Immunological characterization of a Modified Vaccinia Virus Ankara vector expressing the Human Papilloma Virus 16 E1 protein.
Clin Vaccine Immunol. 2013 Dec 4;
Authors: Remy-Ziller C, Germain C, Spindler A, Hoffmann C, Silvestre N, Rooke R, Bonnefoy JY, Préville X
Women showing a normal cytology but diagnosed with a persistent high-risk Human Papilloma Virus (HR-HPV) infection have a higher risk than non-infected women to develop high grade cervical intraepithelial neoplasia and cervical cancer. As no therapeutic management other than surveillance is offered to these women, there is a major challenge to develop novel targeted therapies dedicated to the treatment of these patients. As such, E1 and E2 antigens, expressed early in HPV life cycle, represent very interesting candidates. Both proteins are necessary to maintain coordinated viral replication and gene synthesis during the differentiation process of the epithelium, and are essential for the virus to complete its normal and propagative replication cycle. In the present study, we performed evaluation of a new active targeted immunotherapeutic, a Modified Vaccinia Virus Ankara (MVA) vector containing the E1 sequence of HPV16, aiming at inducing cellular immune responses with potential to help and clear persistent HPV-16 related infection. We carried out an extensive comparative time course analysis of the cellular immune response induced by different schedule of immunization in C57BL/6 mice. We showed that multiple injections of MVA-E1 allowed sustained HPV16E1-specific cellular immune responses in vaccinated mice and had no impact on the exhaustion phenotype of the generated HPV16E1-specific CD8(+) T cells, but led to the differentiation of multi-functional effector T cells with high cytotoxic capacity. This study provides proof of concept that a MVA expressing HPV16E1 can induce robust and long lasting E1-specific responses and warrants further development of this candidate.
PMID: 24307238 [PubMed - as supplied by publisher]
A Review of Therapeutic Aptamer Conjugates with Emphasis on New Approaches.
Pharmaceuticals (Basel). 2013;6(3):340-357
Authors: Bruno JG
The potential to emulate or enhance antibodies…
Development of a domain-specific genetic language to design Chlamydomonas reinhardtii expression vectors.
Bioinformatics. 2013 Nov 8;
Authors: Wilson ML, Okumoto S, Adam L, Peccoud J
High-throughput, cost-effective verification of structural DNA assembly.
Nucleic Acids Res. 2013 Nov 6;
Authors: Dharmadi Y, Patel K, Shapland E, Hollis D, Slaby T, Klinkner N, Dean J, Chandran SS
DNA ‘assembly’ from ‘building blocks’ remains a cornerstone in synthetic biology, whether it be for gene synthesis (∼1 kb), pathway engineering (∼10 kb) or synthetic genomes (>100 kb). Despite numerous advances in the techniques used for DNA assembly, verification of the assembly is still a necessity, which becomes cost-prohibitive and a logistical challenge with increasing scale. Here we describe for the first time a comprehensive, high-throughput solution for structural DNA assembly verification by restriction digest using exhaustive in silico enzyme screening, rolling circle amplification of plasmid DNA, capillary electrophoresis and automated digest pattern recognition. This low-cost and robust methodology has been successfully used to screen over 31 000 clones of DNA constructs at <$1 per sample.
PMID: 24203706 [PubMed - as supplied by publisher]
Development of Artificial Nucleic Acid That Recognizes a CG Base Pair in Triplex DNA Formation.
Yakugaku Zasshi. 2013;133(11):1201-8
Authors: Hari Y
An oligonucleotide that can form a triplex with double-stranded DNA is called a triplex-forming oligonucleotide (TFO). TFOs have gained considerable attention because of their potential as gene targeting tools. However, triplex DNA formation involves inherent problems for practical use. The most important problem is that natural nucleotides in TFO do not have sufficient affinity and base pair-selectivity to pyrimidine-purine base pair, like a CG or TA base pair, within dsDNA. This suggests that dsDNA region including a CG or TA base pair cannot be targeted. Therefore, artificial nucleotides, especially with non-natural nucleobases, capable of direct recognition of a CG or TA base pair via hydrogen bond formation have been developed; however, nucleotides with better selectivity and stronger affinity are necessary for implementing this dsDNA-targeting technology using TFOs. Under such a background, we considered that facile and efficient synthesis of various nucleobase derivatives in TFOs would be useful for finding an ideal nucleobase for recognition of a CG or TA base pair because detailed and rational exploration of nucleobase structures is facilitated. Recently, to develop a nucleobase recognizing a CG base pair, we have used post-elongation modification, i.e., modification after oligonucleotide synthesis, for the facile synthesis of nucleobase derivatives. This review mainly summarizes our recent findings on the development of artificial nucleobases and nucleotides for recognition of a CG base pair in triplexes formed between dsDNA and TFOs.
PMID: 24189561 [PubMed - in process]
The cellular peptidyl-prolyl cis/trans isomerase Pin1 regulates reactivation of Kaposi’s sarcoma-associated herpesvirus from latency.
J Virol. 2013 Oct 30;
Authors: Guito J, Gavina A, Palmeri D, Lukac DM
Ethynyl Side Chain Hydration During Synthesis and Work-up of “Clickable” Oligonucleotides: Bypassing Acetyl Group Formation by Triisopropylsilyl Protection.
J Org Chem. 2013 Oct 18;
Authors: Ingale SA, Mei H, Leonard P, Seela F
Clickable oligonucleotides with ethynyl residues in the 5-position of pyrimidines (ethdC and ethdU) or the 7-position of 7-deazaguanine (ethc7Gd) are hydrated during solid-phase oligonucleotide synthesis and work-up conditions. The side products were identified as acetyl derivatives by MALDI-TOF mass spectra of oligonucleotides and by detection of modified nucleosides after enzymatic phosphodiester hydrolysis. Ethynyl → acetyl group conversion was also studied on ethynylated nucleosides under acidic and basic conditions. It could be shown that side chain conversion depends on the nucleobase structure. Triisopropylsilyl residues were introduced to protect ethynyl residues from hydration. Pure, acetyl group free oligonucleotides were isolated after desilylation in all cases.
PMID: 24138578 [PubMed - as supplied by publisher]
Increasing Platelet Concentrations in Leukocyte-Reduced Platelet-Rich Plasma Decrease Collagen Gene Synthesis in Tendons.
Am J Sports Med. 2013 Oct 17;
Authors: Boswell SG, Schnabel LV, Mohammed HO, Sundman EA, Minas T, Fortier LA
BACKGROUND:Platelet-rich plasma (PRP) is used for the treatment of tendinopathy. There are numerous PRP preparations, and the optimal combination of platelets and leukocytes is not known. HYPOTHESIS:Within leukocyte-reduced PRP (lrPRP), there is a plateau effect of platelet concentration, with increasing platelet concentrations being detrimental to extracellular matrix synthesis. STUDY DESIGN:Controlled laboratory study. METHODS:Different formulations of lrPRP with respect to the platelet:leukocyte ratio were generated from venous blood of 8 horses. Explants of the superficial digital flexor tendon were cultured in lrPRP products for 96 hours. Platelet-derived growth factor-BB (PDGF-BB), tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), and interleukin-1β (IL-1β) concentrations were determined in the media by enzyme-linked immunosorbent assay. Gene expression in tendon tissue for collagen type I and III (COL1A1 and COL3A1, respectively), matrix metalloproteinase-3 and -13 (MMP-3 and MMP-13, respectively), cartilage oligomeric matrix protein (COMP), and IL-1β was determined. Data were divided into 3 groups of lrPRP based on the ratio of platelets:leukocytes and evaluated to determine the effect of platelet concentration. RESULTS:Complete blood counts verified leukocyte reduction and platelet enrichment in all PRP preparations. In the lrPRP preparation, the anabolic growth factors PDGF-BB and TGF-β1 were increased with increasing platelet concentrations, and the catabolic cytokine IL-1β was decreased with increasing platelet concentrations. Increasing the platelet concentration resulted in a significant reduction in COL1A1 and COL3A1 synthesis in tendons. CONCLUSION:Increasing the platelet concentration within lrPRP preparations results in the delivery of more anabolic growth factors and less proinflammatory cytokines, but the biological effect on tendons is diminished metabolism as indicated by a decrease in the synthesis of both COL1A1 and COL3A1. Together, this information suggests that minimizing leukocytes in PRP is more important than maximizing platelet numbers with respect to decreasing inflammation and enhancing matrix gene synthesis. CLINICAL RELEVANCE:This study suggests that reducing leukocytes to minimize catabolic signaling appears to be more important than increasing platelets in an effort to maximize anabolic signaling. Further, a maximum biological threshold of benefit was demonstrated with regard to the number of platelets beyond which further increases in platelet concentration did not result in further anabolic upregulation. In vivo investigations documenting the use of platelets for the treatment of tendinopathy are justified as well as further in vitro characterization of the ideal PRP product for the treatment of tendinopathy and other musculoskeletal applications.
PMID: 24136860 [PubMed - as supplied by publisher]
Targeting and Eradicating Hepatic Cancer Cells with a Cancer-Specific Vector Carrying the Buforin II Gene.
Cancer Biother Radiopharm. 2013 Oct;28(8):623-30
Authors: Wang Y, Qu L, Gong L, Sun L, Gong R, Si J
The Role of Nuclear Bodies in Gene Expression and Disease.
Biology (Basel). 2013 Jul 9;2(3):976-1033
Authors: Morimoto M, Boerkoel CF
This review summarizes the current understanding of the role of n…