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A synthetic analog of ceramide (AD2725) elevated ceramide levels in MDA-MB-435 breast cancer cells inducing apoptosis and resulted in cell death. In animal studies, using a model of xenograft breast cancer cancer cells in nude mice, treatment with AD2725 resulted in a significant reduction of the tumor volume and weight, and a significantly prolonged survival of the treated mice. The analog also significantly elevated INF gamma and IL-12 cytokine levels and reduced the level of IL-10 in the plasma. These data indicate that use of suitable synthetic analogs that most probably elevate the ceramide levels may have beneficial anti-cancer therapeutic effects.

Ninety-one children and adolescents 18 years or younger after allogeneic hematopoietic stem cell transplantation (HSCT) for relapsed or refractory Hodgkin lymphoma (HL) were analyzed. Fifty-one patients received reduced intensity conditioning (RIC); 40 patients received myeloablative conditioning (MAC). Nonrelapse mortality (NRM) at 1 year was 21% (± 4%), with comparable results after RIC or MAC. Probabilities of relapse at 2 and 5 years were 36% (± 5%) and 44% (± 6%), respectively. RIC was associated with an increased relapse risk compared with MAC; most apparent beginning 9 months after HSCT (P = .01). Progression-free survival (PFS) was 40% (± 6%) and 30% (± 6%) and overall survival (OS) was 54% (± 6%) and 45% (± 6%) at 2 and 5 years, respectively. Disease status at HSCT was predictive of PFS in multivariate analysis (P < .001). Beyond 9 months, PFS after RIC was lower compared with MAC (P = .02). Graft-versus-host disease did not affect relapse rate and PFS. In conclusion, children and adolescents with recurring HL show reasonable results with allogeneic HSCT. Especially patients allografted in recent years with good performance status and chemosensitive disease show highly encouraging results (PFS: 60% ± 27%, OS: 83% ± 15% at 3 years). Because relapse remains the major cause of treatment failure, additional efforts to improve disease control are necessary.

After allogeneic bone marrow transplantation (BMT) for leukemia, beneficial graft-vs-leukemia (GVL) effects are usually accompanied by potentially serious graft-vs-host disease (GVHD). Because T cell depletion is the only effective way to prevent GVHD it seems important to understand whether effective GVL can develop after BMT with T cell depletion in GVHD-free recipients. Well-established C57BL/6-->BALB/c chimeras that were free of GVHD, reconstituted with T cell-depleted allogeneic bone marrow cells, and inoculated 3 mo after BMT with a high inoculation of murine B cell leukemia (BCL1) showed no evidence of disease, whereas all control mice developed leukemia and died within 58 days. Results from adoptive transfer experiments in secondary naive BALB/c recipients indicated that all BCL1 cells were eliminated in the chimeras within 14 days. Hence, complete resistance to BCL1 developed in the chimeras despite complete tolerance to host alloantigens. The GVL effects observed in tolerant chimeras were further amplified by administration of immunocompetent allogeneic C57BL/6 spleen cells, low dose rIL-2, or both for 5 days. Our data suggest that GVL effects can develop even after T cell depletion in the absence of clinically overt GVHD and that GVL can be further amplified by rIL-2, either with or without use of additional immunocompetent donor T cells. Our data may provide the basis for new approaches to induce effective GVL after allogeneic BMT with cell therapy and rIL-2 at the stage of minimal residual disease, while avoiding early GVHD induced by the BMT procedure.

We present initial results of WIYN spectroscopic observations of selected objects detected in the Chandra Multiwavelength Plane (ChaMPlane) Survey in fields towards the Galactic anti-center. ChaMPlane is designed to identify low luminosity X-ray sources, both accretion-powered and stellar coronal, in the Galaxy. It also includes a wider-fields optical imaging Survey conducted with the NOAO Mosaic cameras to identify optical counterparts as well as H alpha-selected objects in the ~5 times larger field. We report spectra for 1048 objects in galactic anti-center (i.e. northern) fields, resulting in 609 type determinations. These include 5 new cataclysmic variables, 4 Be stars, 14 lithium-absorption stars, 180 stellar coronal sources (primarily dMe stars), and 29 new quasars. Bright optical counterparts of Chandra sources in this sample are most frequently dMe stars whereas a majority of the faintest (R ~21) spectroscopically classified Chandra source counterparts are quasars. The bulk of H alpha-selected sources appears to be roughly evenly divided between dMe stars and M stars at all magnitudes., Comment: 30 pages including 9 figures

A propos d'une observation: malade de 16 ans atteint d'anemie aplasique severe resistant aux traitements classiques, ayant recu une greffe de moelle d'un frere HLA et DR-identique, presente 9 mois plus tard une maladie GVH chronique se presentant comme une sclerodermie avec ulcerations. Des greffes de peau du donneur de moelle donnent de tres bons resultats

Patients with coincidental rheumatoid arthritis (RA) treated by allogeneic hematopoietic stem cell transplantation (HSCT) for drug induced aplastic anemia have been fortuitously cured of RA. Other than these examples with allogeneic HSCT, there is no known curative therapy for RA. Despite its potential to cure, allogeneic transplantation is not being offered to patients with RA due to transplant related mortality. Advances in HSCT conditioning regimens and better prevention of graft-versus-host disease should allow consideration of allogeneic HSCT as therapy for severe RA. We propose a new, well tolerated, nonmyeloablative allogeneic stem cell transplant regimen as treatment for RA.

During the last couple of years, observers have started to make plans for a Virtual Observatory, as a federation of existing data bases, connected through levels of software that enable rapid searches, correlations, and various forms of data mining. We propose to extend the notion of a Virtual Observatory by adding archives of simulations, together with interactive query and visualization capabilities, as well as ways to simulate observations of simulations in order to compare them with observations. For this purpose, we have already organized two small workshops, earlier in 2001, in Tucson and Aspen. We have also provided concrete examples of theory data, designed to be federated with a Virtual Observatory. These data stem from a project to construct an archive for our large-scale simulations using the GRAPE-6 (a 32-Teraflops special purpose computer for stellar dynamics). We are constructing interfaces by which remote observers can observe these simulations. In addition, these data will enable detailed comparisons between different simulations., Comment: 4 pages, ADASS-XI conference, http://bima.astro.umd.edu/nemo/nvo/

OBJECTIVE: The goals of this work were to develop animal models of lymphedema and tissue flap transfer, and to observe physiologic changes in lymphatic function that occur in these models over time, both systemically with lymphoscintigraphy (LS) and locally using fluorescence microlymphangiography (FM). SUMMARY BACKGROUND DATA: Although lymphedema has been managed by a combination of medical and surgical approaches, no effective long-term cure exists. Surgical attempts aimed at reconnecting impaired lymphatic channels or bypassing obstructed areas have failed. METHODS: The tails of rats (A groups) and mice (B groups) were used because of their different features. Lymphedema was created by ligation of the lymphatics at the tail base and quantified by diameter measurements there. In the experimental group, rectus abdominis myocutaneous flap was transferred across the ligation. In addition to the ligation (A1 and B1) and ligation + flap (A2 and B2) groups, three control groups were included: sham flap with ligation (B4), sham flap alone (B5), and normal (A3 and B3) animals. Observations were made at weekly time points for lymphatic function and continuity. RESULTS: Lymphedema was successfully created in the mouse ligation groups (B1 and B4) and sustained for the entire length of observation (up to 14 weeks). Lymphatic continuity was restored in those animals with transferred flaps across the ligation site (A2 and B2), as seen both by LS and FM. Sham flaps did not visibly affect lymphatic function nor did they cause any visible swelling in the tail. CONCLUSIONS: Acute lymphedema developing after ligation of tail lymphatics in mice can be prevented by myocutaneous flap transfer. Restored lymphatic continuity and function were demonstrable using lymphoscintigraphy and fluorescence microlymphangiography.

An sIL-6R/IL-6 chimera, directly fusing the natural forms of soluble IL-6 receptor and IL-6, as found in human body fluids, was produced in transfected human cells. The secreted p85 glycoprotein was active at a concentration of 120 pM to produce growth-arrest and spindleoid differentiation of murine melanoma F10.9 cells, which do not respond to IL-6 alone. This fusion protein was as active as the yeast-produced p56 fusion protein containing a shortened sIL-6R, linked through a flexible peptide chain to IL-6 (Hyper IL-6). The concentration of Hyper IL-6 needed to arrest the growth of F10.9 cells was much lower than that needed of a combination of IL-6 and sIL-6R, added separately. Hyper IL-6 was also more active than IL-6 in stimulating growth of murine plasmacytoma T1165 cells, the half maximal stimulation being obtained at 2 pM Hyper IL-6 versus 23 pM for IL-6. In order to evaluate the effect of the fused sIL-6R/IL-6 proteins on human hematopoietic primitive progenitor cells, they were added to suspension cultures of CD34+ cells from human cord blood in addition to both flt3/flk2 ligand (FL) and stem cell factor (SCF). Fused sIL-6R/IL-6 produced a marked stimulation of cell expansion and a marked increase in the number of colony forming units when subsequently plated in semi-solid medium with IL-3, GM-CSF, SCF and erythropoietin. Ex-vivo maintenance and expansion of early progenitor cells in bone marrow transplantation protocols may be a potential application for the sIL-6R/IL-6 chimeric glycoproteins.

Background: Gene therapy (GT) with hematopoietic stem cells is a promising treatment for inherited immunodeficiencies. Objectives: Limited information is available on the relative contribution of de novo thymopoiesis and peripheral expansion to T-cell reconstitution after GT as well as on the potential effects of gene transfer on hematopoietic stem cells and lymphocyte replicative lifespan. We studied these issues in patients affected by adenosine deaminase severe combined immune deficiency after low-intensity conditioning and reinfusion of retrovirally transduced autologous CD34(+) cells. Methods: Immunophenotype, proliferative status, telomere length, and T-cell receptor excision circles were investigated at early and late time points (up to 9 years) after GT treatment. Control groups consisted of pediatric healthy donors and patients undergoing allogeneic bone marrow transplantation (BMT). Results: We observed no telomere shortening in the bone marrow compartment and in granulocytes, whereas peripheral blood naive T cells from both GT and BMT patients showed a significant reduction in telomere length compared with healthy controls. This was in agreement with the presence of a high fraction of actively cycling naive and memory T cells and lower T-cell receptor excision circles. Conclusion: These data indicate that T-cell homeostatic expansion contributes substantially to immune reconstitution, like BMT, and is not associated with senescence in the stem cell compartment. (J Allergy Clin Immunol 2011;127:1368-75.) BACKGROUND:Gene therapy (GT) with hematopoietic stem cells is a promising treatment for inherited immunodeficiencies.OBJECTIVES:Limited information is available on the relative contribution of de novo thymopoiesis and peripheral expansion to T-cell reconstitution after GT as well as on the potential effects of gene transfer on hematopoietic stem cells and lymphocyte replicative lifespan. We studied these issues in patients affected by adenosine deaminase severe combined immune deficiency after low-intensity conditioning and reinfusion of retrovirally transduced autologous CD34(+) cells.METHODS:Immunophenotype, proliferative status, telomere length, and T-cell receptor excision circles were investigated at early and late time points (up to 9 years) after GT treatment. Control groups consisted of pediatric healthy donors and patients undergoing allogeneic bone marrow transplantation (BMT).RESULTS:We observed no telomere shortening in the bone marrow compartment and in granulocytes, whereas peripheral blood naive T cells from both GT and BMT patients showed a significant reduction in telomere length compared with healthy controls. This was in agreement with the presence of a high fraction of actively cycling naive and memory T cells and lower T-cell receptor excision circles.CONCLUSION:These data indicate that T-cell homeostatic expansion contributes substantially to immune reconstitution, like BMT, and is not associated with senescence in the stem cell compartment.

T-cell depletion of allogeneic hematopoietic stem cell graft (HSCT) represents the most powerful approach to prevent graft-versus-host disease (GvHD), thus allowing to overcome HLA barriers in patients with high risk malignancies, lacking a conventional donor. We hypothesized that early add-back of suicide-gene transduced donor lymphocytes (TK cells) to leukemic patients undergoing haploidentical HSCT (haplo-HSCT) could provide early immune-reconstitution and selective control of GvHD. In a phase II clinical trial (protocol MMTK007), 17 of 29 enrolled pts, (median age 52), received add-backs of 10^7/kg TK cells 42 days after haplo-HSCT. TK cells engraftment, observed in 14 patients, was necessary and sufficient for a rapid and effective immunereconstitution (IR), with a median of 144 (101–336) CD3+, 59 (28–149) CD4+ and 86 (52–279) CD8+ cells/mcl at day 100 after HSCT. Accordingly, engraftment of TK cells was tightly correlated with clinical outcome: while 3/3 pts who failed TK cells engraftment died of infections, only 1/14 TK engrafted patients died from infections (last event at day +166). As shown in Table I, the immune repertoire of treated patients improved significantly at 6 months post transplant and normalized completely in 12 months. High numbers of T cell precursors specific for CMV and EBV were detected at immune reconstitution (median of 86 and 69 gIFN specific spots/10^5 PBMC respectively) and predicted subsequent freedom from viral reactivation (p=0.002). Six pts developed acute (GvHD), (grade I to IV) that was always completely abrogated by the suicide system. Overall survival of TK cells treated patients is 50% at three years. These results indicate that TK-DLI abolish late mortality after CD34+ haplo-SCT in adults. A phase III multicentric study will start in 2007 to validate prospectively the advantage of TK-DLI in haplo-SCT.

Haplo-identical stem cell transplantation (haplo-SCT) is a therapeutic option for pts with high risk hematologic malignancies lacking an HLA matched donor. The full exploitation of haplo-SCT is limited by delayed immune reconstitution and prolonged risk of life-threatening infections. We previously identified in an academic haplo-SCT trial, effective doses of donor lymphocytes genetically engineered to express a suicide gene (the herpes simplex thymidine kinase (TK-DLI) as an effective tool for preventing disease relapse and promoting immune reconstitution. In case of GvHD, TK-DLI could be selectively eliminated by ganciclovir. A phase II multicenter trial (MM TK007), aimed at verifying the therapeutic activity of early add-backs of TK+ cells after haplo-SCT in inducing a stable immune reconstitution, started in 2002. 20 pts were recruited and transplanted for AML (16, 8 of whom post MDS), ALL (2), advanced NHL and HD (2); disease status at SCT was CR1(5), CR2(7), refractory(8). Median time from diagnosis to SCT was 394 d (95-3752), median age was 52 (17–62). A median of 12.3x106/kg (7.3–15.5) CD34+ selected cells (Clinimacs) and 1.05x104/kg (0.8–1.4) CD3+ cells were infused after a myeloablative conditioning regimen. GvHD prophylaxis was never administered post-SCT. 18/20 pts engrafted with a median time of 13 d (8–21), for ANC >1.0x109/l and 13 d (11–24) for plt >50x109/l. No immune reconstitution and no GvHD were observed in absence of TK-DLI. The statistical end-point of the study was to achieve immune reconstitution in 7/18 treated pts. So far, 10 pts received TK-DLI at a median dose of 107/kg with 1st infusion at d +42. Although this study will continue up to 18 treated pts, the primary end-point has been already achieved since 7/9 evaluable pts obtained CD3+ >100/μl at a median time of 76 d (61–99) from SCT and 21 d (14–31) from TK-DLI. Transduced cells were documented ex vivo in all pts; median counts at peak of immune reconstitution were 483 CD3+/μl, 304 CD8+/μl, 130 CD4+/μl. Prompt immune reconstitution resulted in complete control of viral infections and virtually eliminated late infections frequently reported after haplo-SCT. In 1 pt a grade II biopsy-proven acute GvHD, involving skin and liver was observed and completely abrogated by ganciclovir (10 mg/kg/day) in the absence of immunosuppressive drugs. One pt relapsed at d 300; overall, 6 pts are alive in CR at d +423, +363, +317, +255, +109, +104. The preliminary results of this multicenter phase II study confirm that TK-DLI is an effective tool for promoting immune reconstitution and protecting pts from infectious mortality after haplo-SCT while providing an effective and selective treatment for GvHD. The potential of this strategy in preventing disease relapse is promising and will remain the main focus of the long-term follow-up.