have been well analyzed in the field of tumor immunology. Several studies12 13 have reported that the establishment of lung cancer cell lines was difficult in serum-supplemented growth medium with a low success rate (< 10%). However, it was reported that the use of chemically defined medium such as ACL-3, ACL-4, and HITES could definitely increase the establishment rate of a lung cancer cell line (approximately 20 to 72%).14 15 16 17 First, we tried to establish lung cancer cell lines in a serum-free medium and succeeded in maintaining them for a few passages, but failed to cultivate them for long periods of time as stable cell lines. We, therefore, attempted to establish cell lines with serum-supplemented growth medium, and 15 lung cancer cell lines (2.6%) were thus obtained from 570 specimens, including tumor tissues, pleural and pericardial effusions, and metastatic lymph nodes. Since the success rate of establishing such cell lines was very low, the optimal growth factors for the primary culture of such tumor cells need to be further elucidated.

There have so far been several reports concerning the prognostic impact of ability to establish cell lines in vitro on survival. The ability to establish cell lines was reported to correlate with a low survival rate in patients with lung cancer from whom tumor cells were derived.17 18 Our findings are that the success rate of establishment of lung cancer cell lines was higher in patients at such advanced stages as III and IV than in those at early stages as I and II (Table 1) . Ten of 15 patients (67%) whose tumor cells were established as in vitro culture cell line have already died, and 11 of 15 patients (73%) were at extended stage (stage III or stage IV) at diagnosis (Table 2) . A review19 of the literature reported an absolute lack of MHC class I antigens in approximately 15% and 55% of the surgically removed primary and metastatic melanoma lesions, respectively, stained in immunohistochemical reactions by mAb to monomorphic determinants of HLA class I antigens. The explanation for this mechanism is considered to be loss of transporter molecules,20 mutation localized within ß2-microglobulin gene,21 and the absence of binding of transacting factors involved in MHC class I regulation.22 Although all of lung cancer cell lines in the present study had MHC class I genes by polymerase chain reaction (Table 4) , the loss of MHC class I molecules was found in only 3 of 15 cell lines (20%) on their surface by flow cytometry (Table 3) . A haplotype loss of MHC class I antigens was found in 6 of 15 lung cancer cell lines (40%) [Table 4 ]. Hiraki et al23 reported that a haplotype loss of MHC class I antigens was found in three of seven non-small cell lung cancer cell lines (43%). Grandis et al24 reported that the haplotype loss of MHC class I antigen in squamous cell carcinoma of the head and neck was found in 13 of 26 cell lines (50%). Although a haplotype loss of MHC class I antigens was found in cancer cell lines reported here, CTLs that may recognize tumor antigens presented by the remaining HLA molecules could be successfully derived from all four cell lines (A110 L, A129 L, A904 L, and A529 L) with a haplotype loss (Table 5) . These findings suggest that tumor-specific T cells existed even in patients whose tumors showed a haplotype loss of MHC class I antigens, indicating a possibility of T-cell-mediated immunotherapy for lung cancer patients.

It is well accepted that CTLs require two signals to activate. The first signal is interaction of T-cell receptor with MHC class I antigen complex. The second signal is interaction of T-cell receptor and costimulatory molecules such as CD80.25 We previously reported that CD80 on tumor cells could be a beneficial costimulatory molecule to elicit CTLs against lung cancer.9 In this report, CTLs were successfully derived from all eight patients tested by using CD80 transfectants as stimulator cells (Fig 1 , 2 ). CD80 transfectants thus were good tools to detect tumor-specific CTL responses in lung cancer patients. These results suggest that tumor antigens recognized by CTLs might exist in most of lung cancer cells by means of CD80 transfectants, as previously reported in melanoma.3 26

In melanoma, some of the tumor antigens recognized by CD4 T lymphocytes have been identified.27 28 29 In our study, three lung cancer cell lines expressed MHC class II molecules on their cell surfaces (A925 L, A529 L, and E522 L). For this reason, these cell lines will be suitable tools for identifying MHC class II-restricted antigens recognized by CD4 T lymphocytes. Moreover, immunities specific for tumor cells have been reported to be mediated by either antibodies or CTLs, but both of immune responses also required the help of CD4 T lymphocytes.30 If CTLs and antibodies specific for tumor cells are detected in patients with lung cancer, then MHC class II-restricted CD4 T lymphocytes may be potentially useful for clinical applications.

Immunization with both MHC class I- and class II-restricted tumor antigens may induce antigen-specific CTLs and CD4 T lymphocytes. Therefore, these cell lines could be useful as a valuable in vitro model for further biological and experimental studies, and as identification of tumor antigens recognized by MHC class I-restricted CD8 T cells, MHC class II-restricted CD4 T cells, and antibodies. The cell lines are therefore considered to be important tools for identifying tumor antigens.


   Acknowledgements
 
We thank Mrs. Yumiko Hase, Ms. Kahoru Noda, and Mrs. Miki Shimada for their expert help.


   Footnotes
 
Abbreviations: CM = culture medium; CTL = cytotoxic T lymphocyte; EBV-B = B cells immortalized by the Epstein-Barr virus; HLA = human leukocyte antigen; IFN = interferon; mAb = monoclonal antibody; MHC = major histocompatibility complex; PBL = peripheral blood lymphocyte; RLNL = regional lymph node lymphocyte

Received for publication August 21, 2001. Accepted for publication January 29, 2002.


   References
TOP
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Grili, R, Oxman, AD, Julian, JA (1993) Chemotherapy for advanced non-small-cell lung cancer: how much benefit is enough? J Clin Oncol 11,1855-1872
  2. Damstrup, J, Poulson, HS (1994) Review of the curative role of radiotherapy in treatment of non-small-cell lung cancer. Lung Cancer 11,153-178[ISI][Medline]
  3. van der Bruggen, P, Traversari, C, Chomez, P, et al (1991) A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science 254,1643-1647[Abstract/Free Full Text]
  4. Boel, P, Wildmann, C, Sensi, ML, et al (1995) BAGE: a new gene encoding an antigen recognized on human melanomas by cytolytic T lymphocytes. Immunity 2,167-175[CrossRef][ISI][Medline]
  5. Coulie, PG, Brichard, V, Van Pel, A, et al (1994) A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas. J Exp Med 180,35-42[Abstract/Free Full Text]
  6. Rosenberg, SA, Yang, JC, Schwartzentruber, DJ, et al (1998) Immunologic and therapeutic evaluation of a synthetic peptide vaccine for the treatment of patients with metastatic melanoma. Nat Med 4,321-327[CrossRef][ISI][Medline]
  7. Nestle, FO, Alijagic, S, Gilliet, M, et al (1998) Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med 4,328-332[CrossRef][ISI][Medline]
  8. Takenoyama, M, Yoshino, I, Fujie, H, et al (1998) Autologous tumor-specific cytotoxic T lymphocytes in a patient with lung adenocarcinoma: implications of the shared antigens expressed in HLA-A24 lung cancer cells. Jpn J Cancer Res 89,60-66[CrossRef][ISI]
  9. Takenoyama, M, Yoshino, I, Eifuku, R, et al (2001) Successful induction of tumor-specific cytotoxic T lymphocytes from patients with non-small cell lung cancer using CD80-transfected autologous tumor cells. Jpn J Cancer Res 92,309-315[CrossRef][ISI]
  10. Takenoyama, M, Yasumoto, K, Harada, M, et al (1996) Expression of activation-related molecules on regional lymph node lymphocytes in human lung cancer. Immunobiology 195,140-151[ISI][Medline]
  11. Rivoltini, L, Barracchini, KC, Viggiano, V, et al (1995) Quantitative correlation between HLA class I allele expression and recognition of melanoma cells by antigen-specific cytotoxic T lymphocytes. Cancer Res 55,3149-3157[Abstract/Free Full Text]
  12. Duchesne, GM, Eady, JJ, Peacock, HJ, et al (1987) A panel of human lung carcinoma lines: establishment, properties and common characteristics. Br J Cancer 56,287-293[ISI][Medline]
  13. Bepler, G, Koehler, A, Kiefer, P, et al (1988) Characterization of the state of differentiation of six newly established human non-small-cell lung cancer cell lines. Differentiation 37,158-171[CrossRef][ISI][Medline]
  14. Carney, DN, Gazdar, AF, Bepler, G, et al (1985) Establishment and identification of small cell lung cancer cell lines having classic and variant features. Cancer Res 45,2913-2923[Abstract/Free Full Text]
  15. Brower, M, Carry, DN, Oie, HK, et al (1986) Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. Cancer Res 46,798-806[Abstract/Free Full Text]
  16. Oie, HK, Russell, EK, Carney, DN, et al (1996) Cell culture methods for the establishment of NCI series of lung cancer cell lines. J Cell Biochem Suppl 24,24-31[Medline]
  17. Stevenson, H, Gazdar, AF, Phelps, R, et al (1990) Tumor cell lines established in vitro: an independent prognostic factor for survival in non-small-cell lung cancer. Ann Intern Med 113,764-770[ISI][Medline]
  18. Masuda, N, Fukuoka, M, Takada, M, et al (1991) Establishment and characterization of 20 human non-small cell lung cancer cell lines in a serum-free defined medium (ACL-4). Chest 100,429-438[Medline]
  19. Ferrone, S, Marincola, FM (1995) Loss of HLA class I antigens by melanoma cells: molecular mechanisms, functional significance and clinical relevance. Immunol Today 16,487-494[CrossRef][ISI][Medline]
  20. Versteeg, R, Kruse-Wolters, KM, Plomp, AC, et al (1989) Suppression of class I human histocompatibility leukocyte antigen by c-myc is locus specific. J Exp Med 170,621-635[Abstract/Free Full Text]
  21. Gattoni-Celli, S, Kirsch, K, Timpane, R, et al (1992) ß2-Microgloblingene is mutated in human colon cancer cell line (HCT) deficient in the expression of HLA class I antigens on the cell surface. Cancer Res 52,1201-1204[Abstract/Free Full Text]
  22. Blanchet, O, Bourge, JF, Zinszner, H, et al (1991) DNA binding of regulatory factors interacting with MHC-class-I gene enhancer correlates with MHC-class-I transcriptional level in class-I-defective cell lines. Int J Cancer Suppl 6,138-145[Medline]
  23. Hiraki, A, Kaneshige, T, Kiura, K, et al (1999) Loss of HLA haplotype in lung cancer cell lines: implications for immunosurveillance of altered HLA class I/II phenotypes in lung cancer. Clin Cancer Res 5,933-936[Abstract/Free Full Text]
  24. Grandis, JR, Falkner, DM, Melhem, MF, et al (2000) Human leukocyte antigen class I allelic and haplotype loss in squamous cell carcinoma of the head and neck: clinical and immunogenetic consequences. Clin Cancer Res 6,2794-2802[Abstract/Free Full Text]
  25. Chen, L, Linsley, PS, Hellstrom, KE (1993) Costimulation of T cells for tumor immunity. Immunol Today 14,483-486[CrossRef][ISI][Medline]
  26. Kawakami, Y, Eliyahu, S, Delgado, CH, et al (1994) Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. Proc Natl Acad Sci U S A 91,2829-2833[Abstract/Free Full Text]
  27. Touloukian, CE, Leitner, WW, Topalian, SL, et al (2000) Identification of a MHC class II-restricted human gp100 epitope using DR4-IE transgenic mice. J Immunol 164,3535-3542[Abstract/Free Full Text]
  28. Wang, RF, Wang, X, Atwood, AC, et al (1999) Cloning genes encoding MHC class II-restricted antigens: mutated CDC27 as a tumor antigen. Science 284,1351-1354[Abstract/Free Full Text]
  29. Wang, RF, Wang, X, Rosenberg, SA (1999) Identification of a novel major histocompatibility complex class II-restricted tumor antigen resulting from a chromosomal rearrangement recognized by CD4 (+) T cells. J Exp Med 189,1659-1668[Abstract/Free Full Text]
  30. Bowne, WB, Srinivasan, R, Wolchok, JD, et al (1999) Coupling and uncoupling of tumor immunity and autoimmunity. J Exp Med 190,1717-1722[Abstract/Free Full Text]



This article has been cited by other articles:


Home page
 Clin. Cancer Res.Home page
Y. Nagata, T. Hanagiri, M. Takenoyama, T. Fukuyama, M. Mizukami, T. So, Y. Ichiki, M. Sugaya, K. Sugio, and K. Yasumoto
Identification of the HLA-Cw*0702-Restricted Tumor-Associated Antigen Recognized by a CTL Clone from a Lung Cancer Patient
Clin. Cancer Res., July 15, 2005; 11(14): 5265 - 5272.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
T. So, M. Takenoyama, M. Mizukami, Y. Ichiki, M. Sugaya, T. Hanagiri, K. Sugio, and K. Yasumoto
Haplotype Loss of HLA Class I Antigen as an Escape Mechanism from Immune Attack in Lung Cancer
Cancer Res., July 1, 2005; 65(13): 5945 - 5952.
[Abstract] [Full Text] [PDF]

Home page
 J. Thorac. Cardiovasc. Surg.Home page
R. E. Merritt, R. E. Yamada, R. G. Crystal, and R. J. Korst
Augmenting major histocompatibility complex class I expression by murine tumors in vivo enhances antitumor immunity induced by an active immunotherapy strategy
J. Thorac. Cardiovasc. Surg., February 1, 2004; 127(2): 355 - 364.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
M. Tanabe, H. Izumi, T. Ise, S. Higuchi, T. Yamori, K. Yasumoto, and K. Kohno
Activating Transcription Factor 4 Increases the Cisplatin Resistance of Human Cancer Cell Lines
Cancer Res., December 15, 2003; 63(24): 8592 - 8595.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF) Free
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Article Archive
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (15)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sugaya, M.
Right arrow Articles by Yasumoto, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sugaya, M.
Right arrow Articles by Yasumoto, K.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS