Cytokine expression assessed by flow cytometry in 53 acquired aplastic anemia patients before and after combined immunosuppression (EBMT WPSAA protocols) showed that CD3(+) marrow cells containing ...TNF-alpha, IFN-gamma and IL4 were similar in subjects with disease at onset (DO) and responsive to treatment who had more CD3(+)/TNF-alpha(+) and CD 3(+)/IFN-gamma(+) cells than normal controls. In vitro block of TNF-alpha and/or IFN-gamma significantly increased BFU-e over baseline in 28 patients. In responsive to treatment patients only TNF-alpha block significantly incremented colonies over normal controls. Absolute marrow CD3(+)/TNF-alpha(+) and CD3(+)/IFN-gamma(+) cells prospectively tested in a group of 21 subjects declined significantly more in Responders than in Non Responders to immunosuppression at Response Evaluation Time respect to Diagnosis. Both in Responders and in Non Responders these cells remained higher than in normal controls. This study suggests that immunosuppression does not fully clear excess TNF-alpha and IFN-gamma from marrow of patients with good outcome and raises the hypothesis that additional cytokine blockade might be useful in immunosuppression for acquired aplastic anemia.
Purpose: Few data are available in the literature on chemokine receptor expression and migratory capability of mantle cell lymphoma
(MCL) B cells. Information on these issues may allow us to identify ...novel mechanisms of chemokine-driven tumor cell migration.
Experimental Design: The research was designed to investigate: ( a ) expression of CCR1 to CCR7 and CXCR1 to CXCR5 chemokine receptors; and ( b ) chemotaxis to the respective ligands in MCL B cells and in their normal counterparts, i.e. , CD5 + B cells.
Results: Malignant B cells from MCL patients and normal counterparts displayed similar chemokine receptor profiles. MCL B cells were
induced to migrate by CXCL12 and CCL19, whereas normal CD5 + B cells migrated to the former, but not the latter chemokine. Overnight culture of MCL B cells and their normal counterparts
with CXCL12 cross-sensitized other chemokine receptors to their ligands in some tumor samples but not in CD5 + B cells.
Conclusions: CCR7 and CXCR4 ligands may play a key role in tumor cell migration and spreading in vivo . CXCL12 may additionally contribute by sensitizing MCL B cells to respond to the ligands of other chemokine receptors.
The aim of this study was to evaluate the agricultural reuse of the digestate products (DPs) obtained from mesophilic anaerobic co-digestion of different organic wastes (sludge, cattle slurries and ...organic fraction of municipal solid wastes). At this scope, the content of faecal indicators and pathogens as well as the heavy metal concentration of DPs was monitored. The fertilizing performance of the DPs was also investigated. Co-digestion trials were performed using laboratory-scale (LRs) and pilot-scale reactors (PRs). The microbiological analysis of DPs showed the common presence of Salmonella and an inadequate reduction of indicator organisms during the digestion process, both in the LRs and the PRs. Moreover, the presence of pathogens (e.g. Listeria monocytogenes) in some DP samples highlighted the importance of the microbiological quality evaluation of the DPs to study the possible health risks for consumer. In several samples of DPs, the Cu, Ni and Zn contents exceeded the maximum admissible concentration for fertilizer, as specified by Italian law, suggesting possible environmental contamination if the DPs are used for agricultural purposes. Considering the fertilizing performance, significant differences of growth parameters were observed only for the DPs that were produced by LRs. In conclusion, this work can be considered as a preliminary study to evaluate the possible agricultural reuse of the digestate obtained from different organic wastes.
Neuroblastoma (NB) tumor substantially contributes to childhood cancer mortality. The design of novel drugs targeted to specific molecular alterations becomes mandatory, especially for high‐risk ...patients burdened by chemoresistant relapse. The dysregulated expression of MYCN, ALK, and LIN28B and the diminished levels of miR‐34a and let‐7b are oncogenic in NB. Due to the ability of miRNA‐mimics to recover the tumor suppression functions of miRNAs underexpressed into cancer cells, safe and efficient nanocarriers selectively targeted to NB cells and tested in clinically relevant mouse models are developed. The technology exploits the nucleic acids negative charges to build coated‐cationic liposomes, then functionalized with antibodies against GD2 receptor. The replenishment of miR‐34a and let‐7b by NB‐targeted nanoparticles, individually and more powerfully in combination, significantly reduces cell division, proliferation, neoangiogenesis, tumor growth and burden, and induces apoptosis in orthotopic xenografts and improves mice survival in pseudometastatic models. These functional effects highlight a cooperative down‐modulation of MYCN and its down‐stream targets, ALK and LIN28B, exerted by miR‐34a and let‐7b that reactivate regulatory networks leading to a favorable therapeutic response. These findings demonstrate a promising therapeutic efficacy of miR‐34a and let‐7b combined replacement and support its clinical application as adjuvant therapy for high‐risk NB patients.
Targeted nanocarriers entrapping microRNAs (miRNA)‐mimics are selectively delivered to neuroblastoma cells. The technology exploits the nucleic acids negative charges to build coated cationic liposomes, which are functionalized with antibodies against GD2 receptor. The combined replenishment of miR‐34a and let‐7b by neuroblastoma‐targeted nanoparticles exerts a cooperative down‐modulation of key oncogenes. The reactivated regulatory networks lead to a favorable therapeutic response in preclinical models.
Multiple myeloma (MM) is characterized by the tight dependence of plasma cells (PCs) to the bone marrow (BM) microenvironment that activates the production of several pro-inflammatory and growth ...factors involved in MM cell survival, bone remodeling alterations and increased angiogenesis. CX3CL1/Fractalkine is a unique chemokine synthetized as a membrane-bound protein and released as soluble protein after proteolytic cleavage by two metalloproteases as ADAM10 and ADAM17. CX3CL1 interacts with its single receptor CX3CR1 being involved in several physiological and pathophysiological processes including inflammation and angiogenesis. The role CX3CL1/CX3CR1 axis in MM is still unexplored and unknown.
In this study, firstly we analyzed BM plasma levels of CX3CL1 in MM patients as compared to indolent monoclonal gammopathies and healthy controls. BM plasma was obtained from 38 MM patients (median age 70 years; 53% female and 47% male; ISS I= 32%, II= 34%, III=34%) 25 of them with newly diagnosed and 13 relapsed MM, and from sex-aged matched 14 Smoldering MM (SMM) patients and 11 monoclonal gammopathy of undetermined significance (MGUS) patients. 10 healthy donors (HD) (median age 70 years; 20% female and 80% male) were used as controls. We found that plasma levels were significantly increased in the BM plasma from MM patients compared to HD (median levels 0.88 ng/ml vs 0.56 ng/ml, p=0.004). Furthermore, increased CX3CL1 levels were observed in patients with active MM as compared to patients with asymptomatic disease, as MGUS and SMM (median levels MM vs MGUS: 0.88 ng/ml vs 0.67 ng/ml, p=0.0176; median levels MM vs SMM: 0.88 ng/ml vs 0.65ng/ml, p= 0.0329). Moreover, CX3CL1 levels increased in the advanced stages of disease (median levels ISS III vs ISS I: 1.18 ng/ml vs 0.72 ng/ml, p=0.0115; median levels ISS III vs ISS II: 1.18 ng/ml vs 0.74 ng/ml, p=0.0157). Accordingly, BM soluble CX3CL1 levels were positively correlated with the percentage of BM plasma cells in the cohort of MM patients (p=0.002). Subsequently, the possible link between soluble CX3CL1/fractalkine and BM angiogenesis has been investigated. CD34 expression was evaluated by immunohistochemistry on bone biopsies obtained from 28 MM patients, showed that BM plasma levels of CX3CL1 significantly correlated with both the number of CD34 positive vessels (p= 0.002) and the micro-vessels density (p=0.002). Thereafter, we performed experiments on chorioallantoic membranes to demonstrate the role of CX3CL1 in MM-induced angiogenesis. The incubation of BM plasma of MM patients for 12 days resulted in a significant angiogenic response in the form of numerous allantoic neovessels that were significantly inhibited by blocking anti-CX3CL1 antibody. Finally, to clarify the source of CX3CL1 into the BM of MM patients, we explored CX3CL1 expression either by CD138+ cells on a dataset of 133 MM patients at diagnosis (GSE16122) and 23 human myeloma cell lines (HMCLs) (GSE6205), or on a proprietary dataset of primary mesenchymal stromal cells (MSCs) and osteoblasts (OBs) of 16 MM and 7 HD. Isolated BM endothelial cells from 4 MM patients and controls were also analysed. Purified CD138+ cells, HMCLs and BM endothelial cells were negative for CX3CL1 mRNA expression whereas MSCs and OBs were positive for CX3CL1 even if any statistically significant difference was not observed across the different groups. On the other hand, we found that CD138+ cells and HMCLs expressed both ADAM10 and ADAM17, leading to the hypothesis that the expression of these metalloproteinases by MM cells is accountable for the increased levels of CX3CL1 into the BM microenvironment and consequently increase of the neo-vascularization.
In conclusion, our data indicate that CX3CL1, present at high level into the BM of MM patients, is involved in MM-induced BM neo-vascularization and potentially in MM progression.
No relevant conflicts of interest to declare.
Interleukin (IL)-17A belongs to IL-17 superfamily and binds the heterodimeric IL-17 receptor (R)(IL-17RA/IL-17RC). IL-17A promotes germinal center (GC) formation in mouse models of autoimmune or ...infectious diseases, but the role of IL-17A/IL-17AR complex in human neoplastic GC is unknown. In this study, we investigated expression and function of IL-17A/IL-17AR in the microenvironments of 44 B cell non-Hodgkin lymphomas (B-NHL) of GC origin (15 follicular lymphomas, 17 diffuse large B cells lymphomas and 12 Burkitt lymphomas) and 12 human tonsil GC. Furthermore, we investigated the role of IL-17A in two in vivo models of GC B cell lymphoma, generated by s.c. injection of SU-DHL-4 and OCI-Ly8 cell lines in Severe combined immunodeficiency (SCID)/Non Obese Diabetic (NOD) mice. We found that: (i) B-NHL cell fractions and tonsil GC B cells expressed IL-17RA/IL-17RC, (ii) IL-17A signaled in both cell types through NF-kBp65, but not p38, ERK-1/2, Akt or NF-kBp50/105, phosphorylation, (iii) IL-17A was expressed in T cells and mast cells from neoplastic and normal GC microenvironments, (iv) IL-17A rendered tonsil GC B cells competent to migrate to CXCL12 and CXCL13 by downregulating RGS16 expression; (v) IL-17A stimulated in vitro proliferation of primary B-NHL cells; (vi) IL-17A (1 μg/mouse-per dose) stimulated B-NHL growth in two in vivo models by enhancing tumor cell proliferation and neo-angiogenesis. This latter effect depended on IL-17A-mediated induction of pro-angiogenic gene expression in tumor cells and direct stimulation of endothelial cells. These data define a previously unrecognized role of human IL-17A in promoting growth of GC-derived B-NHL and modulating normal GC B cell trafficking.
Abstract Acute myeloid leukemia is a haematopoietic malignancy originating from the transformation of myeloid progenitors that proliferate and accumulate in the bone marrow. In AML patients the ...survival rate at 5 years is 40–50% highlighting the need for novel therapies. In this study we have asked whether IL-12, an immuno-modulatory cytokine with anti-tumor activity, may inhibit directly AML cell growth. We show that the human AML cell lines U937, K562 and THP-1 expressed both chains of the IL-12 receptor (R), i.e. IL-12Rβ1 and IL-12Rβ2. IL-12 inhibited the angiogenic potential of AML cells in vitro , but did not affect their survival or proliferation. In vivo experiments were performed using SCID–NOD mice injected intraperitoneally (i.p.) with the human U937 AML cell line and subsequently treated with human recombinant IL-12 or PBS i.p. Histological, immunohistochemical and flow cytometric analyses on explanted tumors revealed that IL-12 reduced new vessel formation, induced apoptosis and inhibited tumor cell proliferation. Studies on a panel of angiogenesis related genes in explanted tumors using PCR arrays showed significantly down-regulated expression of numerous pro-angiogenic genes including VEGF-C, IL-6, IL-8, CXCL1, CXCL6 and alanyl aminopeptidase in IL-12 vs PBS treated mice. This study shows for the first time that IL-12 targets directly AML cell growth and paves the way to further investigation of IL-12 as potential drug for AML treatment.
Background Fractalkine/CX3CL1, a surface chemokine, binds to CX3CR1 expressed by different lymphocyte subsets. Since CX3CL1 has been detected in the germinal centres of secondary lymphoid tissue, in ...this study we have investigated CX3CR1 expression and function in human naïve, germinal centre and memory B cells isolated from tonsil or peripheral blood. Methodology/Principal Findings We demonstrate unambiguously that highly purified human B cells from tonsil and peripheral blood expressed CX3CR1 at mRNA and protein levels as assessed by quantitative PCR, flow cytometry and competition binding assays. In particular, naïve, germinal centre and memory B cells expressed CX3CR1 but only germinal centre B cells were attracted by soluble CX3CL1 in a transwell assay. CX3CL1 signalling in germinal centre B cells involved PI3K, Erk1/2, p38, and Src phosphorylation, as assessed by Western blot experiments. CX3CR1+ germinal centre B cells were devoid of centroblasts and enriched for centrocytes that migrated to soluble CX3CL1. ELISA assay showed that soluble CX3CL1 was secreted constitutively by follicular dendritic cells and T follicular helper cells, two cell populations homing in the germinal centre light zone as centrocytes. At variance with that observed in humans, soluble CX3CL1 did not attract spleen B cells from wild type mice. OVA immunized CX3CR1−/− or CX3CL1−/− mice showed significantly decreased specific IgG production compared to wild type mice. Conclusion/Significance We propose a model whereby human follicular dendritic cells and T follicular helper cells release in the light zone of germinal centre soluble CX3CL1 that attracts centrocytes. The functional implications of these results warrant further investigation.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Fractalkine/CX(3)CL1, a surface chemokine, binds to CX(3)CR1 expressed by different lymphocyte subsets. Since CX(3)CL1 has been detected in the germinal centres of secondary lymphoid tissue, in this ...study we have investigated CX(3)CR1 expression and function in human naïve, germinal centre and memory B cells isolated from tonsil or peripheral blood.
We demonstrate unambiguously that highly purified human B cells from tonsil and peripheral blood expressed CX(3)CR1 at mRNA and protein levels as assessed by quantitative PCR, flow cytometry and competition binding assays. In particular, naïve, germinal centre and memory B cells expressed CX(3)CR1 but only germinal centre B cells were attracted by soluble CX(3)CL1 in a transwell assay. CX(3)CL1 signalling in germinal centre B cells involved PI3K, Erk1/2, p38, and Src phosphorylation, as assessed by Western blot experiments. CX(3)CR1(+) germinal centre B cells were devoid of centroblasts and enriched for centrocytes that migrated to soluble CX(3)CL1. ELISA assay showed that soluble CX(3)CL1 was secreted constitutively by follicular dendritic cells and T follicular helper cells, two cell populations homing in the germinal centre light zone as centrocytes. At variance with that observed in humans, soluble CX(3)CL1 did not attract spleen B cells from wild type mice. OVA immunized CX(3)CR1(-/-) or CX(3)CL1(-/-) mice showed significantly decreased specific IgG production compared to wild type mice.
We propose a model whereby human follicular dendritic cells and T follicular helper cells release in the light zone of germinal centre soluble CX(3)CL1 that attracts centrocytes. The functional implications of these results warrant further investigation.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
PDF
