Ung of case 3 patient. A: Before vaccination (May 15, 2003). B: After the third vaccination (June 18, 2003). Rapid growth of the metastatic tumors and pleural effusion were seen.vaccination.Safety and DTH skin test One patient (case 5) experienced slight fever (grade 1) after the first vaccination. No other adverse events were observed during vaccination. DTH skin test was performed at each vaccination and assessed 48 hr later. Reactions were determined as negative in all patients. Clinical response Recognized disease progression occurred in five out of six patients during the vaccination period (Table 1, Fig. 1). In contrast, one patient (case 5) showed no such rapid progression (Table 1, Fig. 2). These patients, except in case 1, had received systemic multidrug chemotherapy from one to four months before enrolling on this study. Tetramer analysis and CTL induction Peptide-specific immunological responses were evaluated in five patients by using HLA-A24/peptide tetramer analysis and in vitro CTL induction. As determined by flowcytometric analysis using HLA/peptide tetramers (Table 2), frequencies of CTLs specific for SYT-SSX B peptide were shown to be at background levels (less than PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/29072704 0.1 ) in three patients prior to vaccination. Those frequencies increased after the first (cases 4 and 6) and the third vaccination (case 2) (Fig. 3). In the remaining two patients (cases 3 and 5), SYT-SSX B peptide-specific CTLs existed beyond the background levels before vaccination. Ofthese, B peptide-specific CTL frequencies increased slightly in case 2 upon a series of vaccinations. On the contrary, the CTL frequencies in peripheral blood decreased to the background level after the third vaccination in case 5, whose metastatic diseases remained stable during the vaccination period. For comparison, tetramers with irrelevant peptides were constructed as internal controls and utilized in four patients. Notably, CTL frequencies reacting to those irrelevant tetramers remained under the background level during the course of vaccinations in all patients. Table 3 depicts the results of CTL induction by in vitro stimulations with SYT-SSX B peptide. Before vaccination, CTLs specific for SYT-SSX B peptide were successfully induced from one patient (case 2) who showed a high frequency of CTL precursors. After the first or third vaccination, CTLs were induced from four of five patients. Figure 4 represents the results of cytotoxicity assay. As shown, CTLs induced from the case 4 patient exhibited cytotoxic activities against T2-A*2402 cells pulsed with SYT-SSX B peptide, and synovial sarcoma cell lines expressing HLAA24 and SYT-SSX (Fuji and HS-SY-II) in various effecter/ target order BMS-214662 ratios examined. In contrast, the cytotoxity was less than 10 against T2-A*2402 cells without peptide pulsation, those pulsed with irrelevant HIV peptide, and tumor cells lacking HLA-A24 and SYT-SSX (SW982 and K562). These findings suggest that induction of peptide-specificPage 4 of(page number not for citation purposes)Journal of Translational Medicine 2005, 3:http://www.translational-medicine.com/content/3/1/ABCDFigure image of the lung of case 5 patient CT scan2 CT scan image of the lung of case 5 patient. A: Before vaccination (July 8, 2003). B: After the first vaccination (July 22, 2003). C: After the third vaccination (August 19, 2003). D: After the sixth vaccination (September 16, 2003). The metastatic tumors appeared to be dormant after the first vaccination.Table 2: HLA-A24/peptid.
