Cationic Liposomes Loaded with a Synthetic Long Peptide and Poly(I:C): a Defined Adjuvanted Vaccine for Induction of Antigen-Specific T Cell Cytotoxicity

For effective cancer immunotherapy by vaccination, co-delivery of tumour antigens and adjuvants to dendritic cells and subsequent activation of antigen-specific cytotoxic T cells (CTLs) is crucial. In this study, a synthetic long peptide (SLP) harbouring the model CTL epitope SIINFEKL was encapsulated with the TLR3 ligand poly(inosinic-polycytidylic acid) (poly(I:C)) in cationic liposomes consisting of DOTAP and DOPC. The obtained particles were down-sized to about 140 nm (measured by dynamic light scattering) and had a positive zeta-potential of about 26 mV (according to laser Doppler electrophoresis). SLP loading efficiency was about 40% as determined by HPLC. Poly(I:C) loading efficiency was about 50%, as assessed from the fluorescence intensity of fluorescently labelled poly(I:C). Immunogenicity of the liposomal SLP vaccine was evaluated in vitro by its capacity to activate dendritic cells (DCs) and present the processed SLP to SIINFEKL-specific T cells. The effectiveness of the vaccine to activate CD8⁺ T cells was analysed in vivo after intradermal and subcutaneous immunisation in mice, by measuring antigen-specific T cells in blood and spleens and assessing their functionality by cytokine production and in vivo cytotoxicity. The liposomal formulation efficiently delivered the SLP to DCs in vitro and induced a functional CD8⁺ T cell immune response in vivo to the CTL epitope present in the SLP. The SLP-specific CD8⁺ T cell frequency induced by the poly(I:C)-adjuvanted liposomal SLP formulation showed an at least 25 fold increase over the T cell frequency induced by the poly(I:C)-adjuvanted soluble SLP. In conclusion, cationic liposomes loaded with SLP and poly(I:C) have potential as a powerful therapeutic cancer vaccine formulation.

Electronic supplementary material

The online version of this article (doi:10.1208/s12248-014-9686-4) contains supplementary material, which is available to authorized users.KEY WORDS: cancer immunotherapy, cationic liposomes, CTL epitope, peptide antigen, Poly(I:C)     

Physical activity, dipping and haemodynamics

OBJECTIVE: To determine the effect of physical activity on diurnal blood pressure (BP) and haemodynamic variation. METHODS: Ambulatory measurements were performed during 24 h in 36 subjects (18 hypertensive, 13 male), aged 49.7 +/- 13.5 years. BP was recorded in the brachial artery. Physical activity and posture were measured with five acceleration sensors. RESULTS: Of the subjects 50% were dippers (nocturnal decrease in systolic or diastolic BP >/= 10%). Dippers and non-dippers had similar daytime BP, daytime, night-time, and day-night difference in physical activity, subjective sleep quality, and nocturnal cardiac output decrease (14.9 +/- 9.6 and 16.0 +/- 5.9%). In non-dippers vascular resistance increased from day to night by 9.7 +/- 8.3%, while it remained unchanged (-1.0 +/- 13.9%) in dippers. Day-night changes in heart rate and cardiac output were correlated with day-night changes in physical activity (r = 0.39 and 0.43), whereas day-night changes in systolic BP were correlated with night-time activity (r = -0.34). By selection of the active (i.e. walking) and inactive (i.e. not walking) periods during the day, we showed that physical activity has a large potential effect on dipping status and diurnal haemodynamic variation underlying BP variation. Depending on the BP taken (systolic or diastolic, respectively) the proportion of dippers increased to 81% or decreased to 25% if only the walking period was considered, whereas it decreased to 36% or increased to 53% if only the non-walking period was considered. CONCLUSIONS: Non-dippers differ from dippers by an increase of vascular resistance during the night. The degree of physical activity normally encountered during ambulatory monitoring has little influence on the diurnal BP profile or dipping status, but significantly influences underlying haemodynamics. Related to the different effects of posture and activity on systolic and diastolic BP, dipping classification may vary with the BP index taken.     

Expression profile and thyroid hormone responsiveness of transporters and deiodinases in early embryonic chicken brain development

We used the chick embryo to study the mechanisms regulating intracellular TH availability in developing brain. TH-transporters OATP1C1 and MCT8, and deiodinases D1, D2, and D3 were expressed in a region-specific way, well before the onset of endogenous TH secretion. Between day 4 and 10 of development MCT8 and D2 mRNA levels increased, while OATP1C1 and D3 mRNA levels decreased. D2 and D3 mRNAs were translated into active protein, while no D1 activity was detectable. Injection of THs into the yolk 24 h before sampling increased TH levels in the brain and resulted in decreased OATP1C1 and increased MCT8 expression in 4-day-old embryos. A compensatory response in deiodinase activity was only observed at day 8. We conclude that THs are active in the early embryonic brain and TH-transporters and deiodinases can regulate their availability. However, the absence of clear compensatory mechanisms at day 4 makes the brain more vulnerable for changes in maternal TH supply.     

Long-term outcome of kidney transplantation in patients with a urinary conduit: a case–control study

Purpose

To study the short- and long-term outcomes of kidney transplantation in patients with a bladder augmentation or urinary diversion compared to patients with a kidney transplantation in a normal functional bladder.

Patients and methods

Between January 2000 and March 2011, 13 patients received 16 grafts into a reconstructed urinary tract. We performed a retrospective case–control study and matched each patient to 4 controls for donor and recipient gender and year of transplantation.

Results

Short- and long-term complications of kidney transplantation occurred in 12 patients, varying from urinary tract infections to medical hospitalization with or without surgical or radiological intervention. In 5 patients, a percutaneous nephrostomy (PCN) was placed followed by surgical re-intervention. In three patients, the grafts failed as a result of chronic rejection and were re-transplanted. There was no graft loss as a result of surgical complications or the reconstructed urinary tract. One-year patient and graft survival was 100 %. After five years, all patients were alive and seven of nine grafts (77.8 %) were functioning. Mean follow-up time was 4.3 years. Among the controls, 55 grafts were transplanted in 52 patients. Ten patients received a PCN. Five patients needed surgical re-intervention. In three patients, transplantectomy was performed for ongoing rejection. Three patients were re-transplanted. One patient had a failing graft 7.5 years post-transplantation and became dialysis dependent.

Conclusion

Kidney transplantation in patients with a reconstructed urinary tract has an increased complication rate. Nevertheless, the long-term results are comparable to patients with a normal urinary bladder.     

Head‐pelvis coupling is increased during turning in patients with Parkinson's disease and freezing of gait

Turning is the most important trigger for freezing of gait (FOG). The aim of this study was to investigate the relationship between impaired head‐pelvis rotation during turning and FOG. Head, trunk, and pelvic rotation were measured at onset and throughout a 180‐degree turn in 13 freezers and 14 nonfreezers (OFF medication). We also studied 14 controls at preferred and slow speed to investigate the influence of turn velocity on axial rotation. Location and duration of FOG episodes were defined during the turn. At turning onset, head rotation preceded thorax and pelvic rotation in all groups, but this craniocaudal sequence disappeared when FOG occurred. Maximum head‐pelvis separation was significantly greater in controls, compared to freezers and nonfreezers (35.4 versus 25.7 and 27.3 degrees; P < 0.01), but this finding was speed dependent. Timing of maximum head‐pelvis separation was delayed in freezers, compared to nonfreezers and controls, irrespective of turn velocity. This delay was correlated with increased neck rigidity (R = 0.62; P = 0.02) and worsened during FOG trials. FOG occurred more often at the end of the turn, when difference in rotation velocity between head and pelvis was greatest. Even after controlling for speed and disease severity, turning in freezers was characterized by delayed head rotation and a closer coupling between head and pelvis, especially in turns where FOG occurred. These changes may be attributed to delayed preparation for the change in walking direction and, as such, contribute to FOG. © 2013 Movement Disorder Society     

Systemic inflammatory immune signatures in a patient with CRB1 linked retinal dystrophy

Background: In this report we describe, for the first time, the activation of the peripheral immune compartment in a patient with a CRB1 linked retinal degenerative disease, masquerading as intermediate uveitis.

Methods: To monitor the immune system during systemic immunosuppressive treatment, given for the initial diagnosis of intermediate uveitis, blood samples were taken before and during therapy, for analysis of peripheral blood mononuclear cell-subsets and circulating immune mediators.

Results: The levels of various pro-inflammatory immune mediators (including MIF, TSLP, CCL2/MCP-1, CXCL9, CXCL10, IFN-β, IL-6, IL-17, IL-21, IL-22, and IL-23) were elevated in serum at the first time point, and decreased under immunosuppressive treatment. In parallel, the frequency of activated (CD86+) CD1c+ myeloid dendritic cells in blood was proportional to the central foveal thickness measured by optical coherence tomography.

Conclusions: These observations challenge the current view on the distinct pathophysiology of retinal degenerative and retinal inflammatory conditions in this patient.