Imaging trapped quantum gases by off-axis holography.

We present a dispersive imaging method for trapped quantum gases based on digital off-axis holography. Both phase delay and intensity of the probe field are determined from the same image. Due to the heterodyne gain inherent to the holographic method, it is possible to retrieve the phase delay induced by the atoms at probe beam doses two orders of magnitude lower than phase-contrast imaging methods. Using the full field of the probe beam, we numerically correct for image defocusing.

Observation of a Space-Time Crystal in a Superfluid Quantum Gas.

Time crystals are a phase of matter, for which the discrete time symmetry of the driving Hamiltonian is spontaneously broken. The breaking of discrete time symmetry has been observed in several experiments in driven spin systems. Here, we show the observation of a space-time crystal using ultracold atoms, where the periodic structure in both space and time is directly visible in the experimental images. The underlying physics in our superfluid can be described ab initio and allows for a clear identification of the mechanism that causes the spontaneous symmetry breaking. Our results pave the way for the usage of space-time crystals for the discovery of novel nonequilibrium phases of matter.

The calreticulin (CALR) exon 9 mutations are promising targets for cancer immune therapy.

The calreticulin (CALR) exon 9 mutations are found in ∼30% of patients with essential thrombocythemia and primary myelofibrosis. Recently, we reported spontaneous immune responses against the CALR mutations. Here, we describe that CALR-mutant (CALRmut)-specific T cells are able to specifically recognize CALRmut cells. First, we established a T-cell culture specific for a CALRmut epitope. These specific T cells were able to recognize several epitopes in the CALRmut C terminus. Next, we established a CALRmut-specific CD4+ T-cell clone by limiting dilution. These CD4+ T cells recognized autologous CALRmut monocytes and hematopoietic stem cells, and T-cell recognition of target cells was dependent on the presence of CALR. Furthermore, we showed that the CALRmut response was human leukocyte antigen (HLA)-DR restricted. Finally, we demonstrated that the CALRmut-specific CD4+ T cells, despite their phenotype, were cytotoxic to autologous CALRmut cells, and that the cytotoxicity was mediated by degranulation of the T cells. In conclusion, the CALR exon 9 mutations are targets for specific T cells and thus are promising targets for cancer immune therapy such as peptide vaccination in patients harboring CALR exon 9 mutations.

Quantum Enhancement of the Index of Refraction in a Bose-Einstein Condensate.

We study the index of refraction of an ultracold bosonic gas in the dilute regime. Using phase-contrast imaging with light detuned from resonance by several tens of linewidths, we image a single cloud of ultracold atoms for 100 consecutive shots, which enables the study of the scattering rate as a function of temperature and density using only a single cloud. We observe that the scattering rate is increased below the critical temperature for Bose-Einstein condensation by a factor of 3 compared to the single-atom scattering rate. We show that current atom-light interaction models to second order of the density show a similar increase, where the magnitude of the effect depends on the model that is used to calculate the pair-correlation function. This confirms that the effect of quantum statistics on the index of refraction is dominant in this regime.

CD19-Chimeric Antigen Receptor T Cells for Treatment of Chronic Lymphocytic Leukaemia and Acute Lymphoblastic Leukaemia.

Adoptive cell therapy (ACT) for cancer represents a promising new treatment modality. ACT based on the administration of cytotoxic T cells genetically engineered to express a chimeric antigen receptor (CAR) recognizing CD19 expressed by B cell malignancies has been shown to induce complete lasting responses in patients with chronic lymphocytic leukaemia (CLL) and acute lymphoblastic leukaemia (ALL). So far, eleven clinical trials including 99 CLL and ALL patients treated with CAR T cells targeting CD19 have been published, and the results from these trials are promising with impressive clinical responses in heavily pretreated patients. Thus, CAR T cell therapy has induced complete responses in both CLL and ALL, and surprisingly, current results indicate that patients with ALL are more prone to respond than are CLL patients. Importantly, the majority of CAR cell studies have observed severe therapy-associated toxicities, which needs attention. Herein we review current data and discuss key aspects of this powerful approach to treat and potentially cure B cell malignancies.

5-Azacytidine treatment sensitizes tumor cells to T-cell mediated cytotoxicity and modulates NK cells in patients with myeloid malignancies.

Treatment with the demethylating agent 5-Azacytidine leads to prolonged survival for patients with myelodysplastic syndrome, and the demethylation induces upregulation of cancer-testis antigens. Cancer-testis antigens are well-known targets for immune recognition in cancer, and the immune system may have a role in this treatment regimen. We show here that 5-Azacytidine treatment leads to increased T-cell recognition of tumor cells. T-cell responses against a large panel of cancer-testis antigens were detected before treatment, and these responses were further induced upon initiation of treatment. These characteristics point to an ideal combination of 5-Azacytidine and immune therapy to preferentially boost T-cell responses against cancer-testis antigens. To initiate such combination therapy, essential knowledge is required about the general immune modulatory effect of 5-Azacytidine. We therefore examined potential treatment effects on both immune stimulatory (CD8 and CD4 T cells and Natural Killer (NK) cells) and immune inhibitory cell subsets (myeloid-derived suppressor cells and regulatory T cells). We observed a minor decrease and modulation of NK cells, but for all other populations no effects could be detected. Together, these data support a strategy for combining 5-Azacytidine treatment with immune therapy for potential clinical benefit.

Characterization and comparison of 'standard' and 'young' tumour-infiltrating lymphocytes for adoptive cell therapy at a Danish translational research institution.

Adoptive cell therapy (ACT) with ex vivo expanded tumour-infiltrating lymphocytes (TILs) in combination with IL-2 is an effective treatment for metastatic melanoma. Modified protocols of cell expansion may allow the treatment of most enrolled patients and improve the efficacy of adoptively transferred cells. The aims of this study were to establish and validate the novel 'Young TIL' method at our institution and perform a head-to-head comparison of clinical-grade products generated with this protocol opposed to the conventional 'Standard TIL', which we are currently using in a pilot ACT trial for patients with melanoma. Our results confirm that 'Young TILs' display an earlier differentiation state, with higher CD27 and lower CD56 expression. In addition, CD8(+) TILs expressing CD27 had longer telomeres compared with the CD27(-). A recently described subset of NK cells, endowed with a high expression of CD56 (CD56(bright)), was detected for the first time in both types of cultures but at a higher frequency on Young TILs. Young and Standard TILs' reactivity against autologous tumours was similar, with significant expression of TNF-α/IFN-γ/CD107a by CD8(+) TILs detected in all cultures analysed. However, either slow expansion with high-dose IL-2 only or large numerical expansion with a rapid expansion protocol, which is required for current therapeutic protocols, significantly modified TIL phenotype by reducing the frequency of less differentiated, cancer-specific TILs. These studies further support the adoption of the Young TIL method in our current ACT trial and highlight the importance of continuous quality control of expansion protocols.

Changes in peripheral blood level of regulatory T cells in patients with malignant melanoma during treatment with dendritic cell vaccination and low-dose IL-2.

In this study, changes in peripheral blood regulatory T cell (Treg) levels were evaluated in 46 progressive patients with melanoma treated with a dendritic cell-based vaccine and concomitant low-dose IFN-α and IL-2. The regulatory subset of CD4 T cells, characterized by CD25(high) , was prospectively analysed in fresh blood, and treatment-associated quantitative and qualitative changes were analysed. By the 4th vaccine, patients showed a marked increase in CD4+ CD25(high) T cell subset from 6% to 22% (P<0.001). At the 6th vaccine, a general decline was observed and a significantly (P=0.01) lower level of CD4+ CD25(high) Treg cells was reached in the group of patients who attained disease stabilization (9.5%) compared to patients with continued progressive disease (14.5%). However, when FoxP3 was employed for retrospective analysis of Tregs on frozen blood, this difference did not reach significance (P=0.09). The vast majority of the Treg produced IL-10 and, to a varying extent, TGF-ß. In addition, sorted CD4+ CD25(high) CD127⁻ Tregs were able to suppress proliferation of peripheral blood mononuclear cells in a dose-dependent manner, thus suggesting a regulatory functionality. These findings emphasize the need for strategies to effectively eliminate Treg cells to optimize the clinical effectiveness of cancer immunotherapy.

Enhanced heat flow in the hydrodynamic collisionless regime.

We study the heat conduction of a cold, thermal cloud in a highly asymmetric trap. The cloud is axially hydrodynamic, but due to the asymmetric trap radially collisionless. By locally heating the cloud we excite a thermal dipole mode and measure its oscillation frequency and damping rate. We find an unexpectedly large heat conduction compared to the homogeneous case. The enhanced heat conduction in this regime is partially caused by atoms with a high angular momentum spiraling in trajectories around the core of the cloud. Since atoms in these trajectories are almost collisionless they strongly contribute to the heat transfer. We observe a second, oscillating hydrodynamic mode, which we identify as a standing wave sound mode.

¿Cómo leen los sordos adultos? La estrategia de palabras clave / How do deaf adults read? The key word strategy

El objetivo principal de este estudio es determinar los mecanismos de lectura utilizados por lectoressordos adultos de lengua española o francesa, quehabían terminado su escolaridad obligatoria yseguían o habían seguido estudios superiores aella. Todos leían regularmente periódicos y librospor información y placer. La hipótesis principal deeste trabajo es que estos lectores utilizan unaestrategia de palabras clave para leer. Esta estrategiaconsiste en identificar algunas de las palabrasde cada frase y elaborar su significado global apartir de éstas. Las palabras identificadas eran, engeneral, sustantivos, adjetivos y/o verbos frecuentesque permiten una interpretación semánticasimple de la frase. La utilización de esta estrategiase debe, probablemente, a habilidades morfosintácticasinsuficientes. Los resultados en ambosgrupos lingüísticos apoyan las predicciones basadasen la hipótesis de estrategia de palabras clave.Además, los participantes realizaron una prueba deortografía léxica en la que tenían que elegir entrevarios ítems homófonos, aquel que estaba correctamenteescrito. Los resultados muestran que, engeneral, los participantes sordos poseen un léxicoortográfico más rico que los oyentes del mismonivel lector. Este hecho es coherente con la estrategiade palabras clave que exige la capacidad deidentificar las palabras principales de una frase.Finalmente, los participantes realizaron una seriede pruebas metafonológicas. Los resultados indicanque su nivel de aciertos es ligeramente inferioral de los oyentes, lo que muestra que poseen representacionesfonológicas de las palabras que conoceny son capaces de manipularlas interiormentepara realizar estas tareas (AU)

The aim of this study is to determine the readingmechanisms used by French or Spanish-speakingdeaf adults, who had finished their compulsorystudies and were attending or had followed secondaryor superior studies. They regularly read newspapersand books for pleasure and information.Our main hypothesis was that they read using akey word strategy. This strategy consists of identifyingsome of the words of the sentence to be readand deriving the general meaning solely on thisbasis. The words selected are basically frequentnames, adjectives and/or verbs allowing a simplesemantic interpretation. The use of this strategyresulted from poor control of morpho-syntacticconstraints of the sentences. The results fulfilledall the predictions derived from this hypothesis inboth language groups. In addition, the participantswere presented with a spelling task in which theyhad to choose the correct spelling of a word fromseveral homophonic items. The results showed thatthe deaf participants possessed an orthographiclexicon richer than hearing controls of the samereading level. That deaf readers possess a hugeorthographic lexicon is in harmony with the keyword strategy, which requires the ability to identifyat least some of the main words of the sentence.Finally, most of the deaf participants reachedscores slightly under the level of the hearing controls in metaphonological tasks. This showedthat they do possess phonological representationsof the words they know and that they are able tomanipulate them in order to do the metaphonologicaltasks (AU)

Damping of superfluid flow by a thermal cloud.

One of the principal signatures of superfluidity is the frictionless flow of a superfluid through another substance. Here, we study the flow of a Bose-Einstein condensate through a thermal cloud and study its damping for different harmonic confinements and temperatures. The damping rates close to the collisionless regime are found to be in good agreement with Landau damping and become smaller for more homogeneous systems. In the hydrodynamic regime, we observe additional damping due to collisions, and we discuss the implications of these findings for superfluidity in this system.

The CIMT-monitoring panel: a two-step approach to harmonize the enumeration of antigen-specific CD8+ T lymphocytes by structural and functional assays.

The interpretation of the results obtained from immunomonitoring of clinical trials is a difficult task due to the variety of methods and protocols available to detect vaccine-specific T-cell responses. This heterogeneity as well as the lack of standards has led to significant scepticism towards published results. In February 2005, a working group was therefore founded under the aegis of the Association for Immunotherapy of Cancer ("CIMT") in order to compare techniques and protocols applied for the enumeration of antigen-specific T-cell responses. Here we present the results from two consecutive phases of an international inter-laboratory testing project referred to as the "CIMT monitoring panel". A total of 13 centers from six European countries participated in the study in which pre-tested PBMC samples, synthetic peptides and PE-conjugated HLA-tetramers were prepared centrally and distributed to participants. All were asked to determine the number of antigen-specific T-cells in each sample using tetramer staining and one functional assay. The results of the first testing round revealed that the total number of cells analyzed was the most important determinant for the sensitive detection of antigen-specific CD8(+) T-cells by tetramer staining. Analysis by ELISPOT was influenced by a combination of cell number and a resting phase after thawing of peripheral blood mononuclear cells. Therefore, the experiments were repeated in a second phase but now the participants were asked to change their protocols according to the new guidelines distilled from the results of the first phase. The recommendations improved the number of antigen-specific T-cell responses that were detected and decreased the variability between the laboratories. We conclude that a two-step approach in inter-laboratory testing allows the identification of distinct variables that influence the sensitivity of different T-cell assays and to formally show that a defined correction to the protocols successfully increases the sensitivity and reduces the inter-center variability. Such "two-step" inter-laboratory projects could define rational bases for accepted international guidelines and thereby lead to the harmonization of the techniques used for immune monitoring.

Large atom number Bose-Einstein condensate of sodium.

We describe the setup to create a large Bose-Einstein condensate containing more than 120 x 10(6) atoms. In the experiment a thermal beam is slowed by a Zeeman slower and captured in a dark-spot magneto-optical trap (MOT). A typical dark-spot MOT in our experiments contains 2.0 x 10(10) atoms with a temperature of 320 microK and a density of about 1.0 x 10(11) atoms/cm(3). The sample is spin polarized in a high magnetic field before the atoms are loaded in the magnetic trap. Spin polarizing in a high magnetic field results in an increase in the transfer efficiency by a factor of 2 compared to experiments without spin polarizing. In the magnetic trap the cloud is cooled to degeneracy in 50 s by evaporative cooling. To suppress the three-body losses at the end of the evaporation, the magnetic trap is decompressed in the axial direction.

Coherent optical transfer of Feshbach molecules to a lower vibrational state.

Using the technique of stimulated Raman adiabatic passage (STIRAP) we have coherently transferred ultracold (87)Rb(2) Feshbach molecules into a more deeply bound vibrational quantum level. Our measurements indicate a high transfer efficiency of up to 87%. Because the molecules are held in an optical lattice with not more than a single molecule per lattice site, inelastic collisions between the molecules are suppressed and we observe long molecular lifetimes of about 1 s. Using STIRAP we have created quantum superpositions of the two molecular states and tested their coherence interferometrically. These results represent an important step towards Bose-Einstein condensation of molecules in the vibrational ground state.

Evidence for involvement of clonally expanded CD8+ T cells in anticancer immune responses in CLL patients following nonmyeloablative conditioning and hematopoietic cell transplantation.

We have analyzed the clonotype composition of CD8+ T cells following nonmyeloablative (NMA) conditioning and hematopoietic cell transplantation (HCT), of patients with chronic lymphocytic leukemia (CLL). Consecutive analyses of blood samples taken up to 2 years following HCT, demonstrated that CD8+ T-cell clonality was highly dynamic in the early phases after HCT, but became more stable after 4-5 months. Moreover, donor lymphocyte infusion (DLI) given for disease progression in one of the patients led to establishment of recurrent as well as new T-cell clonotypes. This coincided with disease remission, strongly suggesting that these T cells were engaged with anti-CLL cytotoxicity. To examine the functional capacity of stable clonally expanded T cells after HCT, CD8+ T cells isolated post-transplant from the recipients were stimulated ex vivo with CLL cells and subsequently analyzed by FACS for surface expression of the marker for cytotoxic activity, CD107a. Stimulation with CLL cells indeed led to surface expression of CD107a, and clonotype analyses of sorted cells demonstrated that CD107a positive T cells were stably expanded following HCT. Our data suggest that clonally expanded CD8+ T-cell clones participate in the ongoing T-cell response against CLL cells following HCT with NMA conditioning.