Virosomes as new carrier system for cancer vaccines

HER-2/neu, a tumor-associated antigen (TAAg), plays a critical role in oncogenesis of various tumor types, and its selective overexpression by malignant tumor cells makes it an ideal target for immunotherapy. A prerequisite for clinical vaccines is the construction of safe and highly immunogenic reagents able to generate efficient immune responses against TAAg. Previous protein vaccines, consisting of the extracellular domain of HER-2/neu (pNeuECD), were shown to elicit an immune response that did not provide protection from transplantable tumors expressing HER-2/neu. Here we showed that virosomes, which consist of reconstituted viral envelopes without viral genetic material, can act as a carrier and an adjuvant for a truncated protein pNeuECD . Mice vaccinated with pNeuECD either encapsulated in virosomes or bound to the virosomal membrane (Vir-pNeuECD), generated rNeu-specific humoral and cytotoxic immune responses. In addition, Vir-pNeuECD induced significant tumor rejection and additionally did not lead to delayed tumor formation when compared with free pNeuECD in complete Freund's adjuvant. There was no difference between the virosomal constructs. Taken together these results suggest that virosomes, as clinically approved safe vaccines, can be used to elicit both humoral and cell-mediated responses against TAAg and induce tumor rejection. Our model is providing important preclinical data to design human vaccination trials for patients with tumors overexpressing HER-2/neu, either as a primary vaccination or as a boost in combination with other vaccines in a context of an adjuvant treatment pla

Targeted and all-sky search for nanosecond optical pulses at Harvard-Smithsonian

We have built a system to detect nanosecond pulsed optical signals from a target list of some 10,000 sun-like stars, and have made some 20,000 observations during its two years of operation. A beamsplitter feeds a pair of hybrid avalanche photodetectors at the focal plane of the 1.5m Cassegrain at the Harvard/Smithsonian Oak Ridge Observatory (Agassiz Station), with a coincidence triggering measurement of pulse width and intensity at sub-nanosecond resolution. A flexible web-enabled database, combined with mercifully low background coincidence rates (approximately 1 event per night), makes it easy to sort through far-flung data in search of repeated events from any candidate star. An identical system will soon begin observations, synchronized with ours, at the 0.9m Cassegrain at Princeton University. These will permit unambiguous identification of even a solitary pulse. We are planning an all-sky search for optical pulses, using a dedicated 1.8m f/2.4 spherical glass light bucket and an array of pixelated photomultipliers deployed in a pair of matched focal planes. The sky pixels, 1.5 arcmin square, tessellate a 1.6 by 0.2 degree patch of sky in transit mode, covering the Northern sky in approximately 150 clear nights. Fast custom IC electronics will monitor corresponding pixels for coincident optical pulses of nanosecond timescale, triggering storage of a digitized waveform of the light flash

Active Antigen-specific Immunotherapy of Melanoma: from Basic Science to Clinical Investigation

Advanced-stage melanoma here dismal prognosis, and novel therapeutic approaches are urgently required. The possibility of taking advantage of the immune response of patients for its treatment has been an appealing concept for almost a century. Only during the last decade, however, has the molecular identification of tumor-associated antigens (TAAs) offered the possibility of vaccinating patients (e.g., active induction of TAA-specific immune responses). Active antigen-specific immunotherapy (AASIT) is currently being investigated in a number of clinical centers as a treatment option for advanced-stage melanoma. A large number of melanoma TAAs have been molecularly characterized and are being used in vaccination trials in various molecular forms and according to various immunization protocols. Here we provide a short overview on melanoma TAAs, the technologies currently in use to induce specific cytotoxic T-lymphocyte (CTL) responses in vivo, and their monitoring. We also propose a tentative AASIT agenda for the next few years, aiming at improving the capacity to induce and monitor TAA-specific immune responses and to verify their clinical effectivenes

Exploring the Validity of the Continuum of Resistance Model for Discriminating Early from Late and Non-uptake of Colorectal Cancer Screening: Implications for the Design of Invitation and Reminder Letters.

This item is under embargo for a period of 12 months from the date of publication, in accordance with the publisher's policy.Background The continuum of resistance model contends that respondents lie at one end of a continuum and non-respondents at the other with respect to factors demonstrated to impact on screening participation. Purpose The aim of this study was to explore the validity of this model for the prediction of participation in colorectal cancer screening. Method People aged 50 to 74 years were asked to complete a survey (n = 1,250). Eligible respondents (n = 376, 30 %) were invited to complete a faecal occult blood test (FOBT). The cutoff period for the determination of participation rates was 12 weeks, with a reminder sent at 6 weeks. Results FOBTs were returned by n = 196 people (132 within 6 weeks, 64 following a reminder). Participation was generally influenced by the same variables in both the first 6 weeks and the second 6 weeks, consistent with the continuum of resistance model. These variables were having known someone with bowel cancer and the social cognitive factor, perceptions of barriers to screening. There is a suggestion, however, that other factors may be differentially associated with early, late and non-participants. Conclusion Participation in screening appears somewhat consistent with the continuum of resistance model in that early and late participants respond to some of the same factors. This suggests that the same messages are relevant to early, late and non-screeners, but further consideration of what other factors may be influencing discrete stages of readiness to participate is necessary.This work was supported by a National Health and Medical Research Council Grant number 324717

Whole blood assessment of antigen specific cellular immune response by real time quantitative PCR: a versatile monitoring and discovery tool

BACKGROUND: Monitoring of cellular immune responses is indispensable in a number of clinical research areas, including microbiology, virology, oncology and autoimmunity. Purification and culture of peripheral blood mononuclear cells and rapid access to specialized equipment are usually required. We developed a whole blood (WB) technique monitoring antigen specific cellular immune response in vaccinated or naturally sensitized individuals. METHODS: WB (300 microl) was incubated at 37 degrees C with specific antigens, in the form of peptides or commercial vaccines for 5-16 hours. Following RNAlater addition to stabilize RNA, the mixture could be stored over one week at room temperature or at 4 degrees C. Total RNA was then extracted, reverse transcribed and amplified in quantitative real-time PCR (qRT-PCR) assays with primers and probes specific for cytokine and/or chemokine genes. RESULTS: Spiking experiments demonstrated that this technique could detect antigen specific cytokine gene expression from 50 cytotoxic T lymphocytes (CTL) diluted in 300 microl WB. Furthermore, the high sensitivity of this method could be confirmed ex-vivo by the successful detection of CD8+ T cell responses against HCMV, EBV and influenza virus derived HLA-A0201 restricted epitopes, which was significantly correlated with specific multimer staining. Importantly, a highly significant (p = 0.000009) correlation between hepatitis B surface antigen (HBsAg) stimulated IL-2 gene expression, as detectable in WB, and specific antibody titers was observed in donors vaccinated against hepatitis B virus (HBV) between six months and twenty years before the tests. To identify additional markers of potential clinical relevance, expression of chemokine genes was also evaluated. Indeed, HBsAg stimulated expression of MIP-1beta (CCL4) gene was highly significantly (p = 0.0006) correlated with specific antibody titers. Moreover, a longitudinal study on response to influenza vaccine demonstrated a significant increase of antigen specific IFN-gamma gene expression two weeks after immunization, declining thereafter, whereas increased IL-2 gene expression was still detectable four months after vaccination. CONCLUSION: This method, easily amenable to automation, might qualify as technology of choice for high throughput screening of immune responses to large panels of antigens from cohorts of donors. Although analysis of cytokine gene expression requires adequate laboratory infrastructure, initial antigen stimulation and storage of test probes can be performed with minimal equipment and time requirements. This might prove important in "field" studies with difficult access to laboratory facilities

Towards Structure-Property-Function Relationships for Eumelanin

We discuss recent progress towards the establishment of important structure-property-function relationships in eumelanins - key functional bio-macromolecular systems responsible for photo-protection and immune response in humans, and implicated in the development of melanoma skin cancer. We focus on the link between eumelanin's secondary structure and optical properties such as broad band UV-visible absorption and strong non-radiative relaxation; both key features of the photo-protective function. We emphasise the insights gained through a holistic approach combining optical spectroscopy with first principles quantum chemical calculations, and advance the hypothesis that the robust functionality characteristic of eumelanin is related to extreme chemical and structural disorder at the secondary level. This inherent disorder is a low cost natural resource, and it is interesting to speculate as to whether it may play a role in other functional bio-macromolecular systems.Comment: 19 pages, 8 figures, Invited highlight article for Soft Matte

Search for Nanosecond Optical Pulses from Nearby Solar‐Type Stars

With "Earth 2000" technology we could generate a directed laser pulse that outshines the broadband visible light of the Sun by 4 orders of magnitude. This is a conservative lower bound for the technical capability of a communicating civilization; optical interstellar communication is thus technically plausible. We have built a pair of systems to detect nanosecond pulsed optical signals from a target list that includes some 13,000 Sun-like stars, and we have made some 16,000 observations totaling nearly 2400 hr during five years of operation. A beam splitter-fed pair of hybrid avalanche photodetectors at the 1.5 m Wyeth Telescope at the Harvard/Smithsonian Oak Ridge Observatory (Agassiz Station) triggers on a coincident pulse pair, initiating measurement of pulse width and intensity at subnanosecond resolution. An identical system at the 0.9 m Cassegrain at Princeton's Fitz-Randolph Observatory performs synchronized observations with 0.1 μs event timing, permitting unambiguous identification of even a solitary pulse. Among the 11,600 artifact-free observations at Harvard, the distribution of 274 observed events shows no pattern of repetition, and is consistent with a model with uniform event rate, independent of target. With one possible exception (HIP 107395), no valid event has been seen simultaneously at the two observatories. We describe the search and candidate events and set limits on the prevalence of civilizations transmitting intense optical pulses

From segment to somite: segmentation to epithelialization analyzed within quantitative frameworks

One of the most visually striking patterns in the early developing embryo is somite segmentation. Somites form as repeated, periodic structures in pairs along nearly the entire caudal vertebrate axis. The morphological process involves short- and long-range signals that drive cell rearrangements and cell shaping to create discrete, epithelialized segments. Key to developing novel strategies to prevent somite birth defects that involve axial bone and skeletal muscle development is understanding how the molecular choreography is coordinated across multiple spatial scales and in a repeating temporal manner. Mathematical models have emerged as useful tools to integrate spatiotemporal data and simulate model mechanisms to provide unique insights into somite pattern formation. In this short review, we present two quantitative frameworks that address the morphogenesis from segment to somite and discuss recent data of segmentation and epithelialization