Which clinical factors delay proper treatment in panic disorder? A cross-sectional multicentric study.

AIM: The aim of the present study was to identify clinical and socio-demographic factors associated with duration of untreated illness (DUI) in patients affected by panic disorder (PD). METHODS: Data were collected from patients' medical records (N = 157) of two mental health services respectively located in Milan and in Monza (Italy). Correlation analyses and analysis of variance (ANOVAs) were run to analyse the relation between DUI and quantitative/qualitative variables respectively. Statistically significant variables in uni- variate analyses were then inserted in a linear multivariable regression model (backward procedure). RESULTS: Mean DUI was 27.33 (±50.56) months. Patients with an earlier age at onset (r = -0.270; p < .01), a longer duration of illness (r = 0.483; p < .01) and who received a lifetime psychotherapy (F = 6.86; p = .01) had a longer DUI. The final global model showed that a longer DUI was associated with pre-onset poly-substance misuse (p = .05) and a longer duration of illness (p < .01). CONCLUSION: The results of our study showed that a longer DUI was predicted by clinical factors such as the presence of a pre-onset poly-substance use disorder and that delayed proper treatment can lead to a chronicization of PD, as indicated by a longer duration of illness. Further studies are needed to in-depth investigate the role of DUI in influencing the course and outcome of anxiety disorders, including PD.

Early colonisation and temporal dynamics of the gut microbial ecosystem in Standardbred foals.

BACKGROUND: Even if horses strictly depend on the gut microbiota for energy homeostasis, only a few molecular studies have focused on its characterisation and none on the perinatal gut microbial colonisation process. OBJECTIVES: To explore the perinatal colonisation process of the foal gut microbial ecosystem and the temporal dynamics of the ecosystem assembly during the first days of life. STUDY DESIGN: Longitudinal study. METHODS: Thirteen Standardbred mare-foal pairs were included in the study. For each pair, at delivery we collected the mare amniotic fluid, faeces and colostrum, and the foal meconium. Milk samples and faeces of both mare and foal were also taken longitudinally, until day 10 post-partum. Samples were analysed by means of next-generation sequencing of the 16S rRNA gene on Illumina MiSeq. RESULTS: Our findings suggest that microbial components derived from the mare symbiont communities establishes in the foal gut since fetal life. After birth, an external transmission route of mare microorganisms takes place. This involves a rapid and dynamic process of assembling the mature foal gut microbiome, in which the founder microbial species are derived from both the milk and the gut microbial ecosystems of the mare. MAIN LIMITATIONS: The inability to discriminate between live and dead cells, the possible presence of contaminating bacteria in low biomass samples (e.g. meconium and amniotic fluid), the limits of the phylogenetic assignment down to species level, and the presence of unassigned operational taxonomic units. CONCLUSIONS: Our data highlight the importance of mare microbiomes as a key factor for the establishment of the gut microbial ecosystem of the foal.

Vaccination to improve the persistence of CD19CAR gene-modified T cells in relapsed pediatric acute lymphoblastic leukemia.

Trials with second generation CD19 chimeric antigen receptors (CAR) T-cells report unprecedented responses but are associated with risk of cytokine release syndrome (CRS). Instead, we studied the use of donor Epstein-Barr virus-specific T-cells (EBV CTL) transduced with a first generation CD19CAR, relying on the endogenous T-cell receptor for proliferation. We conducted a multi-center phase I/II study of donor CD19CAR transduced EBV CTL in pediatric acute lymphoblastic leukaemia (ALL). Patients were eligible pre-emptively if they developed molecular relapse (>5 × 10-4) post first stem cell transplant (SCT), or prophylactically post second SCT. An initial cohort showed poor expansion/persistence. We therefore investigated EBV-directed vaccination to enhance expansion/persistence. Eleven patients were treated. No CRS, neurotoxicity or graft versus host disease (GVHD) was observed. At 1 month, 5 patients were in CR (4 continuing, 1 de novo), 1 PR, 3 had stable disease and 3 no response. At a median follow-up of 12 months, 10 of 11 have relapsed, 2 are alive with disease and 1 alive in CR 3 years. Although CD19CAR CTL expansion was poor, persistence was enhanced by vaccination. Median persistence was 0 (range: 0-28) days without vaccination compared to 56 (range: 0-221) days with vaccination (P=0.06). This study demonstrates the feasibility of multi-center studies of CAR T cell therapy and the potential for enhancing persistence with vaccination.

Gut microbiota and hematopoietic stem cell transplantation: where do we stand?

Advances in biological techniques have potentiated great progresses in understanding the interaction between human beings and the ∼10 to 100 trillion microbes living in their gastrointestinal tract: gut microbiota (GM). In this review, we describe recent emerging data on the role of GM in hematopoietic stem cell transplantation, with a focus on immunomodulatory properties in the immune system recovery and the impact in the development of the main complications, as GvHD and infections.

Wide bandwidth fiber-optic ultrasound probe in MOMS technology: Preliminary signal processing results.

An ultrasonic probe consisting of two optical fiber-based miniaturized transducers for wideband ultrasound emission and detection is employed for the characterization of in vitro biological tissues. In the probe, ultrasound generation is obtained by thermoelastic emission from patterned carbon films in Micro-Opto-Mechanical-System (MOMS) devices mounted on the tip of an optical fiber, whereas acousto-optical detection is performed in a similar way by a miniaturized polymeric interferometer. The microprobe presents a wide, flat bandwidth that is a very attractive feature for ultrasonic investigation, especially for tissue characterization. Thanks to the very high ultrasonic frequencies obtained, the probe is able to reveal different details of the object under investigation by analyzing the ultrasonic signal within different frequencies ranges, as shown by specific experiments performed on a patterned cornstarch flour sample in vitro. This is confirmed by measurements executed to determine the lateral resolution of the microprobe at different frequencies of about 70µm at 120MHz. Moreover, measurements performed with the wideband probe in pulsed-echo mode on a histological finding of porcine kidney are presented, on which two different spectral signal processing algorithms are applied. After processing, the ultrasonic spectral features show a peculiar spatial distribution on the sample, which is expected to depend on different ultrasonic backscattering properties of the analyzed tissues.

Gut microbiota trajectory in pediatric patients undergoing hematopoietic SCT.

Acute GvHD (aGvHD) is the main complication of hematopoietic SCT (HSCT) during the treatment of hematological disorders. We carried out the first longitudinal study to follow the gut microbiota trajectory, from both the phylogenetic and functional points of view, in pediatric patients undergoing HSCT. Gut microbiota trajectories and short-chain fatty acid production profiles were followed starting from before HSCT and through the 3-4 months after transplant in children developing and not developing aGvHD. According to our findings, HSCT procedures temporarily cause a structural and functional disruption of the gut microbial ecosystem, describing a trajectory of recovery during the following 100 days. The onset of aGvHD is associated with specific gut microbiota signatures both along the course of gut microbiota reconstruction immediately after transplant and, most interestingly, prior to HSCT. Indeed, in pre-HSCT samples, non-aGvHD patients showed higher abundances of propionate-producing Bacteroidetes, highly adaptable microbiome mutualists that showed to persist during the HSCT-induced ecosystem disruption. Our data indicate that structure and temporal dynamics of the gut microbial ecosystem can be a relevant factor for the success of HSCT and opens the perspective to the manipulation of the pre-HSCT gut microbiota configuration to favor mutualistic persisters with immunomodulatory properties in the gut.

Cell-based strategies to manage leukemia relapse: efficacy and feasibility of immunotherapy approaches.

When treatment fails, the clinical outcome of acute leukemia patients is usually very poor, particularly when failure occurs after transplantation. A second allogeneic stem cell transplant could be envisaged as an effective and feasible salvage option in younger patients having a late relapse and an available donor. Unmanipulated or minimally manipulated donor T cells may also be effective in a minority of patients but the main limit remains the induction of severe graft-versus-host disease. This clinical complication has brought about a huge research effort that led to the development of leukemia-specific T-cell therapy aiming at the direct recognition of leukemia-specific rather than minor histocompatibility antigens. Despite a great scientific interest, the clinical feasibility of such an approach has proven to be quite problematic. To overcome this limitation, more research has moved toward the choice of targeting commonly expressed hematopoietic specific antigens by the genetic modification of unselected T cells. The best example of this is represented by the anti-CD19 chimeric antigen receptor (CD19.CAR) T cells. As a possible alternative to the genetic manipulation of unselected T cells, specific T-cell subpopulations with in vivo favorable homing and long-term survival properties have been genetically modified by CAR molecules. Finally, the use of naturally cytotoxic effector cells such as natural killer and cytokine-induced killer cells has been proposed in several clinical trials. The clinical development of these latter cells could also be further expanded by additional genetic modifications using the CAR technology.

Miniaturized fiber-optic ultrasound probes for endoscopic tissue analysis by micro-opto-mechanical technology.

A new Micro-Opto-Mechanical System (MOMS) technology for the fabrication of optoacoustic probes on optical fiber is presented. The technology is based on the thermoelastic emission of ultrasonic waves from patterned carbon films for generation and on extrinsic polymer Fabry-Perot acousto-optical transducers for detection, both fabricated on miniaturized single-crystal silicon frames used to mount the ultrasonic transducers on the tip of an optical fiber. Thanks to the fabrication process adopted, high miniaturization levels are reached in the MOMS devices, demonstrating fiber-optic emitters and detectors with minimum diameter around 350 and 250 µm respectively. A thorough functional testing of the ultrasound emitters mounted on 200 and 600 µm diameter optical fibers is presented, in which the fiber-optic emitter with a diameter of 200 µm shows generated acoustic pressures with peak-to-peak value up to 2.8 MPa with rather flat emission spectra extended beyond 150 MHz. The possibility to use the presented optoacoustic sources in conjunction with the fiber-optic acousto-optical detectors within a minimally invasive probe is also demonstrated by successfully measuring the ultrasonic echo reflected from a rigid surface immersed in water with various concentration of scatterers. The resulting spectra highlight the possibility to discriminate the effects due to frequency selective attenuation in a very wide range of frequencies within a biological medium using the presented fiber-optic probes.

Chimeric antigen receptors against CD33/CD123 antigens efficiently target primary acute myeloid leukemia cells in vivo.

As significant numbers of acute myeloid leukemia (AML) patients are still refractory to conventional therapies or experience relapse, immunotherapy using T cells expressing chimeric antigen receptors (CARs) might represent a valid treatment option. AML cells frequently overexpress the myeloid antigens CD33 and CD123, for which specific CARs can be generated. However, CD33 is also expressed on normal hematopoietic stem/progenitor cells (HSPCs), and its targeting could potentially impair normal hematopoiesis. In contrast, CD123 is widely expressed by AML, while low expression is detected on HSPCs, making it a much more attractive target. In this study we describe the in vivo efficacy and safety of using cytokine-induced killer (CIK) cells genetically modified to express anti-CD33 or anti-CD123 CAR to target AML. We show that both these modified T cells are very efficient in reducing leukemia burden in vivo, but only the anti-CD123 CAR has limited killing on normal HSPCs, thus making it a very attractive immunotherapeutic tool for AML treatment.

A probiotics-containing biscuit modulates the intestinal microbiota in the elderly.

OBJECTIVES: Evaluation of the impact of a biscuit containing the probiotics Bifidobacterium longum Bar33 and Lactobacillus helveticus Bar13 on the intestinal microbiota in the elderly. DESIGN: Randomized double-blind placebo-controlled trial. PARTICIPANTS: Thirty-two elderly volunteers living in Italy. The group was composed of 19 women and 13 men aged between 71 and 88 years (mean 76). INTERVENTION: Subjects were randomized in two groups consuming one dose of the probiotics-containing biscuit or placebo once a day for 30 days. MEASUREMENTS: For each subject the intestinal microbiota was characterized using the phylogenetic microarray platform HTF-Microbi.Array before and after intervention. RESULTS: Our data demonstrated that one-month consumption of a probiotics-containing biscuit was effective in redressing some of the age-related dysbioses of the intestinal microbiota. In particular, the probiotic treatment reverted the age-related increase of the opportunistic pathogens Clostridium cluster XI, Clostridium difficile, Clostridium perfringens, Enterococcus faecium and the enteropathogenic genus Campylobacter. CONCLUSION: The present study opens the way to the development of elderly-tailored probiotic-based functional foods to counteract the age-related dysbioses of the intestinal microbiota.

In Vitro and In Vivo Antitumor Effect of Anti-CD33 Chimeric Receptor-Expressing EBV-CTL against CD33 Acute Myeloid Leukemia.

Genetic engineering of T cells with chimeric T-cell receptors (CARs) is an attractive strategy to treat malignancies. It extends the range of antigens for adoptive T-cell immunotherapy, and major mechanisms of tumor escape are bypassed. With this strategy we redirected immune responses towards the CD33 antigen to target acute myeloid leukemia. To improve in vivo T-cell persistence, we modified human Epstein Barr Virus-(EBV-) specific cytotoxic T cells with an anti-CD33.CAR. Genetically modified T cells displayed EBV and HLA-unrestricted CD33 bispecificity in vitro. In addition, though showing a myeloablative activity, they did not irreversibly impair the clonogenic potential of normal CD34(+) hematopoietic progenitors. Moreover, after intravenous administration into CD33(+) human acute myeloid leukemia-bearing NOD-SCID mice, anti-CD33-EBV-specific T cells reached the tumor sites exerting antitumor activity in vivo. In conclusion, targeting CD33 by CAR-modified EBV-specific T cells may provide additional therapeutic benefit to AML patients as compared to conventional chemotherapy or transplantation regimens alone.

Estudio experimental sobre la utilización de células madre humanas en la terapia de los defectos periodontales: resultados preliminares / Experimental study on the use of human stem cell therapy in periodontal defects: Preliminary results

La destrucción del ligamento periodontal y de la cresta alveolar causada por la enfermedad periodontal, constituye la principal causa de pérdida dentaria en pacientes adultos. Resulta difícil que la terapia en la enfermedad periodontal avanzada lleve a una restitutio ad integrum de la estructura que se ha perdido por la imposibilidad de regenerar la correcta anatomía del sistema perdido. Este estudio tiene como objetivo evaluar el potencial y la estabilidad de la utilización de células madre mesenquimales (MSC) en la regeneración periodontal de la estructura original. Su objetivo es determinar un método de regeneración de la estructura periodontal original que se ha perdido después de la enfermedad periodontal, cerca de los elementos dentales vitales sin patologías correlacionadas. Este estudio constituye una experimentación de fase 1 aprobada por el Instituto Superior de Salud sobre un número limitado de pacientes, cuyo objetivo principal será la evaluación de la biocompatibilidad y toxicidad in vivo de las células madre mesenquimales diferenciadas en sentido osteoblástico, sostenidas por andamios de colágeno biomiméticos en pacientes afectados por periodontopatía severa. El protocolo describe el transplante de células madre mesenquimales derivadas de muestras de un aspirado percutáneo de médula ósea. Las células de la médula ósea son manipuladas ex vivo para el aislamiento de la población de células madre mesenquimales. Las células madre adultas son diferenciadas en las líneas celulares que forman el sistema periodontal, sucesivamente inducidas y cultivadas en andamios adecuados definidos en base a características propias con el fin de obtener una distribución uniforme de las líneas celulares, e injertadas en el sitio receptor estimulado adecuadamente hasta la degradación de los andamios, en un tiempo suficiente para garantizar la estabilidad de las diferentes líneas celulares y la recíproca interacción, con el fin de recrear la correcta anatomía estructural del periodonto perdido. Después de una fase de expansión celular, estas células se introducen en una estructura biomimética (andamios en colágeno), y se inducen a la diferenciación en sentido osteogénico. El producto final que consta de andamios y células mesenquimales diferenciadas en sentido osteoblástico, se instala por último en el defecto alveolar periodontal. La experimentación se basa en la posibilidad de utilizar células madre mesenquimales autólogas a partir de la médula ósea humana. La finalidad es la de estimular la regeneración de PDL cerca de la pared radicular y la formación de hueso nuevo sobre el PDL, beneficiándose de las técnicas de regeneración-reconstrucción ósea, de manera que se recree la correcta anatomía periodontal. Los resultados presentados en este trabajo conciernen el primer paciente tratado con esta metodología y afectarán los estudios siguientes para la prueba con una cobertura más amplia (AU)

Destruction of periodontal ligament and alveolar ridge rerorption caused by periodontal disease is the leading cause of tooth loss in adults. Therapy of advanced periodontal disease leads for a total recovery of the structure lost by the inability to regenerate the pristine correct anatomy system. The aim of this study is to assess the potential use of mesenchymal stem cells (MSCs) in the regeneration of original periodontal structure. This study is an experimental phase 1 approved by the Italian Institute of Health on a limited number of patients; main objective will be to assess the biocompatibility and toxicity in vivo of differentiated mesenchymal stem cells into osteoblast sense, supported by biomimetic collagen scaffolds in patients with severe periodontal disease. The protocol describes transplantation of mesenchymal stem cells derived from samples of percutaneous bone marrow aspirate. The bone marrow cells are manipulated ex vivo to isolate mesenchymal stem cell population. After a phase of cell expansion, these cells are induced to differentiate into periodontal system cell lines in a biomimetic structure (collagen scaffold). Induction and culturation in an appropriate scaffold defined on specific characteristics lead to obtain an uniform distribution of cell lines. After 28 days Scs are grafted into the defect site of the patient. Scaffold degradation time will ensure stability of the different cell lines and reciprocal interaction, in order to recreate the correct structural anatomy of periodontium lost. The purpose is to stimulate the regeneration of PDL near the root wall and new bone formation on the PDL, benefiting from the techniques of bone regeneration, reconstruction, as to recreate the pristine periodontal anatomy. The results presented in this paper concern the first patient treated with this method and will affect following studies for test for a wider coverage (AU)

Polyomavirus JC-targeted T-cell therapy for progressive multiple leukoencephalopathy in a hematopoietic cell transplantation recipient.

Progressive multifocal leukoencephalopathy (PML) associated with polyomavirus JC (JCV) infection has been reported to be usually fatal in allogeneic hematopoietic SCT (HSCT) recipients. We present the case of a 19-year-old HSCT patient diagnosed with JCV-associated PML after prolonged immunosuppression for severe GVHD. No short-term neurological improvement was observed after antiviral treatment and discontinuation of immunosuppressive therapy. Donor-derived JCV Ag-specific CTLs were generated in vitro after stimulation with 15-mer peptides derived from VP1 and large T viral proteins. After adoptive CTL infusion, virus-specific cytotoxic cells were shown in the peripheral blood, JCV-DNA was cleared in the cerebrospinal fluid and the patient showed remarkable improvement. Adoptive T-lymphocyte therapy with JCV-specific CTLs was feasible and had no side effects. This case suggests that adoptive transfer of JCV-targeted CTLs may contribute to restore JCV-specific immune competence and control PML in transplanted patients.

Selective elimination of a chemoresistant side population of B-CLL cells by cytotoxic T lymphocytes in subjects receiving an autologous hCD40L/IL-2 tumor vaccine.

Side-population (SP) analysis identifies precursor cells in normal and malignant tissues. Cells with this phenotype have increased resistance to many cytotoxic agents, and may represent a primary drug-resistant population in malignant diseases. To discover whether drug-resistant malignant SP cells are nonetheless sensitive to immune-mediated killing, we first established the presence of a malignant CD5(+)CD19(+) SP subset in the blood of 18/21 subjects with B-cell chronic lymphocytic leukemia (B-CLL). We examined the fate of these cells in six of these individuals who received autologous human CD40 ligand and interleukin-2 (hCD40L/IL-2) gene-modified tumor cells as part of a tumor vaccine study. Vaccinated patients showed an increase in B-CLL-reactive T cells followed by a corresponding decline in circulating CD5(+)CD19(+) SP cells. T-cell lines and clones generated from vaccinated patients specifically recognized B-CLL SP tumor cells. Elimination of SP cells is likely triggered by their increased expression of target antigens, such as receptor for hyaluronan-mediated motility (RHAMM), after stimulation of the malignant cells by hCD40L, as CD8(+) RHAMM-specific T cells could be detected in the peripheral blood of immunized patients and were associated with the decline in B-CLL SP cells. Hence, malignant B cells with a primary drug-resistant phenotype can be targeted by T- cell-mediated effector activity after immunization of human subjects.

A life-threatening paediatric case of acute autoimmune haemolytic anaemia (AIHA) successfully cured by plasma-exchange and combined immunosuppressive treatment.

A 3-year-old boy present with a severe autoimmune haemolytic anaemia, triggered by IgG-class auto-antibodies, with hemoglobin levels decreased to 2, 1 gr/dL. A combined immunosuppressive regimen was begun together with multiple plasma-exchanges and transfusions which sustained the cardio-vascular balance until the specific therapy became effective.

Quantitative variations in the vaginal bacterial population associated with asymptomatic infections: a real-time polymerase chain reaction study.

The real-time polymerase chain reaction (PCR) quantification of several vaginal bacterial groups in healthy women and patients developing asymptomatic bacterial vaginosis (BV) and candidiasis (CA) was performed. Statistical analysis revealed that the BV condition is characterised by a great variability among subjects and that it is associated with a significant increase of Prevotella, Atopobium, Veillonella and Gardnerella vaginalis, and a drop in Lactobacillus. On the contrary, the vaginal microflora of healthy women and patients developing CA was found to be homogeneous and stable over time.

A real-time 2-D vector Doppler system for clinical experimentation.

A real-time hardware software 2-D vector Doppler system has been realized by means of the FEMMINA platform. The system operates by performing two independent 1-D Doppler estimations on the scan plane of a linear array probe along different directions; the probe is connected to a commercial scanner. The reconstructed velocity is presented in real-time as superposition on the conventional B-mode images. Two different scanning techniques have been implemented, in order to carry out the 2-D Doppler investigation in the area of interest. These techniques allow to use the system both in vivo and in vivo. An extensive set of simulations has been performed in order to establish a gold standard regarding vector Doppler 2-D techniques, and to be able to assess the performance of the 2-D Doppler system by comparing simulated and experimental results. The whole real-time 2-D vector Doppler system is fully certified as hospital equipment, and thus it can be employed to carry out an experimental characterization of the 2-D Doppler technique in the clinical environment.

GMP-grade preparation of biomimetic scaffolds with osteo-differentiated autologous mesenchymal stromal cells for the treatment of alveolar bone resorption in periodontal disease.

BACKGROUND: Periodontal disease is a degenerative illness that leads to resorption of the alveolar bone. Mesenchymal stromal cells (MSC) represent a novel tool for the production of biologic constructs for the treatment of degenerative bone diseases. The preparation of MSC differentiated into osteogenic lineage for clinical use requires the fulfillment of strict good manufacturing practice (GMP) procedures. METHODS: MSC were isolated from BM samples and then cultured under GMP conditions. MSC were characterized phenotypically and for their differentiative potential. Cells were seeded onto collagen scaffolds (Gingistat) and induced to differentiate into osteogenic lineages using clinical grade drugs compared with standard osteogenic supplements. Alizarin Red S stain was used to test the deposition of the mineral matrix. Standard microbiologic analysis was performed to verify the product sterility. RESULTS: The resulting MSC were negative for CD33, CD34 and HLA-DR but showed high expression of CD90, CD105 and HLA-ABC (average expressions of 94.3%, 75.8% and 94.2%, respectively). Chondrogenic, osteogenic and adipogenic differentiation potential was demonstrated. The MSC retained their ability to differentiate into osteogenic lineage when seeded onto collagen scaffolds after exposure to a clinical grade medium. Cell numbers and cell viability were adequate for clinical use, and microbiologic assays demonstrated the absence of any contamination. DISCUSSION: In the specific context of a degenerative bone disease with limited involvement of skeletal tissue, the combined use of MSC, exposed to an osteogenic clinical grade medium, and biomimetic biodegradable scaffolds offers the possibility of producing adequate numbers of biologic tissue-engineered cell-based constructs for use in clinical trials.

T cells stimulated by CD40L positive leukemic blasts-pulsed dendritic cells meet optimal functional requirements for adoptive T-cell therapy.

Adoptive T-cell immunotherapy may provide complementary therapy for childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL). In this study, we have analyzed the functional characteristics of anti-BCP-ALL effector T cells generated by co-culturing T lymphocytes and dendritic cells (DC) from allogeneic human stem cell transplantation (HSCT) donors. After 21-day co-culture with DC pulsed with CD40L+ apoptotic BCP-ALL blasts, T cells presented with both effector and central memory phenotype, and showed high and specific cytotoxic activity against leukemic cells (average lysis = 77%), mostly mediated by CD8+ T cells. Noticeably, growth of CD4 T cells was maintained (45% of total cells), which actively produced Th1 cytokines (IFN-gamma, TNF-alpha, IL-2), but not IL-4, IL-5 and IL-10. Anti-BCP-ALL T cells expressed CD49d and CXCR4 (implicated in the recruitment to bone marrow), and CD62L and CCR7 (involved in the migration to lymphoid organs). In accordance with this profile, T cells significantly migrated in response to the chemokines CXCL12 and CCL19. In conclusion, stimulation of T cells with CD40L+BCP-ALL cells-loaded DC not only elicited the generation of potent and specific anti-leukemic cytotoxic effectors, but also the differentiation of specific and functional Th-1 CD4 lymphocytes. These effectors are fully equipped to reach leukemia-infiltrated tissues and have characteristics to support and orchestrate the anti-tumor immune-response.