IN VITRO EFFECTS OF EICOSANOIDS DERIVED FROM DIFFERENT 20-CARBON FATTY ACIDS ON PRODUCTION OF MONOCYTE-DERIVED CYTOKINES IN HUMAN WHOLE BLOOD CULTURES

Cytokine ◽  
2002 ◽  
Vol 20 (5) ◽  
pp. 215-223 ◽  
Author(s):  
E.A Miles ◽  
E Allen ◽  
P.C Calder
Keyword(s):  
Fatty Acids ◽  
Whole Blood ◽  
Blood Cultures ◽  
Human Whole Blood ◽  
In Vitro Effects

scholarly journals Fatty acids modulate cytokine and chemokine secretion of stimulated human whole blood cultures in diabetes

2013 ◽  
Vol 172 (3) ◽  
pp. 383-393 ◽  
Author(s):  
M. C. Simon ◽  
S. Bilan ◽  
B. Nowotny ◽  
T. Dickhaus ◽  
V. Burkart ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Whole Blood ◽  
Blood Cultures ◽  
Human Whole Blood

Elotuzumab (HuLuc63) Activates CD56dim Natural Killer Cells and Monocytes Resulting in the Release of IP-10 and MCP-1

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 108-108 ◽  
Author(s):  
Balaji Balasa ◽  
Mahrukh Huseni ◽  
Jyothi Cherukuri ◽  
Roxanne Steinle ◽  
Amulya Nanisetti ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Nk Cells ◽  
Whole Blood ◽  
Cell Activation ◽  
Nk Cell ◽  
Blood Cultures ◽  
Monocyte Activation ◽  
Human Whole Blood ◽  
Low Levels

Abstract Elotuzumab (formerly known as HuLuc63) is a humanized monoclonal antibody targeting CS1 (CD319, SLAMF7), a protein which is highly and uniformly expressed on myeloma cells. CS1 is also expressed at lower levels on selected lymphocyte subsets in humans, including natural killer (NK) cells and a subset of CD8+ T cells. Elotuzumab mediates killing of multiple myeloma cell lines (OPM-2, L363) via antibody-dependent cellular cytotoxicity in vitro and has significant anti-tumor activity in mouse xenograft models. Elotuzumab is currently being investigated in Phase I safety studies in relapsed multiple myeloma (MM). To further characterize the effects of elotuzumab on different immune cell subsets, we evaluated its effects in human whole blood cultures in vitro. In resting human whole blood, >95% of the CD56dim (CD3−CD56+CD16++) NK population expressed CS1. In contrast, only 50–75% of CD56bright NK cells expressed CS1. The expression level of CS1, as measured by mean fluorescence intensity, was also lower in CD56bright than in CD56dim NK cells. In 24-hr whole blood cultures, elotuzumab selectively activated CD56dim NK cells, as evidenced by up-regulation of CD69, CD11b, CD54, and down-modulation of CD16 expression, while having little or no effect on CD56bright NK cells. Monocytes in the same whole blood culture were also activated as determined by up-regulation of CS1, HLA-DR, and CD54. Monocyte activation was significantly correlated with CD56dim NK cell activation. In contrast, elotuzumab induced little to no activation of either CD4+ or CD8+ T cell subsets. In whole blood cultures, elotuzumab induced low levels of IFN-γ and only negligible levels of TNF-α. However, elotuzumab induced robust levels of the chemokines IP-10 and MCP-1 which correlated significantly with the level of CD56dim NK cell activation. The induction of chemokines was both dose and Fc-dependent, as F(ab′)2 fragments of elotuzumab failed to induce either NK cell activation or release of chemokines. Interestingly, when elotuzumab was added to purified NK cell cultures, no increase in the levels of IP-10 and MCP-1 was observed. Addition of neutralizating anti-IFN-g mAb to elotuzumab-treated whole blood cultures had little effect on NK cell activation but partially reduced monocyte activation and IP-10 release. Finally, addition of anti-CD18 mAb abrogated the elotuzumab-mediated activation of NK cells and monocytes suggesting cell-cell contact plays an important role in elotuzumab-induced activation of CD56dim NK cells and monocytes. In conclusion, the result from this study demonstrated that elotuzumab may activate NK cells via CS1 and CD16 signaling, resulting in the release of low levels of IFN-g and subsequently monocyte-dependant chemokine production including IP-10 and MCP-1. When dosed in a phase I clinical trial, elotuzumab induced a transient elevation of the chemokines IP-10 and MCP-1 in the serum of MM patients, indicating that the whole blood culture system enables the identification of appropriate pharmacodynamic markers.

scholarly journals Sample Aging Profoundly Reduces Monocyte Responses in Human Whole Blood Cultures

2018 ◽  
Vol 2018 ◽  
pp. 1-9
Author(s):  
H. W. Grievink ◽  
M. Moerland
Keyword(s):  
Whole Blood ◽  
Intracellular Cytokine Staining ◽  
Drug Effects ◽  
Blood Cultures ◽  
Blood Collection ◽  
Innate Immune Responses ◽  
Cytokine Release ◽  
Human Whole Blood ◽  
The Public

Human whole blood cultures are widely used for the investigation of physiological pathways and drug effectsin vitro. Detailed information on the effect of “sample aging” (the time span between blood collection and experimental start) on the experimental outcome is not readily available in the public domain. We studied the effect of sample aging on the ability of immune cells to respond to cell-specific immune triggers (LPS, PMA/ionomycin, and SEB). Sample aging at room temperature profoundly inhibited the LPS-induced monocytic cytokine release in minimally diluted whole blood cultures. The reduction ranged from 20–50% after 30 minutes to 80–100% after 10 hours and differed between cytokines (IL-1β, IL-2, IL-6, IFNγ, and TNFα). Sample storage at 4°C or 37°C even worsened this. PMA/ionomycin- and SEB-induced cytokine release, both mainly T-cell-driven, were also reduced by sample aging but to a lesser extent (20–50% after 24 hours). Intracellular cytokine staining revealed that the number of LPS-responding cells was not impacted by sample aging and reduced LPS responsivity could also not be explained by apoptosis or downregulated TLR4 expression. Thus, we speculate that sample aging induces an inhibitory pathway downstream from TLR4 in monocytes. These results underline the importance of quick sample handling when investigating innate immune responses in whole blood, especially for monocyte responses.

scholarly journals AB0095 PRECLINICAL CHARACTERIZATION OF CJ-15314, A HIGHLY SELECTIVE JAK1 INHIBITOR, FOR THE TREATMENT OF AUTOIMMUNE DISEASES

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1347.2-1347
Author(s):  
S. Y. Ki ◽  
H. Shin ◽  
Y. Lee ◽  
H. R. Bak ◽  
H. Yu ◽  
...  
Keyword(s):  
Autoimmune Disease ◽  
Autoimmune Diseases ◽  
Whole Blood ◽  
Inflammatory Diseases ◽  
Selective Inhibition ◽  
Janus Kinase ◽  
Dependent Manner ◽  
Human Whole Blood

Background:Janus kinases (JAK1, JAK2, JAK3, and TYK2) play critical roles in mediating various cytokine signaling, and has been developed as a target for autoimmune diseases such as RA. Tofacitinib, oral Pan-JAK inhibitor, demonstrated efficacy in RA patients, but its widespread use is limited by safety issues. Baricitinib, JAK1/2 inhibitor, is also known to interfere with the hematopoiesis system, such as anemia and thrombocytopenia associated with suppression of JAK2 signals. Therefore, it is necessary to develop a new potent compound that selectively inhibits JAK1 over JAK2, 3Objectives:To identify the pharmacological characteristic based on efficacy of CJ-15314 as potent and selective JAK1 inhibitor for treatment of autoimmune disease.Methods:In vitro, cell-based, kinase panel, Kd value and human whole blood assay were performed to determine the inhibition potency and selectivity for JAK subfamily kinases. In vivo therapeutic potential was evaluated by RA model including rat Adjuvant-Induced Arthritis (AIA) and collagen-induced arthritic (CIA). To confirm the possibility of further expansion into the autoimmune disease, BioMAP® Diversity PLUS® Panel was performed by discoverX.Results:In vitro assay, CJ-15314 inhibited JAK kinase family in a concentration-dependent manner with IC50 values of 3.8 nM against JAK1, Selectivity for JAK1 over JAK2, 3 was approximately 18, 83 fold greater for CJ-15314. In 1mM ATP condition, CJ-15314 has been confirmed to have the highest JAK1 selectivity over competing drugs, under 1 mM ATP condition that reflects the physiological environment in the body. Similarly, Kd values has also confirmed the selectivity of JAK1, which is 10 fold higher than JAK2, 3. Accordingly, in human whole blood assays, CJ-15314 is 11 fold more potent against IL-6 induced pSTAT1 inhibition through JAK1 (IC50 value: 70 nM) than GM-CSF-induced pSTAT5 inhibition (JAK2) whereas baricitinib and filgotinib exhibited only 2 fold and 7 fold respectively.In vivo efficacy model, CJ-15314 inhibited disease severity scores in a dose dependent manner. In the rat AIA model, CJ-15314 at 30 mg/kg dose showed 95.3% decrease in arthritis activity score, 51.2% in figotinib at 30 mg/kg, 97.7% showed baricitinib at 10 mg/kg. CJ-15314 showed superior anti-arthritic efficacy than filgotinib. CJ-15314 also minimally affected anemia-related parameters but not bricitinib end of the 2-week treatment. In the rat CIA model, like 10 mg/kg of bricitinib, 30 mg/kg of CJ-15314 also has a similar effect, with a significant reduction in histopathological scores.In biomap diversity panel, CJ-15314 inhibited the expression of genes such as MCP-1, VCAM-1, IP-10, IL-8, IL-1, sTNF-α and HLA-DR confirming the possibility of expansion into other diseases beyond arthritis.Conclusion:CJ-15314 is a highly selective JAK1 inhibitor, demonstrates robust efficacy in RA animal model and is good candidate for further development for inflammatory diseases.* CJ-15314 is currently conducting a phase I trial in south Korea.References:[1]Clark JD et al. Discovery and development of Janus kinase (JAK) inhibitors for inflammatory diseases. J Med Chem. 2014; 57(12):5023-38.[2]Burmester GR et al. Emerging cell and cytokine targets in rheumatoid arthritis. Nat Rev Rheumatol. 2014; 10(2):77-88[3]Jean-Baptiste Telliez et al. Discovery of a JAK3-selective inhibitor: functional differentiation of JAK3-selective inhibition over pan-JAK or JAK1-selective inhibition. ACS Chem. Biol., 2016; 11 (12):3442-3451Disclosure of Interests:so young Ki Employee of: CJ healthcare, hyunwoo shin Employee of: CJ healthcare, yelim lee Employee of: CJ healthcare, Hyoung rok Bak Employee of: CJ healthcare, hana yu Employee of: CJ healthcare, Seung Chan Kim Employee of: CJ healthcare, juhyun lee Employee of: CJ healthcare, donghyun kim Employee of: CJ healthcare, Dong-hyun Ko Employee of: CJ Healthcare, dongkyu kim Employee of: CJ healthcare

The PstI polymorphism of the endotoxin-inducible heat-shock protein 70-2 gene does not affect messenger RNA level in human whole-blood cultures

2000 ◽  
Vol 26 (8) ◽  
pp. 1139-1143 ◽  
Author(s):  
S. Schroeder ◽  
M. Reck ◽  
L. Eric Lehmann ◽  
M. Book ◽  
A. Hoeft ◽  
...  
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Whole Blood ◽  
Heat Shock Protein 70 ◽  
Messenger Rna ◽  
Blood Cultures ◽  
Human Whole Blood

Chemical Additives in Rumen Fermentations: In Vitro Effects of Various Drugs on Rumen Volatile Fatty Acids and Protozoa

1970 ◽  
Vol 30 (5) ◽  
pp. 812-818 ◽  
Author(s):  
J. J. O'Connor ◽  
G. S. Myers ◽  
D. C. Maplesden ◽  
G. W. Vander Noot
Keyword(s):  
Fatty Acids ◽  
Volatile Fatty Acids ◽  
Chemical Additives ◽  
In Vitro Effects
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