75 research outputs found
Recommended from our members
Longitudinal confocal microscopy imaging of solid tumor destruction following adoptive T cell transfer
A fluorescence-based, high-resolution imaging approach was used to visualize longitudinally the cellular events unfolding during T cell-mediated tumor destruction. The dynamic interplay of T cells, cancer cells, cancer antigen loss variants, and stromal cellsâall color-coded in vivoâwas analyzed in established, solid tumors that had developed behind windows implanted on the backs of mice. Events could be followed repeatedly within precisely the same tumor regionâbefore, during and after adoptive T cell therapyâthereby enabling for the first time a longitudinal in vivo evaluation of protracted events, an analysis not possible with terminal imaging of surgically exposed tumors. T cell infiltration, stromal interactions, and vessel destruction, as well as the functional consequences thereof, including the elimination of cancer cells and cancer cell variants were studied. Minimal perivascular T cell infiltrates initiated vascular destruction inside the tumor mass eventually leading to macroscopic central tumor necrosis. Prolonged engagement of T cells with tumor antigen-crosspresenting stromal cells correlated with high IFNÎł cytokine release and bystander elimination of antigen-negative cancer cells. The high-resolution, longitudinal, in vivo imaging approach described here will help to further a better mechanistic understanding of tumor eradication by T cells and other anti-cancer therapies
The Two-Component Signal Transduction System CopRS of Corynebacterium glutamicum Is Required for Adaptation to Copper-Excess Stress
Copper is an essential cofactor for many enzymes but at high concentrations it is toxic for the cell. Copper ion concentrations â„50 ”M inhibited growth of Corynebacterium glutamicum. The transcriptional response to 20 ”M Cu2+ was studied using DNA microarrays and revealed 20 genes that showed a â„ 3-fold increased mRNA level, including cg3281-cg3289. Several genes in this genomic region code for proteins presumably involved in the adaption to copper-induced stress, e. g. a multicopper oxidase (CopO) and a copper-transport ATPase (CopB). In addition, this region includes the copRS genes (previously named cgtRS9) which encode a two-component signal transduction system composed of the histidine kinase CopS and the response regulator CopR. Deletion of the copRS genes increased the sensitivity of C. glutamicum towards copper ions, but not to other heavy metal ions. Using comparative transcriptome analysis of the ÎcopRS mutant and the wild type in combination with electrophoretic mobility shift assays and reporter gene studies the CopR regulon and the DNA-binding motif of CopR were identified. Evidence was obtained that CopR binds only to the intergenic region between cg3285 (copR) and cg3286 in the genome of C. glutamicum and activates expression of the divergently oriented gene clusters cg3285-cg3281 and cg3286-cg3289. Altogether, our data suggest that CopRS is the key regulatory system in C. glutamicum for the extracytoplasmic sensing of elevated copper ion concentrations and for induction of a set of genes capable of diminishing copper stress
The role of open abdomen in non-trauma patient : WSES Consensus Paper
The open abdomen (OA) is defined as intentional decision to leave the fascial edges of the abdomen un-approximated after laparotomy (laparostomy). The abdominal contents are potentially exposed and therefore must be protected with a temporary coverage, which is referred to as temporal abdominal closure (TAC). OA use remains widely debated with many specific details deserving detailed assessment and clarification. To date, in patients with intra-abdominal emergencies, the OA has not been formally endorsed for routine utilization; although, utilization is seemingly increasing. Therefore, the World Society of Emergency Surgery (WSES), Abdominal Compartment Society (WSACS) and the Donegal Research Academy united a worldwide group of experts in an international consensus conference to review and thereafter propose the basis for evidence-directed utilization of OA management in non-trauma emergency surgery and critically ill patients. In addition to utilization recommendations, questions with insufficient evidence urgently requiring future study were identified.Peer reviewe
Recommended from our members
Research and Design of a Routing Protocol in Large-Scale Wireless Sensor Networks
æ çșżäŒ æćšçœç»ïŒäœäžșć
šçæȘæ„ć性ææŻäčäžïŒéæäșäŒ æćšææŻăć”ć
„ćŒèźĄçźææŻăććžćŒäżĄæŻć€çćèȘç»ç»çœææŻïŒćŻćźæ¶æç„ăééăć€çăäŒ èŸçœç»ććžćșćć
çćç§äżĄæŻæ°æźïŒćšćäșćœéČăçç©ć»çăçŻćąçæ”ăæąé©æçŸăéČæćæăć±é©ćșćèżçšæ§ć¶çéąćć
·æćććčżéçćșçšćæŻă æŹæç 究ćæäșæ çșżäŒ æćšçœç»çć·Čæè·Żç±ćèźźïŒćč¶éćŻč性è§æšĄçæ çșżäŒ æćšçœç»èźŸèźĄäșäžç§æ ç¶è·Żç±ćèźźïŒćźæ čæźèçčć°ć俥æŻæ„ćœąæè·Żç±ïŒä»èçźćäșć€æçčćçè·Żç±èĄšæ„æŸć绎æ€ïŒèçäșäžćż
èŠçćŒéïŒæé«äșè·Żç±æçïŒćźç°äșćż«éææçæ°æźäŒ èŸă äžșæŻææ€è·Żç±ćèźźæŹææćșäșäžç§èȘéćșćšæć°ććé
çźââADARïŒAdaptiveDynamicAddre...As one of the ten high technologies in the future, wireless sensor network, which is the integration of micro-sensors, embedded computing, modern network and Ad Hoc technologies, can apperceive, collect, process and transmit various information data within the region. It can be used in military defense, biomedical, environmental monitoring, disaster relief, counter-terrorism, remote control of haz...ćŠäœïŒć·„ćŠçĄćŁ«éąçł»äžäžïŒäżĄæŻç§ćŠäžææŻćŠéąé俥淄çšçł»_é俥äžäżĄæŻçł»ç»ćŠć·ïŒ2332007115216
Broadband Multi-wavelength Properties of M87 during the 2017 Event Horizon Telescope Campaign
Abstract: In 2017, the Event Horizon Telescope (EHT) Collaboration succeeded in capturing the first direct image of the center of the M87 galaxy. The asymmetric ring morphology and size are consistent with theoretical expectations for a weakly accreting supermassive black hole of mass âŒ6.5 Ă 109 M â. The EHTC also partnered with several international facilities in space and on the ground, to arrange an extensive, quasi-simultaneous multi-wavelength campaign. This Letter presents the results and analysis of this campaign, as well as the multi-wavelength data as a legacy data repository. We captured M87 in a historically low state, and the core flux dominates over HST-1 at high energies, making it possible to combine core flux constraints with the more spatially precise very long baseline interferometry data. We present the most complete simultaneous multi-wavelength spectrum of the active nucleus to date, and discuss the complexity and caveats of combining data from different spatial scales into one broadband spectrum. We apply two heuristic, isotropic leptonic single-zone models to provide insight into the basic source properties, but conclude that a structured jet is necessary to explain M87âs spectrum. We can exclude that the simultaneous Îł-ray emission is produced via inverse Compton emission in the same region producing the EHT mm-band emission, and further conclude that the Îł-rays can only be produced in the inner jets (inward of HST-1) if there are strongly particle-dominated regions. Direct synchrotron emission from accelerated protons and secondaries cannot yet be excluded
Optimierung von retroviralem Gentransfer in T-Lymphocyten fĂŒr die Neuausrichtung von T-ZellspezifitĂ€t
TITLE AND CONTENTS
SUMMARY 1
ZUSAMMENFASSUNG 2
GENERAL INTRODUCTION 3
HIGH-LEVEL GENE EXPRESSION IN T CELLS 30
ENGINEERING RCC-REACTIVE CTL 45
DISCUSSION 58
APPENDIX 78
ABBREVIATIONS 80
REFERENCES 83
ACKNOWLEDGEMENTS 100
PUBLICATIONS 101
EIDESSTATTLICHE ERKLĂRUNG 102Adoptive T cell therapy is a promising approach for the treatment of cancer.
Donor lymphocyte infusions in patients with cytomegalovirus-mediated disease,
Epstein-Barr virus-positive B cell lymphomas, and chronic myelogenous
leukaemia have proven the efficacy of transferred T cells to kill infected or
malignant cells. The main hindrance of using this approach for the cure of
other tumour entities is the often unsuccessful isolation and amplification of
tumour-reactive T cells from patients. A potential method to circumvent this
problem is the transfer of anti-tumour specificity to peripheral blood
lymphocytes by retroviral T cell receptor (TCR) gene transfer. In this thesis
I have compared several retroviral vectors regarding their gene transfer
potential. For this purpose the green fluorescent protein (GFP) was cloned
into vectors containing different cis-regulatory elements. Promoter and
enhancer elements of two different long terminal repeats (LTR) and a cellular
gene were compared. The LTR originating from the myeloproliferative sarcoma
virus (MPSV) was shown to be superior to the Moloney murine leukaemia virus
(MoMLV) LTR and a truncated MoMLV LTR/CD2 enhancer combination. Furthermore,
the impact of a modified leader sequence and the Woodchuck hepatitis virus
post-transcriptional regulatory element (PRE) was analysed. These elements
showed an enhancing effect on transgene expression and retroviral titre,
respectively. Taken together, the MP71 retrovirus containing the MPSV LTR, the
modified leader, and the PRE demonstrated strong and durable transgene
expression in murine and human T lymphocytes. High transduction rates were
obtained when using retroviruses pseudotyped with an ecotropic MoMLV or the
amphotropic 10A1 MLV envelope for murine or human cells, respectively. The
same results were obtained after exchanging GFP with TCR α and ÎČ -chain genes,
linked by an internal ribosomal entry site. The redirected primary human T
lymphocytes showed the same specificity as the tumour infiltrating lymphocyte
(TIL)-26 from which the TCR originated. Furthermore, the intensity of anti-
tumour reactivity, measured as cytokine release and tumour cell lysis, was
comparable for both the TCR-26-grafted T cells and the TIL-26. The TCR
expression, analysed for a period of 100 days, remained stable. This approach
enables us to generate functional renal cell carcinoma (RCC)-specific T
lymphocytes by TCR gene transfer. These data provide the rationale for
adoptive T cell therapy for RCC.Der adoptive T-Zelltransfer ist eine viel versprechende Form der
Tumortherapie. Die Infusion von Donor T-Lymphozyten fĂŒr die Behandlung von
Virusinfektionen (Cytomegalovirus und Epstein-Barr-Virus) und einigen wenigen
Krebsarten hat sich als sehr effektiv erwiesen. FĂŒr die meisten Tumorarten ist
jedoch die Isolierung und in vitro Vermehrung von Tumor-spezifischen T-Zellen
nicht möglich. Eine Abhilfe fĂŒr dieses Problem kann der Transfer von anti-
TumorspezifitÀt schaffen, wie zum Beispiel der retrovirale Transfer von T-Zell
Rezeptor (TCR)-Genen in periphere Blut-Lymphozyten (PBL). In dieser Arbeit
habe ich verschiedene retrovirale Vektoren bezĂŒglich ihrer
Gentransfereffizienz in T-Lymphozyten analysiert. Dazu wurde das grĂŒn
fluoreszierende Protein (GFP) in Vektoren mit verschiedenen cis-
regulatorischen Elementen kloniert. Das Promotor/"Enhancer" Paar des "long
terminal repeats" (LTR) des myeloproliferativen Sarkom-Virus (MPSV) war dabei
dem Paar des LTR des Moloney-Maus-LeukÀmie-Virus (MoMLV) und einem Paar aus
dem MoMLV Promotor und dem "Enhancer" des CD2-Gens ĂŒberlegen. Der Einsatz
einer modifizierten "leader"-Sequenz und des posttranskriptionellen
regulatorischen Elements (PRE) des Waldmurmeltier-Hepatitis-B-Virus fĂŒhrte zu
einer Erhöhung der Transgenexpression bzw. des Virustiters. Die
Expressionsuntersuchungen ergaben, dass die Kombination aus der LTR des MPSV,
der modifizierten "leader"-Sequenz und des PRE geeignet ist, um Gene in Maus-
und humanen T-Zellen in hohem MaĂe und fĂŒr lĂ€ngere Zeit zu exprimieren. Des
weiteren wurde gezeigt, dass sich das ecotrope MoMLV, bzw. das amphotrope 10A1
-MLV-HĂŒllprotein, gut eignen, um Maus-, bzw. humane T-Zellen, zu
transduzieren. Der Austausch des GFP durch TCR α und ÎČ -Ketten-Gene, verbunden
durch eine interne Ribosomen-Eintrittsstelle, ergab Àhnliche
Expressionsergebnisse. Der transferierte TCR-26 wurde aus dem
Nierenzellkarzinom-spezifischen Tumor-infiltrierenden Lymphozyten Klon-26
gewonnen. Die TCR-26-modifizierten T-Zellen zeigten dieselbe SpezifitÀt wie
der Ursprungsklon. Auch war die anti-TumorreaktivitÀt, gemessen als
Zytokinfreisetzung und Zelllyse, der beiden Zellpopulationen vergleichbar. Die
ĂŒber einen Zeitraum von 100 Tagen analysierte TCR-Expression war stabil. Diese
Ergebnisse zeigen, dass der Transfer von anti-NierenzellkarzinomspezifitÀt auf
primÀre T-Zellen möglich ist, und somit ein adoptiver T-Zelltransfer als
Nierenzellkarzinomtherapie denkbar
- âŠ