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Additional file 2: Fig. S2. Representative example of the viability of MDDCs under different maturation protocols and conditions. MDDCs were isolated from PBMCs and induced to maturation according to two protocols based on IL-4 and IFN-�� as depicted in Additional file 1: Fig. S1. Maturation of MDDCs was performed starting from either freshly collected PBMCs (red bars) or PBMCs frozen in liquid nitrogen upon collection and thawed after 2 weeks. Viability was assessed by flow cytometry using a LIVE/DEAD stain. Data were normalized over the fresh-cell condition.

Additional file 11: Table S1. List of peptides used for pulsing MDDCs of each individual. Enneamers (9 mers) were designed according to their predicted immunogenicity in a given individual, as described in the main text. Peptides were then used to pulse MDDCs before their reinfusion in MDDCT recipients according to the workflow depicted in Additional file 1: Fig. S1. Note that some autologous peptides are common to more than one patient.

Additional file 7: Fig. S7. Cytokine expression in unstimulated CD4+ and CD8+ T-lymphocytes following MDDC MDDCT. Data were obtained as described in Fig. 3.

Additional file 6: Fig. S6. Cytokine expression in SEB/brefeldin-stimulated CD4+ and CD8+ T-lymphocytes following MDDC MDDCT. Data were obtained as described in Fig. 3.

Additional file 3: Fig. S3. Representative example of the maturation profile of MDDCs subjected to alternative differentiation and freezing protocols. MDDCs were isolated from PBMCs and induced to maturation according to two protocols based on IL-4 and IFN-�� as depicted in Additional file 1: Fig. S1. Maturation/activation markers CD80 and HLA-DR were analyzed by flow cytometry in cells isolated from fresh PBMCs or from PBMCs thawed after freezing using FBS or a specific freezing medium (as described in the Methods section).

Additional file 5: Fig. S5. Gating strategy employed to analyze cytokine production in T-lymphocytes. Panels A, B. Cytokine production was determined in cells left unstimulated (A) or stimulated ex-vivo with the cell therapy peptides (B).

Additional file 1: Fig. S1. Schematic workflow of the protocols used for the maturation of MDDCs and MDDCT preparation. Maturation of MDDCs from monocytes was induced using either a cytokine cocktail based on IL-4 (longer protocol; Panel A) or a cytokine cocktail based on IFN-�� (shorter protocol; Panel B).

Additional file 4: Fig. S4. Gating strategy employed to analyze maturation markers of MDDCs

Additional file 9: Fig. S9. Impact of MDDCT on the level of IFN- �� in plasma. The Fig. shows the level of IFN-�� in individuals receiving the personalized dendritic-cell therapy (DCT), alone (G5, in black) or following the conditioning regimen consisting of auranofin + nicotinamide (G6, in blue). IFN-gamma was measured by ELISA testing, and data were analyzed by two-way ANOVA. The P-value reported refers to the interaction between time and the previous conditioning regimen.

Additional file 8: Fig. S8. Inter-group comparison of the immunogenicity of MDDCT. Comparison of the levels of IFN-��, IL-2, and TNF production levels by CD4+ and CD8+ T-cells in individuals receiving MDDCT only (G5) and individuals receiving MDDCT with nicotinamide and auranofin (G6). Data were analyzed by one-way ANOVA followed by Dunn��s post-test. *p

Additional file 10: Fig. S10. Impact of the time on ART on cytokine production post-MDDCT. The graph visualizes the relation between the time in days since the first MDDCT dose and the expression of immune response mediating cytokines in CD4+ (Panel A) and CD8+ (Panel B) T-cells. Patients were stratified into two categories based on the overall time of ART administration since their diagnosis (i.e. > 10 years, ���long��� and

Background— Very few studies have measured disease penetrance and prognostic factors of Chagas cardiomyopathy among asymptomatic Trypanosoma cruzi –infected persons. Methods and Results— We performed a retrospective cohort study among initially healthy blood donors with an index T cruzi –seropositive donation and age-, sex-, and period-matched seronegatives in 1996 to 2002 in the Brazilian cities of São Paulo and Montes Claros. In 2008 to 2010, all subjects underwent medical history, physical examination, ECGs, and echocardiograms. ECG and echocardiogram results were classified by blinded core laboratories, and records with abnormal results were reviewed by a blinded panel of 3 cardiologists who adjudicated the outcome of Chagas cardiomyopathy. Associations with Chagas cardiomyopathy were tested with multivariate logistic regression. Mean follow-up time between index donation and outcome assessment was 10.5 years for the seropositives and 11.1 years for the seronegatives. Among 499 T cruzi seropositives, 120 (24%) had definite Chagas cardiomyopathy, and among 488 T cruzi seronegatives, 24 (5%) had cardiomyopathy, for an incidence difference of 1.85 per 100 person-years attributable to T cruzi infection. Of the 120 seropositives classified as having Chagas cardiomyopathy, only 31 (26%) presented with ejection fraction P 3 heart sound. Conclusions— There is a substantial annual incidence of Chagas cardiomyopathy among initially asymptomatic T cruzi –seropositive blood donors, although disease was mild at diagnosis.

BackgroundStudies have shown that human immunodeficiency virus (HIV) residual risk is higher in Brazilian than in US and European blood donors, probably due to failure to defer at-risk individuals in Brazil. This study assessed the impact of an educational brochure in enhancing blood donors' knowledge about screening test window phase and reducing at-risk individuals from donating.Study design and methodsThis trial compared an educational intervention with a blood center's usual practice. The brochure was distributed in alternating months to all donors. After donating, sampled participants completed two questions about their HIV window period knowledge. The impact on HIV risk deferral, leaving without donation, confidential unit exclusion (CUE) use, and test positivity was also analyzed.ResultsFrom August to November 2007 we evaluated 33,940 donations in the main collection center of Fundação Pró-Sangue/Hemocentro de São Paulo in São Paulo, Brazil. A significant (p < 0.001) pamphlet effect was found on correct responses to both questions assessing HIV window phase knowledge (68.1% vs. 52.9%) and transfusion risk (91.1% vs. 87.2%). After adjusting for sex and age, the pamphlet effect was strongest for people with more than 8 years of education. There was no significant pamphlet effect on HIV risk deferral rate, leaving without donation, use of CUE, or infectious disease rates.ConclusionWhile the educational pamphlet increased window period knowledge, contrary to expectations this information alone was not enough to make donors self-defer or acknowledge their behavioral risk.