scholarly journals Evaluation of in vitro immunomodulatory properties of biologically active substances and drugs

Author(s):  
A. V. Duzh ◽  
A. Y. Hancharou

A new combined method for screening of immunomodulatory properties of drugs was developed, including: expression of CD80, HLA-DR, CD32, CD205 and CD197, interleukin-12 production and apoptosis by DCs; production of reactive oxygen species and evaluation of phagocytosis by neutrophils; expression of CD69 and production of tumor necrosis factor-α by Jurkat-tat cells; and CD80 and HLA-DR expression, viability and apoptosis using the Daudi cell line.

Blood ◽  
2004 ◽  
Vol 104 (9) ◽  
pp. 2810-2817 ◽  
Author(s):  
Anna Smed-Sörensen ◽  
Karin Loré ◽  
Lilian Walther-Jallow ◽  
Jan Andersson ◽  
Anna-Lena Spetz

Abstract Dendritic cells (DCs) are antigen-presenting cells with the capacity to prime naive T cells for efficient cellular responses against pathogens such as HIV-1. DCs are also susceptible to HIV-1 infection, which may impair their ability to induce immunity. Here, we examined the ability of HIV-1-infected, in vitro-derived DCs to respond to CD40 ligand (CD40L) stimulation with the aim to study events during early HIV-1 infection. HIV-1BaL-infected p24+ DCs were detected after only 3 days of exposure to highly concentrated virus. We show that HIV-1-infected DCs up-regulated costimulatory molecules, but were skewed in their production of effector cytokines in response to CD40L stimulation. CD40L stimulation induced significant secretion of tumor necrosis factor α (TNFα) and interleukin 12 (IL-12) p70 from both HIV-1-exposed and unexposed DCs. Intracellular stainings of HIV-1-exposed DCs revealed that TNFα could be detected in both the p24- and p24+ DCs, but IL-12 p70 could be found only in the p24- DCs. Thus, although p24+ DCs showed a mature phenotype similar to p24- DCs after CD40L stimulation, they appeared to have an impaired cytokine profile. These observations suggest that HIV-1 infection disables DC function, a phenomenon that may be relevant for optimal induction of HIV-1-specific immune responses. (Blood. 2004;104:2810-2817)


2020 ◽  
Vol 02 (02) ◽  
pp. e109-e116
Author(s):  
Ying Zheng ◽  
Xinyi Xiao ◽  
Zhuoyi Yang ◽  
Meiqi Zhou ◽  
Hui Chen ◽  
...  

AbstractInterleukin-1 receptor antagonist is an important acute-phase protein and an immune mediator, and its expression is associated with the development of hepatitis or acute liver failure. The aim of this study was to investigate whether recombinant human interleukin-1 receptor antagonist directly targets and improves cell survival in a carbon tetrachloride-induced hepatocyte injury model in vitro. A human hepatoma cell line and a mouse hepatocyte cell line were used to establish carbon tetrachloride-induced cell injury models in vitro, and cell viability, apoptosis, and reactive oxygen species level were determined to assess the degree of hepatocellular damage. Quantitative real-time polymerase chain reaction was used to analyze the level of interleukin-1β, interleukin-6, and tumor necrosis factor-α mRNA in cells; extracellular regulated protein kinases 1/2 phosphorylation in hepatocytes was analyzed using western blotting. Recombinant human interleukin-1 receptor antagonist could directly target hepatocytes, improve cell survival, and decrease carbon tetrachloride-induced cell apoptosis in vitro. In hepatocytes, recombinant human interleukin-1 receptor antagonist remarkably downregulated expression of interleukin-1β, interleukin-6, and tumor necrosis factor-α in hepatocytes exposed to carbon tetrachloride. It also decreased accumulation of reactive oxygen species and abrogated the suppression of extracellular regulated protein kinases 1/2 phosphorylation induced by carbon tetrachloride. However, stimulation of cells with an extracellular regulated protein kinases 1/2 inhibitor blocked the recombinant human interleukin-1 receptor antagonist-induced upregulation of extracellular regulated protein kinase1/2 activation and abrogated the improvement in hepatocyte survival following carbon tetrachloride treatment. Collectively, these findings provide new insights into the hepatocyte-protective mechanism of recombinant human interleukin-1 receptor antagonist.


2021 ◽  
Vol 9 (2) ◽  
pp. 151-156
Author(s):  
Larysa M. Strilchuk ◽  
Marta O. Kondratyuk

Toxic excess of biologically active substances named middle molecules (MM), which include byproducts of normal and altered metabolism, products of inflammation and oxidation, bacterial remnants, antibodies and immunoactive substances, plays an important role in pathogenesis of gallbladder diseases and biliary autonomous viscero-visceral cardioneuropathy (BAVVCNP). In order to assess activity of the endogenous intoxication syndrome secondary to BAVVCNP, we examined 20 patients with coronary heart disease to determine the levels of MM in the blood (total and at 238, 254, 266, and 280 nm waves); the levels of MM in urine (at 238, 254, 266, 282, 288, and 310 nm waves) with calculation of aromaticity index (MM 238/280), peptide-nucleotide index (MM 238/266), distribution index (MM 280/254), and L-arginine, and nitrites of the urine. The results were statistically processed. It was revealed that in case of BAVVCNP the severity of the endogenous intoxication syndrome was higher for all specific parameters of endotoxicosis, and especially for the total level of MM in blood (0.77 ± 0.13 units vs. 0.46 ± 0.13 units, p = 0.08), the MM level at 238 nm wave (1.53 ± 0.55 vs. 0.49 ± 0.06, p = 0.08) and hydrophilic MM level in the urine at 288 nm long waves (0.72 ± 0.12 vs. 0.40 ± 0.11, p = 0.05) and 310 nm (0.27 ± 0.08 vs. 0.10 ± 0.03, p <0.05). According to the literature, this may indicate an increase in levels of cholecystokinin, leptin, endothelin, proinflammatory interleukins and tumor necrosis factor α. According to the correlation analysis, activation of endogenous intoxication syndrome was associated with lipid distress syndrome, increased leptin content and accelerated renal filtration.


2021 ◽  
Vol 11 (6) ◽  
pp. 2555
Author(s):  
Lyudmila Asyakina ◽  
Svetlana Ivanova ◽  
Alexander Prosekov ◽  
Lyubov Dyshlyuk ◽  
Evgeny Chupakhin ◽  
...  

This work aims to study the qualitative composition of biologically active substance (BAS) extracts in vitro callus, cell suspension, and root cultures of the medicinal plant Rhaponticum carthamoides. The research methodology is based on high-performance liquid chromatography, and 1H nuclear magnetic resonance (NMR) spectra, to study the qualitative and quantitative analysis of BAS. The results of the qualitative composition analysis of the dried biomass extracts of in vitro callus, cell suspension and root cultures showed that the main biologically active substances in the medicinal plant Rhaponticum carthamoides are 2-deoxy-5,20,26-trihydroxyecdyson (7 mg, yield 0.12%), 5,20,26-trihydroxyecdyson 20,22-acetonide (15 mg, yield 0.25%), 2-deoxy-5,20,26-trihydroxyecdyson 20,22-acetonide (6 mg, yield 0.10%), 20,26-dihydroxyecdyson 20,22-acetonidecdyson 20,22-acetonide (5 mg, yield 0.09%), and ecdyson 20,22-acetonide (6 mg, yield 0.10%). In the future, it is planned to study the antimicrobial, antioxidant, and antitumor activity of BAS of extracts of in vitro callus, cell suspension, and root cultures of the medicinal plant Rhaponticum carthamoides, for the production of pharmaceuticals and dietary supplements with antitumor, antimicrobial and antioxidant effects.


2019 ◽  
Vol 20 (18) ◽  
pp. 4556 ◽  
Author(s):  
Hanna Zielinska-Blizniewska ◽  
Przemyslaw Sitarek ◽  
Anna Merecz-Sadowska ◽  
Katarzyna Malinowska ◽  
Karolina Zajdel ◽  
...  

Obesity is a complex disease of great public health significance worldwide: It entails several complications including diabetes mellitus type 2, cardiovascular dysfunction and hypertension, and its prevalence is increasing around the world. The pathogenesis of obesity is closely related to reactive oxygen species. The role of reactive oxygen species as regulatory factors in mitochondrial activity in obese subjects, molecules taking part in inflammation processes linked to excessive size and number of adipocytes, and as agents governing the energy balance in hypothalamus neurons has been examined. Phytotherapy is the traditional form of treating health problems using plant-derived medications. Some plant extracts are known to act as anti-obesity agents and have been screened in in vitro models based on the inhibition of lipid accumulation in 3T3-L1 cells and activity of pancreatic lipase methods and in in vivo high-fat diet-induced obesity rat/mouse models and human models. Plant products may be a good natural alternative for weight management and a source of numerous biologically-active chemicals, including antioxidant polyphenols that can counteract the oxidative stress associated with obesity. This review presents polyphenols as natural complementary therapy, and a good nutritional strategy, for treating obesity without serious side effects.


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