Purified recombinant proteins and peptides, which are currently under development in various anti-cancer vaccination approaches, lack sufficient immunogenicity. Therefore, potent adjuvants are needed ...to induce strong and persistent anti-tumor immunity. However, currently only few adjuvants are licensed, most of which primarily enhance antibody, but not T cell responses.Here, we demonstrate that a novel, well defined, and thoroughly characterized RNA-based adjuvant mediates balanced and long-lasting humoral and cellular immune responses. Our adjuvant significantly enhances anti-tumor immunity, and even complete tumor rejection can be achieved as shown for the syngeneic TC-1 tumor model, a murine model of human HPV-induced cervical cancer.Our adjuvant acts locally, promoting strong but transient up-regulation of anti-viral and pro-inflammatory cytokines, CXCR3-ligands and cytoplasmic RNA sensors at the injection site, avoiding any systemic cytokine release. These changes are followed by activation of different subsets of immune cells in the draining lymph nodes. In repeated dose toxicity studies carried out in mice and pigs no toxicity events were observed demonstrating an excellent preclinical safety profile. A Phase I first in man clinical trial testing different doses of RNAdjuvant® alone and in combination with reduced doses of the licensed rabies vaccine Rabipur® is currently ongoing. Healthy volunteers receive 2 intramuscular injections on days 0 and 21 either with RNAdjuvant® alone or in combination with 1/20 or 1/10 of the licensed Rabipur® dose. In both groups vaccinations were well tolerated with mild to moderate injection site reactions and flu-like symptoms as main side effects. Virus neutralizing antibody titers (VNTs) are measured on days 14 and 28 and a significant increase in median VNTs is observed after vaccination with 1/10 dose Rabipur® / RNAdjuvant® compared to 1/10 dose Rabipur® alone. In summary, our data suggest that RNAdjuvant® represents a novel, highly efficacious adjuvant candidate that can enhance cellular and humoral immune responses.Preliminary results of a first in human trial show a favorable safety profile and enhancement of immune responses in combination with a licensed rabies vaccine indicating an antigen sparing effect.While the field of cancer vaccination is currently obstructed by a lack of potent and safe adjuvants, RNAdjuvant® has the potential to fill this gap and to improve the efficacy of many cancer vaccines.
A novel, disruptive vaccination technology Kallen, Karl-Josef; Heidenreich, Regina; Schnee, Margit ...
Human vaccines & immunotherapeutics,
06/2013, Letnik:
9, Številka:
10
Journal Article
Recenzirano
Odprti dostop
Nucleotide based vaccines represent an enticing, novel approach to vaccination. We have developed a novel immunization technology, RNActive
®
vaccines, that have two important characteristics: mRNA ...molecules are used whose protein expression capacity has been enhanced by 4 to 5 orders of magnitude by modifications of the nucleotide sequence with the naturally occurring nucleotides A (adenosine), G (guanosine), C (cytosine), U (uridine) that do not affect the primary amino acid sequence. Second, they are complexed with protamine and thus activate the immune system by involvement of toll-like receptor (TLR) 7. Essentially, this bestows self-adjuvant activity on RNActive
®
vaccines. RNActive
®
vaccines induce strong, balanced immune responses comprising humoral and cellular responses, effector and memory responses as well as activation of important subpopulations of immune cells, such as Th1 and Th2 cells. Pre-germinal center and germinal center B cells were detected in human patients upon vaccination. RNActive
®
vaccines successfully protect against lethal challenges with a variety of different influenza strains in preclinical models. Anti-tumor activity was observed preclinically under therapeutic as well as prophylactic conditions. Initial clinical experiences suggest that the preclinical immunogenicity of RNActive
®
could be successfully translated to humans.
The rapid degradation of ribonucleic acids (RNA) by ubiquitous ribonucleases limits the efficacy of new therapies based on RNA molecules. Therefore, our aim was to characterize the natural ...ribonuclease activities on the skin and in blood plasma i.e. at sites where many drugs in development are applied. On the skin surfaces of Homo sapiens and Mus musculus we observed dominant pyrimidine-specific ribonuclease activity. This activity is not prevented by a cap structure at the 5'-end of messenger RNA (mRNA) and is not primarily of a 5'- or 3'-exonuclease type. Moreover, the ribonuclease activity on the skin or in blood plasma is not inhibited by chemical modifications introduced at the 2'OH group of cytidine or uridine residues. It is, however, inhibited by the ribonuclease inhibitor RNasin although not by the ribonuclease inhibitor SUPERase* In. The application of our findings in the field of medical science may result in an improved efficiency of RNA-based therapies that are currently in development.
IntroductionCV9104 is a novel mRNA-based therapeutic vaccine for prostate cancer encoding the prostate cancer-associated antigens PSA, PSMA, PSCA, STEAP, PAP and MUC1 by engineered mRNA molecules ...optimized for protein expression and immunostimulation (RNActive®). CV9103, an mRNA-based vaccine against 4 of these antigens was previously shown to be safe and immunogenic in a pPhase I/IIA trial. Immune responses against multiple antigens were associated with improved survival in patients with mCRPC.Design and methodsSafety and immunogenicity of first human exposure to CV9104 was investigated in the Phase I part. In the Phase II part, chemotherapy-naïve patients with a- or minimally symptomatic mCRPC were randomized to CV9104 or placebo. Primary endpoint is overall survival. Secondary endpoints include progression free survival, quality of life and immunogenicity among others. Upon disease or symptom progression blinded treatment is continued in combination with the subsequent systemic cancer treatment until second progression. Blood samples for immune monitoring and other biomarkers were taken at baseline, week 6 (1 week post fourth vaccination) and week 24 (6 weeks post ninth vaccination). Cellular and humoral immune responses against all vaccine antigens were analyzed by intracellular cytokine staining and IgM and IgG ELISA, respectively. Blood leukocyte phenotyping was performed by flow cytometry. Transcriptome analyses in whole blood were performed and serum samples are currently analyzed for miRNAs.ResultsSeven patients were enrolled in the Phase I part. Six of these patients were evaluable for immune responses. The independent data monitoring board recommended continuation into Phase II since nature and severity of adverse events were favorable and in line with previous results from CV9103 Recruitment of the Phase II part was finished in 12/2013 with 197 patients randomized. The trial is ongoing.5 of 6 patients of the Phase I part exhibited antigen-specific immune responses post vaccination. All CV9104 antigens were immunogenic. Multiple immune responses against ≥2 antigens were observed in 2 patients. In addition, phenotypic changes in CD4+ and CD8+ T cells consistent with an effector phenotype were observed.ConclusionResults from the Phase I part suggest that the self-adjuvanted mRNA vaccine CV9104 constitutes a well-tolerated approach to induce antigen-specific cellular and humoral immune responses against multiple antigens.Trial registration: http://www.clinicaltrials.gov/ct2/show/NCT01817738.
Anti-tumor vaccines targeting the entire tumor antigen repertoire represent an attractive immunotherapeutic approach. In the context of a phase I/II clinical trial, we vaccinated metastatic melanoma ...patients with autologous amplified tumor mRNA. In order to provide the large quantities of mRNA needed for each patient, the Stratagene Creator SMART cDNA library construction method was modified and applied to produce libraries derived from the tumors of 15 patients. The quality of those mRNA library vaccines was evaluated through sequencing and microarray analysis.
Random analysis of bacterial clones of the library showed a rate of 95% of recombinant plasmids among which a minimum of 51% of the clones contained a full-Open Reading Frame. In addition, despite a biased amplification toward small abundant transcripts compared to large rare fragments, we could document a relatively conserved gene expression profile between the total RNA of the tumor of origin and the corresponding in vitro transcribed complementary RNA (cRNA). Finally, listing the 30 most abundant transcripts of patient MEL02's library, a large number of tumor associated antigens (TAAs) either patient specific or shared by several melanomas were found.
Our results show that unlimited amounts of cRNA representing tumor's transcriptome could be obtained and that this cRNA was a reliable source of a large variety of tumor antigens.
This scoping review aimed to generate an overview of existing quality management (QM) models for inpatient healthcare published in peer-reviewed literature.
Peer-reviewed publications published until ...June 2016 were retrieved from the databases Medline, PubMed, CINAHL and Cochrane Library using search terms related to QM and models.
Publications mentioning a QM model for general application in healthcare or inpatient care in their title or abstract were included. Languages considered were: English, French, German, Italian and Spanish.
Data extraction was 3-fold. First, publication characteristics were summarized. Second, the frequency of each identified model was documented and the publications were divided into conceptual and implementation publications. Third, relevant QM models were identified and information regarding the model, including content and relationship with other models, was extracted.
Of 925 retrieved publications, 213 were included. The included publications reported on 64 different QM models that were suitable for or used in inpatient care. Seventeen models were identified as being relevant. The 17 models were then categorized into three different levels: conceptual quality improvement models, concrete application models and country specific adaptations.
This scoping review provides an overview of 17 existing QM models for inpatient care and their relationships with each other. Various types of models with differing aspects and components exist. In searching for QM models, many different concepts like QM system, accreditation or methodologies appeared. For future investigation, concepts of interest should be clarified.
We show that oomycete-derived Nep1 (for necrosis and ethylene-inducing peptide1)-like proteins (NLPs) trigger a comprehensive immune response in Arabidopsis thaliana, comprising posttranslational ...activation of mitogen-activated protein kinase activity, deposition of callose, production of nitric oxide, reactive oxygen intermediates, ethylene, and the phytoalexin camalexin, as well as cell death. Transcript profiling experiments revealed that NLPs trigger extensive reprogramming of the Arabidopsis transcriptome closely resembling that evoked by bacteria-derived flagellin. NLP-induced cell death is an active, light-dependent process requiring HSP90 but not caspase activity, salicylic acid, jasmonic acid, ethylene, or functional SGT1a/SGT1b. Studies on animal, yeast, moss, and plant cells revealed that sensitivity to NLPs is not a general characteristic of phospholipid bilayer systems but appears to be restricted to dicot plants. NLP-induced cell death does not require an intact plant cell wall, and ectopic expression of NLP in dicot plants resulted in cell death only when the protein was delivered to the apoplast. Our findings strongly suggest that NLP-induced necrosis requires interaction with a target site that is unique to the extracytoplasmic side of dicot plant plasma membranes. We propose that NLPs play dual roles in plant pathogen interactions as toxin-like virulence factors and as triggers of plant innate immune responses.
Mitogen-activated protein kinase (MAPK) cascades are universal and highly conserved signal transduction modules in eucaryotes,
including plants. These protein phosphorylation cascades link ...extracellular stimuli to a wide range of cellular responses.
However, the underlying mechanisms are so far unknown as information about phosphorylation substrates of plant MAPKs is lacking.
In this study we addressed the challenging task of identifying potential substrates for Arabidopsis thaliana mitogen-activated protein kinases MPK3 and MPK6, which are activated by many environmental stress factors. For this purpose,
we developed a novel protein microarray-based proteomic method allowing high throughput study of protein phosphorylation.
We generated protein microarrays including 1,690 Arabidopsis proteins, which were obtained from the expression of an almost nonredundant uniclone set derived from an inflorescence meristem
cDNA expression library. Microarrays were incubated with MAPKs in the presence of radioactive ATP. Using a threshold-based
quantification method to evaluate the microarray results, we were able to identify 48 potential substrates of MPK3 and 39
of MPK6. 26 of them are common for both kinases. One of the identified MPK6 substrates, 1-aminocyclopropane-1-carboxylic acid
synthase-6, was just recently shown as the first plant MAPK substrate in vivo , demonstrating the potential of our method to identify substrates with physiological relevance. Furthermore we revealed transcription
factors, transcription regulators, splicing factors, receptors, histones, and others as candidate substrates indicating that
regulation in response to MAPK signaling is very complex and not restricted to the transcriptional level. Nearly all of the
48 potential MPK3 substrates were confirmed by other in vitro methods. As a whole, our approach makes it possible to shortlist candidate substrates of mitogen-activated protein kinases
as well as those of other protein kinases for further analysis. Follow-up in vivo experiments are essential to evaluate their physiological relevance.
To identify classes of functioning trajectories in individuals with spinal cord injury (SCI) undergoing initial rehabilitation after injury and to examine potential predictors of class membership to ...inform clinical planning of the rehabilitation process.
Longitudinal analysis of the individual's rehabilitation stay using data from the Inception Cohort of the Swiss Spinal Cord Injury Cohort Study (SwiSCI).
Initial rehabilitation in specialized centers in Switzerland.
Individuals with newly acquired SCI (N=748; mean age, 54.66±18.38y) who completed initial rehabilitation between May 2013 and September 2019. The cohort was primarily composed of men (67.51%), persons with paraplegia (56.15%), incomplete injuries (67.51%), and traumatic etiologies (55.48%).
Not applicable.
Functioning was operationalized with the interval-based sum score of the Spinal Cord Independence Measure version III (SCIM III). For each individual, the SCIM III sum score was assessed at up to 4 time points during rehabilitation stay. The corresponding time of assessment was recorded by the difference in days between the SCIM III assessment and admission to the rehabilitation program.
Latent process mixed model analysis revealed 4 classes of functioning trajectories within the present sample. Class-specific predicted mean functioning trajectories describe stable high functioning (n=307; 41.04%), early functioning improvement (n=39; 5.21%), moderate functioning improvement (n=287; 38.37%), and slow functioning improvement (n=115; 15.37%), respectively. Out of 12 tested factors, multinomial logistic regression showed that age, injury level, injury severity, and ventilator assistance were robust predictors that could distinguish between identified classes of functioning trajectories in the present sample.
The current study establishes a foundation for future research on the course of functioning of individuals with SCI in initial rehabilitation by identifying classes of functioning trajectories. This supports the development of specifically tailored rehabilitation programs and prediction models, which can be integrated into clinical rehabilitation planning.
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