Immune-Complex Mimics as a Molecular Platform for Adjuvant-Free Vaccine Delivery

Protein-based vaccine development faces the difficult challenge of finding robust yet non-toxic adjuvants suitable for humans. Here, using a molecular engineering approach, we have developed a molecular platform for generating self-adjuvanting immunogens that do not depend on exogenous adjuvants for induction of immune responses. These are based on the concept of Immune Complex Mimics (ICM), structures that are formed between an oligomeric antigen and a monoclonal antibody (mAb) to that antigen. In this way, the roles of antigens and antibodies within the structure of immune complexes are reversed, so that a single monoclonal antibody, rather than polyclonal sera or expensive mAb cocktails can be used. We tested this approach in the context of Mycobacterium tuberculosis (MTB) infection by linking the highly immunogenic and potentially protective Ag85B with the oligomeric Acr (alpha crystallin, HspX) antigen. When combined with an anti-Acr monoclonal antibody, the fusion protein formed ICM which bound to C1q component of the complement system and were readily taken up by antigen-presenting cells in vitro. ICM induced a strong Th1/Th2 mixed type antibody response, which was comparable to cholera toxin adjuvanted antigen, but only moderate levels of T cell proliferation and IFN-γ secretion. Unfortunately, the systemic administration of ICM did not confer statistically significant protection against intranasal MTB challenge, although a small BCG-boosting effect was observed. We conclude that ICM are capable of inducing strong humoral responses to incorporated antigens and may be a suitable vaccination approach for pathogens other than MTB, where antibody-based immunity may play a more protective role

The making of a mammalian peroxisome, version 2.0: mitochondria get into the mix

This is the author accepted manuscript. The final version is available from Nature Publishing Group via the DOI in this record.A recent report from the laboratory of Heidi McBride (McGill University) presents a role for mitochondria in the de novo biogenesis of peroxisomes in mammalian cells (1). Peroxisomes are essential organelles responsible for a wide variety of biochemical functions, from the generation of bile, to plasmalogen synthesis, reduction of peroxides, and the oxidation of very long chain fatty acids (2). Like mitochondria, peroxisomes proliferate primarily through growth and division of pre-existing peroxisomes (3-6). However, unlike mitochondria, peroxisomes do not fuse (5,7); further, and perhaps most importantly, they can also be born de novo, a process thought to occur through the generation of pre-peroxisomal vesicles that originate from the endoplasmic reticulum (reviewed in (8,9). De novo peroxisome biogenesis has been extensively studies in yeast, with a major focus on the role of the ER in this process. Comprehensive studies in mammalian cells are, however, scarce (5,10-12). By exploiting patient cells lacking mature peroxisomes, Sugiura et al. (1) now assign a role to ER and mitochondria in de novo mammalian peroxisome biogenesis by showing that the formation of immature preperoxisomes occurs through the fusion of Pex3- / Pex14-containing mitochondriaderived vesicles with Pex16-containing ER-derived vesicles

An Adaptive Thresholding Method for BTV Estimation Incorporating PET Reconstruction Parameters: A Multicenter Study of the Robustness and the Reliability

Objective. The aim of this work was to assess robustness and reliability of an adaptive thresholding algorithm for the biological target volume estimation incorporating reconstruction parameters. Method. In a multicenter study, a phantom with spheres of different diameters (6.5–57.4 mm) was filled with 18F-FDG at different target-to-background ratios (TBR: 2.5–70) and scanned for different acquisition periods (2–5 min). Image reconstruction algorithms were used varying number of iterations and postreconstruction transaxial smoothing. Optimal thresholds (TS) for volume estimation were determined as percentage of the maximum intensity in the cross section area of the spheres. Multiple regression techniques were used to identify relevant predictors of TS. Results. The goodness of the model fit was high (R2: 0.74–0.92). TBR was the most significant predictor of TS. For all scanners, except the Gemini scanners, FWHM was an independent predictor of TS. Significant differences were observed between scanners of different models, but not between different scanners of the same model. The shrinkage on cross validation was small and indicative of excellent reliability of model estimation. Conclusions. Incorporation of postreconstruction filtering FWHM in an adaptive thresholding algorithm for the BTV estimation allows obtaining a robust and reliable method to be applied to a variety of different scanners, without scanner-specific individual calibration

Minimum acceptable sensitivity of intraoperative gamma probes used for sentinel lymph node detection in melanoma patients

The aim of this study was to determine the suspension level for the sensitivity of an intraoperative scintillation gamma probe in the detection of the sentinel lymph node (SLN) in melanoma patients. Thirty-eight consecutive patients with melanoma were enrolled in the study during a 12-month period and underwent lymphatic scintigraphy after the peritumoral intradermal administration of about 14\ua0MBq of (99m)Tc-nanocolloids. The SLNs were successfully removed during the surgical intervention about 4\ua0h later. To identify and localize the SLN, a scintillation NaI(Tl) collimated probe was used. Predictably, the probe sensitivity decreased as the photopeak energy window was progressively narrowed, from 6.9\ua0\ub1\ua00.7 counts per second (cps)/kBq (designated as the 'optimum,' or 'OPT,' sensitivity) to 2.5\ua0\ub1\ua00.3 cps/kBq (LOW sensitivity) and to 1.4\ua0\ub1\ua00.2 cps/kBq (VLOW sensitivity). Maximum lymph node count rates (cps) were determined for the foregoing energy windows prior to skin incision (PREOPT, PRELOW, PREVLOW, respectively) and in\ua0vivo after incision (INVOPT, INVLOW, INVVLOW). Forty-three SLNs were removed with a mean source-to-detector distance of 46\ua0\ub1\ua024\ua0mm (min 12\ua0mm, max 92\ua0mm). Four SLNs could not have been detected using PRELOW. This figure would have decreased to 34, with nine undetectable lymph nodes, with PREVLOW. One SLN could not have been identified using INVLOW and four could not have be identified using INVVLOW. In the clinical scenario of SLN detection in melanoma patients, a system sensitivity of 2.5\ua0cps/kBq represents a suspension level, that is, a level under which the equipment must be suspended from clinical use and the poor performance must be investigated

Influence of reconstruction settings on the performance of adaptive thresholding algorithms for FDG-PET image segmentation in radiotherapy planning.

Abstract The purpose of this study was to analyze the behavior of a contouring algorithm for PET images based on adaptive thresholding depending on lesions size and target-to-background (TB) ratio under different conditions of image reconstruction parameters. Based on this analysis, the image reconstruction scheme able to maximize the goodness of fit of the thresholding algorithm has been selected. A phantom study employing spherical targets was designed to determine slice-specific threshold (TS) levels which produce accurate cross-sectional areas. A wide range of TB ratio was investigated. Multiple regression methods were used to fit the data and to construct algorithms depending both on target cross-sectional area and TB ratio, using various reconstruction schemes employing a wide range of iteration number and amount of postfiltering Gaussian smoothing. Analysis of covariance was used to test the influence of iteration number and smoothing on threshold determination. The degree of convergence of ordered-subset expectation maximization (OSEM) algorithms does not influence TS determination. Among these approaches, the OSEM at two iterations and eight subsets with a 6-8 mm post-reconstruction Gaussian three-dimensional filter provided the best fit with a coefficient of determination R\ub2 = 0.90 for cross-sectional areas 64 133 mm\ub2 and R\ub2 = 0.95 for cross-sectional areas > 133 mm\ub2. The amount of post-reconstruction smoothing has been directly incorporated in the adaptive thresholding algorithms. The feasibility of the method was tested in two patients with lymph node FDG accumulation and in five patients using the bladder to mimic an anatomical structure of large size and uniform uptake, with satisfactory results. Slice-specific adaptive thresholding algorithms look promising as a reproducible method for delineating PET target volumes with good accuracy