Dimethyl Sulfoxide (DMSO) Decreases Cell Proliferation and TNF-α, IFN-γ, and IL-2 Cytokines Production in Cultures of Peripheral Blood Lymphocytes

Umbilical cord blood (CB) transplantation is thought to be associated with a reduced risk of severe graft-versus-host-disease (GVHD) compared with bone marrow transplantation (BMT). The cytokine cascade is known to be important in the pathogenesis of GVHD; however, previous studies investigating the cytokine secretion pattern of CB cells have been contradictory because of variations in experimental techniques. In this study, the cytokine profile of cord and adult blood lymphocytes and lymphocyte subsets has been assessed at the single-cell level by flow cytometry, using CD4/CD8 and CD45RA/CD45RO markers. Cord and adult blood mononuclear cells were stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of monensin. After 4 to 24 hours of incubation, interleukin-2 (IL-2), IL-4, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) production was measured by three-color flow cytometry. The results show that cord blood lymphocytes (CBL) produce less IL-2, IL-4, IFN-γ, and TNF-α than adult peripheral blood lymphocytes (ABL). Further subset analysis showed that in CBL the majority of cytokine producing cells were CD4+CD45RA+, whereas in ABL the cytokine-producing cells were both CD4+CD45RO+ and CD8+CD45RO+. These results suggest that the reduced incidence of GVHD in CB transplantation may partly due to the altered cytokine profile seen in CBL.

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Integration Analysis of MicroRNA and mRNA Expression Profiles in Human Peripheral Blood Lymphocytes Cultured in Modeled Microgravity

BioMed Research International ◽

10.1155/2014/296747 ◽

2014 ◽

Vol 2014 ◽

pp. 1-16 ◽

Cited By ~ 23

Author(s):

C. Girardi ◽

C. De Pittà ◽

S. Casara ◽

E. Calura ◽

C. Romualdi ◽

...

Keyword(s):

Cell Proliferation ◽

Inflammatory Response ◽

Mrna Expression ◽

Peripheral Blood ◽

Human Peripheral Blood ◽

Expression Profiles ◽

Microgravity Condition ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Human Peripheral Blood Lymphocytes

We analyzed miRNA and mRNA expression profiles in human peripheral blood lymphocytes (PBLs) incubated in microgravity condition, simulated by a ground-based rotating wall vessel (RWV) bioreactor. Our results show that 42 miRNAs were differentially expressed in MMG-incubated PBLs compared with 1 g incubated ones. Among these, miR-9-5p, miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p were the most dysregulated. To improve the detection of functional miRNA-mRNA pairs, we performed gene expression profiles on the same samples assayed for miRNA profiling and we integrated miRNA and mRNA expression data. The functional classification of miRNA-correlated genes evidenced significant enrichment in the biological processes of immune/inflammatory response, signal transduction, regulation of response to stress, regulation of programmed cell death, and regulation of cell proliferation. We identified the correlation of miR-9-3p, miR-155-5p, miR-150-3p, and miR-378-3p expression with that of genes involved in immune/inflammatory response (e.g., IFNG and IL17F), apoptosis (e.g., PDCD4 and PTEN), and cell proliferation (e.g., NKX3-1 and GADD45A). Experimental assays of cell viability and apoptosis induction validated the results obtained by bioinformatics analyses demonstrating that in human PBLs the exposure to reduced gravitational force increases the frequency of apoptosis and decreases cell proliferation.

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Effect of myelopeptides on IFN-γ and IL-4 production by peripheral blood lymphocytes

Doklady Biological Sciences ◽

10.1134/s0012496607020214 ◽

2007 ◽

Vol 413 (1) ◽

pp. 161-163

Author(s):

S. V. Gein ◽

T. V. Gavrilova ◽

V. A. Chereshnev

Keyword(s):

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Ifn Γ

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Correlation between the expression of Fas, Bcl-2 in peripheral blood lymphocytes and the level of IFN-γ, IL-4 in serum of patients with condyloma acuminata

Journal of Huazhong University of Science and Technology [Medical Sciences] ◽

10.1007/bf02830716 ◽

2004 ◽

Vol 24 (1) ◽

pp. 90-91 ◽

Cited By ~ 1

Author(s):

Liu Houjun ◽

Lin Nengxing ◽

Tu Yating ◽

Liu Zhixiang ◽

Huang Changzheng

Keyword(s):

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Condyloma Acuminata ◽

Blood Lymphocytes ◽

Ifn Γ

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TNF-α-Induced Secretion of C-C Chemokines Modulates C-C Chemokine Receptor 5 Expression on Peripheral Blood Lymphocytes

The Journal of Immunology ◽

10.4049/jimmunol.164.12.6180 ◽

2000 ◽

Vol 164 (12) ◽

pp. 6180-6187 ◽

Cited By ~ 41

Author(s):

Felicita Hornung ◽

Giuseppe Scala ◽

Michael J. Lenardo

Keyword(s):

Chemokine Receptor ◽

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes ◽

Tnf Α ◽

Chemokine Receptor 5

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S. PYOGENES M49-16 ARGININE DEIMINASE INHIBITS PROLIFERATIVE ACTIVITY IN HUMAN PERIPHERAL BLOOD LYMPHOCYTES

Russian Journal of Infection and Immunity ◽

10.15789/2220-7619-adf-1363 ◽

2019 ◽

Author(s):

E. Starickova ◽

T. Leveshko ◽

D. Churakina ◽

I. Kudryavtsev ◽

L. Burova ◽

...

Keyword(s):

Immune Response ◽

Cell Proliferation ◽

Peripheral Blood ◽

Parental Strain ◽

Human Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Arginine Deiminase ◽

Intact Cells ◽

Blood Lymphocytes ◽

Human Peripheral Blood Lymphocytes

Arginine deiminase is one of three enzymes constituting the arginine deiminase system in bacteria. It was demonstrated that arginine deiminase exerts anti-proliferative effects on some primary and immortalized mouse and human cells. It is assumed that the inhibitory effect of arginine deiminase on cell proliferation might be related to its ability to result in the arginine exhaustion. T lymphocytes depend on arginine for proliferation, T-cell receptor complex expression, and the differentiation of memory cells. The aim of the current study was to investigate an impact streptococcal arginine deiminase on functions of human peripheral blood lymphocytes. For this, we comparatively analyzed effects of Supernatant of Destroyed Streptococcal Cells (SDSCs) derived from parental strain S. pyogenes M49-16 and its isogenic mutant S. pyogenes M49-16delArcA bearing inactivated arginine deiminase gene (ArcA) on immune cell functions.An impact of supernatants on cell viability was estimated by staining with DAPI dye. Cell proliferation was assessed by MTT-test and flow cytometry by using the method based on intracellular protein staining with vital fluorescent CFSE (carboxyfluoresceinsuccinimidylester) dye. In addition, the level of lymphocyte tyrosine phosphatase CD45 expression in various culturing conditions was evaluated. It was demonstrated that S. pyogenes M49-16 SDSCs had no impact on cells viability. Parental strain-derived SDSC exerted virtually no effect on intact cells proliferation, but considerably suppressed ConA-induced cell proliferation.At the same time, mutant strain-derived SDSC significantly stimulated spontaneous cell proliferation, but not that one after mitogen exposure. It was observed that increased proliferation was accompanied by upregulated CD45 expression, although it was not significant in all cases. These data allow to conclude that bacterial arginine deiminase could be one of pathogenicity factors able to limit lymphocyte proliferation and immune response and could be a part of pathogen strategy to suppress immune response in order to improve bacterial growth and dissemination.

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The effect of deoxynucleosides on cell proliferation of peripheral blood lymphocytes treated with deferoxamine or hydroxyurea

Transfusion Science ◽

10.1016/s0955-3886(00)00094-1 ◽

2000 ◽

Vol 23 (3) ◽

pp. 243-244

Author(s):

Tjomme van der Bruggen ◽

Niki A Georgiou ◽

Maroeska Oudshoorn ◽

Hans S.L.M Nottet ◽

Joannes J.M Marx ◽

...

Keyword(s):

Cell Proliferation ◽

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Blood Lymphocytes

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HLA-A*0201 Restricted Killing Activity of WT1-Specific CTL Generated From the Peripheral Blood Lymphocytes of Normal Healthy Donors.

Blood ◽

10.1182/blood.v114.22.3026.3026 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3026-3026

Author(s):

Deepa Kolaseri Krishnadas ◽

Mindy Stamer ◽

Kim Dunham ◽

Lei Bao ◽

Kenneth Lucas

Keyword(s):

T Cells ◽

Cell Line ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Target Cells ◽

Cd8 Cells ◽

Blood Lymphocytes ◽

Healthy Donors ◽

Ifn Γ

Abstract Abstract 3026 Poster Board II-1002 The Wilms' tumor antigen (WT1) is over-expressed on several human leukemia and solid tumors, and thus is considered as a potential target for cancer immunotherapy. Combating leukemia by targeting WT1 expressing leukemic cells using in vitro generated WT1-specific CTL is one potential approach, but it is difficult to generate an immune response against WT1 due to low T cell precursor frequency in normal healthy individuals. Earlier studies have shown the generation of WT1-A*0201 peptide specific CTL from CD8+ T cells by cloning. Another study reported the production of IFN- γ by WT-1 specific CD8+ T cells. However, the cytolytic killing ability of these IFN- γ producing cells was not further characterized. Here, we demonstrate the generation of WT1-A*0201 specific CTL from the peripheral blood lymphocytes (PBL) of normal healthy donors using CD137 selection. The PBL were stimulated once with RMFPNAPYL (WT1-A*0201 peptide) pulsed autologous dendritic cells and twice with WT1-A*0201 peptide pulsed irradiated peripheral blood mononuclear cells (PBMC). Following three stimulations, the PBL were selected for CD137+ expression and rapidly expanded with OKT3 and IL-2. The WT1-A*0201 specific CTL showed killing of target cells and production of IFN-γ in an antigen-specific manner. The percent killing of WT1-A*0201 peptide pulsed T2 cells (TAP−, HLA- A2+) and autologous B blast (BB) were significantly higher when compared with their control targets. T2 cells and BB either pulsed with an irrelevant A*0201 peptide or un-pulsed served as the control. We have observed similar results with WT1-A*0201 specific CTL generated from normal donor CD8+ cells. However, the efficiency of WT1-A*0201 CTL generated from PBL to kill target cells and produce IFN- γ was higher than CTL from CD8+ cells. The CTL generated from PBL killed BA25, a WT1 expressing A2+ leukemia cell line but failed to kill Molt-4, a WT1 expressing A2− cell line, clearly indicating HLA-A2 restricted CTL activity. The specificity of the generated CTL were further confirmed by staining with WT1-HLA-A*0201 tetramer. The percentage of WT1-specific CD3+CD8+Tetramer+ cells either remained same or higher in CTL generated from PBL when compared with those generated from CD8+ cells. CD137 selection leads to the generation of significant number of CTL in a shorter time when compared to conventional cloning methods. In addition, generation of WT1-A*0201 specific CTL from PBL avoids CD8+ selection. Currently, we are aiming to generate WT1-specific CTL using an overlapping WT1 peptide-mix in order to widen our ability to treat patients with different HLA types. This study has implications for cellular immunotherapy in leukemia patients who relapse following allogeneic stem cell transplantation. Disclosures No relevant conflicts of interest to declare.

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Inhibition of interferon-γ expression by osmotic shrinkage of peripheral blood lymphocytes

AJP Cell Physiology ◽

10.1152/ajpcell.00259.2002 ◽

2003 ◽

Vol 284 (1) ◽

pp. C200-C208 ◽

Cited By ~ 14

Author(s):

K. S. Lang ◽

C. Weigert ◽

S. Braedel ◽

S. Fillon ◽

M. Palmada ◽

...

Keyword(s):

Peripheral Blood ◽

Human Leukocyte ◽

Renal Medulla ◽

Interferon Γ ◽

Peripheral Blood Lymphocytes ◽

Activated T Cells ◽

Stimulatory Action ◽

Blood Lymphocytes ◽

Combined Application ◽

Ifn Γ

A hypertonic environment, as it prevails in renal medulla or in hyperosmolar states such as hyperglycemia of diabetes mellitus, has been shown to impair the immune response, thus facilitating the development of infection. The present experiments were performed to test whether hypertonicity influences activation of T lymphocytes. To this end, peripheral blood lymphocytes (PBL) of cytomegalovirus (CMV)-positive donors were stimulated by human leukocyte antigen (HLA)-A2-restricted CMV epitope NLVPMVATV to produce interferon (IFN)-γ at varying extracellular osmolarity. As a result, increasing extracellular osmolarity during exposure to the CMV antigen indeed decreased IFN-γ formation. Addition of NaCl was more effective than urea. A 50% inhibition was observed at 350 mosM by addition of NaCl. The combined application of the Ca2+ionophore ionomycin (1 μg/ml) and the phorbol ester phorbol 12-myristate 13-acetate (PMA; 5 μg/ml) stimulated IFN-γ production, an effect again reversed by hyperosmolarity. Moreover, hyperosmolarity abrogated the stimulating effect of ionomycin (1 μg/ml) and PMA (5 μg/ml) on the transcription factors activator protein (AP)-1, nuclear factor of activated T cells (NFAT), and NF-κB but not Sp1. In conclusion, osmotic cell shrinkage blunts the stimulatory action of antigen exposure on IFN-γ production, an effect explained at least partially by suppression of transcription factor activation.

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