Dynamic helical CT of T1 and T2 glottic carcinomas: predictive value for local control with radiation therapy.

BACKGROUND AND PURPOSE: Tumor volume and cartilage invasion have been suggested as prognostic factors of glottic carcinomas following definitive radiation therapy. Radiologic examinations provide additional information regarding the deep extension of tumor. We determined whether dynamic helical CT can predict local control of early (T1 and T2 stage) glottic carcinomas treated with definitive radiation therapy. METHODS: Sixty-eight patients with early glottic carcinoma evaluated on pretreatment dynamic helical CT were treated with definitive radiation therapy. Tumor detectability, maximum dimension, tumor volume, and involvement of anatomic subsites (anterior commissure, ventricle, subglottic region, and thyroid and arytenoid cartilages) were determined by consensus by three radiologists without previous knowledge of the clinical information. The CT findings were correlated with local control. RESULTS: The two-year local control rate was 76%; 91% for T1 and 60% for T2 lesions. Univariate analysis revealed clinical T stage, tumor detectability, maximum dimension, tumor volume, anterior commissure involvement, ventricle involvement, and thyroid cartilage involvement as significant prognostic factors. Thyroid cartilage involvement was an independent predictor by multivariate analysis. The lesions separate from the thyroid cartilage had a 95% probability of local control, whereas the lesions adjacent to the cartilage had only a 42% control rate. CONCLUSION: Dynamic helical CT provides prognostic information for the results of definitive radiation therapy. Patients with a tumor adjacent to the thyroid cartilage had an increased risk of local failure.

Analysis of individual specific cytotoxic T lymphocytes for two MAGE-3-derived epitopes presented by HLA-A24.

BACKGROUND: The human MAGE-3 gene encodes tumor-specific antigens that are recognized by cytotoxic T lymphocytes (CTLs) and expressed in a high percentage of various malignant tumors. Of the five MAGE-3-derived CTL epitopes identified to date, two nonapeptides (TFPDLESEF and IMPKAGLLI, designated MAGE-3.A24a and MAGE-3.A24b, respectively) can be expressed on the tumor surface by binding to the HLA-A24 molecule, which is the most frequent HLA class I molecule in Asian populations. To compare the immunogenecities of the two peptides, individual specific CTL lines were generated for each peptide (MAGE-3.A24a and MAGE-3.A24b). METHODS: Peripheral blood mononuclear cells (PBMCs) from four HLA-A24+ healthy donors were stimulated in vitro with autologous dendritic cells pulsed with MAGE-3.A24a, MAGE-3.A24b or both and were subsequently cultivated with a cytokine combination including interleukin-2. RESULTS: We succeeded in generating peptide-specific CTL lines in two of the four donors. The two CTL lines showed similar cytolytic levels against three MAGE-3+/HLA-A24+ cancer cell lines and also target cells pulsed with the corresponding peptide. Cytolytic activities were blocked by either anti-CD8 or anti-HLA-A24 monoclonal antibodies. CONCLUSIONS: The results suggest that MAGE-3.A24a and MAGE-3.A24b peptides have equal potential in inducing MAGE-3-specific and HLA-A24-restricted CTLs.

A wild-type sequence p53 peptide presented by HLA-A24 induces cytotoxic T lymphocytes that recognize squamous cell carcinomas of the head and neck.

Evidence has accumulated indicating that HLA-A2-restricted CTLs specific for human wild-type sequence p53 epitopes lyse tumor cells expressing mutant p53. To explore the possibility that wild-type sequence p53 peptides could also be used in vaccines for patients expressing HLA-A24 antigen, another frequent HLA class I allele, we investigated the induction of HLA-A24-restricted p53-specific CTLs from the peripheral blood lymphocytes of normal donors. Of six p53-derived peptides possessing an HLA-A24 binding motif, the p53 peptide 125-134 (p53(125-134)) was found to have a high binding capacity and induced peptide-specific CTLs from peripheral blood mononuclear cells, using peptide-pulsed autologous dendritic cells and subsequent cultivation with cytokines interleukin 2 and interleukin 7. Bulk CTL populations lysed peptide-pulsed HLA-A24+ targets as well as HLA-A24+ squamous cell carcinoma of the head and neck (SCCHN) cell lines. However, IFN-gamma pretreatment of HLA-A24+ SCCHN cell lines was necessary for lysis, suggesting that a ligand density higher than that normally expressed by tumor cells is required for these CTLs to mediate lysis. Moreover, a cloned CTL, designated TH#99, isolated from the bulk population by limiting dilution, lysed HLA-A24+ SCCHN targets more efficiently than the bulk CTL population. Lysis was inhibited by anti-HLA class I monoclonal antibody but not by anti-HLA-DR monoclonal antibody. These results indicate that HLA-A24-restricted CTLs recognizing the wild-type sequence p53(125-134) can be generated using autologous dendritic cells from precursors present in peripheral blood lymphocytes obtained from normal HLA-A24+ donors. This finding suggests that vaccine strategies targeting wild-type sequence p53 epitopes can be extended to a wider range of cancer patients.

Early glottic squamous cell carcinoma. Predictive value of MR imaging for the rate of 5-year local control with radiation therapy.

PURPOSE: To evaluate MR findings in early (T1 and T2 stages) glottic carcinomas and the predictive value of MR imaging for the rate of 5-year local control with radiation therapy. MATERIAL AND METHODS: Eighty-three patients with early glottic carcinomas were prospectively examined with MR at 1.5 T. MR investigation included unenhanced T1-weighted, T2-weighted, dynamic and contrast-enhanced T1-weighted images. Three patients with presumed advanced diseases on MR were initially treated with total laryngectomy and were excluded from the study. The remaining 80 patients were treated with radiation therapy with curative intent. Tumor detectability, size and relationship to the thyroid cartilage were determined on MR images. The MR findings were then correlated with the rate of local control. RESULTS: Forty-eight of 80 lesions (60%) were detected on MR imaging. All detected lesions but 1 demonstrated increased signal on T2-weighted images. The lesions were best delineated on dynamic images (statistically significant). The 5-year local control rate with radiation therapy was 72%. Univariate analysis revealed clinical T stage, MR detectability, tumor size and relationship to the thyroid cartilage as significant predictors. Multivariate analysis revealed that the relationship to the thyroid cartilage was an independent factor. CONCLUSION: MR provides prognostic information about the results of definitive radiation therapy. To evaluate the tumor extension in lesions detected on precontrast MR images, contrast-enhanced dynamic images should be obtained.

A newly identified MAGE-3-derived epitope recognized by HLA-A24-restricted cytotoxic T lymphocytes.

Five MAGE-3-derived peptides carrying an HLA-A24-binding motif were synthesized. Binding capacity of these peptides was analyzed by an HLA-class-I stabilization assay. Two of the 5 peptides bound to HLA-A*2402 molecule with high affinity, and 3 peptides with low affinity. Peripheral-blood mononuclear cells (PBMC) depleted of CD4+T cells were stimulated with the peptides to determine whether these peptides would induce cytotoxic T lymphocytes (CTL) from PBMCs obtained from 7 healthy HLA-A*2402+ donors. Peptide M3-p97 (TFPDLESEF; corresponding to amino-acid residues 97-105 of MAGE-3), with high binding capacity to the HLA-A*2402 molecule, elicited the peptide-specific and HLA-A24-restricted CD8+CTL lines in 2 of the 7 donors, while none of the 4 other peptides induced CTL specific for the corresponding peptide in any of the donors. CTL lines induced by stimulation with peptide M3-p97 exhibited cytolytic activities against HLA-A*2402 transfectant cell lines (C1R-A*2402) in the presence of peptide M3-p97, but not in unloaded or irrelevant peptide-pulsed C1R-A*2402 cells. The CTL lines and a cloned CD8+CTL isolated from one of the bulk populations by limiting dilution could lyse MAGE-3+/HLA-A*2402+ squamous-cell-carcinoma(SCC) lines but neither MAGE-3-/HLA-A*2402+ nor MAGE-3+/HLA-A*2402- SCC lines, indicating that M3-p97 can be naturally processed and presented on the tumor-cell surface in association with HLA-A*2402 molecules. Combined with the 4 currently reported CTL epitopes derived from MAGE-3 and presented by HLA-A1, HLA-A2, HLA-A24 or HLA-B44, identification of this CTL epitope presented by the HLA-A*2402 molecule will extend the application of MAGE-3-derived peptides for immunotherapy for cancer patients.

Frequency of HLA-A alleles in Japanese patients with head and neck cancer.

BACKGROUND: Association between certain human leukocyte antigen (HLA) types such as HLA-A1 and -A3 and squamous cell carcinoma of the head and neck (SCCHN) has been demonstrated in the Caucasian population. HLA typings in these studies were performed by conventional serological methods. However, recent comparison studies between serological and molecular typings have revealed that the former are often inaccurate. METHODS: The frequency of HLA-A alleles in 100 Japanese patients with SCCHN and 100 control subjects was determined by the polymerase chain reaction, with primers specific for the HLA-A locus, in combination with dot-blot hybridization with 31 sequence-specific oligonucleotides. RESULTS: The frequencies of HLA-A*2602 and HLA-A*3303 were higher and those of HLA-A*2603 and HLA-A*3101 were lower in the patients with SCCHN than in healthy controls, but these differences were not statistically significant. In the 39 male patients with laryngeal carcinoma, the most common malignancies in Japanese patients with SCCHN, the frequency of HLA-A*2402 was significantly lower than that in the 80 male controls; however, after correction of the P value, statistical significance was not confirmed. In oral carcinoma patients, the frequency of HLA-A*2402 was significantly higher than that in healthy controls. CONCLUSIONS: These results suggest that the contribution of certain HLA-A alleles to susceptibility to SCCHN may differ between sites in the head and neck regions, despite these cancers being of an identical histological type, and that HLA-A*2402 may influence the development of oral carcinoma in Japanese patients.