Purpose: The aim of this study is to find a laboratory marker for overall survival in advanced cancer patients who were vaccinated with peptides based on pre-existing, peptide-specific CTL precursors in the circulation.Experimental Design: A group of 113 patients with advanced cancer (28 colorectal, 22 prostate, 15 lung, 14 gastric, and 34 other cancers) was enrolled in a Phase I clinical study of peptide vaccination in which peptidespecific CTL precursors of prevaccination peripheral blood mononuclear cells were measured, followed by vaccination with these peptides (maximum of four). For cellular responses, pre and postvaccination (sixth) peripheral blood mononuclear cells were provided for measurement of both peptide-specific CTL precursors by IFN-␥ release assay and tumor reactivity by 51 Cr release assay. Delayed type hypersensitivity was also measured. For humoral response, pre and postvaccination (sixth) sera were provided for measurement of peptide-reactive IgG by an ELISA.Results: The median survival time and 1-year survival rate of the total cases were 346 ؎ 64.9 days and 44.6%, respectively, and those of patients vaccinated more than six times (n ‫؍‬ 91) were 409 ؎ 15 days and 54.4%, respectively. In these 91 patients, the overall survival of patients whose sera showed increased levels of peptide-reactive IgG (n ‫؍‬ 60) was significantly more prolonged (P ‫؍‬ 0.0003) than that of patients whose sera did not (n ‫؍‬ 31), whereas none of cellular responses correlated with overall survival.Conclusions: Peptide-specific IgG in postvaccination sera could be a suitable laboratory maker for the prediction of prolonged survival in advanced cancer patients vaccinated with peptides based on pre-existing CTL precursors.

Recent clinical trials of peptide vaccine for cancer patients have rarely resulted in tumor regression. One of the reasons for this failure could be an insufficient induction of anti-tumor responses in these regimens, in which peptide-specific memory cytotoxic T lymphocytes (CTLs) were not measured prior to vaccination. We investigated in this study whether pre-vaccination measurement of peptide-specific CTLs can provide any advantages in lung cancer patients receiving peptide vaccination with regard to safety and immunological responses. Ten patients with advanced lung cancer received vaccination with peptides under a regimen of CTL precursor-oriented vaccination, in which pre-vaccination peripheral blood mononuclear cells (PBMCs) were at first screened for reactivity in vitro to each of 14 peptides, followed by in vivo administration of only the reactive peptides. Profiles of the vaccinated peptides varied markedly among the 10 patients. This regimen was generally well-tolerated, although local skin reactions, diarrhea, and colitis were observed in 8, 2, and 1 patient, respectively. Increased CTL responses against the immunized peptides and tumor cells were observed in the post-vaccination PBMCs from 4 of 8 and 3 of 10 patients tested, respectively. Peptidespecific IgG became detectable in post-vaccination sera in 4 of 10 patients tested, and these 4 patients had a long progression-free survival. Furthermore, the median survival time of 9 patients with non-small cell lung cancer was 668.0 ± ± ± ±164.2 days. These results encourage further development of CTL precursor-oriented peptide vaccination for lung cancer patients. (Cancer Sci 2003; 94: 548-556) dentification of a large number of antigenic epitopes 1-9) recognized by cytotoxic T lymphocytes (CTLs) reacting to tumor cells has opened the door to clinical trials of peptide-based immunotherapy for cancer patients. [10][11][12][13][14][15][16] Many clinical trials of peptide-based immunotherapy for malignant melanoma and other epithelial tumors have been conducted in the past decade, but major clinical responses were rarely obtained in these clinical studies, including our trial of a cyclophilin B (CypB) peptide vaccination for advanced lung cancer patients 17) and a SART3 peptide vaccination for advanced colorectal cancer patients.18) One reason for this failure to obtain tumor regression could be an insufficient induction of anti-tumor responses in these vaccine regimens, in which peptide-specific memory T cells were not measured in pre-vaccination peripheral blood mononuclear cells (PBMCs). We have speculated that vaccination based on information from pre-vaccination measurement of peptide-specific CTLs in the circulation might induce potent anti-tumor immune responses in cancer patients. This hypothesis is based on the assumption that initiation of immune-boosting of CTLs through peptide vaccination could be more effective than immune-priming of naive T cells with regard to induction of prompt and strong immunity to both the peptide and tumor cells. We ha...

SUMMARYWe investigated surface antigens and spontaneous cytokine production of T cells from bronchoalveolar lavage fluid (BALF) and aqueous humor (AH) from pulmonary sarcoidosis patients for a better understanding of the role of T cells ingranuloma formation. The levels of CD3., CD I Ib. and CD28 antigen expression on freshly isolated T cells in the BALF of patients were significantly lower than those in peripheral blood lymphocytes (PBL) of cither sarcoidosis patients or healthy donors (HD). In contrast, the levels of CD80 (B7/B7-1) and CD86 (B7O/B7-2) antigen expression were significantly higher on these T cells and alveolar macrophages in the BALF of patients. Fiftythree T cell clones (TCC) established from the BALF and AH of the three sarcoidosis patients displayed primarily either CD4^ CDllb+ CD28^ or CD4^ CDllb^ CD28" phenotypes. Most (61-90%) of these TCC spontaneously produced greater amounts of IL-la, IL-10. tumour necrosis factor (TNF), and granulocyte-macrophage colony-stimulating factor (GM-CSF) than did TCC from the PBL from sarcoidosis patients or HD (/'<0 05). Interferon-gamma (IFN-7), IL-6. and IL-2, but not IL-4, were also produced by 40-48% of these TCC. These results suggest that CD4^ T cells of the affected organs of sarcoidosis patients are activated and involved in the immunopathogenesis of sareoidosis through production of various eytokines.

SUMMARYSarcoidosis is a systemic granulomatous disease of unknown etiology characterized by the pronounced accumulation of CD4 þ T cells and macrophages in the affected organs. TCR variable (V)a and Vb gene usage in patients with sarcoidosis is still a matter of discussion. In this investigation, we analysed TCRVa and -Vb gene usage in bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells (PBMC) of 30 patients with active pulmonary sarcoidosis using an adapter ligation method, reverse transcriptase-polymerase chain reaction (RT-PCR), and sequence-specific oligonucleotide probe (SSOP) analyses. There was no significant difference in TCR-Va or -Vb gene usage between BALF (n = 12) or PBMC (n = 27) of patients and PBMC of healthy subjects (n = 10). Neither selective TCR-Va nor -Vb expansion was observed in the paired BALF and PBMC from seven of nine patients. However, selective expansions were observed in a few TCR-Va or -Vb subsets in the BALF or PBMC of some individuals. Although a modest increase in a few TCR-Va or -Vb subsets was observed in the BALF or PBMC of some individuals, the increased TCR-Va or -Vb subsets were not closely associated with the HLA-DRB1, DQA1, DQB1, and DPB1 alleles of these patients. These results suggest that TCR-Va or -Vb gene usage is not restricted in both lung and peripheral blood in the majority of patients with active pulmonary sarcoidosis.

In order to examine ultrasonic absorption mechanisms and dynamical properties in aqueous solutions of heterocyclic amines, the absorption coefficients in a frequency range from 3.0 to 220 MHz were measured in solutions of pyrrolidine and piperidine at 25 °C. In the solution of pyrrolidine, a single relaxational absorption associated with a proton transfer reaction was observed, and the rate and thermodynamic parameters were determined from the concentration dependence of the relaxation frequency and the maximum absorption per wavelength. In the solution of piperidine, another relaxational absorption in addition to that due to the proton transfer reaction was observed in the relatively concentrated solution. It was attributed to a perturbation of an equilibrium associated with an aggregation reaction of nonionized molecules. The mean aggregation number and the rate parameters were estimated from the solute concentration dependence of the relaxation parameters. The results were compared with those observed in aliphatic amine solutions.