Anti-HBs re-seroconversion after liver transplantation in a patient with past HBV infection receiving a HBsAg positive graft

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Synergistic Proapoptotic Activity of Recombinant Trail Plus the AKT Inhibitor Perifosine in Acute Myelogenous Leukemia Cells

To potentiate the response of acute myelogenous leukemia (AML) cells to TNF-Related Apoptosis- Inducing Ligand (TRAIL) cytotoxicity, we have examined the efficacy of a combination with perifosine, a novel phosphatidylinositol 3-kinase (PI3K)/Akt signaling inhibitor. The rationale for using such a combination is that perifosine was recently described to increase TRAIL-R2 receptor expression and decrease the cellular FLICE-Inhibitory Protein (cFLIP) in human lung cancer cell lines. Perifosine and TRAIL both induced cell death by apoptosis in the THP-1 AML cell line, which is characterized by constitutive PI3K/Akt activation, but lacks functional p53. Perifosine, at concentrations below IC50, dephosphorylated Akt and increased TRAIL-R2 levels, as demonstrated by western blot, RT-PCR, and flow cytometric analysis. Perifosine also decreased the long isoform of cFLIP (cFLIP-L) and the X-linked Inhibitor of Apoptosis Protein (XIAP) expression. Perifosine and TRAIL synergized to activate caspase-8 and induce apoptosis, which was blocked by a caspase- 8 selective inhibitor. Upregulation of TRAIL-R2 expression was dependent on a protein kinase Cα/ c-Jun-NH2-kinase 2/c-Jun signaling pathway activated by perifosine through reactive oxygen species production. Perifosine synergized with TRAIL also in primary AML cells displaying constitutive activation of the Akt pathway, by inducing apoptosis, Akt dephosphorylation, TRAIL-R2 upregulation, cFLIP-L and XIAP downregulation, and c-Jun phosphorylation. The combined treatment negatively affected the clonogenic activity of CD34+ cells from AML patients. In contrast, CD34+ cells from healthy donors were resistant to perifosine and TRAIL treatment. Our findings suggest that the combination perifosine and TRAIL might offer a novel therapeutic strategy for AML. Originally published Cancer Research, Vol. 68, No. 22, Nov 200

Glutathione transferase-A2 S112T polymorphism predicts survival, transplant-related mortality, busulfan and bilirubin blood levels after allogeneic stem cell transplantation

Busulfan liver metabolism depends on glutathione, a crucial mediator of cellular and systemic stress. Here we investigated 40 polymorphisms at 27 loci involved in hepatic glutathione homeostasis, with the aim of testing their impact on the clinical outcome of 185 busulfan-conditioned allogeneic transplants. GSTA2 S112T serine allele homozygosity is an independent prognostic factor for poorer survival (RR=2.388), for increased any time- and 100-day transplant-related mortality (RR=4.912 and RR=5.185, respectively). The genotype also predicts a wider busulfan area under the concentration-time curve (1214.36 \ub1 570.06 vs. 838.10 \ub1 282.40 mMol*min) and higher post-transplant bilirubin serum levels (3.280 \ub1 0.422 vs. 1.874+0.197 mg/dL). In vitro, busulfan elicits pro-inflammatory activation (increased NF-KappaB activity and interleukin-8 expression) in human hepatoma cells. At the same time, the drug down-regulates a variety of genes involved in bilirubin liver clearance: constitutive androstane receptor, multidrug resistance-associated protein, solute carrier organic anion transporters, and even GSTA2. It is worthy of note that GSTA2 also acts as an intra-hepatic bilirubin binding protein. These data underline the prognostic value of GSTA2 genetic variability in busulfan-conditioned allotransplants and suggest a patho-physiological model in which busulfan-induced inflammation leads to the impairment of post-transplant bilirubin metabolis

Distribution of killer cell immunoglobulin-like receptors genes in the Italian Caucasian population

BACKGROUND: Killer cell immunoglobulin-like receptors (KIRs) are a family of inhibitory and activatory receptors that are expressed by most natural killer (NK) cells. The KIR gene family is polymorphic: genomic diversity is achieved through differences in gene content and allelic polymorphism. The number of KIR loci has been reported to vary among individuals, resulting in different KIR haplotypes. In this study we report the genotypic structure of KIRs in 217 unrelated healthy Italian individuals from 22 immunogenetics laboratories, located in the northern, central and southern regions of Italy. METHODS: Two hundred and seventeen DNA samples were studied by a low resolution PCR-SSP kit designed to identify all KIR genes. RESULTS: All 17 KIR genes were observed in the population with different frequencies than other Caucasian and non-Caucasian populations; framework genes KIR3DL3, KIR3DP1, KIR2DL4 and KIR3DL2 were present in all individuals. Sixty-five different profiles were found in this Italian population study. Haplotype A remains the most prevalent and genotype 1, with a frequency of 28.5%, is the most commonly observed in the Italian population. CONCLUSION: The Italian Caucasian population shows polymorphism of the KIR gene family like other Caucasian and non-Caucasian populations. Although 64 genotypes have been observed, genotype 1 remains the most frequent as already observed in other populations. Such knowledge of the KIR gene distribution in populations is very useful in the study of associations with diseases and in selection of donors for haploidentical bone marrow transplantation

Long-Term Outcome After Adoptive Immunotherapy With Natural Killer Cells: Alloreactive NK Cell Dose Still Matters

open26noRecently, many reports were published supporting the clinical use of adoptively transferred natural killer (NK) cells as a therapeutic tool against cancer, including acute myeloid leukemia (AML). Our group demonstrated promising clinical response using adoptive immunotherapy with donor-derived alloreactive KIR-ligand-mismatched NK cells in AML patients. Moreover, the antileukemic effect was correlated with the dose of infused alloreactive NK cells (“functional NK cell dose”). Herein, we update the results of our previous study on a cohort of adult AML patients (median age at enrollment 64) in first morphological complete remission (CR), not eligible for allogeneic stem cell transplantation. After an extended median follow-up of 55.5 months, 8/16 evaluable patients (50%) are still off-therapy and alive disease-free. Overall survival (OS) and disease-free survival (DFS) are related with the dose of infused alloreactive NK cells (≥2 × 105/kg).openParisi S.; Ruggeri L.; Dan E.; Rizzi S.; Sinigaglia B.; Ocadlikova D.; Bontadini A.; Giudice V.; Urbani E.; Ciardelli S.; Sartor C.; Cristiano G.; Nanni J.; Zannoni L.; Chirumbolo G.; Arpinati M.; Lewis R.E.; Bonifazi F.; Marconi G.; Martinelli G.; Papayannidis C.; Paolini S.; Velardi A.; Cavo M.; Lemoli R.M.; Curti A.Parisi, S.; Ruggeri, L.; Dan, E.; Rizzi, S.; Sinigaglia, B.; Ocadlikova, D.; Bontadini, A.; Giudice, V.; Urbani, E.; Ciardelli, S.; Sartor, C.; Cristiano, G.; Nanni, J.; Zannoni, L.; Chirumbolo, G.; Arpinati, M.; Lewis, R. E.; Bonifazi, F.; Marconi, G.; Martinelli, G.; Papayannidis, C.; Paolini, S.; Velardi, A.; Cavo, M.; Lemoli, R. M.; Curti, A

PREPARATO TOPICO DA SIERO CORDONALE PER LA TERAPIA DI DIFETTI EPITELIALI CORNEALI PERSISTENTI SIMTI

Introduzione. La risoluzione di danni epiteliali corneali estesi e persistenti, che inficiano la qualit\ue0 della visione, richiedono fattori di crescita specifici, presenti nelle lacrime umane, in particolare l\u2019Epidermal Growth Factor (EGF), ed il Trasforming Growth Factor-beta (TGF-β). Alla base di questo, sta il razionale della terapia mediante somministrazione topica di siero autologo, preparato dal siero periferico del paziente stesso. Le patologie corneali che pi\uf9 di frequente si sono avvalse della terapia topica con siero autologo sono presenti in pazienti affetti da Sindrome di Sjogren primaria (SS-I) o secondaria e Graft Versus Host Disease (GVHD), due affezioni che inducono secchezza degli epiteli, da cui pu\uf2 svilupparsi una cheratopatia puntata, con erosioni medie e gravi persistenti della cornea E\u2019 stato di recente proposto l\u2019uso di preparati derivati dal siero di sangue cordonale per la cura dell\u2019occhio secco di grado severo, basandosi sulla presenza anche in questo siero di fattori di crescita, fattori neurotrofici e i componenti essenziali delle lacrime. Scopo del presente lavoro \ue8 confrontare le concentrazioni di EGF e TGF-b nel siero e nel plasma di pazienti possibili candidati alla terapia topica, vs siero e plasma cordonali. Metodi EGF e TGFb sono stati determinati utilizzando kit ELISA (Quantikine R&D System, Minneapolis, MN, USA) nei seguenti campioni: 1. plasma e siero di pazienti SS-I (8 pazienti) e GVHD (8 pazienti); 2. Plasma e siero di sangue cordonale (4 unit\ue0) sia appena prelevato che dopo stoccaggio di 6 mesi a \u2013 80\ub0C (periodo finestra per l\u2019effettuazione degli esami infettivologici); 3. prove pilota di preparato topico ottenuto per filtrazione dal siero dei pazienti o dal siero cordonale dopo il periodo finestra di 6 mesi a \u201380\ub0C. I dati sono stati analizzati statisticamente applicando Mann-Whitney test, significativit\ue0 p< 0.05. Risultati . I dati ottenuti sono raccolti nella tabella. In tutti i campioni i fattori di crescita analizzati sono risultati in maggior concentrazione nel siero rispetto al plasma (p sempre < 0.05). In particolare, il siero cordonale ha evidenziato concentrazioni da tre a dieci volte superiori al plasma e significativamente superiori rispetto al siero dei pazienti. Il preparato topico mantiene inalterati le concentrazioni anche dopo stoccaggio. Discussione e commento. Il siero da sangue cordonale appare un promettente emoderivato da cui ottenere preparati topici per la cura di epiteliopatie severe, stabili in efficacia anche dopo stoccaggio e filtrazione. Il preparato ottenuto presenta notevoli vantaggi rispetto all\u2019autosiero terapia in quanto: a. non richiede il prelievo di sangue da pazienti in condizioni di salute compromessa; b. pu\uf2 essere ottenuto in quantit\ue0 concettualmente illimitata; c. contiene concentrazioni significativamente superiori di fattori di crescita necessari alla riepitelizzazione; d. \ue8 sicuro in quanto controllato dopo periodo finestra; e. contrariamente al siero dei pazienti non contiene citokine pro-infiammatorie