Biomedical subjects

Sebastian Tuve

Publications and source records attributed to Sebastian Tuve.

5 recordsLinked to original sources

PubMed2006

A new group B adenovirus receptor is expressed at high levels on human stem and tumor cells.

CD46 is used by human group B adenoviruses (Ads) as a high-affinity attachment receptor. Here we show evidence that several group B Ads utilize an additional receptor for infection of human cells, which is different from CD46. We tentatively named this receptor receptor X. Competition studies with unlabeled and labeled Ads, recombinant Ad fiber knobs, and soluble CD46 and CD46 antibodies revealed three different subgroups of group B Ads, in terms of their receptor usage. Group I (Ad16, -21, -35, and -50) nearly exclusively uses CD46. Group II (Ad3, -7p, and -14) utilizes receptor X and not CD46. Group III (Ad11p) uses both CD46 and the alternative receptor X. Interaction of group II and III Ads with receptor X occurs via the fiber knob. Receptor X is an abundantly expressed glycoprotein that interacts with group II and III Ads at relatively low affinity in a Ca(2+)-dependent manner. This receptor is expressed at high levels on human mesenchymal and undifferentiated embryonic stem cells, as well as on human cancer cell lines. These findings have practical implications for stem cell and gene therapy.

Adenoviruses, Human↗

PubMed2006

Evaluation of adenovirus vectors containing serotype 35 fibers for vaccination.

In contrast to commonly used serotype 5-based adenovirus (Ad) vectors, Ad's containing fibers derived from B-group serotype 35 (Ad5/35) efficiently transduce human DCs ex vivo and appear to target antigen-presenting cells after intravenous injection into baboons. Based on this, Ad5/35 vectors could be valuable tools for immunotherapy and vaccination. On the other hand, a number of studies indicate that signaling through the B-group Ad receptor, CD46, can cause tolerance or immunosuppression. Since mice do not express CD46 in a human-like pattern, we studied the in vivo properties of Ad5/35 in transgenic mice that express CD46 in a pattern and at a level similar to those of humans. Hypersensitivity assays and analyses of frequencies of regulatory T cells and T cell responses did not indicate that Ad5/35 injection exerts detrimental effects on the host's immune system. An Ad5/35 vector expressing a model antigen was able to trigger a strong T cell response against the test antigen after intramuscular injection. Overall, compared to Ad5 vectors, Ad5/35 vectors had a better safety profile, reflected by lower serum levels of proinflammatory cytokines.

Adenoviridae↗

PubMed2006

Effect of adenovirus-mediated heat shock protein expression and oncolysis in combination with low-dose cyclophosphamide treatment on antitumor immune responses.

Heat shock proteins such as gp96 have the ability to chaperone peptides and activate antigen-presenting cells. In this study, we tested whether adenovirus-mediated overexpression of secreted or membrane-associated forms of gp96 in tumor cells would stimulate an antitumor immune response. Studies were carried out in C57Bl/6 mice bearing aggressively growing s.c. tumors derived from syngeneic TC-1 cells, a cell line that expresses HPV16 E6 and E7 proteins. We found that secreted gp96 can induce protective and therapeutic antitumor immune responses. Our data also indicate that the antitumor effect of sgp96 expression seems to be limited by the induction of suppressive regulatory T cells (Treg). TC-1 tumor transplantation increased the number of splenic and tumor-infiltrating Tregs. Importantly, treatment of mice with low-dose cyclophosphamide decreased the number of Tregs and enhanced the immunostimulatory effect of sgp96 expression. We also tested whether an oncolytic vector (Ad.IR-E1A/TRAIL), that is able to induce tumor cell apoptosis and, potentially, release cryptic tumor epitopes in immunogenic form, could stimulate antitumor immune responses. Although tumor cells infected ex vivo with Ad.IR-E1A/TRAIL had no antitumor effect when used as a vaccine alone, the additional treatment with low-dose cyclophosphamide resulted in the elimination of pre-established tumors. This study gives a rationale for testing approaches that suppress Tregs in combination with oncolytic or immunostimulatory vectors.

Adenoviridae↗

PubMed2004

Quantitative TP73 transcript analysis in hepatocellular carcinomas.

PURPOSE: The p53 family member p73 displays significant homology to p53, but data from primary tumors demonstrating increased expression levels of p73 in the absence of any gene mutations argue against a classical tumor suppressor function. A detailed analysis of the p73 protein in tumor tissues has revealed expression of two classes of p73 isoforms. Whereas the proapoptotic, full-length, transactivation-competent p73 protein (TA-p73) has a putative tumor suppressor activity similar to p53, the antiapoptotic, NH(2)-terminally truncated, transactivation-deficient p73 protein (DeltaTA-p73) has been shown to possess oncogenic activity. The oncogenic proteins can be generated by the following two different mechanisms: (a) aberrant splicing (p73Deltaex2, p73Deltaex2/3, DeltaN'-p73) and (b) alternative promoter usage of a second intronic promoter (DeltaN-p73). The purpose of our study was to elucidate the origin of DeltaTA-p73 isoforms in hepatocellular carcinomas. EXPERIMENTAL DESIGN: We analyzed the underlying mechanisms of p73 overexpression in cancer cells by quantification of p73 transcripts from 10 hepatocellular carcinoma patients using isoform-specific real-time reverse transcription-PCR. RESULTS: Our data demonstrate that only aberrantly spliced DeltaTA-p73 transcripts from the TA promoter show significantly increased expression levels in the tumor whereas the DeltaN-p73 transcript generated from the second promoter is not significantly up-regulated. CONCLUSIONS: Although we only analyzed 10 patient samples the results strongly suggest that the elevated activity of the first promoter (TA promoter) accounts for high-level expression of both full-length TA-p73 and aberrantly spliced DeltaTA-p73 isoforms in hepatocellular carcinoma tissues.

Aged↗

PubMed2004

Alterations of DeltaTA-p 73 splice transcripts during melanoma development and progression.

In the last 2 years, it has become apparent that the p53-family members p53 and p73 play fundamentally different roles in human malignancies. In contrast to p53, many studies on cancer patients failed to detect mutational inactivation of p73 and reported overexpression of wild-type p73 instead. A possible explanation was provided by the recent discovery of N-terminal truncated isoforms of p73 (DeltaTA-p73) that act as dominant-negative inhibitors of wild-type p53 and TA-p73 and result in tumor growth in nude mice. We investigated the role of DeltaTA-p73 in the development and progression of human melanomas, which lack p53 mutations. We analyzed 8 benign melanocytic nevi, 8 primary melanomas and 19 melanoma metastases for alterations of TA-p73 and DeltaTA-p73 expression using isoform-specific real-time RT-PCR. Based on our results, p73Deltaex2 and Deltaex2/3 spliced transcripts derived from the first promoter were significantly up-regulated in melanoma metastases, whereas DeltaN-p73 generated from the second promoter was the predominant isoform in benign nevi. Moreover, increased expression of p73Deltaex2 and p73Deltaex2/3 correlated with high-levels of both TA-p73 and E2F1. Our data suggest a potential function of DeltaTA-p73 splice isoforms in melanoma progression.

DNA-Binding Proteins↗