Coculture of human articular chondrocytes with peripheral blood mononuclear cells as a model to study cytokine-mediated interactions between inflammatory cells and target cells in the rheumatoid joint

AbstractContextAndrogens are important modulators of immune cell function impacting proliferation, differentiation and cytokine production. The local generation of active androgens from circulating androgen precursors is an important mediator of androgen action in peripheral androgen target cells or tissue.ObjectiveTo characterize the activation of classic and 11-oxygenated androgens in human peripheral blood mononuclear cells (PBMCs).MethodsPBMCs were isolated from healthy male donors and incubated ex vivo with precursors and active androgens of the classic and 11-oxygenated androgen pathways. Steroids were quantified by liquid chromatography-tandem mass spectrometry. The expression of genes encoding steroid-metabolizing enzymes was assessed by quantitative PCR.ResultsPBMCs generated 8-fold higher amounts of the active 11-oxygenated androgen 11-ketotestosterone than the classic androgen testosterone from their respective precursors. We identified the enzyme AKR1C3 as the major reductive 17β-hydroxysteroid dehydrogenase in PBMCs responsible for both conversions and found that within the PBMC compartment natural killer cells are the major site of AKRC13 expression and activity. Steroid 5α-reductase type 1 catalyzed the 5α-reduction of classic but not 11-oxygenated androgens in PBMCs. Lag time prior to the separation of cellular components from whole blood increased 11KT serum concentrations in a time-dependent fashion, with significant increases detected from two hours after blood collection.Conclusions11-oxygenated androgens are the preferred substrates for androgen activation by AKR1C3 in PBMCs, primarily conveyed by natural killer cell AKR1C3 activity, yielding 11KT the major active androgen in PBMCs. Androgen metabolism by PBMCs can affect the measurement results of serum 11-ketotestosterone concentrations, if samples are not separated in a timely fashion.

Download Full-text

Exo-D-MAPPS Attenuates Production of Inflammatory Cytokines and Promoted Generation of Immunosuppressive Phenotype in Peripheral Blood Mononuclear Cells

Serbian Journal of Experimental and Clinical Research ◽

10.2478/sjecr-2019-0045 ◽

2019 ◽

Vol 0 (0) ◽

Author(s):

Carl Randall Harrell ◽

Bojana Simovic Markovic ◽

Crissy Fellabaum ◽

Dragica Miloradovic ◽

Aleksandar Acovic ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Peripheral Blood Mononuclear Cells ◽

Immune Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Biological Properties ◽

Target Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Abstract Mesenchymal stem cells (MSCs) produce immunomodulatory factors that regulate production of cytokines and chemokines in immune cells affecting their functional properties. Administration of MSCs-sourced secretome, including MSC-derived conditioned medium (MSC-CM) and MSC-derived exosomes (MSC-Exos), showed beneficial effects similar to those observed after transplantation of MSCs. Due to their nano-size dimension, MSC-Exos easily penetrate through the tissue and in paracrine and endocrine manner, may deliver MSC-sourced factors to the target immune cells modulating their function. MSCs derived from amniotic fluid (AF-MSCs) had superior cell biological properties than MSCs derived from bone marrow. We recently developed “Exosomes Derived Multiple Allogeneic Proteins Paracrine Signaling (Exo-d-MAPPS)”, a biological product in which the activity is based on AF-MSC-derived Exos capable to deliver immunomodulatory molecules and growth factors to the target cells. Herewith, we analyzed immunosuppressive capacity of Exo-d-MAPPS against human peripheral blood mononuclear cells (pbMNCs) and demonstrated that Exo-d-MAPPS efficiently suppressed generation of inflammatory phenotype in activated pbMNCs. Exo-d-MAPPS attenuated production of inflammatory cytokines and promoted generation of immunosuppressive phenotype in Lipopolysaccharide-primed pbMNCs. Exo-d-MAPPS treatment reduced expansion of inflammatory Th1 and Th17 cells and promoted generation of immunosuppressive T regulatory cells in the population of Concanavalin A-primed pbMNCs. Similarly, Exod-MAPPS treatment suppressed pro-inflammatory and promoted anti-inflammatory properties of α-GalCer-primed pbMNCs. In summing up, due to its capacity for suppression of activated pbMNCs, Exo-d-MAPPS should be further explored in animal models of acute and chronic inflammatory diseases as a poten-tially new remedy for the attenuation of detrimental immune response.

Download Full-text

I. The morphology of interleukin-2 stimulated human peripheral blood mononuclear cells killing tumor Cells in vitro

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100127980 ◽

1987 ◽

Vol 45 ◽

pp. 734-735

Author(s):

G. R. Hook ◽

M. A. Greenwood ◽

D. Barba ◽

B. Ikejiri ◽

S. N. Chen ◽

...

Keyword(s):

Tumor Cells ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Interleukin 2 ◽

Lak Cells ◽

Target Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Culture of peripheral blood mononuclear cells (PBM) with recombinant interleukin-2 (rIL-2) results in lymphokine activated killer cells (LAK) that can kill fresh non-cultured, natural killer cell resistant tumor cells. Clinically, human LAK and rIL-2 immunotherapy can cause regression of some tumors in humans and preliminary trials using LAK cells to treat glioma patients are beginning . LAK effector cells are a subset of the LAK population that must physically contact target cells in order to kill them. In this and the following abstract, we report the morphology of LAK cells killing human glioma derived cells in vitro.PBM cells were obtained from normal adult volunteers and stimulated in culture for 4 days with rIL-2 (Cetus, Emeryville, CA). Glioma-derived cells were obtained at surgery from a recurrent glioma patient and established as a cell line (SNB-92). SNB-92 cells were positive for glial fibrillary acidic protein and were used at passage 8.

Download Full-text