Interferon-α Abrogates Tolerance Induction by Human Tolerogenic Dendritic Cells

BACKGROUND: Administration of interferon-α (IFN-α) represents an approved adjuvant therapy as reported for malignancies like melanoma and several viral infections. In malignant diseases, tolerance processes are critically involved in tumor progression. In this study, the effect of IFN-α on tolerance induction by human tolerogenic dendritic cells (DC) was analyzed. We focussed on tolerogenic IL-10-modulated DC (IL-10 DC) that are known to induce anergic regulatory T cells (iTregs). METHODOLOGY/PRINCIPAL FINDINGS: IFN-α promoted an enhanced maturation of IL-10 DC as demonstrated by upregulation of the differentiation marker CD83 as well as costimulatory molecules. IFN-α treatment resulted in an increased capacity of DC to stimulate T cell activation compared to control tolerogenic DC. We observed a strengthened T cell proliferation and increased IFN-γ production of CD4(+) and CD8(+) T cells stimulated by IFN-α-DC, demonstrating a restoration of the immunogenic capacity of tolerogenic DC in the presence of IFN-α. Notably, restimulation experiments revealed that IFN-α treatment of tolerogenic DC abolished the induction of T cell anergy and suppressor function of iTregs. In contrast, IFN-α neither affected the priming of iTregs nor converted iTregs into effector T cells. CONCLUSIONS/SIGNIFICANCE: IFN-α inhibits the induction of T cell tolerance by reversing the tolerogenic function of human DC

Differential Effects of Mutations on the Transport Properties of the Na⁺/H⁺ Antiporter NhaA from Escherichia coli

Na+/H+ antiporters show a marked pH dependence, which is important for their physiological function in eukaryotic and prokaryotic cells. In NhaA, the Escherichia coli Na+/H+ antiporter, specific single site mutations modulating the pH profile of the transporter have been described in the past. To clarify the mechanism by which these mutations influence the pH dependence of NhaA, the substrate dependence of the kinetics of selected NhaA variants was electrophysiologically investigated and analyzed with a kinetic model. It is shown that the mutations affect NhaA activity in quite different ways by changing the properties of the binding site or the dynamics of the transporter. In the first case, pK and/or KDNa are altered, and in the second case, the rate constants of the conformational transition between the inside and the outside open conformation are modified. It is shown that residues as far apart as 15–20 Å from the binding site can have a significant impact on the dynamics of the conformational transitions or on the binding properties of NhaA. The implications of these results for the pH regulation mechanism of NhaA are discussed

Netzwerk BioMeT Dresden Schlussbericht

SIGLEAvailable from TIB Hannover: F04B1179 / FIZ - Fachinformationszzentrum Karlsruhe / TIB - Technische InformationsbibliothekBundesministerium fuer Bildung und Forschung (BMBF), Bonn (Germany)DEGerman

Nachweis gentechnischer Veränderungen in Mais mittels PCR

Ein PCR-Nachweis für gentechnisch veränderten Mais «Event 176» der Fa. Ciba-Geigy wurde etabliert. Der Mais enthält Gene, die Selbstschutz gegen den Maiszünsler (Delta-Endotoxin-Gen ausBacillus thuringiensis) und Toleranz gegen das Herbizid Basta (Phosphinothricin-Resistenz-Gen ausStreptomyces hygroscopicus) vermitteln. Zudem enthält der Mais ein Ampicillin-Resistenz-Gen. Für die Amplifikation von Bereichen aus allen drei Genen wurden PCR-Primer entworfen. Mit Hilfe dieser Primer und mit «Event 176»-Mais-DNA als Template konnten die entsprechenden Genbereiche in der PCR amplifiziert werden. Die PCR-Produkte wurden sequenziert, um ihre Identität zu bestätigen. Mit Hilfe der Delta-Endotoxin-PCR wurden, auch in Gegenwart von 104fachem Überschuß nicht gentechnisch veränderter Mais-DNA, fünf haploide Genome der «Event 176»-DNA nachgewiesen.A PCR-test for the genetically modified maize «Event 176» of Ciba-Geigy was established. The maize contains genes conferring resistance to the European corn borer (delta-endotoxin gene fromBacillus thuringiensis) and tolerance to the herbicide Basta (phosphinothricin resistance gene fromStreptomyces hygroscopicus). The maize contains also an ampicillin resistance gene. Primers were designed and using «Event 176»-maize-DNA as template internal regions of the three genes were amplified with PCR. The PCR products were sequenced to confirm their identity. Using the deltaendotoxin primers in PCR down to 5 haploid genomes of «Event 176»-DNA could be detected, even in the presence of a 104fold excess of DNA from non-modified maize