Leukemia cell microvesicles promote survival in umbilical cord blood hematopoietic stem cells

Microvesicles can transfer their contents, proteins and RNA, to target cells and thereby transform them. This may induce apoptosis or survival depending on cell origin and the target cell. In this study, we investigate the effect of leukemic cell microvesicles on umbilical cord blood hematopoietic stem cells to seek evidence of apoptosis or cell survival. Microvesicles were isolated from both healthy donor bone marrow samples and Jurkat cells by ultra-centrifugation and were added to hematopoietic stem cells sorted from umbilical cord blood samples by magnetic associated cell sorting (MACS) technique. After 7 days, cell count, cell viability, flow cytometry analysis for hematopoietic stem cell markers and qPCR for P53 gene expression were performed. The results showed higher cell number, higher cell viability rate and lower P53 gene expression in leukemia group in comparison with normal and control groups. Also, CD34 expression as the most important hematopoietic stem cell marker, did not change during the treatment and lineage differentiation was not observed. In conclusion, this study showed anti-apoptotic effect of leukemia cell derived microvesicles on umbilical cord blood hematopoietic stem cells

MicroRNA-21 over expression in umbilical cord blood hematopoietic stem progenitor cells by leukemia microvesicles

Microvesicles are able to induce the cell of origin's phenotype in a target cell. MicroRNA-21, as an oncomir, is up-regulated in almost all cancer types such as leukemia which results in cell proliferation. In this study, we examine the ability of leukemia microvesicles to induce proliferation in hematopoietic stem progenitor cells (HSPCs) via microRNA-21 dysregulation. Herein, leukemia microvesicles were isolated from HL-60 and NB-4 cell lines by ultracentrifugation, and then their protein content was measured. Normal HSPCs were isolated from umbilical cord blood samples by a CD-34 antibody. These cells were treated with 20 and 40 μg/mL leukemia microvesicles for 5 and 10 days, respectively. Cell count, CD-34 analysis, and a microRNA-21 gene expression assay were done at days 5 and 10. HSPCs showed a significant increase in both microRNA-21 gene expression and cell count after treating with leukemia microvesicles compared with the control group. CD-34 analysis as stemness proof did not show any difference among the studied groups. This data suggests that HSPC proliferation followed by microRNA-21 gene over expression can be another evidence of a leukemia-like phenotype induction in a healthy target cell by leukemia microvesicles

LY86, LRG1 and PDE9A genes overexpression in umbilical cord blood hematopoietic stem progenitor cells by acute myeloid leukemia (M3) microvesicles

Background Microvesicles as a new device of cell–cell communication are potentially able to induce some phenotypes and genotypes of an origin cell in a target cell. We evaluate the role of leukemia microvesicles on the leukemia stem cells (LSCs)-specific genes expression in healthy hematopoietic stem progenitor cells (HSPCs). Methods HL-60 and NB-4 cell lines were selected for microvesicles isolation by ultracentrifugation. Healthy HSPCs were obtained by magnetic association cell sorting (MACS) and CD-34 micro-beads from umbilical cord blood samples and then, were treated with 20 and 40 μg/ml leukemia microvesicles for 10 days, respectively. LY86, LRG1 and PDE9A genes expression as LSC specific genes were analyzed by QRT-PCR. Surface CD-34 antigen as stemness marker was measured by flow cytometry technique. Results Healthy HSPCs showed a significant increase in LSC specific genes expression after treatment with both 20 and 40 μg/ml leukemia microvesicles at day 10. All studied groups showed more than 70% surface CD-34 antigen at the last day of experiment which proved HSPCs stemness. Conclusion Our results suggest that healthy HSPCs can be transformed genetically by leukemia microvesicles to over express LSC specific genes. This may be further evidence of leukemia-like transformation by leukemia microvesicles. Keywords: Microvesicles, Hematopoietic stem progenitor cells, Leukemia, Cell communicatio

Increased Plasma Levels of Soluble CD27 among HIV/HCV Co-infected and HIV/HCV/GBV-C Triply Infected Subjects

CD27 is a biomarker associated with both T-cells and B-cells activation .Plasma soluble CD27 (sCD27) was identified as  a marker of disease outcome in Human Immunodeficiency Virus (HIV) infection .Testing of plasma sCD27 represents a good tool to monitor the change of immune activation during HIV infection.We sought to analyses role of Hepatitis C Virus (HCV) and also GB Virus type C (GBV-C) co-infections on HIV-related immune activation, through measuring sCD27 plasma levels.Blood samples from a total of 86 patients with HIV infection were taken. Plasmas were analyzed for HCV using serologic test and GBV-C by reverse transcriptase polymerase chain reaction (RT-PCR). CD4+ and CD8+T-cell counts were evaluated by CD3/CD4+ and CD3/CD8+ double staining of whole blood followed by flow cytometric analysis .Then  Cross-sectional comparison of sCD27 plasma levels was carried out among patients : HIV (n=20), HIV/ GBV-C (n=14), HIV/ (HCV) (n=26) and HIV/HCV/GBV-C (n=26).Plasma level of sCD27 was higher in HIV/HCV/GBV-C patients as compared to HIV mono-infected patients (p= 0.006) and based on results there was significant differences in the plasma levels of sCD27 between HIV-infected individuals with and without HCV coinfection (P=0.017) and also correlation between sCD27 and percent of CD4+T-cells was in highest level among HIV/HCV co-infected patients group [r= -0.59 (p=0.001)]. High levels of sCD27 among HIV/HCV patients argues in favor of sCD27 plasma level determination for monitoring of clinical features among HIV/HCV coinfected patients

The antitumor efficiency of combined electrochemotherapy and single dose irradiation on a breast cancer tumor model

Background. The aim of this study was to investigate the antitumor effectiveness of electrochemotherapy with cisplatin combined with suboptimal radiotherapy doses. Tumor radiosensitization was evaluated on large invasive ductal carcinoma tumors in Balb/C mice. Materials and methods. Tumors of an average volume of 630 mm3 were treated with cisplatin, electric pulses, radiotherapy, electrochemotherapy, alone as well as in appropriate combinations. Tumors were irradiated with Cobalt-60 γ-rays at doses 3 Gy and 5 Gy in combination with electrochemotherapy using cisplatin. Controls included each of the treatments alone as well as the combination of the radiotherapy with electric pulses alone or with cisplatin alone. Antitumor effectiveness was evaluated by tumor growth delay, tumor-doubling time, inhibition ratio and the objective response rates. Results. As anticipated, electrochemotherapy was more effective than the treatment with cisplatin alone or the application of the electric pulses alone. When treatments were combined with tumor irradiation at either 3 or 5 Gy, the combination with electrochemotherapy was more effective: at 5 Gy, 2 animals out of 8 were in complete remission 100 days later. In general the higher 5 Gy dose of γ-radiation was more effective than the lower one of 3 Gy. Conclusions. The results of our study demonstrate that irradiation doses, 3 Gy or 5 Gy, increase the antitumor effectiveness of electrochemotherapy with cisplatin on invasive ductal carcinoma tumors. Good antitumor results were achieved in experimental tumors with a size comparable to clinical lesions, demonstrating that this three-modality combined treatment is useful for the treatment of large lesions even at sub-optimal radiotherapy doses

Hepatitis B Virus Infection In An Anti-Hbc Negative Patient: A Case Report

One of the best reliable markers of hepatitis B virus infection is antibodies to the core antigen (Anti-HBc). A first-time blood donor with HBsAg positivity was identified as an HBV carrier that was anti-HBc negative. The patient was followed for 24 months in order to investigate the evolution of his HBV serological profiles and HBV-DNA (PCR). In the follow-up for 24 months, HBsAg, HBeAg and HBV-DNA (PCR) were positive but all the time anti-HBc remained negative. HBV DNA viral load was 3.4x106 copies per mL. In the immunohistochemical study on the needle liver biopsy, the hepatocytes were positive for HBcAg and HBsAg. For this immunological situation, the most probable hypothesis is an immunotolerance to HBV due to an in utero HBV infection. This situation does not impose a risk of HBV transmission by blood transfusion, because HBsAg positive donations are excluded and discarded by HBsAg screening tests