Article Figures & Data
Figures
- Fig. 1.
A, cytotoxic activities of the effector cells from patient F. Y. against target FY cells and inhibition of CTL response by anti-HLA class I mAbs. PBMCs from patient F. Y. were collected and cultured as described in “Materials and Methods.” Effectors were stimulated under four different conditions, as follows: (a) PBMCs only; (b) PBMCs + irradiated FY cells (10:1); (c) PBMCs + nonpulsed autologous DCs (10:1); and (d) PBMCs and DCs pulsed with MAGE-3-encoded HLA-A24-binding peptide (10:1). After 3 weeks of in vitro stimulation, a standard 51Cr release CTL assay was carried out. Target FY cells (MAGE-3 and HLA-A24+) were either nontreated or preincubated with anti-HLA class I mAbs and then added at an effector:target ratio of 5:1 and incubated for 4 h. ∗, P < 0.0001 (Student’s t test). B, cytotoxic activities of the effector cells from patient F. Y. against various target bladder cancer cells. Effector cells were prepared, and CTL assay was conducted as the same method described in Fig. 1A. Four different established bladder cancer cell lines were used as target cells. The characteristics of each cell line are shown in Table 2<$REFLINK> . ∗, P < 0.0001 (Student’s t test).
- Fig. 2.
Phenotype of the effector cells stimulated by autologous DCs pulsed with MAGE-3-encoded HLA-A24 binding peptide. Adherent PBMCs were collected from patient F. Y. and stimulated as described in “Materials and Methods.” Flow cytometric analysis of effector cells was performed weekly during the induction of effector cells. Single representative data are shown.
- Fig. 3.
Complete response of a metastatic lesion in liver by MAGE peptide-pulsed DC immunotherapy in a patient with advanced bladder cancer. CT scan shows a solitary liver metastatic lesion (arrow in Fig. 3A) completely disappeared after 18 × biweekly DC vaccination (B).
- Fig. 4.
Effectiveness of the MAGE peptide-pulsed DC immunotherapy against para-aortic lymph node metastasis of bladder cancer. A bulky para-aortic lymph node metastasis was observed by computed tomography scan before the MAGE peptide-pulsed DC vaccination (A; arrow) in the same patient as in Fig. 3<$REFLINK> . A >50% reduction in the size of metastasis was evident after the DC vaccination (B, arrow).
- Fig. 5.
Histopathological evaluation of representative biopsy specimens taken from inguinal lymph node metastasis of a bladder cancer patient before and after the MAGE peptide-pulsed DC immunotherapy. A typical transitional cell carcinoma was evident before the DC vaccination (A). Significant necrotic changes (arrows) were demonstrated 3 months after the DC vaccination (B; H&E, original magnification ×100).
Tables
- Table 1
The expression of MAGE genes according to pathological stages in bladder and upper tract urothelial cancersa,b
Pathological stage No. of tumors No. of MAGE + tumors MAGE-1 MAGE-2 MAGE-3 Any of three Bladder cancer <pT1 17 1 1 1 1 (6%) pT2≤ 11 4 3 6 8 (73%)c Total 28 5 4 7 9 (32%) Upper tract urothelial cancer <pT1 4 2 1 1 2 (50%) pT2≤ 4 3 4 4 4 (100%) Total 8 5 5 5 6 (75%) -
a Clinical tissue samples were obtained from the patients at the time of surgery performed at Keio University Hospital.
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b Expression of MAGE genes was detected by RT-PCR amplification of total RNA isolated from each sample using specific oligonucleotide primers.
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c Statistical significance between stages <pT1 and pT2≤ was analyzed by Mann-Whitney nonparametric U test. P = 0.0003.
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- Table 2
The expression of MAGE genes and HLA typing in established bladder cancer cell linesa,b
Cell line Pathology MAGE-1 MAGE-2 MAGE-3 HLA-A2 HLA-A24 KU-1 TCC, G2 − − − − − KU-7 TCC, G1 ++ ± +++ + − KU-19-19 TCC, G3 − + +++ + − T24 TCC, G2 − + ± + + FY TCC, G3 ++ +++ +++ − + -
a Expression of MAGE genes was detected by RT-PCR amplification of total RNA isolated from cell lysates of each cell line using specific oligonucleotide primers.
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b The relative levels of gene expression were determined as follows: the strongest positive band was defined as 100% and graded “−” for no expression; ±, <2% positive; +, 2–25% positive; ++, 26–50% positive; +++, >50% positive.
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- Table 3
Patients’ profiles and responses to DC vaccination
HLA-A24 MAGE-3 Sites of tumors No. of DC vaccinations Clinical response Previous treatment Survivala Right inguinal LNb Radical cystectomy F.Y. + + Para-aorta LN 6 Complete responsec Chemotherapy 2d 75 yr F Right external iliac LN Radiation G.K. 65 yr M + + Iliac LN Para-aorta LN Liver Braine 18 Partial response of para-aorta LN and complete response of liver metastasis overall progression Radical cystectomy Chemotherapy Radiation 8 N.H. 64 yr M + + Bilateral inguinal LN Perineum 6 Partial response TURBT Chemotherapy Radiation 5 N.T. + + Local recurrence 6 Progressive disease Radical cystectomy 2 56 yr M Pleural dissemination Chemotherapy Radiation -
a Survival is defined as months to death after the DC vaccination.
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b LN, lymph node.
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c Determined by autopsy.
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d Death attributable to perforation of small intestine.
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e Solitary brain metastasis developed 6 months after the DC vaccination and was surgically removed.
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Volume 7, Issue 1
