The effects of oxygen and medicines on T cells in hypoxic co-culture

2021 ◽  
Keyword(s):  
T Cells ◽  
Cell Viability ◽  
Oxygen Supply ◽  
Interleukin 2 ◽  
Hypoxic Condition ◽  
Severe Trauma ◽  
Multiple Organ ◽  
Trauma Patients ◽  
Low Oxygen ◽  
Western Blots

Objectives: Many patients with massive hemorrhage, respiratory failure due to trauma admit the emergency department, and further that the experience can fall into shock, inducing to sepsis, multiple organ failure due to hyperinflammation or immunosuppression. In the these patients, the low oxygen flow with immunosuppression is believed to play a significant role. Hence, oxygen supply and medicines is essential in severe trauma patients. Therefore, this study aims to investigate the effects of oxygen and variable medicines in hypoxic condition. Methods: T cells and macrophages were plated into trans-well plate for co-culture for 30 minutes in hypoxia. After that, the cells were stimulated with lipopolysaccharide (LPS) followed by variable medicines by normoxia or oxygen supply for 2 hrs and cells were inculated overnight under normoxic conditions. The T cell viability was measured by MTT, and the expression of interleukin-2 (IL-2), interleukin-8 (IL-8) and macrophage migration inhibitory factor (MIF) were measured by western blots using the T cells with co-culture with inflammatory maccrophages. Also, the concentration of MIF was analyzed by ELISA. Results: The T cells viability was decreased in hypoxia with LPS stimulation, however, pentoxifylline (PTX) effectively restored cell viability regardless of oxygen state (p < 0.05). Besides, PTX in oxygen supply status restored the decreases in IL-2 expression of T cells and the increases MIF in the LPS stimulation with hypoxia (p < 0.05). Conclusions: PTX has more effectively restored the T cells immunosuppression in hypoxia during oxygen supply, and has an immunomodulation effect by controlling hyperinflammation.

scholarly journals The impacts of oxygen and pentoxifylline in hypoxic condition

2022 ◽  
Vol 20 ◽  
pp. 205873922110565
Author(s):  
Young-Duck Cho ◽  
Sung-Hyuk Choi ◽  
Sung-Jun Park ◽  
Jung-Youn Kim ◽  
Chae-Seung Lim ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Protease Inhibitor ◽  
Oxygen Supply ◽  
Hypoxic Condition ◽  
Severe Trauma ◽  
Massive Hemorrhage ◽  
Trauma Patients ◽  
Low Oxygen ◽  
Inflammatory Conditions

Introduction:Among major trauma patients in the emergency department, the leading cause of morbidity and mortality is a hemorrhagic shock. The low oxygen flow with hypovolemia in trauma patients is believed to play a significant role. Hence, oxygen supply is essential in severe trauma patients with massive hemorrhage. This study aimed to investigate the effect of oxygen supply in hypoxic condition and variable treatments such as pentoxifylline (PTX), glycerol, hypertonic saline (HTS), protease inhibitor, and dexamethasone (DEXA) in macrophage and T cells. Method:Nitric oxide synthase (iNOS) and macrophage migration inhibitory factor (MIF) were measured for macrophage. MIF, interleukin (IL)-2, and IL-8 were measured for T cells. T cell viability was measued by MTT assay. Results: Pentoxifylline decreased iNOS expression mostly followed by glycerol under hypoxia. Under the hyperoxia, PTX and other treatments decreased iNOS expressions in macrophage. MIF expression was lowered with PTX under hypoxia. PTX, glycerol, HTS, and protease inhibitor were effective under hyperoxia in macrophage. PTX increased T cell survival under hypoxia. Under the hyperoxia, IL-2 expressions were upregulated with PTX, glycerol, and HTS. PTX and other treatments were effective for IL-8. Our results indicate that the PTX and the other agents tested reversed the effects of stimulation of lipopolysaccharide, PGE2 in hypoxia or hypoxia. Conclusion:Our study demonstrated potential usefulness in improving immune systems during severe inflammatory conditions similar to septic shock possibly caused by massive hemorrhage.

scholarly journals The Impact of Oxygen and Pentoxifylline in Hypoxic Condition

2020 ◽  
Author(s):  
Young-Duck Cho ◽  
Sung-Hyuk Choi ◽  
Sung-Jun Park ◽  
Woo-Sung Yu ◽  
Jung-Youn Kim ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Protease Inhibitor ◽  
Hypoxic Condition ◽  
Severe Trauma ◽  
Massive Hemorrhage ◽  
Trauma Patients ◽  
Low Oxygen ◽  
Inflammatory Conditions ◽  
The Impact

Abstract Background: Among major trauma patients in the emergency department (ED), the leading cause of morbidity and mortality is a hemorrhagic shock. The low oxygen flow with hypovolemia in trauma patients is believed to play a significant role. Hence, oxygen supply is essential in severe trauma patients with massive hemorrhage. This study aimed to investigate the effect of different oxygen environments (hypoxia 1%, normoxia 20%, hyperoxia 80% oxygen) and variable treatments such as pentoxifylline (PTX), glycerol, hypertonic saline (HTS), protease inhibitor and dexamethasone (DEXA) in macrophage and T cells. Methods: Nitric oxide synthase (iNOS) and macrophage migration inhibitory factor (MIF) were measured for macrophage. MIF, interleukin (IL)-2 and 8 were measured for T cells. MTT assay was done for measuring T cell viability. Coculture was done on T cells on top of macrophages and the same protocols were carried out. Results: PTX decreased iNOS expression mostly followed by Glycerol under hypoxia. Under the hyperoxia, PTX and other treatments decreased iNOS expressions. MIF expression was lowered with PTX under hypoxia. PTX, glycerol, HTS, and protease inhibitor were effective under hyperoxia. PTX increased T cell survival under hypoxia. Under the hyperoxia, IL-2 expressions were upregulated with PTX, Glycerol, and HTS. PTX and other treatments were effective for IL-8, too. Our results indicate that PTX, HTS, and other medications with oxygen showed an effect on macrophage and T cells under various insults and stimulations. Conclusion: Our study demonstrated potential usefulness in improving immune systems during severe inflammatory conditions similar to septic shock possibly caused by massive hemorrhage.

scholarly journals Pentoxifylline Restores T Cell Viability in Hyper-inflammatory Conditions

2020 ◽  
Author(s):  
Sung-Jun Park ◽  
Sung-Hyuk Choi ◽  
Young-Duck Cho ◽  
Jung-Youn Kim ◽  
Han-Jin Cho ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cell Viability ◽  
Interleukin 2 ◽  
Cell Interaction ◽  
Jurkat Cells ◽  
Cell Mediated Immunity ◽  
Western Blots ◽  
Inflammatory Conditions ◽  
Tnf Α

Abstract Background: Immunity is the state of having sufficient biological defenses to avoid infection, trauma, or other unwanted biological invasions. T-cells and macrophages play important role in cell-mediated immunity. Pentoxifylline (PTX) is known to decrease pro-inflammatory cytokine and tumor necrosis factor (TNF-α). However, the effect on the immune system is not known well. This study aims to investigate the effect of PTX on inflammation. Methods: THP-1 derived macrophages were incubated with lipopolysaccharide (LPS) and/or indicated concentration of PTX for 8hr and wash with PBS to eliminate the effect of LPS. In this media, Jurkat cells were plated into trans-well plate and co-culture was done at 12hr. The T cell viability was measured by MTT. Also, the expression of Interleukin-2 (IL-2) was analyzed by RT-PCR and western blots. Results: PTX restored the increased concentration of MIF, TLR4 protein level and mRNA expression of TLR4 in LPS stimulated THP-1 derived macrophages. However, PTX did not restore the decreased T cell proliferation with PGE2 in Jurkat cells. In the co-culture study, The T cell viability was decreased in the THP-1 derived macrophage cells stimulated with LPS. The additional PTX restored the T cell's viability. Besides, PTX restored the decrease in the IL-2 expression of Jurkat cells in the LPS stimulated THP-1 derived macrophages. Conclusion: LPS stimulated THP-1 derived macrophages inhibited the T cell viability in hyper-inflammation conditions. However, PTX restored the T cells viability with increase IL-2. PTX influenced the cell-cell interaction, therefore, had its immunomodulatory effects.

Combined Activation of Coagulation and Inflammation has an Important Role in Multiple Organ Dysfunction and Poor Outcome after Severe Trauma

2002 ◽  
Vol 88 (12) ◽  
pp. 943-949 ◽  
Author(s):  
Takashi Kameue ◽  
Naoyuki Matsuda ◽  
Mineji Hayakawa ◽  
Toshiteru Ishitani ◽  
Yuji Morimoto ◽  
...  
Keyword(s):  
Organ Dysfunction ◽  
Neutrophil Elastase ◽  
Cell Interaction ◽  
Severe Trauma ◽  
Multiple Organ ◽  
Trauma Patients ◽  
Multiple Organ Dysfunction ◽  
Endothelial Cell Interaction ◽  
Poor Outcome ◽  
Close Relationship

SummaryWe tested the hypothesis that activated neutrophil-endothelial cell interaction in DIC can cause endothelial injury contributing to multiple organ dysfunction syndrome (MODS) and a poor outcome after trauma. Fifty-eight severe trauma patients, 29 with DIC and 29 without DIC were studied. Serial levels of soluble L-, P-, and E-selectins, ICAM-1, VCAM-1, thrombomodulin, and neutrophil elastase were measured on days 0-4 after trauma. The numbers of systemic inflammatory response syndrome (SIRS) criteria that patients met were determined, simultaneously. In the DIC patients, higher DIC scores, lower platelet counts, and a longer duration of SIRS were found compared with the non-DIC patients. The incidence of ARDS and MODS were higher in patients with DIC than in those patients without DIC, and the DIC patients had poor outcome. Soluble L-selectin (sL-selectin) level on Day 1 in the DIC patients who died was markedly lower than those in the non-DIC patients. The levels of sPand sE-selectins, sICAM-1, and sVCAM-1 were more elevated in the patients with DIC than in those without DIC on days 2 to 4. Neutrophil elastase and sThrombomodulin levels in the DIC patients persistently increased during the study period compared to those in the non-DIC patients. Maximum DIC scores in the DIC group showed good correlations with peak levels of sICAM-1, sVCAM-1, neutrophil elastase, sThrombomodulin, and the number of dysfunctioning organs. Highly activated and sustained inflammation caused by neutrophil-endothelium interaction in DIC gives rise to MODS and poor outcome in patients with severe trauma. These results suggest a close relationship between inflammation and thrombosis in posttrauma DIC.

scholarly journals Splenectomy is Associated with Altered Leukocyte Kinetics after Severe Trauma

2020 ◽  
Author(s):  
Michel Teuben ◽  
Arne Hollman ◽  
Taco J. Blokhuis ◽  
Roman Pfeifer ◽  
Roy Spijkerman ◽  
...  
Keyword(s):  
Immune Response ◽  
Immune System ◽  
Peripheral Blood ◽  
Cellular Immune Response ◽  
Severe Trauma ◽  
Multiple Organ ◽  
Liver Trauma ◽  
Trauma Patients ◽  
Severely Injured ◽  
Cellular Immune

Abstract Background Inadequate activation of the innate immune system after trauma can lead to severe complications such as Acute Respiratory Distress Syndrome and Multiple Organ Dysfunction Syndrome. The spleen is thought to modulate the cellular immune system. Furthermore, splenectomy is associated with improved outcome in severely injured trauma patients. We hypothesized that a splenectomy alters the cellular immune response in polytrauma.Methods All adult patients with an ISS ≥ 16 and suffering from splenic or hepatic injuries were selected from our prospective trauma database. Absolute leukocyte numbers in peripheral blood were measured. White blood cell kinetics during the first 14 days were compared between splenectomized patients, patients treated surgically for liver trauma and nonoperatively treated individuals.Results A total of 129 patients with a mean ISS of 29 were included. Admission characteristics and leukocyte numbers were similar in all groups, except for slightly impaired hemodynamic status in patients with operatively treated liver injuries. On admission, leukocytosis occurred in all groups. During the first 24 hours, leukopenia developed gradually, although significantly faster in the operatively treated patients. Thereafter, leukocyte levels normalized in all nonoperatively treated cases whereas leukocytosis persisted in operatively treated patients. This effect was significantly more prominent in splenectomized patients than all other conditions. Conclusions This study demonstrates that surgery for intra-abdominal injuries is associated with an early drop in leucocyte numbers in peripheral blood. Moreover, splenectomy in severely injured patients is associated with an altered cellular immune response reflected by a persistent state of prominent leukocytosis after trauma.

scholarly journals Effects of methanolic plant extracts on cell proliferation and HIF activity under hypoxic condition in vitro

2019 ◽  
pp. 1-9
Author(s):  
Pheik-Sheen Cheow ◽  
Norazizah Shafee ◽  
Sien-Yei Liew ◽  
Muhajir Hamid
Keyword(s):  
Cell Viability ◽  
Plant Extracts ◽  
Hypoxia Inducible Factor ◽  
Hypoxic Condition ◽  
Luciferase Reporter ◽  
Transcriptional Level ◽  
Low Oxygen ◽  
Melastoma Malabathricum ◽  
Clinacanthus Nutans

Low oxygen tension is termed as hypoxia. Hypoxia will lead to transcription of hypoxia-inducible factor (HIF) and regulation of downstream gene expression. Underexpression or overexpression of HIF was found to be responsible for various diseases. Proper regulation of this transcriptional factor will aid in treatment of the related diseases. Nowadays, many different approaches are used to modulate HIF, including the usage of naturally-derived plant extracts. Plant extracts are widely accepted compared to other treatments as they are less harmful to the patient and are widely available. In this study, the cytotoxicity of eight different plant extracts under two different gaseous conditions, hypoxic and normoxic, were examined. We also examined the HIF activity shown by the cells under treatment of various concentrations of plant extracts. All eight plants were dried, blended, extracted using methanol, and evaporated to form crude plant extracts. MTT assay was performed by treating the cells with different concentrations of plant extracts and cell viability was determined. Meanwhile, HIF activity of the cells was evaluated by using single luciferase reporter assay. Relative cytotoxicity shown by the cells was different for each plant extract under the various concentration. Pereskia bleo, Orthosiphon aristatus, and Clinacanthus nutans showed high cell viability, 80% of cell viability, within the range of concentration tested. In contrast, Gynura procumbens, Hydrocotyle sibthorpiodies, Pereskia grandifolia, Strobilanthes cripus, and Melastoma malabathricum showed low cell viability. Most of the cells showed activation of HIF activity when treated with different concentrations of the plant extracts. When cells were treated with high concentrations of plant extracts, inhibition of HIF activity were seen and was correlated with low cell viability after treatment. The most notable part of the study was that more than 100% HIF activation was observed for Clinacanthus nutans. However, the cell viability remained high. This might indicate that Clinacanthus nutans is a promising candidate to activate HIF at a transcriptional level with minimal cytotoxicity.

scholarly journals Splenectomy is associated with altered leukocyte kinetics after severe trauma

2021 ◽  
Vol 26 (1) ◽  
Author(s):  
Michel Paul Johan Teuben ◽  
Arne Hollman ◽  
Taco Blokhuis ◽  
Roman Pfeifer ◽  
Roy Spijkerman ◽  
...  
Keyword(s):  
Immune Response ◽  
Immune System ◽  
Peripheral Blood ◽  
Cellular Immune Response ◽  
Severe Trauma ◽  
Multiple Organ ◽  
Liver Trauma ◽  
Trauma Patients ◽  
Severely Injured ◽  
Cellular Immune

Abstract Background Inadequate activation of the innate immune system after trauma can lead to severe complications such as Acute Respiratory Distress Syndrome and Multiple Organ Dysfunction Syndrome. The spleen is thought to modulate the cellular immune system. Furthermore, splenectomy is associated with improved outcome in severely injured trauma patients. We hypothesized that a splenectomy alters the cellular immune response in polytrauma. Methods All adult patients with an ISS ≥ 16 and suffering from splenic or hepatic injuries were selected from our prospective trauma database. Absolute leukocyte numbers in peripheral blood were measured. White blood cell kinetics during the first 14 days were compared between splenectomized patients, patients treated surgically for liver trauma and nonoperatively treated individuals. Results A total of 129 patients with a mean ISS of 29 were included. Admission characteristics and leukocyte numbers were similar in all groups, except for slightly impaired hemodynamic status in patients with operatively treated liver injuries. On admission, leukocytosis occurred in all groups. During the first 24 h, leukopenia developed gradually, although significantly faster in the operatively treated patients. Thereafter, leukocyte levels normalized in all nonoperatively treated cases whereas leukocytosis persisted in operatively treated patients. This effect was significantly more prominent in splenectomized patients than all other conditions. Conclusions This study demonstrates that surgery for intra-abdominal injuries is associated with an early drop in leucocyte numbers in peripheral blood. Moreover, splenectomy in severely injured patients is associated with an altered cellular immune response reflected by a persistent state of prominent leukocytosis after trauma.

Sign in / Sign up

Export Citation Format

Share Document