Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Vol. 1, 501-507, May 1995 Clinical Prolonged Disease-free Epithelial Ovarian Period Cancer Tumor-infiltrating Kazuyuki Fujita, Koichi and Adoptive Shoji two Kodama, Takeshi Tanaka2 Department of Obstetrics Niigata and Gynecology, was (5) ited that more patients Thirteen did not show any with detectable epithelial lesion ovarian after cancer, who hardly cisplatin-containing chemotherapy following primary operation, were treated with adoptive transfer of tumor-infiltrating lymphocytes (TIL group). Eleven patients with almost equivalent conditions of disease, who were treated with only chemotherapy following primary operation, served as a control group. The median time of follow-up was 36 (range, 23-44) months in the TIL group and 33 (range, 14-48) months in the control group. The estimated 3-year overall survival rate of diseasefree patients in the TIL group and in the control group was 100% and 67.5%, respectively. A significant difference was noticed between the overall survival rate of the ilL group and the control group (P < 0.01). Furthermore, the estimated 3-year disease-free survival rate of the patients in the TIL group and in the control group was 82.1 % and 54.5%, respectively. The disease-free survival rate of patients in the TIL group and in the control group was significantly different (P < 0.05). These results suggest that the adoptive transfer of TILs after all chemotherapy has been finished might be one promising method to achieve complete cure of advanced epithelial ovarian cancer. to treat can cancers achieve lymphoma, agents as a supplement cures and of certain prolong ( 1-4). However, even vanced condition cannot lives now, have been used to surgery diseases, be completely 20 years and radiotherapy such of patients epithelial for over as leukemia with ovarian ovarian improve cancer overcome with by the primary PATIENTS 7/19/94; This was work accepted supported 2/3/95. in part by a grant-in-aid from the Ministry Health and Welfare for the Comprehensive 10-Year Strategy Control. 2 To whom requests for reprints should be addressed. use lasts the infusion This 3-5 with alone exhib- result to metastatic the duration only adoptive 5 of 7 patients of tumor. rate of patients, suggests melanoma of response months, we treated ilLs can 13 patients cancer with the adoptive transfer cisplatin-containing chemotherapy AND of advanced Gynecology They stage Stage were to with of cryoprefollowing a METHODS of either Center, or April 1989 documented and March 1992, epithelial ovarian 43 pacancer (International Federation of Obstetrics II, III, or IV) were enrolled in this treated cology in the Department Niigata Niigata of Obstetrics University City Hospital, Hospital. and Niigata Eligibility and study. criteria GyneCancer were as follows: age not less than 18 years; Eastern Cooperative Oncology Group performance status of 3 or lower (able to perform minimal self-care); life expectancy more than 2 months; no chemotherapy or radiotherapy for 4 weeks prior to entry into protocol; adequate bone marrow function (WBC count >4000/ mm3; platelet count > 100,000/mm3), hepatic function (total bilirubin mine concentration >2.0 concentration gave Good were excluded chemotherapy. tumor < informed Clinical mass was following from ination, after mg/dl), 1.5 mg/dl); consent cryopreservation considered renal function (creat- infection. to the Japanese Guidelines. All Gov- In 19 patients, re- clinically detected after completion of a primary operation. These patients other and were 24 patients detected by CF scan, and/or MRI, 1 1 patients were and and no active according Practice this study In the was obtained I TIL by et a!. melanoma with (6), cancer is similar However, since Patients. Between tients with histologically cells Received with decrease tumors operation. and con- treated in single epithelial ovarian served TILs after and ad- cancer a 50% solid Rosenberg of metastatic trials ovarian to ilLs. from rIL-2. treated clinical the survival sidual tumor chemotherapy cancer with In our by ilLs ernment Chemotherapeutic generated in (1-4). improve the survival rate of patients in this manner. In an attempt to eliminate minimal residual tumor and patients INTRODUCTION cells patients is detected 3 years suspensions regression than obtained ABSTRACT about objective ovarian that epithelial in sensitivity of the disease within in 1 1 of 20 of ilLs. epithelial Niigata recurrence are lymphoid observed transfer 951 [T. T.], Japan and single-cell reported [A. T.]; and of of the patients culturing of Obstetrics and Gynecology, Niigata University School 1, Asahimachi-dori [K. F., H. I., Ko. T., S. K., Ke. T.]; of Obstetrics and Gynecology, Niigata City Hospital Center, treatment, thirds ilLs3 Takahashi, Department of Medicine, Department Cancer 501 Advanced Transfer ventional Ikarashi, Hirokazu Tokunaga, Kenichi after with Research Lymphocytes1 Takakuwa, Akiteru in Patients Cancer treated with in whom ultrasonogram, whose TILs or consent as a control no residual internal examwere < 1 X i0 to receiving group, second-line ilLs was not and the remaining of for Cancer The abbreviations used are: ilL, tumor-infiltrating lymphocyte; rIL-2, recombinant interleukin 2; C-FDA, carboxyfluorescein diacetate; CT, computed tomography; MRI, magnetic resonance imaging. 3 Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. 502 Adoptive Immunotherapy Using Cryopreserved TIL .13 patients performed until 4 Primar or Incompleted) Viable cells were at the density containing 5% 24 patients residual the tumor DMSO, 4 1 1 patients control 13 patients group NoTILs TIL 1 x 10 > ery rate as group the cells number Clinical protocol for ovarian of patients cancer 3-h in stages II, III, and IV. three another before cells exceeded received (TIL group; Treatment sues with treatment Fig. 1). Design. resected reached TIL 100 in liquid tion, TILs were at a primary approximately after operation. 5 X units/mI rIL-2, nitrogen for several the obtained When 10 after TILs were from the 2-3 months. of chemo- completion weeks frozen of and After cancer number tis- C-FDA mented with patients for 20 ml 25% mm 15-30 HBSS opera- 5-fluorouracil-cyclophosphamide-Adriamycin-cisplatin or cyclophosphamide-Adniamycin-cisplatin was administered all patients. 5-Fluorouracil-cyclophosphamide-Adniamycin-cisplatin 350 was mg/m2 on day administered according to the following cyclophosphamide on day 1, 40 mg/m2 1, 50 mg/m2 orouracil cisplatin as a continuous on day infusion phamide-Adriamycin-cisplatin administration. to five serum, were All performed and did not show of chemotherapy for this was not any study. After visited the rence, and internal serum, and hospital examination detectable lesion the primary recovery to the regularly examination, toxicity by patients. for ultrasonogram after due infusions for TIL After early tumor or CT last of pa- scan in were For a recov- and cultured S weeks after before of thawing), administra- administered i.v. were and washed 0.2% to FITC-labeled in phenol sodium azide mAb, incubated anti-HLA-DR (Becton at Dickinson In vitro cytotoxicity of TILs fresh, frozen targets of autologous and 1(562 cells. Cryopreserved digestion were thawed 10% human AB serum was prepared incubation with labeled with Sigma C-FDA three (50 Co., times with and cultured in for 24 h. Single- from monolayers of viable trypsin at 37#{176}C and washed Chemical incubated was or allogetumor cells with various ig/ml diluted St. Louis, HBSS. with MO) RPM! for 60 mm The cells numbers tumor in the (5 X of effector Japan), and an automated Compact MT). as C-FDA in the surviving fluorescence microscope Results follows: C-FDA count - test spontaneous C-FDA C-FDA licates were three-step were <10%. Target of Conversion using effector The units count - in medium to determine respectively. triplicate of (maximum = incubated cells. to lytic cells was photometer C-FDA cells at was to percentage cytotoxicity X-100 were used C-FDA counts, determined titrations converted count)/(maximum count). alone or with 2% Triton spontaneous and maximum with were percentage 10) cells 37#{176}C for 3 h. After incubation, C-FDA in the supernatant masked with 5 mM calf hemoglobin (Wako Pharmaceuticals, percentages recur- marker then cytotoxicity of thawed cultivation discharge, detection for consid- to the and ( Reitz completion were were FCS Cytotoxicity. medium; Ltd., Tokyo, counted with chemo- described operation from check and eligibility the patients were given rIL-2 was used only administered tients the and at 37#{176}C and washed were three for tumor marker in CT scan, and/or MRI of operation matched following chemotherapy cycle, TILs simultaneously. and the effect who for After albumin 1% obtained by enzymatic RPM! 1640 containing 1640 5-fluorouracil and Japan). Vitro medium 5-flu- cyclophos- chemotherapy examination, check by ultrasonogram, patients mg/m2 The HBSS, and 2 weeks an appropriate assessed by using neic tumor cells cell suspension cells by brief schedule: Adniamycin 350 1-5. excluded underwent to evaluate Only above ered regimen patients cycles. Internal and examination therapy. 1, and on days to at 37#{176}C (with TILs (1 X 10) anti-CD56, Tokyo, In cryopreserved the primary Ltd.), months. at 4#{176}C for 45 mm, washed twice, and resuspended in 0.5 ml medium for fluorescence-activating cell sorting analysis. The mAbs used were anti-CD3, anti-CD4, anti-CD8, anti-CD16, Japan, cultivation Product several iO#{176}. Just the 7.5% with simultaneously. containing with anti-CD25, of cells cells; 10k’ the phenotype of TILs was determined by flow and in vitro cytotoxicity of TILs was assessed by a assay. The TILs in 100 ml sterile saline supple- 4#{176}C, stained patients X serum with 1 X in of cultivation, 5 AB to plain cells/ml i05 2 weeks. freezing Flow Cytometry. 13 X weeks for The (7). removed (Plancer times cells. elsewhere 2 rapidly isolated i07 of TILs, red-free therapy of human thawed for (3 weeks X were 2-3 nitrogen culture atmosphere described freezer liquid washed above 2-6 approximately were 70-95%), cytometry from they cultivation or No consent 1 in (7). of initially was After in 30% from complete number the cells reached isolated elsewhere in anti-CD3-coated with was rIL-2. frozen performed in a humidified sample a programmed described cells/ml, used were placed at a density TILs was described IL-2, activation, cryopreserved tion Fig. of administration, excluded iO tumor with then using were and (Costar) medium were counted at 37#{176}C.The of CD3 number TILs (-) TILs methods 2 X each therapy of TILs. to the medium plates complete (+) CO2 from 1 week 6-well tumor of additional detected. exogenous culture After 19 patients then from cells complete residual according free viable No was Infusion tissue medium 4 and cancer plates Chemotherapy 4 weeks. lesion Culture Operation (Completed every a recurrent the All measurements SDs was of the performed Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. repas Clinical Table I Phenotypic flow cytometry analysis and characteristics of killing of fresh cancer tissues The phenotype of TILs was determined by fluorescence-activated cell Dickinson fluorescence-activated cell sorter. Fluonescein isothiocyanate-labeled appropriate dilution. Leu-4 recognizes mature T cells (CD3); Leu-3 recognizes class Il-restricted helper/inducer T cells (CD4); HLA-DR recognizes B cells, cells and neutrophils (CD16); anti-IL-2 receptor recognizes low affinity interleukin associated with natural killer activity (CDS6). One lytic unit is defined as the % of Positive Patient CD3 TIL cultures CD4 CD8 with CD3CD4 CD25 No. of cells injected (X lOu) CD56 dominant epithelial Killing activity (LUS(J107 Autologous cells) K562 Allogeneic group) 10.7 99.1 2.3 38.1 ND” 1.0 52.1 ND 53.6 43.2 99.4 2.6 44.3 ND 2.8 83.3 17.8 64.5” 15.2’ 7 99.3 80.3 16.7 99.2 1.2 92.8 9.0 3.5 16.2 10.5 10.7” 20.6 11 98.3 80.5 14.7 98.0 8.0 14.3 32.5”56.2’ with CD3CD8 0.8 cells in excess of 50% 1 99.1 36.0 68.3 92.9 2 5 6 99.9 99.1 98.3 90.8 98.3 95.2 34.9 8.5 21.9 9.2 10.3 28.7 68.2 91.7 76.8 80.5 87.2 66.3 81.8 90.6 99.0 99.9 98.9 98.0 12 99.2 20.4 82.1 13 99.9 37.6 60.8 not done. tumor tumor tumor tumor ND, Fresh C Fresh d Fresh e Fresh 1Fresh Fresh I’ Fresh Fresh J Tumor tumor tumor tumor tumor cells g I described cells cells cells cells from from from from patient patient patient patient cells cells cells cells from from from from from patient patient patient patient patient I 1. previously number (8). of effector cells 65.4 (CD8 1.6 dominant 9.4 1.5 45.2 group) ND 4.4 52.3 ND ND 2.9 2.4 1.8 3.5 2.0 2.0 66.7 100.2 90.9 95.2 121.0 104.9 16.5 28.1 22.3 28.3 18.4 26.4 54.l 22.7 43.5” 66.7’ 58.1’ 8.2k 45.0’ 217h 48.6’ 19.4’ 72.6” 52.9’ 0.9 1.1 4.0 10.6 3.5 7.0 66.5 92.0 50.5 90.8 90.3 33.9 ND ND 3.5 21.2 4.1 9.1 99.9 1.0 65.2 20.0 1.0 98.9 21.3 58.0’ 84.8 0.1 26.7 11.4 2.6 69.3 18.6 736h542j 7. 10. 4. 8. 6. 9. One lytic unit 50% was lysis defined as of 5 X i03 the variety of allogeneic target target cells, tumor of the same killer-sensitive Survival of TILs when the patient between survival Kaplan-Meier data until is still curves were obtained the death alive. were of patients or curves using and the Survival calculated higher two against autologous in the CD8 in the RESULTS CD4 cytotoxicity r!L-2 and from advanced stage Infusion freshly ovarian markers, rate, and cytotoxic observed with preparations 70-95% by the dye exclusion After recovery was against in cell two test from against activity of (10.5-28.3 compared surface group were the group), cells lymphocytes expression cells were were mainly culture in excess TIL period activated HLA-DR 50% cytotoxic cells 13 (CD8’ ilLs (CD4 were (Table In 9 of 4 preparations, of preparations for T lymphocytes, antigen. CD3CD8 and in the remaining 13 propagated the were of the Phenotypes with cinoma dominant tested activity preparations lytic units/107 The serous a nomas, to those group mucinous The years. cystadenocarcinomas, and two endometnioid patient as K562 cell (Table cytotoxic cells generally ages In the 13 patients two low fresh in the in the control serous of the patients group two endo- adenocar- ranged (Table undifferentiated adenocarcinomas TIL group cystadeno- undifferentiated control varied it was and allogeneic 2), seven as The cytotoxic The cystadenocarcinomas, one 7, well types. high types. K562 of 1 1 patients and included. 55) of preparations cells autologous of prepara- tumor however, both at least killing of eight showed to was from cell characteristics similar with Preferential against cells); adenocarcinomas, were 13 tumor tested that against to 67 (median, at least against all three 1 1 and allogeneic 4). In the TIL carcinomas, metrioid patients regard compared preparation and natural With and one of three cell fresh the activity autologous tumor some with cells. Patients. activity. allogeneic from activity tumor the All 13 patients TILs of the by during to be viable cryopreservation. change and these CD3CD4 of tumors major as judged dominant propagated In all 13 preparations, no phocytes, preparations, We against line. in seven In the tumor, and cytotoxic types. group group. low cells cell observed dominant was in the 13 patients are presented in Table 1. All TIL cultures consisted of mainly CD3 T lym- growth cultured TILs rIL-2 expanded resected cancer. of TILs were confirmed when recovered from cryopreservation, of TILs. was dominant the tumor tumor cells type, cell study, autologous ovarian autologous leukemia in this tumor fresh histological cultured killing allogeneic tions including K562 preferential from method. Culture 65.3’ 2. cells. last contact differences ND 5. mediating Statistical Analysis. the day of administration 503 ovarian 88.1 b once from 99.9 9 10 group). propagated 99.9 8 with TILs 3 4 TIL cultures a CD16 of 50% (CD4 of cultured Research sorting analysis performed with a 488-nm argon laser on a Becton mAbs were purchased from Becton Dickinson Japan and used at the class I-restricted cytotoxic/suppressor T cells (CD8); Leu-2 recognizes macrophages, and activated T cells; Leu-1 ic recognizes natural killer 2 receptor(CD2S); and Leu-19 recognizes the major subset of cells number of effector cells mediating 50% lysis of 5 X tO3 target cells. cells HLA-DR cells in excess cells Cancer from 3), seven adenocarci- were included. Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. 28 504 Adoptive Immunotherapy Using T able 2 ilL Cryopreserved Characteni sties of the patients treated with adoptive transf en of cultu red ilLs after chemotherapy Prognosis Age No. of chemotherapy courses (yr)/ performance Patient status Clinical diagnosis 1 58/0 Ovarian Histopathology cancer stage Previous treatment Serous Regimen Outcome Outcome disease Observed period (mo) of Surgery (N’) FCAP” 5 NED NED 44 cystoadenocarcinoma Surgery (B1C) CAP 5 NED NED 39 cystoadenocarcinoma lie 48/0 Ovarian cancer Mucinous 3 56/0 stage Ovarian stage IV cancer lIe Endometrioid adenocarcmoma Surgery (Bi) FCAP 3 NED NED 36 5 49/0 Ovarian cancer Endometrioid adenocarcinoma Surgery (Bi) FCAP 5 NED REC 39 6 28/0 stage lie Ovarian cancer stage lib Mucinous cystoadenocarcinoma Surgery (A) FCAP 3 NED NED 36 7 51/0 Ovarian cancer Undifferentiated Surgery (Bi) FCAP 5 NED NED 37 stage Ovarian stage Ovarian stage Ovarian IlIc cancer ha cancer IV cancer 2 8 55/0 10 48/0 12 31/1 adenocarcinoma Serous cystoadenocarcinoma Surgery (A) FCAP 3 NED NED 31 Serous cystoadenocarcinoma Surgery (Bi) FCAP 5 NED REC 33 Serous cystoadenocarcinoma Surgery (Bi) CAP 5 NED NED 32 Serous cystoadenocarcinoma Surgery(A) CAP 5 NED NED 25 stage IlIc 14 67/0 15 64/0 16 55/1 19 Ovarian stage Ovarian stage Ovarian stage Ovarian 57/1 cancer lIe cancer Ilic cancer Mucinous cystoadenocarcinoma Surgery (B2”) CAP 5 NED NED 38 Serous cystoadenocarcinoma Surgery (Bi) CAP 5 NED NED 23 Serous cystoadenocarcinoma Surgery (B2) CAP 5 NED NED 24 lib cancer stage Ilic REC, a No I, FCAP, macroscopic tumor residuum. 5-fluorouracil-cyclophosphamide-Adniamycin-cisplatin; recurrence of disease. Largest diameter of tumor residuum was less than 2 cm. Largest diameter of tumor residuum was larger than 2 cm. NED, no evidence of disease; d The ages years. of the Primary patients tumor ranged from reduction 42 surgery to 68 (median, revealed that 49) residual ( Fig. 3). The difference patients in the tumor mass remained in 9 of 13 patients in the TIL group and in 6 of 11 patients in the control group (“Residuum,” Table 4). However, by completion of cisplatin-containing chemotherapy, group was were detected group after macroscopic completion these by residual 15 patients examinations CT scan, and/or infused into plications i3 such tumor mass and they such as internal MRI. When patients were as nausea, entirely diagnosed disappeared to be free examination, the of had the of disease autologous group, vomiting, in in the ultrasonography, cultured TIL no hepatitis, sion, or respiratory distress due ability and loss of intravascular Survival Analysis. patients. erage, The 33.6) (average, TILs were remarkable com- oliguria, to increased fluid were 34.6) overall group and tively (Fig. of patients group 3-year Survival observation months 3-year and cyclophosphamide-Adniamycin-cisplatin; hypoten- capillary observed permein any in months survival 2). The the rate and between group and that 23 and the overall of patients 14 The patients 100% for to 44 from group. of disease-free was available from group control group difference in the TIL were ranged ilL in the in the control data period all control group was 82.1% and 54.5%, the disease-free and that significant in the vaginal 22 and TIL and residual ‘ Table disease-free 4). was ence between 76.2% TIL group these rate in the and 33.3%, that control the respectively patients group had operation (‘ ‘ Re- estimated group survival of the significant TIL Before nine the primary patients, the disease-free and statistically For the ilL administration. in the after lesions from chemotherapy, patients tumor survival group six ilL control Recurrent of two patients from rate of in the (P < 0.05). stump 32 weeks group survival of patients and (Fig. 3-year in the control 4). The differ- rate of the patients patients in the control in the group was (P < 0.05). (av- to 48 in the TIL respec- survival rate in the control was statistically significant (P < 0.01). The estimated disease-free survival rate of the patients in the TIL group in the group of cisplatin-containing siduum,’ DISCUSSION Although estimated 67.5%, between TIL statistically macroscopic patient. 24 CAP, C respectively imen chemotherapy is reported epithelial ovarian an advanced isolated with tumors sion (7). the median about solid tumors by culturing The shown to express administration with a variety cisplatin-containing 2 years treatment rate for patients (4). specific of TILs of human reg- in the survival is only and in patients cancer, the effective stage from rIL-2 with to be relatively ilLs have of at been single-cell suspensions lysis of autologous mediates tumors tumor (9); regres- however, Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. in Clinical Tab le 3 Characteristi cs of the patients treated withou t adoptive tran sfer of cul tured ilLs Cancer Research 505 afte r chemotherapy Prognosis Age (yr)/ No. of performance Previous diagnosis Histopathology Observed chemotherapy Regimen courses Outcome of period Patient status Clinical 1 58/0 Ovarian cancer stage lie Serous cystoadenocarcinoma Surgery (A”) FCAP” 5 NED NED 2 46/0 Ovarian Serous cystoadenocarcinoma Surgery (B2’) CAP 5 NED REC, Serous eystoadenoeareinoma Surgery (A) CAP 3 NED NED 33 cancer treatment Outcome disease (mo) 48 DOD 30 stage Ilie 3 43/0 4 49/0 5 49/1 6 42/0 7 42/0 Ovarian cancer stage Ovarian stage Ovarian stage Ovanna stage lIe cancer lie cancer lib cancer 11th Endometrioid adenocarcinoma Surgery (A) FCAP 5 NED NED 28 Endometnioid adenocareinoma Surgery (A) FCAP 5 NED NED 47 Serous eystoadenocareinoma Surgery (BV’) FCAP 5 NED NED 42 cancer Serous eystoadenocarcinoma Surgery (Bi) FCAP 5 NED DOD 39 Serous eystoadenocareinoma Surgery (Bi) FCAP 5 NED REC, DOD 24 Undifferentiated adenocareinoma Serous cystoadenocareinoma Surgery (B2) FCAP 5 NED REC, DOD 29 Surgery (B2) FCAP 5 NED REC 47 Undifferentiated adenocareinoma Surgery (A) CAP 5 NED NED 14 Ovarian stage 8 68/0 9 68/0 10 47/0 11 50/0 REC, a No b FCAP, macroscopic IV Ovarian cancer stage Ovarian stage Ovarian stage Ovarian stage Ilie cancer Ilie cancer IIIe cancer lie tumor residuum. 5-fluorouraeil-cyelophosphamide-Adriamycin-eisplatin; Largest d diameter Table 4 to tumor Characteristics residuum was o f patients ilL No. of patients macroscopic tumor CMacroscopic clinical critical trials, point chemotherapy with regard injection agglutinin, (n=13) 7 4 2 5 5 4 5 9 6 7 3 2 1 7 0 2 2 P<O.Ol ‘___i control 6o group (n=ll) 1 3 Y_I faa Srt 4 Pr.o1 2 Overall survival rate of patients with ovarian cancer stage II, III, and IV who were diagnosed as having no evidence of disease after completion of chemotherapy. Fig. of ilLs is limited (6). The in combination with especially and the drugs. reported killer group group 11 51.1 ± 9.5 of responses transfer (10) that the of cellular by delayed-type natural TIL residuum. immunoactivation as demonstrated cyclophosphamide-Adriamyein-eisplatin; cancer Control is to establish adequate conditions, to timing for infusion of lymphocytes we CAP, operation. the duration of anticancer induces ov arian residuum. of adoptive Recently, TILs tumor of disease; 2 em. operation Histopathology Serous Mucinous Endometrioid Undifferentiated a Diagnosed at the primary No with no evidence than 2 cm. than group Completed” Residuumc b less 13 51.4 ± 10.8 Age (yr) Clinical stage” II III IV Status of primary our NED, recurrence of disease; DOD, dead of disease. CLargest diameter of tumor residuum was larger cytolytic adoptive transfer immunity in patients, hypersensitivity activity of to phytohemagainst K562 cells, and blood. percentages These of cells findings bearing suggest mor regression was mediated immunity. Cisplatin-containing tered after noactivation Additionally, adoptive transfer of cellular drug-resistant the CD8 antigen in peripheral the possibility that in part by the chemotherapy, activated cellular if it is adminis- of ilLs, immunity tumor interferes observed with tu- the immu- in patients induced by ilLs. cells possess increased often Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. 506 Adoptive Immunotherapy Using Cryopreserved TIL so L, 80 C C 3 60 60 L, C control group (n=l 1) 40 S I control 20 0 3 Years Fig. 3 Disease-free survival completion Start with as having and sensitivity examined investigate the setting, cytotoxicity TILs ovarian cancer no evidence stage of disease II, after when major the change markers; slightly against autologous TILs period was but still tumor of patients the completion after As shown with and valuable cytotoxie was recognized in patients with tumors in the was detected lose transfer 1, ilLs demonstrated surface out patients operation, after re- CD3CD8 (c) cells of CD56 Ten or mixtures or CD16 of 1 1 preparations, had some well as autologous cytotoxic cells in addition A significant of both, TILS were with activity tumor the exception against cells, present allogeneic and several ilLs a small number to anticancer cells substantial num- in some of preparation tumor could 7, cells as kill K562 part of the clinical with measurable treated gous TILs new study, no significant injected, and nomodulation in patients and that of sitivity to phytohemagglutin patients in the control group. One of our interests was to evaluate whether adoptive transfer of ilLs prolongs the diseasefree survival time of patients. In this study the difference be- transfer of ilLs tween rate of disease-free the disease-free patients survival and that of patients in the control icant. It has been well documented of a primary operation epithelial ovarian cancer rate of patients group in the TIL group group was statistically signifthat the degree of completion influences prognosis of advanced stages, tients receive several courses of chemotherapy clinical study of patients with stage III epithelial 5-year survival rate in cases with no macroscopic of patients with although the pa(14-16). ovarian residual In our cancer, tumor protocol degree tumor cells. Therefore, by their characteristics. in the TIL survival ease, in primary TILs which ilLs mass, rate can elim- are refractory were administered and tumor regres- direct tumor few reports our rence. One of two ilL group showed and was trials, tumor with allogeneic tumor cells. noted between the overall to autologous difference of carried survival that cells patients was tumor lysis mediated have dealt with combined differences among patients in the TIL group despite different transferred ilLs, such as the ratio of CD4 :CD84 of cells cases. five at the time by the analysis on ilL-treated patients. This article is the of the survival rate of patients with epithelial ovarian chemotherapy. In this or in patients drugs. prognostic first report cancer and disease-free tumor after of recurrence the analysis indicating of residual and tumor residual group, sion was assumed to reflect infused ilLs (4, 5). However, killing in the CD8 in the CD4 CD3CD4 while also mate in the ilL 41%, group when residual of disease no sign no residual increased preference since (a) histocompatible cells were not targeted in our assay, we could not clarify the cell type responsible for the cytotoxic- bers observed cm, study, macroscopic a significantly was <2 Furthermore, with had macroscopic in the TIL had In a large to the patients considered the cytotoxicity of the mixed cell activities, of mainly who of cryopreserved dominant group. However, we against tumor cells to be a measure consisted on who In this patients group activity of eight cultures three cultures TILs in four in the control tumors 22%. 4 tPrsSocol and had no evidence with cm, operation. of autologous tumor cells in seven dominant group and in one of (b) Cultured >2 primary of chemotherapy. in Table 84%, fro. Slat of patients operation was (7). These findings, along with of TILs (6), led us to initiate the adoptive survival primary to 3 months. in cell Disease-free after We might exceeds 4 tumor of chemotherapy. ilLs timing and rate cells covery from cryopreservation the results of the clinical trial TILs suitable grow in growth decreased treatment not of cultivation noted fresh with could of Fig. completion (1 1-13). of cryopreservation can be used since 3 Y_1 to immunotherapy influence whether clinical ity. (n=6) (,O_ 4 Protocol of chemotherapy. immunogenieity also from of patients III, and IV who were diagnosed No group ‘20 while of killing activity with ilLs and were observed conditions of cells, number against autolo- we could not predict the effect of We also estimated whether immuinduced by ilLs influences the recur- patients with recurrent disease that was in the very little change in delayed-type hypersenand compared in the other with patient, natural patients killer with activity nonrecurrent immunoactivation was disclearly induced relation (data not shown). An explanation between the clinical outcome transfused a variety ilLs might be that infused ilLs are still a mixture of of cells and that many unsolved issues remain regard- ing 4 the function K. Tanaka, of CD4 unpublished and CD8 for after and cells the and the lack of corphenotype of of class data. Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. I and Clinical class II MHC-restricted quite lation small to evaluate and disease-free investigate the parameter This TILs study after method cells. suggests all chemotherapy to obtain Since the number the correlation survival rate, favorable of patients was between immunomodufurther study is needed to to predict effectiveness of ilLs. transfer of cryopreserved has been finished is one feasible outcomes. 1934-1939, manuscript. 9. Rosenberg, Clin. 10. Oneol., REFERENCES 1. Shelley, W. E., Carmichael, J. C., Brown, L. B., Fraser, R. C., Kirk, M. E., Krepart, G. V., Levitt, M., Roy, M., Willan, A. R., and Wilson, K. S. Adriamycin and cisplatin in the treatment of stage III and IV epitherial ovarian carcinoma. Gynecol. Oncol., 29: 208-221, 1988. 2. Nguyen, H. N., Averette, H. E., Hoskins, W., Sevin, B., Penalver, M., and Steren, A. National survey of ovarian carcinoma VI. Cancer (Phila.), 72: 3007-3011, 1993. 3. Kobayashi, H., Maeda, M., Hayata, T., and Kawashima, Y. Clinical study of combination chemotherapy with CDDP, ADM and CPM for ovarian cancer. J. Jpn. Soc. Cancer Then., 23: 829-836, 1988. de-Gramont, A., Drolet, Y., Lavoie, A., Painchaud, M., Blouin, R., Tessier, C., and Ouellet, P. Adroamycin and cis-platinum in advanced ovarian cancer. Eur. J. Cancer Clin. Oncol., 21: 665-669, 1985. 4. Rosenberg, S. A., Packard, B. S., Aebersold, P. M., Solomon, D., Topaliam, S. L., Toy, S. T., Simon, P., Lotze, M. T., Yang, J. G., Seipp, C. A., Simpson, C., Carter, C., Bock, C., Sehwartzentruber, D., Wei, J. P., and White, D. E. Use of tumor-infiltrating lymphocytes and 5. in the N. Engl. immunotherapy of patients 25: 1676-1680, 1988. with metastatie mera- J. Med., 6. Aoki, Y., Takakuwa, K., Kodama, S., Tanaka, Tokunaga, A., and Takahashi, T. Use of adoptive K., Takahashi, M., transfer of tumor- S. A. The immunotherapy 180-199, 1992. and gene therapy of cancer. J. 10: Ikarashi, H., Fujita, Takahashi, K., Takakuwa, T., and Tanaka, ovarian lymphocytes. noma. 1991. Ikarashi, H., Aoki., Y., Fujita, K., Kodama, S., and Tanaka, K. Solid-phase anti-CD3 antibody activation and eryopreservation of human t tumor infiltrating lymphocytes derived from epithelial ovarian cancer. Jpn. J. Cancer Res., 83: 1359-1365, 1992. epithelial interleukin-2 cisplatin-containcancer. Cancer Bruning, J. W., Kardol, M. J., and Arentzen, R. Carboxyfluorescein fluorochromasia assays. 1. Non-radioactivity labeled cell mediated lympholysis. J. Immumol. Methods, 33: 33-44, 1980. We thank Minako Kimura, Blood Transfusion Division, Niigata University Hospital, for analyzing lymphocytes on the fluorescenceactivating cell sorting and Motoko Koshikawa for assistance in preparthis 51: with ovarian 507 8. ACKNOWLEDGMENTS ing Res., or in combination with epithelial Research 7. that adoptive clinical infiltrating lymphocytes alone ing chemotherapy in patients Cancer cancer Cancer K., Kodama, K. Immunomodulation after adoptive 190-196, Res., 54: transfer of S., Tokunaga, A., in patients with tumor-infiltrating 1994. 11. Biedler, J. L., and Riehm, H. Reduced tumor producing capacity of Chinese hamster cells resistant to actinomycin D and daunomycin. Proc. Am. Assoc. Cancer Res., 11: 8-il, 1970. 12. Allavena, P., Damia, G., Colombo, T., Maggioni, and Mantovani, A. Lymphokine-aetivated mediated eytotoxieity on tumor cell lines Cell. Immunol., 120: 250-258, 1989. 13. Gambacorti-Passerini, M., Fossati, moresistant murine and Radrizzani, lymphocytes. 14. tive Cancer Gniffiths, surgical Res., 48: Cancer Treat. Rep., 63: 235, 15. Diaz-Rubio, E., Eseudero, 2372-2376, 171-173, 1988. Fuller A. F. Role pf cytoredueof advanced ovarian cancer. 1979. J. L., Herraiz, F. Treatment of advanced ovarian cyclophosphamide (PAC). Eur. 1989. 16. Rakar, Prognostic M., C., Rivoltini, L., Supino, R., Rodolfo, M., G., and Parminani, G. Susceptibility of ehehuman cells to lysis by interleukin 2-activated C. T., Parker, L. M., and treatment in the management Gonzalez-Larniba, D., D’Inealei, killer (LAK) and monocyteresistant to antitumor agents. M., Martin-Jimenez, M. A., Lopez-Vega, M., Vidart, J. A., J. M., and Bullom, cancer with cisplatin, Adriamycin and J. Gynaeeol. Oneol., 10: 424-432, S., Kovacic, J., Cavic, M., Lukanovie, A., and Mozina, factors in ovarian cancer. Eur. J. Gynaeeol. Oneol., 1990. Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. A. II: Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research. Prolonged disease-free period in patients with advanced epithelial ovarian cancer after adoptive transfer of tumor-infiltrating lymphocytes. K Fujita, H Ikarashi, K Takakuwa, et al. Clin Cancer Res 1995;1:501-507. Updated version E-mail alerts Reprints and Subscriptions Permissions Access the most recent version of this article at: http://clincancerres.aacrjournals.org/content/1/5/501 Sign up to receive free email-alerts related to this article or journal. To order reprints of this article or to subscribe to the journal, contact the AACR Publications Department at [email protected]. To request permission to re-use all or part of this article, contact the AACR Publications Department at [email protected]. Downloaded from clincancerres.aacrjournals.org on April 28, 2017. © 1995 American Association for Cancer Research.