Treatment of relapse after allogeneic stem cell transplantation in children and adolescents with ALL: the Frankfurt experience.

Therapy for post-transplant relapse of paediatric ALL is limited. Standardised curative approaches are not available. We hereby describe our local procedure in this life-threatening situation. A total of 101 ALL patients received their first allogeneic stem cell transplantation (SCT) in our institution. After relapse, our primary therapeutic goal was to cure the patient with high-dose chemotherapy or specific immunotherapy (HDCHT/SIT) followed by a second SCT from a haploidentical donor (transplant approach). If this was not feasible, low-dose chemotherapy and donor lymphocyte infusions (LDCHT+DLI) were offered (non-transplant approach). A total of 23 patients suffered a post-transplant relapse. Eight patients received HDCHT/SIT, followed by haploidentical SCT in 7/8. Ten received LDCHT+DLI. The eight patients treated with a second transplant and the ten treated with the non-transplant approach had a 4-year overall survival of 56% and 40%, respectively (P=0.232). Prerequisites for successful treatment of post-transplant relapse by either a second transplant or experimental non-transplant approaches are good clinical condition and the capacity to achieve haematological remission by the induction treatment element.

Nilotinib combined with interleukin-2 mediates antitumor and immunological effects in a B16 melanoma model.

The immune system contributes to tumor cell killing which can be enhanced by cancer chemotherapeutics and immune modulatory pharmaceuticals such as tyrosine kinase inhibitors (TKIs). Recently, the beneficial effect of natural killer (NK) cells was demonstrated when combining interleukin-2 (IL-2) with the TKI imatinib. The aim of the present study was to address the antitumor and immunological effects of recently approved TKIs. Therefore, we focused on the comparison of the efficacy between imatinib and nilotinib in combination with IL-2 in a murine B16F10 melanoma model. Both TKIs possessed antitumor activity in vivo. However, the combination of nilotinib and IL-2 showed a superior outcome. Importantly, both the use of immunodeficient Rag2γc-/- mice, which lack T-lymphocytes, B-lymphocytes and NK cells, as well as NK cell-depletion in C57Bl/6 mice reduced the therapeutic effect of nilotinib. Flow cytometry revealed a significant increase in the IFN-γ-producing CD27+ NK cell subpopulation following treatment with nilotinib and IL-2. Furthermore, the therapeutic antitumor effect of nilotinib/IL-2 was completely lost in IFN-γ-/- mice. In summary, we suggest that nilotinib combined with IL-2 confers high antitumor activity involving the subset of IFN-γ-producing CD27+ NK cells. These new insights are of high importance for the understanding and development of immunotherapeutic protocols using TKIs.

The interaction of NK cells and dendritic cells in the tumor environment: how to enforce NK cell & DC action under immunosuppressive conditions?

The crosstalk of natural killer (NK) and dendritic cells (DCs) plays an important role in the induction of the tumor-specific immune response against cancer. During the last decade, our advanced understanding of the immune system led to the development of new therapeutic strategies in the field of immunotherapy and cellular immunology. However, these immunotherapeutic concepts have not been as successful as initially expected because of their inability to counteract cancer-induced immunosuppressive pathways. Some of the major difficulties of effective cellular immunotherapy are the highly immunosuppressive factors induced by tumor cells themselves or by their microenvironment. Therefore, one major challenge in immunotherapy is the question: "How to enforce NK cell & DC action under immunosuppressive conditions?" This review focuses on the current knowledge on the tumor microenvironment, the crosstalk of NK cells and DCs, as well as their deregulation in the complex interplay with the immunosuppressive tumor microenvironment. We further discuss possible strategies to minimize the negative impact of the tumor microenvironment on the immune system.

Killer dendritic cells and their potential role in immunotherapy.

Tumor immunosurveillance is mediated by innate and adaptive components of cellular immunity. A complex network of cellular interactions is needed to elicit protective antitumoral CD4+and CD8+T cell responses. Thereby dendritic cells (DCs) play a central role as professional antigen presenting cells (APCs) that take up antigens, process, and present them to prime naïve T cells. Recognition and lysis of tumor cells has been attributed to innate effectors such as natural killer (NK), NKT and gammadeltaT cells. Recently, novel subsets of cytotoxic DCs, called "killer DCs" (KDCs), have been reported in rodents and humans. Killer dendritic cells could directly link innate and adaptive immunity. This review aims at comparing the different KDC populations, their phenotypes, killer function, and their potential application for anticancer immunotherapy.

Exosomes for the treatment of human malignancies.

Exosomes are nanometer particles (50-100 nm) secreted by most living cells. The first description of exosomes was made in 1987 by Rose Johnstone, who described a vesicle formation during the maturation process of reticulocytes. At this time it has been suggested that exosome release could represent a major route for the externalization of obsolete membrane proteins. A renewed vision of exosome function was raised when Graça Raposo demonstrated in 1996 that exosomes derived from B cells could have immunogenic capacities. Since then, exosomes have been described in numerous cell types IN VITRO, including hematopoietic and nonhematopoietic cells. The physiological relevance of exosomes IN VIVO still remains unclear. Studies have demonstrated that exosomes can play a role in the physiology of originating cells (i.e., reticulocyte-derived exosomes). Furthermore, exosomes can act on intercellular communication by allowing exchange of proteins, lipids, and also mRNA between cells. Finally, exosomes have been shown to modulate the immune system (i.e., dendritic cells, B cells, and tumor cells). In the present review, we have focused on the potential therapeutic role of exosomes as a cell free vaccine in cancer.

Tumor stress, cell death and the ensuing immune response.

A cornucopia of physiological and pathological circumstances including anticancer chemotherapy and radiotherapy can induce cell death. However, the immunological consequences of tumor cell demise have remained largely elusive. The paradigm opposing 'apoptosis versus necrosis' as to their respective immunogenicity does not currently hold to predict long-term immunity. Moreover, the notion that tumor cells may be 'stressed' before death to be recognized by immune cells deserves to be underlined. 'Eat-me', 'danger' and 'killing' signals released by stressed tumor under the pressure of cytotoxic compounds may serve as links between the chemotherapy-elicited response of tumor cells and subsequent immune responses. This review will summarize the state-of-the-art of cancer immunity and describe how tumor cell death dictates the links between innate and acquired immunity.

Interferon-induced antiviral Mx1 GTPase is associated with components of the SUMO-1 system and promyelocytic leukemia protein nuclear bodies.

Mx proteins are interferon-induced large GTPases, some of which have antiviral activity against a variety of viruses. The murine Mx1 protein accumulates in the nucleus of interferon-treated cells and is active against members of the Orthomyxoviridae family, such as the influenza viruses and Thogoto virus. The mechanism by which Mx1 exerts its antiviral action is still unclear, but an involvement of undefined nuclear factors has been postulated. Using the yeast two-hybrid system, we identified cellular proteins that interact with Mx1 protein. The Mx1 interactors were mainly nuclear proteins. They included Sp100, Daxx, and Bloom's syndrome protein (BLM), all of which are known to localize to specific subnuclear domains called promyelocytic leukemia protein nuclear bodies (PML NBs). In addition, components of the SUMO-1 protein modification system were identified as Mx1-interacting proteins, namely the small ubiquitin-like modifier SUMO-1 and SAE2, which represents subunit 2 of the SUMO-1 activating enzyme. Analysis of the subcellular localization of Mx1 and some of these interacting proteins by confocal microscopy revealed a close spatial association of Mx1 with PML NBs. This suggests a role of PML NBs and SUMO-1 in the antiviral action of Mx1 and may allow us to discover novel functions of this large GTPase.

Spatial determination of elements in green leaves of oak trees (Quercus robur) by laser ablation-ICP-MS.

Laser ablation inductively coupled plasma mass spectrometry (laser ablation-ICP-MS) has been applied to the spatially resolved determination of the elements Mg, Ca, Cu, Ni, Ba, Al, Pb, Sr and Mn in green leaves of oak trees. Instrument operating parameters such as the laser wavelength and the pulse energy have been optimized to provide the sensitivity and reproducibility required for the analysis. The method provides spatial resolution down to 300 microm with the use of the 355 nm wavelength (3rd harmonic of the 1,064 nm Nd:YAG laser wavelength) and the pulse energy of 50 mJ. Plant standards and cellulose, doped with multi element solution standards, dried and pressed to pellets were used as calibration samples. To compensate for signal fluctuations caused by the variation of the ablated sample mass 13C was used as a "natural" internal standard. The accuracy of the calibration was verified with selected samples analyzed by ICP-MS (high pressure digestion, 170 degrees C, 10(7) Pa, HNO3, 2 h) and by laser ablation-ICP-MS. Recovery rates between 93% (Cu) and 108% (Mn) were obtained. Leaves taken from oak trees (Quercus robur) were analyzed.

Solar blind photocell--a simple element specific detector in Hg, As and Se speciation.

An element-specific detection method, based on atomic absorption spectrometry (AAS) using solar blind photocells instead of a dispersion system, is described for the determination of Hg-, As-, and Se-species. Spectrometric investigations of AAS background lamps for As and Se measured with a CsI-cathode photocell shows its quality as narrow band detector. Species determination can be carried out subsequently to prior separation by HPLC or GC. The LODs for alkylated Hg species were below 1 ng/L, and for methylated As species below 1 microg/L. The relative standard deviation was < 10%. With the components described the production of cheap and automated dedicated speciation spectrometers is possible.

Molecular and functional characterization of the small Ca(2+)-regulated K+ channel (rSK4) of colonic crypts.

Colonic crypt cells possess basolateral Ca(2+)-regulated K+ channels which support Cl- secretion by providing the necessary driving force. The pharmacological characteristics of these channels were examined in Ussing chamber experiments of rat and rabbit colon mucosa by the use of blockers. The chromanol 293B, a blocker of KVLQT1 channels, and clotrimazole (CTZ), a blocker of small Ca(2+)-activated K+ channels, blocked stimulated Cl- secretion completely. Small-conductance Ca(2+)-activated K+ channels (SK) in excised basolateral patches of rat colonic crypts were inhibited concentration dependently by the imidazoles CTZ, NS004 and NS1619 and activated by 1-EBIO. These properties are similar to those of the known human SK channel (hSK4). hSK4-expressing Xenopus laevis oocytes showed ionomycin-activated and CTZ-inhibited K+ currents. When P2Y2 receptors were coexpressed these currents were also activated by ATP. The concentration/response curve was identical to that of rat SK channels. In human colonocytes (T84) exposed to hSK4 antisense probes, but not to sense probes, carbachol-induced K+ currents were attenuated. With RT-PCR an hSK4 could be demonstrated in human colon and in T84 colonocytes. By homology cloning the SK of the rat colon (rSK4) was identified. This protein has a high homology to hSK4 and mouse IK1. These data indicate that the Ca(2+)-activated and imidazole-inhibited basolateral K+ current in the colon is caused by SK4 channels.

Case report. A disseminated infection due to Chrysosporium queenslandicum in a garter snake (Thamnophis)

A male garter snake (Thamnophis) from a private terrarium was spontaneously and simultaneously infected with Chrysosporium queenslandicum and Geotrichum candidum. The autopsy revealed disseminated mycotic alterations in skin, lungs and liver. Chrysosporium queenslandicum grew well at 28 degrees C, the optimal temperature of the animal. This is the first description of a Chrysosporium queenslandicum infection in a garter snake.

[Problems of orthodontic care in a patient with Wiedemann-Beckwith-syndrome]. / Probleme bei der kieferorthopädischen Betreuung eines Patienten mit Widemann-Beckwith-Syndrom.

A review of cases with Wiedemann-Beckwith-syndrome in the literature and a comparison with our case is presented. The specific craniofacial and stomatognathical findings are highlighted. Aspects and problems of orthodontic treatment are discussed.

[Possibilities of clinico-cytological diagnosis in contagious equine metritis (CEM)]. / Möglichkeiten der klinisch-zytologischen Diagnostik bei der contagiösen equinen Metritis (CEM).

Clinical, bacteriological and serological examinations on a 6 years old pony mare were performed. Cytological alterations in the genital tract were also recorded. A cellular reaction was seen after infection with T. equigenitalis. This reaction is an evidence for infection but it is not specific for this organism. Cytological studies should be performed on mares especially in cases of latent infections to complete bacteriological examination and to prevent false positive or negative results.

[Histologic and biochemical studies on the uteri of ovariectomized gilts during the first two months of pregnancy after different hormone substitutions. 1. Changes in the structure of the placenta during pregnancy weeks 3-8 after constant high doses of progesterone and estradiol benzoate]. / Histologische und biochemische Untersuchungen an Uteri ovarektomierter Jungsauen während der ersten zwei Graviditätsmonate nach unterschiedlicher Hormonsubstitution. 1. Mitteilung: Veränderungen der Struktur der Plazenta materna während der 3.-8. Graviditätswoche bei gleichbleibend hoher Applikation von Progesteron und Ostradiolbenzoat.

Sixteen pregnant gilts were ovariectomised (Groups 1 and 2: 11th to 15th days of gravidity, Group 3: 19th to 22nd days of gravidity, Group 4: 34th to 45th days of gravidity). Daily intramuscular injections of 120 mg of progesterone and 250 micrograms of oestradiol benzoate were applied to them, beginning on the day of ovariectomy to preserve gravidity. All experimental animals were sacrificed between the 21st and 25th days of gravidity (Group 1) and between the 31st and 37th days (Groups 2 and 3) as well as between the 52nd and 61st days (Group 4). Living and dead embryos were numerically recorded, and histological als well as biochemical studies were conducted in all uteri. Embryo survival rates were normal, that is between 62.8 and 80 percent in Groups 1, 2, and 3, with the numbers of living embryos being 9.5, 7.5, and 8.0. Gravidity-specific uterus alterations were typical, despite constant hormone substitution. The thickness of endometrium declined with significance over the period under review. Plication was intensified, while the surface epithelium was flattened. Vascularisation increased, particularly in the subepithelial region, and the endometrial stroma became more oedematous. Full functionality was retained by the uterine glands in the period under review (increase in glandular epithelium as well as rising activity of alkaline phosphatase). Significant increase was recorded from the activity of acid phosphatase, whereas glycogen concentrations went down in the myometrium.

[Histologic and biochemical studies of the uteri of ovariectomized gilts during the first two months of pregnancy after different hormone substitutions. 2. Effects of reducing the progesterone dosage on the survival rate of embryos and the structure of the placenta and the influence of oral administration of norgestrel]. / Histologische und biochemische Untersuchungen an Uteri ovarektomierter Jungsauen während der ersten zwei Graviditätsmonate nach unterschiedlicher Hormonsubstitution. 2. Mitteilung: Auswirkungen einer Reduzierung der Progesterondosis auf die Uberlebensrate der Embryonen und die Struktur der Plazenta materna sowie Einfluss einer oralen Verabreichung von Norgestrel.

Studies were conducted into ovariectomised gilts in early gravidity, with the view to finding out if reduction of daily progesterone doses had an impact upon embryo survival rate and on the structure of the placenta and if experimentally induced progesterone deficit could be offset by oral administration of norgestrel, a synthetic progestagen. Embryo survival were found to drop from 80 to 26.1 percent in response to reduced progesterone dosage from 120 mg to 40 mg, average numbers of living embryos per animal being 9.6 or 3.0. Oral administration of 12 mg of norgestrel, in addition to injection of 40 mg of progesterone, enhanced the survival rate to 64.6 percent, the average number of living embryos coming to 8.9. Reduction of progesterone doses compared to animals with sufficient progesterone supply. Depressed the beginning decrease of the thickness endometrium and surface epithelium oedematisation of the endometrial stroma was mitigated, and subepithelial hyperaemia disappeared altogether. The secretory activity of uterine glands declined, and so did the endometrial activities of acid and alkaline phosphatases. Administration of norgestrel proved helpful in substantive removal of manifestations observed in the progesterone deficit group.

[Histologic and biochemical studies of the uteri of ovariectomized gilts during the first two months of pregnancy after different hormone substitutions. 3. Local effects of the conceptus on the structure of the placenta and its modification by different progesterone and norgestrel administration]. / Histologische und biochemische Untersuchungen an Uteri ovarektomierter Jungsauen während der ersten zwei Graviditätsmonate nach unterschiedlicher Hormonsubstitution. 3. Mitteilung: Lokale Effekte des Konzeptus auf die Struktur der Plazenta materna und ihre Beeinflussung durch unterschiedliche Progesteron- bzw. Norgestrelapplikation.

Optical light microscopy was used in investigations of ovariectomised gravid gilts to which progesterone doses between 120 mg and 40 mg as well as 250 micrograms of oestradiol benzoate had been daily applied to preserve gravidity, with 12 mg of norgestrel being additionally administered to some of them. These investigations were conducted for the purpose of studying locally delimited effects of conception on the placental structure. Uterus tissue was sampled from living and dead embryos (centre and sides of ampullae) as well as from uterus regions free of foetal membranes (in-between ampullae). With adequate progesterone supply, embryos were shown to clearly affect the endometrial structures. Endometrium in the centre of ampullae, with living embryos, was lower than at points without embryos. Surface epithelium was flattened, and endometrial stroma was more strongly oedematised. Strongly pronounced hyperaemia occurred to subepithelial stroma in the centre of ampullae, and uterine glandular function was unambiguously stimulated. These embryo-triggered effects were much less or no longer detectable at all under conditions of inadequate progesterone supply (40 mg/die). Administration of 12 mg of norgestrel, in addition to 40 mg/die of progesterone, enabled embryos to exercise gravidity-specific influence upon the endometrium, as in cases of sufficient progesterone supply.

[Histological and biochemical studies of the uterus of ovariectomized gilts during the first two months of pregnancy after different hormone substitutions. 4. Effect of the conceptus on the activities of alkaline and acid phosphatase and the concentration of glycogen in the uterus during different supplies of progesterone and norgestrel]. / Histologische und biochemische Untersuchungen an Uteri ovarektomierter Jungsauen während der ersten zwei Graviditätsmonate nach unterschiedlicher Hormonsubstitution. 4. Mitteilung: Einfluss des Konzeptus auf die Aktivitäten der alkalischen und sauren Phosphatase sowie die Konzentration des Glykogens im Uterus bei unterschiedlichem Angebot von Progesteron sowie Norgestrel.

Forty-five ovariectomised gilts in early gravidity received injections of progesterone doses between 120 mg and 40 mg as well as 250 micrograms/die of oestradiol-benzoate and oral application of 12 mg/die of norgestrel, the latter being administered in addition to the 40 mg dose of progesterone. The animals were killed between the 20th and 24th or on the 35th days of gravidity, before endometrial activities were determined of alkaline and acid phosphatases at various points of the uterus (centre and sides of ampullae and in-between ampullae). Under conditions of sufficient progesterone supply (120 mg/die and 100 mg/die), endometrial activities of both alkaline and acid phosphatases in the centre of ampullae were found to be higher than those in-between them. Activities were lower with inadequate progesterone supply (40 mg/die), and differences between points of sampling were less strongly pronounced for alkaline phosphatase. Endometrial glycogen concentrations in the centre of ampullae were lower than those in-between. Values in response to inadequate progesterone administration were lower than those following sufficient supply. With regard to glycogen concentrations, differences between points of sampling in the myometrium were below those in the endometrium.

[Infant hypotonia, obesity, hypogenitalism and oligophrenia--new viewpoints on the etiology and symptoms of Prader-Willi syndrome]. / Säuglingshypotonie, Adipositas, Hypogenitalismus und Oligophrenie--neue Gesichtspunkte zur Atiologie und Symptomatik des Prader-Willi-Syndroms.

We present four cases of Prader-Willi syndrome. Two of them have an abnormality of a chromosome 15, the other both show different chromosomal abnormalities. Translocations or deletions were found recently in the bands 15q11/12 in about 60% of the cases of Prader-Willi syndrome. The consequences for diagnosis, symptomatology and genetic counselling of the syndrome are discussed.