scholarly journals The Effects of IL-2 Concentration and Adding Manner on Proliferation and Function of CIKs

2020 ◽  
Author(s):  
Yali Liu ◽  
Jicheng Li ◽  
Liangtao Zhao ◽  
Jiarui Zhu ◽  
Hongxia Wang ◽  
...  

Abstract BackgroundCytokine-induced killer cells (CIKs) adoptive cell transfer (ACT) is a common malignant tumor treatment method. Interleukin-2 (IL-2) is one of essential cytokines for CIKs culture. In most studies, only the effect of IL-2 concentration on function of CIKs was studied, but the difference between multifarious adding manner of IL-2 was not explored. MethodsThis study established a novel sequential adding manner of IL-2. Different concentration of IL-2 was added in different CIKs induction phase. Then, proliferation ability of CIKs was evaluated using cell proliferation curves, immune phenotype was analyzed by flow cytometry (FCM), IFN-γ secretion ability and cytotoxicity were detected by ELISA Kit and CCK-8 Kit respectively . Multiple comparison tests were conducted between each group to compare function of CIKs in 12 experimental groups. ResultsAs the IL-2 concentration increased, the number of CIKs continued to increase in each group, but its function was not positively related with its number: CD3+ CD56+ subpopulation ratio, INF-γ secretion ability and cytotoxicity presented irregular changes. During quiescent phase and logarithmic growth phase, adding 300 and 1000 U/mL IL-2 respectively could obtain amounts of powerful CIKs (CD3+ CD56+ subpopulation ratio: 40.9%, INF-γ secretion ability: 542pg/mL, cytotoxicity: 40:1, 74.22) . ConclusionsDifferent concentration of IL-2 had a greater influence on biological function of CIKs in different growth phase, it was better to add IL-2 sequentially in quiescent phase and exponential growth phase of CIKs.

2009 ◽  
Vol 9 (3) ◽  
pp. 337-342
Author(s):  
Tetsuji Okuda ◽  
Yuki Uehara ◽  
Tsung-Yueh Tsai ◽  
Satoshi Nakai ◽  
Michihiro Akiba ◽  
...  

Bacteria and picophytoplankton can be decomposed by chlorine in disinfection process of drinking water treatment, and organic matters released from them would contribute to assimilable organic carbon (AOC). Therefore, the specific AOC formation potential of each picoplankton, i.e. bacteria and picophytoplankton, in the chlorination were evaluated and compared in this research. The nutrient state of bacteria affects onto its specific AOC formation potential. The specific AOC formation potential of bacteria starved, which expected to be similar state to natural bacteria in a reservoir, was 0.8 × 10−13 g/cell. It was half of that of bacteria in the logarithmic growth phase. The difference of specific AOC formation potential of bacteria between in the logarithmic growth phase and in the starving condition would be caused by the amount of cellular organic carbons. The specific AOC formation potential of picophytoplankton was 6.4 × 10−13 g/cell, and 8 times higher than that of bacteria starved. The AOC formed by chlorination of picoplankton should be considered for AOC control in summer season when both bacteria and picophytoplankton multiply.


Author(s):  
B. L. Soloff ◽  
T. A. Rado

Mycobacteriophage R1 was originally isolated from a lysogenic culture of M. butyricum. The virus was propagated on a leucine-requiring derivative of M. smegmatis, 607 leu−, isolated by nitrosoguanidine mutagenesis of typestrain ATCC 607. Growth was accomplished in a minimal medium containing glycerol and glucose as carbon source and enriched by the addition of 80 μg/ ml L-leucine. Bacteria in early logarithmic growth phase were infected with virus at a multiplicity of 5, and incubated with aeration for 8 hours. The partially lysed suspension was diluted 1:10 in growth medium and incubated for a further 8 hours. This permitted stationary phase cells to re-enter logarithmic growth and resulted in complete lysis of the culture.


2019 ◽  
Vol 20 (11) ◽  
pp. 1046-1051 ◽  
Author(s):  
Przemysław Gajda-Morszewski ◽  
Klaudyna Śpiewak-Wojtyła ◽  
Maria Oszajca ◽  
Małgorzata Brindell

Lactoferrin was isolated and purified for the first time over 50-years ago. Since then, extensive studies on the structure and function of this protein have been performed and the research is still being continued. In this mini-review we focus on presenting recent scientific efforts towards the elucidation of the role and therapeutic potential of lactoferrin saturated with iron(III) or manganese(III) ions. The difference in biological activity of metal-saturated lactoferrin vs. the unmetalated one is emphasized. The strategies for oral delivery of lactoferrin, are also reviewed, with particular attention to the metalated protein.


Author(s):  
Marc Permanyer ◽  
Berislav Bošnjak ◽  
Silke Glage ◽  
Michaela Friedrichsen ◽  
Stefan Floess ◽  
...  

AbstractSignaling via interleukin-2 receptor (IL-2R) is a requisite for regulatory T (Treg) cell identity and function. However, it is not completely understood to what degree IL-2R signaling is required for Treg cell homeostasis, lineage stability and function in both resting and inflammatory conditions. Here, we characterized a spontaneous mutant mouse strain endowed with a hypomorphic Tyr129His variant of CD25, the α-chain of IL-2R, which resulted in diminished receptor expression and reduced IL-2R signaling. Under noninflammatory conditions, Cd25Y129H mice harbored substantially lower numbers of peripheral Treg cells with stable Foxp3 expression that prevented the development of spontaneous autoimmune disease. In contrast, Cd25Y129H Treg cells failed to efficiently induce immune suppression and lost lineage commitment in a T-cell transfer colitis model, indicating that unimpaired IL-2R signaling is critical for Treg cell function in inflammatory environments. Moreover, single-cell RNA sequencing of Treg cells revealed that impaired IL-2R signaling profoundly affected the balance of central and effector Treg cell subsets. Thus, partial loss of IL-2R signaling differentially interferes with the maintenance, heterogeneity, and suppressive function of the Treg cell pool.


2021 ◽  
Vol 29 (1) ◽  
pp. 230949902199606
Author(s):  
Takeshi Mochizuki ◽  
Koichiro Yano ◽  
Katsunori Ikari ◽  
Ken Okazaki

Purpose: This study investigated the clinical effects of different patellar components without being affected by the femoral component design in total knee arthritis (TKA) for patients with knee osteoarthritis (OA). Methods: In total, 48 patients with OA who met the criteria of the American College of Rheumatology for OA were enrolled and randomly assigned in a 1:1 ratio to two groups according to the usage of patellar component design for TKA (medialized dome type [dome group] or medialized anatomic type [anatomic group]). To evaluate the clinical outcomes for TKA, knee range of motion (ROM), pain intensity of 0–100 mm visual analog scale (pain VAS), and the Japanese Knee Osteoarthritis Measure (JKOM) score were obtained at baseline and year 1. Results: The difference in knee ROM, pain VAS, or total JKOM score at year 1 was not significant between the dome and anatomic groups ( p = 0.398, 0.733 and 0.536, respectively). Moreover, similar results were obtained for changes in knee ROM, pain VAS, or total JKOM scores from baseline. In both groups, the pain VAS and total JKOM scores were significantly improved at year 1. Conclusion: Both dome and anatomic groups in TKA are significantly effective for pain and function using the JKOM score. However, their efficacy did not differ, according to the JKOM score. Results of this study are rare information focusing on the patellar component design and provide one of the insights into the TKA clinical management.


2021 ◽  
Vol 11 (9) ◽  
pp. 4048
Author(s):  
Javier A. Linares-Pastén ◽  
Lilja Björk Jonsdottir ◽  
Gudmundur O. Hreggvidsson ◽  
Olafur H. Fridjonsson ◽  
Hildegard Watzlawick ◽  
...  

The structures of glycoside hydrolase family 17 (GH17) catalytic modules from modular proteins in the ndvB loci in Pseudomonas aeruginosa (Glt1), P. putida (Glt3) and Bradyrhizobium diazoefficiens (previously B. japonicum) (Glt20) were modeled to shed light on reported differences between these homologous transglycosylases concerning substrate size, preferred cleavage site (from reducing end (Glt20: DP2 product) or non-reducing end (Glt1, Glt3: DP4 products)), branching (Glt20) and linkage formed (1,3-linkage in Glt1, Glt3 and 1,6-linkage in Glt20). Hybrid models were built and stability of the resulting TIM-barrel structures was supported by molecular dynamics simulations. Catalytic amino acids were identified by superimposition of GH17 structures, and function was verified by mutagenesis using Glt20 as template (i.e., E120 and E209). Ligand docking revealed six putative subsites (−4, −3, −2, −1, +1 and +2), and the conserved interacting residues suggest substrate binding in the same orientation in all three transglycosylases, despite release of the donor oligosaccharide product from either the reducing (Glt20) or non-reducing end (Glt1, Gl3). Subsites +1 and +2 are most conserved and the difference in release is likely due to changes in loop structures, leading to loss of hydrogen bonds in Glt20. Substrate docking in Glt20 indicate that presence of covalently bound donor in glycone subsites −4 to −1 creates space to accommodate acceptor oligosaccharide in alternative subsites in the catalytic cleft, promoting a branching point and formation of a 1,6-linkage. The minimum donor size of DP5, can be explained assuming preferred binding of DP4 substrates in subsite −4 to −1, preventing catalysis.


Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 162
Author(s):  
Yohei Shimasaki ◽  
Koki Mukai ◽  
Yuki Takai ◽  
Xuchun Qiu ◽  
Yuji Oshima

Peroxiredoxin (Prx) is a relatively recently discovered antioxidant enzyme family that scavenges peroxides and is known to be present in organisms from biological taxa ranging from bacteria to multicellular eukaryotes, including photosynthetic organisms. Although there have been many studies of the Prx family in higher plants, green algae, and cyanobacteria, few studies have concerned raphidophytes and dinoflagellates, which are among the eukaryotic algae that cause harmful algal blooms (HABs). In our proteomic study using 2-D electrophoresis, we found a highly expressed 2-Cys peroxiredoxin (2-CysPrx) in the raphidophyte Chattonella marina var. antiqua, a species that induces mass mortality of aquacultured fish. The abundance of the C. marina 2-CysPrx enzyme was highest in the exponential growth phase, during which photosynthetic activity was high, and it then decreased by about a factor of two during the late stationary growth phase. This pattern suggested that 2-CysPrx is a key enzyme involved in the maintenance of high photosynthesis activity. In addition, the fact that the depression of photosynthesis by excessively high irradiance was more severe in the 2-CysPrx low-expression strain (wild type) than in the normal-expression strain (wild type) of C. marina suggested that 2-CysPrx played a critical role in protecting the cell from oxidative stress caused by exposure to excessively high irradiance. In the field of HAB research, estimates of growth potential have been desired to predict the population dynamics of HABs for mitigating damage to fisheries. Therefore, omics approaches have recently begun to be applied to elucidate the physiology of the growth of HAB species. In this review, we describe the progress we have made using a molecular physiological approach to identify the roles of 2-CysPrx and other antioxidant enzymes in mitigating environmental stress associated with strong light and high temperatures and resultant oxidative stress. We also describe results of a survey of expressed Prx genes and their growth-phase-dependent behavior in C. marina using RNA-seq analysis. Finally, we speculate about the function of these genes and the ecological significance of 2-CysPrx, such as its involvement in circadian rhythms and the toxicity of C. marina to fish.


2001 ◽  
Vol 281 (5) ◽  
pp. H1938-H1945 ◽  
Author(s):  
Chari Y. T. Hart ◽  
John C. Burnett ◽  
Margaret M. Redfield

Anesthetic regimens commonly administered during studies that assess cardiac structure and function in mice are xylazine-ketamine (XK) and avertin (AV). While it is known that XK anesthesia produces more bradycardia in the mouse, the effects of XK and AV on cardiac function have not been compared. We anesthetized normal adult male Swiss Webster mice with XK or AV. Transthoracic echocardiography and closed-chest cardiac catheterization were performed to assess heart rate (HR), left ventricular (LV) dimensions at end diastole and end systole (LVDd and LVDs, respectively), fractional shortening (FS), LV end-diastolic pressure (LVEDP), the time constant of isovolumic relaxation (τ), and the first derivatives of LV pressure rise and fall (dP/d t max and dP/d t min, respectively). During echocardiography, HR was lower in XK than AV mice (250 ± 14 beats/min in XK vs. 453 ± 24 beats/min in AV, P < 0.05). Preload was increased in XK mice (LVDd: 4.1 ± 0.08 mm in XK vs. 3.8 ± 0.09 mm in AV, P < 0.05). FS, a load-dependent index of systolic function, was increased in XK mice (45 ± 1.2% in XK vs. 40 ± 0.8% in AV, P < 0.05). At LV catheterization, the difference in HR with AV (453 ± 24 beats/min) and XK (342 ± 30 beats/min, P < 0.05) anesthesia was more variable, and no significant differences in systolic or diastolic function were seen in the group as a whole. However, in XK mice with HR <300 beats/min, LVEDP was increased (28 ± 5 vs. 6.2 ± 2 mmHg in mice with HR >300 beats/min, P < 0.05), whereas systolic (LV dP/d t max: 4,402 ± 798 vs. 8,250 ± 415 mmHg/s in mice with HR >300 beats/min, P < 0.05) and diastolic (τ: 23 ± 2 vs. 14 ± 1 ms in mice with HR >300 beats/min, P < 0.05) function were impaired. Compared with AV, XK produces profound bradycardia with effects on loading conditions and ventricular function. The disparate findings at echocardiography and LV catheterization underscore the importance of comprehensive assessment of LV function in the mouse.


2005 ◽  
Vol 71 (10) ◽  
pp. 6008-6013 ◽  
Author(s):  
Domitille Fayol-Messaoudi ◽  
Cédric N. Berger ◽  
Marie-Hélène Coconnier-Polter ◽  
Vanessa Liévin-Le Moal ◽  
Alain L. Servin

ABSTRACT The mechanism(s) underlying the antibacterial activity of probiotic Lactobacillus strains appears to be multifactorial and includes lowering of the pH and the production of lactic acid and of antibacterial compounds, including bacteriocins and nonbacteriocin, non-lactic acid molecules. Addition of Dulbecco's modified Eagle's minimum essential medium to the incubating medium delays the killing activity of lactic acid. We found that the probiotic strains Lactobacillus johnsonii La1, Lactobacillus rhamnosus GG, Lactobacillus casei Shirota YIT9029, L. casei DN-114 001, and L. rhamnosus GR1 induced a dramatic decrease in the viability of Salmonella enterica serovar Typhimurium SL1344 mainly attributable to non-lactic acid molecule(s) present in the cell-free culture supernatant (CFCS). These molecules were more active against serovar Typhimurium SL1344 in the exponential growth phase than in the stationary growth phase. We also showed that the production of the non-lactic acid substance(s) responsible for the killing activity was dependent on growth temperature and that both unstable and stable substances with killing activity were present in the CFCSs. We found that the complete inhibition of serovar Typhimurium SL1344 growth results from a pH-lowering effect.


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