Stem Cell Graft Cellular Composition, Hematological and Immune Recovery in NHL Patients Mobilized with or without Plerixafor: A Prospective Comparison

Abstract Introduction: Plerixafor, a reversible CXCR4 antagonist, may be used to enhance mobilization of CD34+ cells after G-CSF or chemotherapy plus G-CSF. There is a paucity of prospective data regarding cellular composition of grafts collected after plerixafor in chemomobilized patients. Also the data concerning hematopoietic or immune recovery after high-dose chemotherapy in plerixafor treated lymphoma patients is limited. Patients and methods: Thirty-one patients with NHL were included into this prospective study. There were 14 males and 17 females with a median age of 62 years (range 19 - 73). Altogether fourteen patients received plerixafor (plerixafor group) for poor or late mobilization whereas 17 did not (control group). All patients were mobilized with chemotherapy plus G-CSF. Cryopreserved graft samples were analyzed with flow cytometry for T and B cells (CD3/CD8/CD45/CD19) as well as for NK cells (CD3/CD16+CD56). Also CD34+ cell subclasses were analyzed (CD34/CD38/CD133). Complete blood counts were evaluated at +15 days, 1, 3, 6 and 12 months post-transplant. To evaluate immune reconstitution, flow cytometry of lymphocyte subsets (T, B, NK) was performed at 1, 3 and 6 months after the graft infusion with the same antibody panel as for graft analysis. Results: The median number of infused viable CD34+ cells was higher in the control group (3.1 x 106/kg vs. 2.0 x 106/kg, p = 0.036) (Table 1). The median percentage of the most primitive stem cells (CD34+CD133+CD38-) in the grafts was higher in the plerixafor group (3.5 % vs 1.2 %, p = 0.001), but there was no significant difference in the absolute counts (0.07 x 106/kg vs 0.05 x 106/kg, p = 0.620). The median amounts of CD3+CD4+ and CD3+CD8+ T cell subsets, CD3+ and NK (CD3-CD16/56+) cells were all significantly higher in the plerixafor group (Table 1). The neutrophil counts at +15 days after the graft infusion were lower in the plerixafor group (2.1 x 109/l vs. 4.8 x 109/l, p = 0.013). Otherwise there was no significant difference in the hematological reconstitution between the groups. The immune reconstitution was comparable except for the higher number of NK cells in the plerixafor group at one month (0.4 x 109/l vs. 0.1 x 109/l, p = 0.001). Also a trend towards faster recovery of blood CD4+ T cells was observed after one month in the plerixafor group (0.2 x 109/l vs. 0.1 x 109/l, p = 0.097). Conclusions: This prospective study evaluating cellular composition of grafts confirms that the apheresis products collected from plerixafor-treated NHL patients contain a greater proportion of the more primitive stem cells and a greater number of T lymphocytes and NK cells compared to patients mobilized without plerixafor. Hematopoietic reconstitution was comparable between the groups except for slower neutrophil recovery in the plerixafor-group. Immune reconstitution was comparable but NK cell as well as CD4+ T cell recovery was faster in the plerixafor group. These results will be further evaluated in a larger set of patients. Also the possible effect of graft composition on the progression free and overall survival will be evaluated in the ongoing GOA (Graft and Outcome in Autologous transplantation) study. Table 1. Cellular composition of freezed grafts of NHL patients mobilized with or without plerixafor. Mobilization with plerixafor (n = 14) Mobilization without plerixafor (n = 17) P - value Blood graft analysis (x106/kg) CD34 w/a 7AADCD34 w 7AADCD34+CD38-Proportion of CD34+ CD133+CD38- cells from all CD34+ cells (%)CD3+CD4+CD8+CD19+NK 2.1 (0.8 – 5.3)2.0 (0.6 – 5.5)0.07 (0.01 – 0.17)3.5 (0.80 – 10.80)178.3 (49.2 – 454.4)82.1 (29.1 – 267.1)75.4 (16.5 – 279.1)0.0 (0.0 – 0.0)21.5 (0.4 – 39.5) 3.4 (1.9 – 7.2)3.1 (1.5 – 6.7)0.05 (0.11 – 0.18)1.2 (0.44 – 5.3)66.2 (16.6 – 415.4)35.6 (8.0 – 114.3)23.3 (8.4 – 301.8)0.0 (0.0 – 3.2)6.6 (0.6 – 20.7) 0,0070.0360.6200.0010.0010.0010.0030.1920.001 7AAD = 7-Aminoactinomycin D w/a = without w = with Disclosures Jantunen: Sanofi: Employment.

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Faster Engraftment and Immune Reconstitution after Haploidentical Allogenic Hematopoietic Cell Transplantation with CD3/CD19 Depleted as Compared to CD34 Selected Grafts.

Blood ◽

10.1182/blood.v106.11.2200.2200 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2200-2200

Author(s):

Haegele Matthias ◽

Christoph Faul ◽

Peter Lang ◽

Michael Schumm ◽

Rupert Handgretinger ◽

...

Keyword(s):

Stem Cells ◽

Nk Cells ◽

Myeloid Leukaemia ◽

Cell Transplantation ◽

Bacterial Infections ◽

Immune Reconstitution ◽

Hematopoietic Cell Transplantation ◽

Hematopoietic Cell ◽

Cd34 Cells ◽

Significant Difference

Abstract Haploidentical allogeneic hematopoietic cell transplantation (HHCT) using megadoses of CD34-selected hematopoietic stem cells (HSC) is complicated by slow engraftment and delayed immune reconstitution leading to intensive transfusional support, high risk of rejection and high incidence of life threatening infections. Graft CD3/CD19 depletion on a CliniMACS device results in grafts not only containing CD34+ stem cells but also CD34 negative progenitors, natural killer-, dendritic- and facilitating-cells, potentially leading to improved engraftment and immune reconstitution. We report a retrospective comparative study of eleven HHCTs with CD34 selected HSC (CD34) and seven HHCTs with CD3/CD19 depleted grafts (CD3/CD19) performed sequentially at a single institution with a median follow-up > 1 year. Conditioning was 12 Gy TBI, cyclosphosphamide (120 mg/kg) and ATG with or without either etoposide (40 mg/kg) or thiotepa (10mg/kg) in the CD34 group. All patients in the CD3/CD19 group received fludarabine (200 mg/m2), thiotepa (10 mg/kg), melphalan (120 mg/m2) and OKT-3 (5 mg/day, day −5 to +14). No postgrafting immunosuppression was used. Only the CD34 group received G-CSF posttransplant. Median age was 30 (range, 19–47) years in the CD34 group compared to 37 (range, 31–58) years in the CD3/CD19 group. Diagnoses in the CD34 group included acute myeloid leukaemia (AML, n=3), acute lymphoblastic leukaemia (ALL, n=3), chronic myeloid leukaemia (CML, n=2), and one patient each with Non-Hodgkin’s lymphoma, myelodysplastic syndrome and paroxysmal nocturnal hemoglobinuria. Diagnoses in the CD3/CD19 group were AML (n=3), ALL(n=3), and multiple myleoma (n=1). Graft composition was comparable with a median of 9,6x10E6 CD34+ cells/kg and 2,2x10E4 CD3+ cells/kg in the CD34 group versus 7,6x10E6 CD34+ cells/kg and 1,3 x10E4 CD3+ cells/kg in the CD3/CD19 group. The grafts in the CD3/CD19 group, however, contained significantly more CD34 negative cells (99 versus 3 %, p=<0.0001) such as NK-cells, monocytes and granulocytes. Engraftment in the CD3/19 group was significantly faster than in the CD34 group, with a median time to >500 neutrophils/μL of 13 versus 19 days (p=<0.0265) and a median time to >20000 platelets/μL of 13 versus 35 days (p=<0.0001). There was also a significant faster recovery of NK-cells with a median day 100 CD56+ count of 1808 versus 209 cells/μL (p=0.0413). The number of CD3+ cells on day 100 showed no significant difference. No cases of graft rejection have been observed in both groups. Transfusional requirements in the CD3/19 group were lower with a median of 11 versus 35 days with platelet support (p=0.039). There was no significant difference in the incidence of GVHD which was ≤ II in both groups. We observed a lower incidence of bacterial infections in the CD3/C19 group. In conclusion, CD3/19 depleted grafts allow haploidentical hematopoietic cell transplantation with faster engraftment and NK-cell reconstitution leading to a lower rate of bacterial infections and less transfusional requirements.

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Improved Immune Recovery after Transplantation of TCRαβ/CD19 Depleted Allografts from Haploidentical Donors in Pediatric Patients

Blood ◽

10.1182/blood.v124.21.852.852 ◽

2014 ◽

Vol 124 (21) ◽

pp. 852-852

Author(s):

Peter Lang ◽

Tobias Feuchtinger ◽

Heiko-Manuel Teltschik ◽

Wolfgang Schwinger ◽

Patrick Schlegel ◽

...

Keyword(s):

Stem Cells ◽

T Cells ◽

T Lymphocytes ◽

Nk Cells ◽

Pediatric Patients ◽

Conflicts Of Interest ◽

Γδ T Cells ◽

Control Group ◽

Unrelated Donor ◽

Immune Recovery

Abstract Transplantation of haploidentical stem cells has become an accepted option for pediatric patients and adults with high risk malignancies who lack a matched related or unrelated donor. In recent years, the majority of pediatric transplant centers chose the CD34 positive selection of peripheral stem cells, which allowed minimizing GvHD by effective reduction of T cells in the graft. However, infectious complications caused by delayed immune recovery were a major reason for transplant related mortality (TRM). In order to improve the immune recovery, we have established a new T-cell depletion method which removes αβ+ T-lymphocytes via a biotinylated anti-TcRαβ antibody followed by an anti-biotin antibody conjugated to magnetic microbeads while retaining γδ+ T-lymphocytes, natural killer (NK) cells and other cells in the graft. In addition, CD19+ B-lymphocytes were concomitantly depleted for the prevention of post-transplant EBV-associated lymphoproliferative disease. Immune recovery was retrospectively analyzed in a cohort of 41 patients with acute leukemia, MDS and non-malignant diseases, who received αβ T and B cell depleted allografts from haploidentical family donors. Conditioning regimens consisted of fludarabine or clofarabine, thiotepa, melphalan and serotherapy with OKT3 or ATG-Fresenius®. Graft manipulation was carried out with anti TCRαβ and anti CD19 antibodies and immunomagnetic microbeads. γδ T cells and NK cells remained in the grafts. Primary engraftment occurred in 88%, acute graft versus host disease (aGvHD) grade II and III-IV occurred in 10% and 15%. Immune recovery data were available in 26 patients and comparable after OKT3 (n=7) or ATG-F® (n=19). Median time to reach > 100 CD3+/µl, > 200 CD19+ cells/µl and > 200 CD56+ cells/µl for the whole group was 13, 127 and 12.5 days. Compared to a historical control group of patients with CD34 positive selected grafts, significantly higher cell numbers were found for CD3+ at days +30 and +90 (267 vs. 27 and 397 vs. 163 cells/µl), for CD3+4+ at day +30 (58 vs. 11 cells/µl) and for CD56+ at day +14 (622 vs. 27 cells/µl). The clinical impact of this accelerated immune recovery will be evaluated in an ongoing prospective multi-center trial. Disclosures No relevant conflicts of interest to declare.

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Effect of Mesenchymal Stem Cell-derived Microvesicles on Megakaryocytic Differentiation of CD34+ Hematopoietic Stem Cells

Advanced Pharmaceutical Bulletin ◽

10.34172/apb.2020.038 ◽

2020 ◽

Vol 10 (2) ◽

pp. 315-322

Author(s):

Sara Aqmasheh ◽

Karim Shamsasenjan ◽

Elham Khalaf Adeli ◽

Aliakbar Movassaghpourakbari ◽

Parvin Akbarzadehlaleh ◽

...

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Control Group ◽

Hematopoietic Stem ◽

Cd34 Cells ◽

Significant Difference ◽

Significant Expression ◽

Late Stages ◽

Megakaryocyte Differentiation

Purpose: Mesenchymal stem cells (MSCs) release hematopoietic cytokines, growth factors, and Microvesicles (MVs) supporting the hematopoietic stem cells (HSCs). MVs released from various cells, playing a crucial role in biological functions of their parental cells. MSC-derived MVs contain microRNAs and proteins with key roles in the regulation of hematopoiesis. Umbilical cord blood (UCB) is a source for transplantation but the long-term recovery of platelets is a main problem. Therefore, we intend to show that MSC-MVs are able to improve the differentiation of UCB-derived CD34+ cells to megakaryocyte lineage. Methods: In this descriptive study, MSCs were cultured in DMEM to collect the culture supernatant, which was ultracentrifuged for the isolation of MVs. HSCs were isolated from UCB using MACS method and cultured in IMDM supplemented with cytokines and MVs in three different conditions. Megakaryocyte differentiation was evaluated through the expression of specific markers and genes after 72 hours, and the data was analyzed by t test (P<0.05). Results: The expression of specific megakaryocyte markers (CD41 and CD61) in the presence of different concentrations of MSC-MVs did not show any significant difference. Also, the expression of specific genes of megakaryocyte lineage was compared with control group. The expression of GATA2 and c-Mpl was significantly increased, GATA1 was not significantly decreased, and FLI1 was significantly decreased. Conclusion: MSC-MVs could improve the expression of specific megakaryocyte genes; however, there was no significant expression of CD markers. Further studies, including the evaluation of late stages of megakaryocyte differentiation, are required to evaluate platelet production and shedding

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Study of Human Amniotic Membrane Mesenchymal Stem Cells Using Gelatin and Alginate as Nontoxic Scaffolds

Recent Advances in Biology and Medicine ◽

10.18639/rabm.2019.877306 ◽

2019 ◽

Vol 5 ◽

pp. 1

Author(s):

Nike Hendrijantini ◽

Poedjo Hartono ◽

Helen Susilowati ◽

Cindy K. Hartono ◽

Reni P. Daniati ◽

...

Keyword(s):

Stem Cells ◽

Flow Cytometry ◽

Mesenchymal Stem Cells ◽

Amniotic Membrane ◽

Colorimetric Assay ◽

Control Group ◽

Human Amniotic Membrane ◽

Average Percentage ◽

Significant Difference

Perinatal mesenchymal stem cells (MSCs), for example, from amniotic membrane, have advantages over adult sources, such as bone marrow, in terms of ease of availability, cell naivety, tissue stem cell abundance, high capacity of proliferation, and less donor site morbidity, because it does not require invasive procedures. Natural polymer scaffolds, such as gelatin and alginate, are biocompatible. Combination of stem cells from amniotic membrane (hAMSCs) and gelatin or alginate as scaffold can be promising. However, cytotoxicity comparison of gelatin and alginate to hAMSCs has not been widely studied. This study was aimed to compare cytotoxicity of gelatin and alginate on hAMSCs in vitro. Isolation and culture were performed on hAMSCs of the healthy full-term pregnancy. In passage 4, Flow Cytometry CD90, CD105, and CD73 phenotype characterization was done. Colorimetric assay of 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was performed to measure the cytotoxicity. There were three sample groups: (control group) hAMSCs with alpha-minimum essential medium (α-MEM) solution as control; (gelatin group) hAMSCs with gelatin; (alginate group) hAMSCs with alginate. Each group consisted of 12 samples. Flow cytometry of hAMSCs expressed 28.78% CD90, 36.95% CD105, and 44.41% CD73 surface markers. No sample depicted toxicity in either gelatin or alginate group, and this is indicated by the average percentage of living cells in gelatin 97.26% and in alginate 98.43%. No statistically significant difference (ρ=0.057) of cytotoxicity was found between gelatin and alginate to hAMSCs. Gelatin and alginate were nontoxic to hAMSCs in vitro.

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Effect of Placental Extract on Omentum Mesenchymal Stem Cells Differentiation in NMRI Mice

Journal of Molecular Biology Research ◽

10.5539/jmbr.v7n1p176 ◽

2017 ◽

Vol 7 (1) ◽

pp. 176

Author(s):

Maryam Sadat Nezhadfazel ◽

Kazem Parivar ◽

Nasim Hayati Roodbari ◽

Mitra Heydari Nasrabadi

Keyword(s):

Cell Cycle ◽

Stem Cells ◽

Flow Cytometry ◽

Endothelial Cells ◽

Mesenchymal Stem Cells ◽

Control Group ◽

Fat Cell ◽

Nmri Mice ◽

Differentiated Cells ◽

Placental Extract

Omentum mesenchymal stem cells (OMSCs) could be induced to differentiate into cell varieties under certain conditions. We studied differentiation of OMSCs induced by using placenta extract in NMRI mice. Mesenchymal stem cells (MSCs) were isolated from omentum and cultured with mice placenta extract. MSCs, were assessed after three passages by flow cytometry for CD90, CD44, CD73, CD105, CD34 markers and were recognized their ability to differentiate into bone and fat cell lines. Placenta extract dose was determined with IC50 test then OMSCs were cultured in DMEM and 20% placenta extract.The cell cycle was checked. OMSCs were assayed on 21 days after culture and differentiated cells were determined by flow cytometry and again processed for flow cytometry. CD90, CD44, CD73, CD105 markers were not expressed, only CD34 was their marker. OMSCs were morphologically observed. Differentiated cells are similar to the endothelial cells. Therefore, to identify differentiated cells, CD31 and FLK1 expression were measured. This was confirmed by its expression. G1 phase of the cell cycle shows that OMSCs compared to the control group, were in the differentiation phase. The reason for the differentiation of MSCs into endothelial cells was the sign of presence of VEGF factor in the medium too high value of as a VEGF secreting source.

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Impact of Induction Treatment on Stem Cell Collection: A COVID Related Study

Blood ◽

10.1182/blood-2021-149812 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 4848-4848

Author(s):

Brad Rybinski ◽

Ashraf Z. Badros ◽

Aaron P. Rapoport ◽

Mehmet Hakan Kocoglu

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Research Funding ◽

Median Number ◽

Cell Transplant ◽

Cd34 Cells ◽

Significant Difference ◽

Number Of Cycles ◽

Time Required ◽

Stem Cell Collection

Abstract Introduction: Standard induction therapy for multiple myeloma consists of 3-6 cycles of bortezomib, lenalidomide, and dexamethasone (VRd) or carfilzomib, lenalidomide and dexamethasone (KRd). Receiving greater than 6 cycles of a lenalidomide containing regimen is thought to negatively impact the ability to collect sufficient CD34+ stem cells for autologous stem cell transplant (Kumar, Dispenzieri et al. 2007, Bhutani, Zonder et al. 2013). Due to the COVID-19 pandemic, at least 20 patients at University of Maryland Greenebaum Comprehensive Cancer Center (UMGCC) had transplant postponed, potentially resulting in prolonged exposure to lenalidomide containing induction regimens. Here, in the context of modern stem cell mobilization methods, we describe a retrospective study that suggests prolonged induction does not inhibit adequate stem cell collection for transplant. Methods: By chart review, we identified 56 patients with multiple myeloma who received induction with VRd or KRd and underwent apheresis or stem cell transplant at UMGCC between 10/1/19 and 10/1/20. Patients were excluded if they received more than 2 cycles of a different induction regimen, had a past medical history of an inborn hematological disorder, or participated in a clinical trial of novel stem cell mobilization therapy. We defined 1 cycle of VRd or KRd as 1 cycle of "lenalidomide containing regimen". In accordance with routine clinical practice, we defined standard induction as having received 3-6 cycles of lenalidomide containing regimen and prolonged induction as having received 7 or more cycles. Results: 29 patients received standard induction (Standard induction cohort) and 27 received prolonged induction (Prolonged induction cohort) with lenalidomide containing regimens. The median number of cycles received by the Standard cohort was 6 (range 4-6), and the median number of cycles received by the Prolonged cohort was 8 (range 7-13). The frequency of KRd use was similar between patients who received standard induction and prolonged induction (27.58% vs. 25.93%, respectively). Standard induction and Prolonged induction cohorts were similar with respect to clinical characteristics (Fig 1), as well as the mobilization regimen used for stem cell collection (p = 0.6829). 55/56 patients collected sufficient stem cells for 1 transplant (≥ 4 x 10 6 CD34 cells/kg), and 40/56 patients collected sufficient cells for 2 transplants (≥ 8 x 10 6 CD34 cells/kg). There was no significant difference in the total CD34+ stem cells collected at completion of apheresis between standard and prolonged induction (10.41 and 10.45 x 10 6 CD34 cells/kg, respectively, p = 0.968, Fig 2). Furthermore, there was no significant correlation between the number of cycles of lenalidomide containing regimen a patient received and total CD34+ cells collected (R 2 = 0.0073, p = 0.5324). Although prolonged induction did not affect final stem yield, prolonged induction could increase the apheresis time required for adequate collection or result in more frequent need for plerixafor rescue. There was no significant difference in the total number of stem cells collected after day 1 of apheresis between patients who received standard or prolonged induction (8.72 vs. 7.96 x 10 6 cells/kg, respectively, p = 0.557). However, patients who received prolonged induction were more likely to require 2 days of apheresis (44% vs. 25%, p = 0.1625) and there was a trend toward significance in which patients who received prolonged induction underwent apheresis longer than patients who received standard induction (468 vs 382 minutes, respectively, p = 0.0928, Fig 3). In addition, longer apheresis time was associated with more cycles of lenalidomide containing regimen, which neared statistical significance (R 2 = 0.0624, p = 0.0658, Fig 4). There was no significant difference between standard and prolonged induction with respect to the frequency of plerixafor rescue. Conclusions: Prolonged induction with lenalidomide containing regimens does not impair adequate stem cell collection for autologous transplant. Prolonged induction may increase the apheresis time required to collect sufficient stem cells for transplant, but ultimately clinicians should be re-assured that extending induction when necessary is not likely to increase the risk of collection failure. Figure 1 Figure 1. Disclosures Badros: Janssen: Research Funding; J&J: Research Funding; BMS: Research Funding; GlaxoSmithKline: Research Funding.

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The Role of 5-HT on Proplatelet Formation and Thrombopoietin Production

Blood ◽

10.1182/blood-2020-140183 ◽

2020 ◽

Vol 136 (Supplement 1) ◽

pp. 14-14

Author(s):

Mo Yang ◽

Enyu Liang ◽

Jieyu Ye ◽

Beng H Chong ◽

Liang Li

Keyword(s):

Stem Cells ◽

Flow Cytometry ◽

Traumatic Stress ◽

Conflicts Of Interest ◽

Control Group ◽

Mice Model ◽

Synthesis Inhibitor ◽

Rhodamine Phalloidin ◽

Proplatelet Formation ◽

Group 5

Background: Our previous work confirmed that serotonin (5-HT) promotes the proliferation of hemopoietic stem cells and megakaryocytes (Yang M et al, Stem Cells, 2007; 2014). However, the mechanisms remain indefinite. Methods: Q-PCR, Flow Cytometry, Western Blot, or Immunofluorescence microscope were used in the receptor and TPO study. MTT/CCK-8, Proplatelet assay, and Flow Cytometry were also used in cell proliferation and apoptosis study. The relationship between 5-HT and TPO was studied in a traumatic stress mice model. Results: In-vitro study, there was a stimulating effect of 5-HT on proplatelet formation in human bone marrow megakaryocytes. Human BM MK progenitors cultured in serum-free medium with either 5-HT (200nM) or TPO (100 ng/ml) had more proplatelet bearing MKs than the control group (5-HT (12.3 ± 5.0)% vs. Control (6.2 ± 3.5)%, P=0.025; TPO (15.6 ± 2.5)% vs. Control, P=0.04; n=4). The 5-HT treatment group showed more mature and more in the final stage MK cells as compared to the TPO group. 5-HT2A, 2B, 2C receptors were detected in the surface of megakaryocytes. The effect of 5-HT on proplatelet formation in MK cells was via 5-HT2 receptors and this effect was reduced by 5-HT2 receptor inhibitor ketanserin. 5-HT acted on cytoskeleton reorganization in MKs via 5-HT2 receptors and ERK1/2 pathway. Using an immunofluorescence microscope with F-actin specific binder rhodamine-phalloidin staining, the polymerized actin level was lower in the control group than the 5-HT group and actin distributed diffusely throughout the cytoplasm. In contrast, the polymerization actin level was higher in the 5-HT group. Adding ketanserin and ERK1/2 inhibitor PD98059 to 5-HT treatment, the fluorescence intensity was correspondingly reduced. Our data also demonstrated that ERK1/2 was activated in MKs treated with 5-HT for 30 minutes. In a traumatic stress mice model, both of 5-HT and TPO were increased, but the increasing of TPO is posterior to 5-HT. After added LX1606, the synthesis inhibitor of 5-HT, 5-HT was reduced markedly, as well as TPO. The expression of TPO mRNA and the production of TPO protein were increased as compared with the control in this model. Conclusions: This study suggests that 5-HT promotes thrombopoiesis from two aspects: one is the direct effect on megakaryocytes. 5-HT could promote the proplatelet formation from megakaryocytes. The second is the indirect effect by promoting the production of TPO, which is a paracrine secretion to influence thrombopoiesis. Disclosures No relevant conflicts of interest to declare.

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Clinical efficacy on glycemic control and safety of mesenchymal stem cells in patients with diabetes mellitus: Systematic review and meta-analysis of RCT data

PLoS ONE ◽

10.1371/journal.pone.0247662 ◽

2021 ◽

Vol 16 (3) ◽

pp. e0247662

Author(s):

Jingjing He ◽

Desheng Kong ◽

Zhifen Yang ◽

Ruiyun Guo ◽

Asiamah Ernest Amponsah ◽

...

Keyword(s):

Diabetes Mellitus ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Treatment Group ◽

Random Effects ◽

Meta Analysis ◽

Cochrane Library ◽

Control Group ◽

Efficacy And Safety ◽

Significant Difference

Background Diabetes mellitus as a chronic metabolic disease is threatening human health seriously. Although numerous clinical trials have been registered for the treatment of diabetes with stem cells, no articles have been published to summarize the efficacy and safety of mesenchymal stem cells (MSCs) in randomized controlled trials (RCTs). Methods and findings The aim of this study was to systematically review the evidence from RCTs and, where possible, conduct meta-analyses to provide a reliable numerical summary and the most comprehensive assessment of therapeutic efficacy and safety with MSCs in diabetes. PubMed, Web of Science, Ovid, the Cochrane Library and CNKI were searched. The retrieval time was from establishment of these databases to January 4, 2020. Seven RCTs were eligible for analysis, including 413 participants. Meta-analysis results showed that there were no significant differences in the reduction of fasting plasma glucose (FPG) compared to the baseline [mean difference (MD) = -1.05, 95% confidence interval (CI) (-2.26,0.16), P<0.01, I2 = 94%] and the control group [MD = -0.62, 95%CI (-1.46,0.23), P<0.01, I2 = 87%]. The MSCs treatment group showed a significant decrease in hemoglobin (Hb) A1c [random-effects, MD = -1.32, 95%CI (-2.06, -0.57), P<0.01, I2 = 90%] after treatment. Additionally, HbA1c reduced more significantly in MSC treatment group than in control group [random-effects, MD = -0.87, 95%CI (-1.53, -0.22), P<0.01, I2 = 82%] at the end of follow-up. However, as for fasting C-peptide levels, the estimated pooled MD showed that there was no significant increase [MD = -0.07, 95%CI (-0.30, 0.16), P<0.01, I2 = 94%] in MSCs treatment group compared with that in control group. Notably, there was no significant difference in the incidence of adverse events between MSCs treatment group and control group [relative risk (RR) = 0.98, 95%CI (0.72, 1.32), P = 0.02, I2 = 70%]. The most commonly observed adverse reaction in the MSC treatment group was hypoglycemia (29.95%). Conclusions This meta-analysis revealed MSCs therapy may be an effective and safe intervention in subjects with diabetes. However, due to the limited studies, a number of high-quality as well as large-scale RCTs should be performed to confirm these conclusions.

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Role and effects of zinc supplementation in HIV-infected patients with immunovirological discordance: A randomized, double blind, case control study

PLoS ONE ◽

10.1371/journal.pone.0244823 ◽

2021 ◽

Vol 16 (1) ◽

pp. e0244823

Author(s):

Macarena Silva ◽

Carmen G. Montes ◽

Andrea Canals ◽

Maria J. Mackenna ◽

Marcelo Wolff

Keyword(s):

Zinc Supplementation ◽

Group Versus ◽

Cd4 Count ◽

Control Group ◽

Immune Recovery ◽

Plasma Zinc ◽

Absolute Increase ◽

Number Of Patients ◽

Significant Difference ◽

Zinc Levels

Introduction It has been estimated that between 15% and 18% of patients who start antiretroviral therapy (ART) do not achieve a successful immune recovery despite complete virological suppression. In the literature this phenomenom is known as poor immune recovery or immunovirological discordance (IVD). Zinc has an immunomodulatory role associated with T lymphocytes and its supplementation could enhance immune recovery. Objective To determine if zinc supplementation on IVD patients prevents immune failure after 12 months of supplementation. Secondary objectives were to determine serum zinc levels in HIV patients with and without IVD and the frequency of hypozincemia in discordant patients. Method We reviewed the historical record of patients under care at Arriarán Foundation. Following inclusion criteria were defined: 1) age ≥ 18 years, 2) standard ART (three effective drugs) for at least 18 months, 3) virologically suppressed for 12 months, 3) persistence of CD4 count ≤200 cells/mm3 and/or increase ≤ 80 cells/mm3 after one year of viral undetectability. A control group was assigned paired 1:1 by sex, age (± 2 years) that did achieved an increase of CD4> 350 cells/ mm3. In both groups plasma zinc levels were determined. In a later phase, patients with IVD were randomized to receive zinc (15 mg daily) versus placebo. Patients were followed for 12 months with CD4 count, viral load and zinc levels determinations every 4–6 months. Results A total of 80 patients, 40 patients with IVD criteria and 40 controls were included. 92.5% were men, and age average was 47.5 years. The median baseline CD4 was 189 cells/mm3 (71–258) in the cases vs. 552.5 cells/ mm3 (317–400) in the control group with a median increase at the end of the study of 39 cell/mm3 and 19 cell/mm3 respectively. There was no difference in baseline plasma zinc levels between both groups (81.7 + 18.1 in cases versus 86.2 + 11.0 in controls). In the 40 patients with IVD, the median absolute increase in CD4 after annual zinc supplementation was 31.5 cells/mm3 in the treated group versus 50 cells/mm3 in the placebo group, this difference being statistically not significant (p = 0.382). Conclusions Patients with IVD have plasma zinc levels similar to those who achieve adequate immune recovery. Zinc supplementation in IVD patients showed a statistically non-significant difference in in CD4 levels between cases and controls. The results warrant a comparative study with a larger number of patients.

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Suppression of transforming growth factor-β by mesenchymal stem-cells accelerates liver regeneration in liver fibrosis animal model

Zinc supplementation improves heme biosynthesis in rats exposed to lead ◽

10.18051/univmed.2021.v40.29-35 ◽

2021 ◽

Vol 40 (1) ◽

pp. 29

Author(s):

Nur Anna C Sa’dyah ◽

Agung Putra ◽

Bayu Tirta Dirja ◽

Nurul Hidayah ◽

Salma Yasmine Azzahara ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Growth Factor ◽

Liver Fibrosis ◽

Transforming Growth Factor ◽

Healing Process ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Significant Difference ◽

T1 And T2

Introduction Liver fibrosis (LF) results from the unregulated chronic wound healing process in liver tissue. Transforming growth factor-beta (TGF-β) is the major contributing cytokine of LF promotion through activation of quiescent hepatic stellate cells (HSCs) into myofibroblasts (MFs) and increased extracellular matrix (ECM) deposition such as collagen leading to scar tissue development. Mesenchymal stem cells (MSCs) have an immunomodulatory capability that could be used as a new treatment for repairing and regenerating LF through suppression of TGF-β. This study aimed to examine the role of MSCs in liver fibrosis animal models through suppression of TGF-β levels without scar formation particularly in the proliferation phase. Methods In this study, a completely randomized design was used with sample size of 24. Male Sprague Dawley rats were injected intraperitoneally (IP) with carbon tetrachloride (CCl4), twice weekly, for eight weeks to induce LF. Rats were randomly assigned to four groups: negative control, CCl4 group, and CCL4 + MSC-treated groups T1 and T2, at doses of 1 x 106 and 2x106 cells, respectively. TGF-β levels were analyzed by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and a least significant difference (LSD) was used to analyse the data. Results The TGF levels of LF rat models decreased on day 7 after MSC administration. The levels of TGF-β in both MSC groups T1 and T2 decreased significantly compared with the control group (p<0.05). The TGF-β suppression capability of T2 was optimal and more significant than that of T1. Conclusion MSCs can suppress TGF levels in liver fibrosis induced rats.

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