ENDOTHELIAL PROGENITOR CELLS IN MYELOPROLIFERATIVE NEOPLASMS – PRELIMINARY REPORT

The aim of this study was to assess the number of endothelial progenitor cells in patients with chronic myeloid leukemia (CML), polycythemia vera (PV), essential thrombocythemia (ET). The study involved 21 patients (mean age 61.77) with myeloproliferative neoplasms, hospitalized and diagnosed at the Hematology Clinic of Dr. J. Biziel University Hospital No. 2 in Bydgoszcz, Poland. Material and methods. The study group included 12 patients with ET, 5 with PV, 4 with CML. The control group consisted of 25 healthy volunteers, age- and sex- matched. The material for the study was venous blood collected from the elbow vein into tube containing K2EDTA. The number of endothelial progenitor cells was measured with FACSCalibur flow cytometer (Becton Dickinson, San Diego, USA) using monoclonal antibodies directed against antigens specific for endothelial progenitor cells (EPCs). Results. We observed significantly increased number of EPCs in patients with myeloproliferative neoplasms (MPNs) in comparison to the control group. Detailed analysis showed slightly higher number of EPCs in patients with PV and ET than in the controls, but the differences were not statistically significant. The highest statistically significant number of EPCs was observed in patients with CML. Conclusions. Increased number of EPCs in patients with myeloproliferative neoplasms may indicate increased angiogenesis in these diseases and participation of EPCs in the process of neovascularization.Celem niniejszej pracy była ocena liczby i funkcji komórek progenitorowych śródbłonka w przewlekłej białaczce szpikowej (PBS), czerwienicy prawdziwej (CzP), nadpłytkowości samoistnej (NS). Badaniem objęto 21 pacjentów z nowotworami mieloproliferacyjnymi (średnia wieku 61,77), hospitalizowanych w Oddziale Klinicznym Hematologii i Chorób Rozrostowych Układu Krwiotwórczego Szpitala Uniwersyteckiego nr 2 im. Jana Biziela w Bydgoszczy. Materiał i metody. Badania przeprowadzono u 12 chorych na ET, 4 chorych na CML i 5 chorych na PV. Grupę kontrolną stanowiło 25 zdrowych ochotników. Materiałem do badań była krew pobrana w godzinach porannych z nakłucia żyły łokciowej do probówki zawierającej wersenian dwupotasowy (EDTA). Po inkubacji z odpowiednimi odczynnikami dokonana została analiza cytometryczna przy użyciu cytometru przepływowego FACS Calibur (Becton Dickinson, San Diego, USA) z zastosowaniem programu komputerowego CellQuest. Wyniki. U chorych na przewlekłe nowotwory mieloproliferacyjne stwierdzono istotnie statystyczną zwiększoną liczbę EPCs w porównaniu z grupą kontrolną. U chorych z PV i ET stwierdzono nieznacznie zwiększoną liczbę EPCs w porównaniu do grupy kontrolnej, a różnica ta okazała się nieistotna statystycznie. Najwyższą liczbę EPCs stwierdzono u pacjentów z CML i różnica ta była istotna statystycznie. Wnioski. Zwiększenie liczby EPCs w grupie chorych na przewlekłe nowotwory mieloproliferacyjne świadczy o aktywacji procesów angiogenezy w tych nowotworach i prawdopodobnie czynnym udziale tych komórek w procesie nowotworzenia naczyń

KOMÓRKI PROGENITOROWE ŚRÓDBŁONKA W NOWOTWORACH MIELOPROLIFERACYJNYCH – DONIESIENIA WSTĘPNE

The aim of this study was to assess the number of endothelial progenitor cells in patients with chronic myeloid leukemia (CML), polycythemia vera (PV), essential thrombocythemia (ET). The study involved 21 patients (mean age 61.77) with myeloproliferative neoplasms, hospitalized and diagnosed at the Hematology Clinic of Dr. J. Biziel University Hospital No. 2 in Bydgoszcz, Poland. M a t e r i a l a n d m e t h o d s . The study group included 12 patients with ET, 5 with PV, 4 with CML. The control group consisted of 25 healthy volunteers, age- and sex- matched. The material for the study was venous blood collected from the elbow vein into tube containing K2EDTA. The number of endothelial progenitor cells was measured with FACSCalibur flow cytometer (Becton Dickinson, San Diego, USA) using monoclonal antibodies directed against antigens specific for endothelial progenitor cells (EPCs). R e s u l t s . We observed significantly increased number of EPCs in patients with myeloproliferative neoplasms (MPNs) in comparison to the control group. Detailed analysis showed slightly higher number of EPCs in patients with PV and ET than in the controls, but the differences were not statistically significant. The highest statistically significant number of EPCs was observed in patients with CML. C o n c l u s i o n s . Increased number of EPCs in patients with myeloproliferative neoplasms may indicate increased angiogenesis in these diseases and participation of EPCs in the process of neovascularization.Celem niniejszej pracy była ocena liczby i funkcji komórek progenitorowych śródbłonka w przewlekłej białaczce szpikowej (PBS), czerwienicy prawdziwej (CzP), nadpłytkowości samoistnej (NS). Badaniem objęto 21 pacjentów z nowotworami mieloproliferacyjnymi (średnia wieku 61,77), hospitalizowanych w Oddziale Klinicznym Hematologii i Chorób Rozrostowych Układu Krwiotwórczego Szpitala Uniwersyteckiego nr 2 im. Jana Biziela w Bydgoszczy. M a t e r i a ł i m e t o d y . Badania przeprowadzono u 12 chorych na ET, 4 chorych na CML i 5 chorych na PV. Grupę kontrolną stanowiło 25 zdrowych ochotników. Materiałem do badań była krew pobrana w godzinach porannych z nakłucia żyły łokciowej do probówki zawierającej wersenian dwupotasowy (EDTA). Po inkubacji z odpowiednimi odczynnikami dokonana została analiza cytometryczna przy użyciu cytometru przepływowego FACS Calibur (Becton Dickinson, San Diego, USA) z zastosowaniem programu komputerowego CellQuest. Wy n i k i . U chorych na przewlekłe nowotwory mieloproliferacyjne stwierdzono istotnie statystyczną zwiększoną liczbę EPCs w porównaniu z grupą kontrolną. U chorych z PV i ET stwierdzono nieznacznie zwiększoną liczbę EPCs w porównaniu do grupy kontrolnej, a różnica ta okazała się nieistotna statystycznie. Najwyższą liczbę EPCs stwierdzono u pacjentów z CML i różnica ta była istotna statystycznie. Wn i o s k i . Zwiększenie liczby EPCs w grupie chorych na przewlekłe nowotwory mieloproliferacyjne świadczy o aktywacji procesów angiogenezy w tych nowotworach i prawdopodobnie czynnym udziale tych komórek w procesie nowotworzenia naczyń

Immunomodulatory effects of inosine pranobex on cytokine production by human lymphocytes

Inosine pranobex (inosine dimepranol acedoben, isoprinosine) (Inos) is an immunomodulatory and antiviral drug used in some viral infections, especially in patients with weakened immunity. In the present study, effects of Inos on the production of cytokines attributable to Th1 (IL-2, IFN-γ, and TNF-α) or Th2 cells (IL-4, IL-5, and IL-10) were tested in human peripheral blood lymphocyte cultures stimulated with phytohemagglutinin (PHA). Inos enhanced TNF-α secretion significantly (in short-term – 24-hour, and prolonged term – 72-hour cultures) and IFN-γ (in 72-hour cultures). Surprisingly, production of IL-10 by PHA-stimulated lymphocytes was suppressed by Inos in a dose-dependent manner in both 24-hour and 72-hour cultures. These results shed some light on immunomodulatory properties of Inos and suggest applicability of this agent in patients with a depressed function of the immune system

Mn-based methacrylated gellan gum hydrogels for MRI-guided cell delivery and imaging

This work aims to engineer a new stable injectable Mn-based methacrylated gellan gum (Mn/GG-MA) hydrogel for real-time monitored cell delivery into the central nervous system. To enable the hydrogel visualization under Magnetic Resonance Imaging (MRI), GG-MA solutions were supplemented with paramagnetic Mn2+ ions before its ionic crosslink with artificial cerebrospinal fluid (aCSF). The resulting formulations were stable, detectable by T1-weighted MRI scans and also injectable. Cell-laden hydrogels were prepared using the Mn/GG-MA formulations, extruded into aCSF for crosslink, and after 7 days of culture, the encapsulated human adipose-derived stem cells remained viable, as assessed by Live/Dead assay. In vivo tests, using double mutant MBPshi/shi/rag2 immunocompromised mice, showed that the injection of Mn/GG-MA solutions resulted in a continuous and traceable hydrogel, visible on MRI scans. Summing up, the developed formulations are suitable for both non-invasive cell delivery techniques and image-guided neurointerventions, paving the way for new therapeutic procedures.Sílvia Vieira acknowledges the FCT Ph.D. scholarship (SFRH/BD/102710/2014). J. Miguel Oliveira and J. Silva-Correia acknowledge the FCT grants under the Investigator FCT program (IF/01285/2015 and IF/00115/2015, respectively). The authors also acknowledge the funds provided under the project NanoTech4ALS, funded under the EU FP7 M-ERA.NET program, and ESF (POWR.03.02.00-00-I028/17-00)

Experimental Strategies of Mesenchymal Stem Cell Propagation: Adverse Events and Potential Risk of Functional Changes

Mesenchymal stem cells (MSCs) are attractive candidates for cell-based tissue repair approaches. Hundreds of clinical trials using MSCs have been completed and many others are still being investigated. For most therapeutic applications, MSC propagation in vitro is often required. However, ex vivo culture condition is not fully physiological and may affect biological properties of MSCs including their regenerative potential. Moreover, both cell cryopreservation and labelling procedure prior to infusion may have the negative impact on their expected effect in vivo. The incidence of MSC transformation during in vitro culture should be also taken into consideration before using cells in stem cell therapy. In our review, we focused on different aspects of MSC propagation that might influence their regenerative properties of MSC. We also discussed the influence of different factors that might abolish MSC proliferation and differentiation as well as potential impact of stem cell senescence and aging. Despite of many positive therapeutic effects of MSC therapy, one has to be conscious about potential cell changes that could appear during manufacturing of MSCs

Immunomodulatory effects of inosine pranobex on cytokine production by human lymphocytes

Inosine pranobex (inosine dimepranol acedoben, isoprinosine) (Inos) is an immunomodulatory and antiviral drug used in some viral infections, especially in patients with weakened immunity. In the present study, effects of Inos on the production of cytokines attributable to Th1 (IL-2, IFN-g, and TNF-a) or Th2 cells (IL-4, IL-5, and IL-10) were tested in human peripheral blood lymphocyte cultures stimulated with phytohemagglutinin (PHA). Inos enhanced TNF-a secretion significantly (in short-term - 24-hour, and prolonged term - 72-hour cultures) and IFN-g (in 72-hour cultures). Surprisingly, production of IL-10 by PHA-stimulated lymphocytes was suppressed by Inos in a dose-dependent manner in both 24-hour and 72-hour cultures. These results shed some light on immunomodulatory properties of Inos and suggest applicability of this agent in patients with a depressed function of the immune system

Challenges and Controversies in Human Mesenchymal Stem Cell Therapy

Stem cell therapy is being intensely investigated within the last years. Expectations are high regarding mesenchymal stem cell (MSC) treatment in translational medicine. However, many aspects concerning MSC therapy should be profoundly defined. Due to a variety of approaches that are investigated, potential effects of stem cell therapy are not transparent. On the other hand, most results of MSC administration in vivo have confirmed their safety and showed promising beneficial outcomes. However, the therapeutic effects of MSC-based treatment are still not spectacular and there is a potential risk related to MSC applications into specific cell niche that should be considered in long-term observations and follow-up outcomes. In this review, we intend to address some problems and critically discuss the complex nature of MSCs in the context of their effective and safe applications in regenerative medicine in different diseases including graft versus host disease (GvHD) and cardiac, neurological, and orthopedic disorders

Modulation of LPS-Induced Neurodegeneration by Intestinal Helminth Infection in Ageing Mice

Parasitic helminths induce a transient, short-term inflammation at the beginning of infection, but in persistent infection may suppress the systemic immune response by enhancing the activity of regulatory M2 macrophages. The aim of the study was to determine how nematode infection affects age-related neuroinflammation, especially macrophages in the nervous tissue. Here, intraperitoneal LPS-induced systemic inflammation resulting in brain neurodegeneration was enhanced by prolonged Heligmosomoides polygyrus infection in C57BL/6 mice. The changes in the brain coincided with the increase in M1 macrophages, reduced survivin level, enhanced APP and GFAP expression, chitin-like chains deposition in the brain and deterioration behaviour manifestations. These changes were also observed in transgenic C57BL/6 mice predisposed to develop neurodegeneration typical for Alzheimer’s disease in response to pathogenic stimuli. Interestingly, in mice infected with the nematode only, the greater M2 macrophage population resulted in better results in the forced swim test. Given the growing burden of neurodegenerative diseases, understanding such interactive associations can have significant implications for ageing health strategies and disease monitoring

Transplantation of Human Glial Progenitors to Immunodeficient Neonatal Mice with Amyotrophic Lateral Sclerosis (SOD1/rag2)

Amyotrophic lateral sclerosis (ALS) is a progressive, fatal disease with no effective therapy. The neurodegenerative character of ALS was an appealing target for stem cell-based regenerative approaches. Different types of stem cells have been transplanted in both preclinical and clinical settings, but no convincing outcomes have been noted. Human glial restricted precursors (hGRPs) transplanted intraventricularly to neonatal, immunodeficient mice rescued lifespan of dysmyelinated mice. Intraspinal injection of hGRPs also provided benefits in the mouse model of ALS. Therefore, we have recently developed an immunodeficient model of ALS (double mutant SOD1/rag2), and, in this study, we tested the strategy previously used in dysmyelinated mice of intraventricular transplantation of hGRPs to immunodeficient mice. To maximize potential therapeutic benefits, the cells were implanted into neonates. We used magnetic resonance imaging to investigate the progression of neurodegeneration and therapeutic responses. A cohort of animals was devoted to survival assessment. Postmortem analysis included immunohistochemistry, Nissl staining, and Western blots. Cell transplantation was not associated with improved animal survival, slowing neurodegeneration, or accumulation of misfolded superoxide dismutase 1. Postmortem analysis did not reveal any surviving hGRPs. Grafting into neonatal immunodeficient recipients did not prevent ALS-induced cell loss, which might explain the lack of positive therapeutic effects. The results of this study are in line with the modest effects of clinical neurotransplantations. Therefore, we urge stem cell and ALS communities to develop and implement cell tracking methods to better understand cell fates in the clinic