ABSTRACT
BACKGROUND: Human adipose tissue-derived mesenchymal stromal cells (AD-MSCs) have been under focus in regenerative medicine since their discovery as a suitable source of MSCs. AD-MSCs are heterogeneous cells and exhibit variations in population doubling time, morphology and proliferative capacity. This study investigated if human AD-MSCs are developing, during in vitro long-term cultivation, in an unwanted or aberrant way. METHODS: This study monitored AD-MSCs during their in vitro culture till the tenth passage investigating proliferation kinetics, DNA index and surface markers expression. Also, periostin gene expression was examined. RESULTS: The proliferation capacity and colony forming unit were decreased after passage 6 and the population doubling time was increased. Flow cytometric analysis revealed that newly cultivated population strongly expressed MSCs markers, furthermore, reduction of CD105 expression appeared in passage 5 onwards, the later was associated with significant increase in expression of CD34 (a hematopoietic cell marker). Also, reduction of CD73 and CD90 expression was observed from passage 8. Furthermore, during the first six passages, periostin expression was significantly unchanged, with significant upregulation in late passages. CONCLUSIONS: Long-term cultivation of human AD-MSCs changed their characters in an aberrant way and the first four passages might be the most appropriate passages for therapy. More investigation and understanding of these variations are needed to help in standardizing the expansion of MSCs-based therapies.
ABSTRACT
The mechanism of action of CD8+CD25High+FOXP3+ T cells in hepatocellular carcinoma (HCC) has not been fully understood. Herein, the role of CD8+CD25High+FOXP3+ T cells in HCC was compared with that of CD4+CD25High+FOXP3+ regulatory T cells (conventional Tregs). Thirty-five patients with HCC and twenty age and sex-matched healthy adults (controls) were enrolled. The percentage of CD8+CD25High+FOXP3+ T cells and conventional Tregs in peripheral blood was measured by flow cytometry. Our results revealed that the percentage of peripheral CD8+CD25High+FOXP3+ T cells in HCC patients was significantly higher than controls (Pâ¯=â¯0.005). The conventional Tregs showed the same trend with a higher level in HCC than controls (Pâ¯<â¯0.0001). FOXP3 expression of CD8+CD25High+ T cells is higher than that of CD8+CD25low+ and CD8+CD25Negative T cells. The percentage of CD8+CD25High+FOXP3+ T cells positively correlated with that of conventional Tregs in HCC patients but not in controls. The higher alpha-fetoprotein positively correlated with the higher CD8+CD25High+FOXP3+ T cells and conventional Tregs (R2â¯=â¯0.481, Pâ¯<â¯0.0001 and R2â¯=â¯0.249, Pâ¯=â¯0.001, respectively). The frequency of both CD8+CD25High+FOXP3+ T cells and conventional Tregs was significantly increased in HCC with multiple lesions compared with those with one or two lesions. In conclusion: CD8+CD25High+FOXP3+ T cells similar to conventional Tregs might be used as biomarkers of HCC progression. Therapy targeting the peripherally expanded CD8+CD25High+FOXP3+ T cells may provide a novel perspective for HCC treatment.
Subject(s)
CD8-Positive T-Lymphocytes/metabolism , Carcinoma, Hepatocellular/immunology , Forkhead Transcription Factors/metabolism , Interleukin-2 Receptor alpha Subunit/metabolism , Liver Neoplasms/immunology , T-Lymphocytes, Regulatory/metabolism , Aged , Aged, 80 and over , Biomarkers, Tumor/metabolism , Carcinoma, Hepatocellular/blood , Case-Control Studies , Female , Humans , Liver Neoplasms/blood , Lymphocyte Count , Male , Middle Aged , Prospective Studies , alpha-Fetoproteins/metabolismABSTRACT
BACKGROUND AND AIM: Myeloid-derived suppressor cells (MDSCs) play a pivotal role in tumor immunity and induction of immune tolerance to a variety of antitumor effectors, including T lymphocytes. Herein, we tried to evaluate the frequency and clinical significance of MDSCs and different lymphocyte subsets in hepatitis C virus (HCV)-related hepatocellular carcinoma (HCC). METHODS: Four groups were enrolled; chronic HCV (CHC; n = 40), HCV-related liver cirrhosis (n = 40), HCV-related HCC (HCV-HCC; n = 75), and healthy control group (n = 20). The percentage of peripheral lymphocytes subsets and total MDSCs with their main two subsets; monocytic (M-MDSCs) and granulocytic (G-MDSCs) was evaluated by flow cytometry. RESULTS: The frequency of total MSDCs and M-MDSCs was significantly elevated in HCV-HCC especially patients with advanced stage HCC compared with those with early-stage HCC. The frequency of total MSDCs and M-MDSCs was positively correlated with ALT, AFP, and HCV viral load and negatively correlated with CD8+ T-cell frequency. CD4 + T cells were significantly decreased in HCV-HCC patients. The frequency of CD4 + T cells and CD8 + T cells was negatively correlated with AFP and AST, but not with albumin or HCV viral load. CONCLUSION: Taken together, our data suggest that MDSCs, M-MDSCs, and lymphocyte subsets are associated with the development and progression of HCV-related HCC.
