Expamers: a new technology to control T cell activation

T cell activation is a cornerstone in manufacturing of T cell-based therapies, and precise control over T cell activation is important in the development of the next generation T-cell based therapeutics. This need cannot be fulfilled by currently available methods for T cell stimulation, in particular not in a time dependent manner. Here, we describe a modular activation reagent called Expamers, which addresses these limitations. Expamers are versatile stimuli that are intended for research and clinical use. They are readily soluble and can be rapidly bound and removed from the cell surface, allowing nearly instantaneous initiation and termination of activation signal, respectively. Hence, Expamers enable precise regulation of T cell stimulation duration and provide promise of control over T cell profiles in future products. Expamers can be easily adopted to different T cell production formats and have the potential to increase efficacy of T cell immunotherapeutics

Novel Serial Positive Enrichment Technology Enables Clinical Multiparameter Cell Sorting

A general obstacle for clinical cell preparations is limited purity, which causes variability in the quality and potency of cell products and might be responsible for negative side effects due to unwanted contaminants. Highly pure populations can be obtained best using positive selection techniques. However, in many cases target cell populations need to be segregated from other cells by combinations of multiple markers, which is still difficult to achieve – especially for clinical cell products. Therefore, we have generated low-affinity antibody-derived Fab-fragments, which stain like parental antibodies when multimerized via Strep-tag and Strep-Tactin, but can subsequently be removed entirely from the target cell population. Such reagents can be generated for virtually any antigen and can be used for sequential positive enrichment steps via paramagnetic beads. First protocols for multiparameter enrichment of two clinically relevant cell populations, CD4high/CD25high/CD45RAhigh ‘regulatory T cells’ and CD8high/CD62Lhigh/CD45RAneg ‘central memory T cells’, have been established to determine quality and efficacy parameters of this novel technology, which should have broad applicability for clinical cell sorting as well as basic research

Infrared Narrow-Band Tomography of the Local Starburst NGC 1569 with LBT/LUCIFER

We used the near-IR imager/spectrograph LUCIFER mounted on the Large Binocular Telescope (LBT) to image, with sub-arcsec seeing, the local dwarf starburst NGC 1569 in the JHK bands and HeI 1.08 micron, [FeII] 1.64 micron and Brgamma narrow-band filters. We obtained high-quality spatial maps of HeI, [FeII] and Brgamma emission across the galaxy, and used them together with HST/ACS images of NGC 1569 in the Halpha filter to derive the two-dimensional spatial map of the dust extinction and surface star formation rate density. We show that dust extinction is rather patchy and, on average, higher in the North-West (NW) portion of the galaxy [E_g(B-V) = 0.71 mag] than in the South-East [E_g(B-V) = 0.57 mag]. Similarly, the surface density of star formation rate peaks in the NW region of NGC 1569, reaching a value of about 4 x 10^-6 M_sun yr^-1 pc^-2. The total star formation rate as estimated from the integrated, dereddened Halpha luminosity is about 0.4 M_sun yr^-1, and the total supernova rate from the integrated, dereddened [FeII] luminosity is about 0.005 yr^-1 (assuming a distance of 3.36 Mpc). The azimuthally averaged [FeII]/Brgamma flux ratio is larger at the edges of the central, gas-deficient cavities (encompassing the super star clusters A and B) and in the galaxy outskirts. If we interpret this line ratio as the ratio between the average past star formation (as traced by supernovae) and on-going activity (represented by OB stars able to ionize the interstellar medium), it would then indicate that star formation has been quenched within the central cavities and lately triggered in a ring around them. The number of ionizing hydrogen and helium photons as computed from the integrated, dereddened Halpha and HeI luminosities suggests that the latest burst of star formation occurred about 4 Myr ago and produced new stars with a total mass of ~1.8 x 10^6 M_sun. [Abridged]Comment: accepted for publication in A

Taste Manipulation during a Food Cue-Reactivity Task

These data were collected to test the primary hypothesis that tasting high-calorie foods during a food cue-reactivity task would evoke stronger craving relative to not tasting (i.e., only observing) high-calorie foods. The data indicated that tasting high-calorie foods suppressed cue-elicited food craving compared to not tasting high-calorie foods (a nonsignificant, though medium-sized effect). The taste manipulation did not affect calorie intake during a subsequent bogus taste test. Results also indicated that cue-elicited food craving to in vivo food cues elicited stronger craving than photographic food cues. The data included in this upload include basic demographic variables, baseline craving, cue-elicited craving ratings in response to food cues, divided by cue type (low-, high-calorie) and cue mode (photographic, in vivo), and total calorie intake during the bogus taste test. Baseline and cue-elicited craving ratings reflect sum scores from the 3-item Intense Desire to Eat subscale of the Food Cravings Questionnaire-State (FCQ-S). Cue-elicited craving was assessed after the administration of each food cue presented during a food cue-reactivity task (8 low-calorie and 8 high-calorie food cues, divided evenly between photographic and in vivo food cues). Craving scores were averaged across the 3-item FCQ-S ratings by cue type and cue mode to produce four cue-elicited craving scores (low-calorie photographic, low-calorie in vivo, high-calorie photographic, high-calorie in vivo). During in vivo food cue trials, participants randomized to the No Taste group closely observed, but were unable to taste food cues, whereas participants randomized to the Taste group closely observed and then consumed the food cues (this was the only difference between groups). After the food cue-reactivity task, participants engaged in a 10-min bogus taste test of 4 high-calorie foods. Total grams of food consumed was converted to total calories consumed based on the manufacturer's nutrition information

Taste Manipulation during a Food Cue-Reactivity Task

These data were collected to test the primary hypothesis that tasting high-calorie foods during a food cue-reactivity task would evoke stronger craving relative to not tasting (i.e., only observing) high-calorie foods. The data indicated that tasting high-calorie foods suppressed cue-elicited food craving compared to not tasting high-calorie foods (a nonsignificant, though medium-sized effect). The taste manipulation did not affect calorie intake during a subsequent bogus taste test. Results also indicated that cue-elicited food craving to in vivo food cues elicited stronger craving than photographic food cues. The data included in this upload include basic demographic variables, baseline craving, cue-elicited craving ratings in response to food cues, divided by cue type (low-, high-calorie) and cue mode (photographic, in vivo), and total calorie intake during the bogus taste test. Baseline and cue-elicited craving ratings reflect sum scores from the 3-item Intense Desire to Eat subscale of the Food Cravings Questionnaire-State (FCQ-S). Cue-elicited craving was assessed after the administration of each food cue presented during a food cue-reactivity task (8 low-calorie and 8 high-calorie food cues, divided evenly between photographic and in vivo food cues). Craving scores were averaged across the 3-item FCQ-S ratings by cue type and cue mode to produce four cue-elicited craving scores (low-calorie photographic, low-calorie in vivo, high-calorie photographic, high-calorie in vivo). During in vivo food cue trials, participants randomized to the No Taste group closely observed, but were unable to taste food cues, whereas participants randomized to the Taste group closely observed and then consumed the food cues (this was the only difference between groups). After the food cue-reactivity task, participants engaged in a 10-min bogus taste test of 4 high-calorie foods. Total grams of food consumed was converted to total calories consumed based on the manufacturer's nutrition information.THIS DATASET IS ARCHIVED AT DANS/EASY, BUT NOT ACCESSIBLE HERE. TO VIEW A LIST OF FILES AND ACCESS THE FILES IN THIS DATASET CLICK ON THE DOI-LINK ABOV

Molecular cloning, DNA sequence and transcriptional analysis of the Rhodospirillum molischianum B800/850 light-harvesting genes

The amino acid sequences of the B800/850 light-harvesting proteins from Rhodospirillum molischianum were determined by Edman degradation. On the basis of these amino acid sequences, two degenerated oligonucleotides were synthesized and used for PCR of genomic DNA. The resulting 150 by DNA fragment was cloned, sequenced and used for subsequent Southern blot analysis of digested genomic DNA. A 2.3 kbp EcoRI fragment strongly hybridized to the probe and a size selected genomic library from genomic DNA was constructed. One clone scored positive during screening of the library with the PCR-fragment and subsequent DNA sequence analysis of the clone revealed the presence of three A-genes (A1A2A3) encoding a-polypetides and of two B-genes (B1B2) encoding β-polypeptides of the 13800/850 complex. The arrangement of the different genes are B1A1, B2A2 and A3 where only B1 and B2 are preceded by typical Shine-Dalgamo sequences. In addition, typical nucleotide sequences for a rho-independent termination of transcription are located downstream of the genes A1 and A2. The deduced amino acid sequences revealed that the α-genes encoded for identical polypeptides, whereas the deduced β-polypeptides differed in their amino acid sequence at four positions. Transcriptional operon analysis revealed that the genes A1B1 and A2B2 are both dicistronically transcribed, whereas the gene A3 is not

Unexpected similarities of the B800-850 light-harvesting complex from Rhodo-spirillum molischianum to the B870 light-harvesting complexes from other purple photosynthetic bacteria

The B800-850 light-harvesting complex (also called LH2) was isolated from photosynthetic membranes of Rhodospirillum molischianum DSM 119 using molecular sieve and ion-exchange chromatography. Its two bacteriochlorophyll a-binding polypeptides (alpha-subunit and beta-subunit) were purified with a reverse-phase HPLC system. The complete amino acid sequences of both subunits have been determined. The alpha- and beta-subunits consist of 56 and 45 amino acids, respectively, corresponding to molecular weights of 5939 and 5133. In contrast to the B800-850 complexes from other photosynthetic bacteria, the native B800-850 complex from Rs. molischianum is most likely an octamer of monomers with a stoichiometry of three bacteriochlorophyll a and 1.5 lycopenes per alpha,beta-subunit. Resonance Raman spectra provide evidence for a 5-coordinated Mg2+ in the BChl, and a carotenoid mainly in the all-trans configuration. A comparison between resonance Raman data from different photosynthetic bacteria indicates that the BChl a-binding site of the B800-850 complex from Rs. molischianum is more similar to the B870 complexes (also called LH1) than to the B800-850 complexes of other photosynthetic bacteria. Sequence similarities especially between the beta-subunits of the B800-850 complex of Rs. molischianum and the B870 and B800-850 complexes of other photosynthetic bacteria agree with this result and provide information on the mode of pigment binding in bacterial antenna complexes