P1623Activation of invariant natural killer T cells by alpha-galactosylceramide ameliorates doxorubicin-induced cardiotoxicity in mice

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
Y Obata ◽  
N Ishimori ◽  
A Saito ◽  
S Kinugawa ◽  
I Nakano ◽  
...  
Keyword(s):  
Body Weight ◽  
Natural Killer ◽  
Intraperitoneal Injection ◽  
Antineoplastic Agent ◽  
Left Ventricular ◽  
Dependent Manner ◽  
Inkt Cells ◽  
Anti Inflammatory ◽  
Invariant Natural Killer ◽  
Natural Killer T

Abstract Objective Doxorubicin (DOX) is an effective antineoplastic agent commonly used to treat many types of cancer but its clinical use is limited because of cardiotoxicity, which might proceed to irreversible cardiac dysfunction in a dose-dependent manner. The precise mechanism of DOX-induced cardiotoxicity is still not fully elucidated but it has been reported that cardiac inflammation is involved in the cardiotoxicity. Invariant natural killer T (iNKT) cells, a unique subset of T lymphocytes that recognize glycolipid antigens and secrete a large amount of Th1 and Th2 cytokines on activation, have been shown to play crucial roles in the regulation of immune responses. However, it remains unclear whether iNKT cells are involved in DOX-induced cardiotoxicity. Methods and results Male C57BL/6J mice were administered DOX (20mg/kg body weight single intraperitoneal injection; n=28) or vehicle (Vehicle; n=6). DOX-administered mice were further divided into 2 groups; α-galactosylceramide (αGC, 0.1μg/g body weight twice intraperitoneal injection; DOX-αGC; n=14), which specifically activates iNKT cells, or phosphate-buffered saline alone (PBS; DOX-PBS; n=14) 4 days before and 3 days after DOX administration. Survival rate at 14 days after DOX/Vehicle administration was significantly lower in DOX-PBS than in Vehicle (71% vs. 100%, P<0.05), and this decrease was completely attenuated in DOX-αGC (100%, P<0.05 vs. DOX-PBS). Echocardiography at 14 days after DOX/Vehicle administration revealed that left ventricular (LV) fractional shortening was significantly reduced in DOX-PBS compared to Vehicle (49.3±0.8% vs. 59.2±1.7%, P<0.05), and this decrease was completely attenuated in DOX-αGC (57.7±1.3%, P<0.05 vs. DOX-PBS) without affecting LV end-diastolic diameter. Picro-sirius red staining revealed that the ratio of fibrosis area to the cardiac tissue was markedly higher in DOX-PBS than in Vehicle (4.3±0.5% vs. 2.2±0.1%, P<0.05), and this increase was completely attenuated in DOX-αGC (2.8±0.1%, P<0.05 vs. DOX-PBS). Real-time PCR analysis revealed that mRNA expression of anti-inflammatory Th2 cytokine IL-4 was enhanced by 7.9-folds in DOX-αGC compared to DOX-PBS, though the difference did not reach statistically significance (P=0.09). Conclusions Activation of iNKT cells by αGC ameliorates DOX-induced cardiotoxicity in mice via up-regulation of anti-inflammatory IL-4 and reducing cardiac fibrosis. iNKT cell activation may be a novel therapeutic strategy against DOX-induced cardiotoxicity. Acknowledgement/Funding Japan Agency for Medical Research and Development (18lm0203001j0002) and JSPS KAKENHI (18K15834)

Abstract 11982: Circulating Invariant Natural Killer T Cells are Decreased in Patients With Chronic Heart Failure

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Akimichi Saito ◽  
Naoki Ishimori ◽  
Mikito Nishikawa ◽  
Shintaro Kinugawa ◽  
Hiroyuki Tsutsui
Keyword(s):  
Heart Failure ◽  
Chronic Heart Failure ◽  
Ejection Fraction ◽  
Natural Killer ◽  
Left Ventricular ◽  
Idiopathic Dilated Cardiomyopathy ◽  
Inkt Cells ◽  
Lv Ejection Fraction ◽  
Invariant Natural Killer ◽  
Natural Killer T

Objective: Inflammatory mediators play a crucial role in the development of chronic heart failure (HF). Invariant natural killer T (iNKT) cells, a unique subset of T lymphocytes, which recognize glycolipid antigens and secrete a large amount of T helper (Th) 1/Th2 cytokines on activation, function as immunomodulatory cells in the various pathological processes. We have demonstrated that iNKT cells have a protective role against the development of left ventricular (LV) remodeling and failure after myocardial infarction in mice. However, it remains unclear whether iNKT cells are involved in the development of HF in humans. Methods and Results: Nine HF patients (NYHA II or III, LV ejection fraction 26.3±3.0%) and 8 healthy controls were studied. The mean age and male gender were comparable between HF and controls (51.2±5.1 vs. 45.1±4.5 years and 77.8 vs. 75.0%). The causes of HF were idiopathic dilated cardiomyopathy in 3, ischemic in 2, and others in 4 patients. Plasma BNP was significantly higher in HF than in controls (739.4±207.2 vs. 19.8±6.5 pg/mL, P <0.01). The number of circulating iNKT cells, identified by the positive-staining of Vα24-Jα18 T Cell Receptor by flow-cytometric analysis, was significantly lower in HF (747±85 vs. 1058±271 counts/mL, P <0.01). Its ratio to the total lymphocyte was also significantly lower (0.111±0.004 vs. 0.146±0.035%, P <0.01). Plasma interleukin-6 and high-sensitivity CRP were significantly higher in HF (3.99±0.86 vs. 0.78±0.14 pg/mL and 0.28±0.10 vs. 0.06±0.02 mg/dL, respectively, both P <0.01). LV ejection fraction ( r =0.72, P <0.05) and plasma log BNP ( r =-0.70, P <0.05) were significantly correlated to the ratio of iNKT cells among HF patients. Conclusions: Circulating iNKT cells were decreased in HF patients, suggesting that they have a potential role in the development of human HF.

scholarly journals Activation of Invariant Natural Killer T Cell Subsets in C57BL/6J Mice by Different Injection Modes of α-galactosylceramide

2020 ◽  
Author(s):  
Ming Meng ◽  
Shengde Chen ◽  
Xiang Gao ◽  
Huifang Liu ◽  
Huijuan Zhao ◽  
...  
Keyword(s):  
Natural Killer ◽  
Subcutaneous Injection ◽  
Intraperitoneal Injection ◽  
T Cell Subsets ◽  
Activation Mechanism ◽  
Inkt Cells ◽  
Healthy Control ◽  
Injection Modes ◽  
Invariant Natural Killer ◽  
Natural Killer T

Whether different injection modes of α-galactosylceramide (α-GalCer) affect the activation of different subsets of invariant natural killer T (iNKT) cells in different tissues and organs of mice is unclear. This study included healthy control, subcutaneous injection, and intraperitoneal injection groups (n=10 in each group). The subcutaneous and intraperitoneal injection groups were injected with α-Galcer (0.1 mg/kg weight), and then the changes in thymus, spleen, and liver iNKT cell frequencies and subsets were observed. The intraperitoneal injection of α-GalCer could increase the frequency of splenic iNKT cells, but the subcutaneous injection did not affect the frequency. Neither injection had any effect on the frequency of iNKT cells in the thymus and liver. The subcutaneous injection of α-GalCer increased the rate of iNKT2 subsets in the thymus but did not affect the rate of iNKT1 subsets. However, the intraperitoneal injection of α-GalCer did not affect thymus iNKT1 and iNKT2 subsets. Interestingly, the subcutaneous injection of α-GalCer significantly increased the proportion of iNKT1 in the spleen and liver but did not significantly change the proportion of iNKT2. The intraperitoneal injection of  α-GalCer significantly increased the rate of iNKT2 in spleen and liver but decreased the rate of iNKT1. Subsets of iNKT1 or iNKT2 cells in the spleen and liver were selectively activated by the subcutaneous or intraperitoneal injection of α-GalCer. It provides a valuable means for treating tumors and certain autoimmune diseases. Further exploration of the activation mechanism may provide new ideas about the development of related vaccines.

scholarly journals Migration, Distribution, and Safety Evaluation of Specific Phenotypic and Functional Mouse Spleen-Derived Invariant Natural Killer T2 Cells after Adoptive Infusion

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Dongzhi Chen ◽  
Wenbin Xu ◽  
Jingfang Teng ◽  
Huifang Liu ◽  
Yuanyuan Wang ◽  
...  
Keyword(s):  
Natural Killer ◽  
Cell Sorting ◽  
Intraperitoneal Injection ◽  
Inflammatory Cytokine ◽  
Inkt Cells ◽  
Anti Inflammatory ◽  
Invariant Natural Killer ◽  
Anti Inflammatory Cytokine ◽  
Magnetic Activated Cell Sorting

Herein, the migration distribution and safety of specific phenotypic and functionally identified spleen-derived invariant natural killer T2 (iNKT2) cells after adoptive infusion in mice were studied. The proliferation and differentiation of iNKT cells were induced by intraperitoneal injection of α-galactosylceramide (α-GalCer) in vivo. Mouse spleens were isolated in a sterile environment. iNKT cells were isolated by magnetic-activated cell sorting columns (MS columns). Cytometric bead array (CBA) assay was used to detect cytokine secretion in the supernatant stimulated by iNKT cells. The basic life status of the mice was observed, and systematic quantitative scoring was conducted after injecting spleen-derived iNKT cells through the tail vein. An in vivo imaging system was used to trace the migration and distribution of iNKT cells in DBA mice. The percentage of the iNKT2 subgroup was the highest in 3 days after intraperitoneal injection of α-GalCer, and iNKT2 subsets accounted for more than 92% after separation and purification by magnetic-activated cell sorting (MACS). Anti-inflammatory cytokine IL-4 was mainly found in the supernatant of cell cultures. The adoptive infusion of iNKT cells into healthy mice resulted in no significant change in the basic life status of mice compared with the noninjected group. iNKT cells were detected in the lung, spleen, and liver, but no fluorescence was detected in lymph nodes and thymus. After dissecting the mice, it was found that there were no significant abnormalities in the relevant immune organs, brain, heart, kidney, lung, and other organs. Intraperitoneal injection of α-GalCer results in a large number of iNKT2 cells, mainly secreting anti-inflammatory cytokine IL-4, from the spleen of mice. After adoptive infusion, the iNKT2 cells mainly settled in the liver and spleen of mice with a satisfactory safety profile.

scholarly journals Preclinical Evaluation of Invariant Natural Killer T Cells Modified with CD38 or BCMA Chimeric Antigen Receptors for Multiple Myeloma

2021 ◽  
Vol 22 (3) ◽  
pp. 1096
Author(s):  
Renée Poels ◽  
Esther Drent ◽  
Roeland Lameris ◽  
Afroditi Katsarou ◽  
Maria Themeli ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Natural Killer ◽  
Therapeutic Potential ◽  
Cytotoxic Activities ◽  
Dependent Manner ◽  
Antigen Receptors ◽  
Inkt Cells ◽  
Chimeric Antigen Receptors ◽  
Invariant Natural Killer ◽  
Natural Killer T

Due to the CD1d restricted recognition of altered glycolipids, Vα24-invariant natural killer T (iNKT) cells are excellent tools for cancer immunotherapy with a significantly reduced risk for graft-versus-host disease when applied as off-the shelf-therapeutics across Human Leukocyte Antigen (HLA) barriers. To maximally harness their therapeutic potential for multiple myeloma (MM) treatment, we here armed iNKT cells with chimeric antigen receptors (CAR) directed against the MM-associated antigen CD38 and the plasma cell specific B cell maturation antigen (BCMA). We demonstrate that both CD38- and BCMA-CAR iNKT cells effectively eliminated MM cells in a CAR-dependent manner, without losing their T cell receptor (TCR)-mediated cytotoxic activity. Importantly, iNKT cells expressing either BCMA-CARs or affinity-optimized CD38-CARs spared normal hematopoietic cells and displayed a Th1-like cytokine profile, indicating their therapeutic utility. While the costimulatory domain of CD38-CARs had no influence on the cytotoxic functions of iNKT cells, CARs containing the 4-1BB domain showed a better expansion capacity. Interestingly, when stimulated only via CD1d+ dendritic cells (DCs) loaded with α-galactosylceramide (α-GalCer), both CD38- and BCMA-CAR iNKT cells expanded well, without losing their CAR- or TCR-dependent cytotoxic activities. This suggests the possibility of developing an off-the-shelf therapy with CAR iNKT cells, which might even be boostable in vivo by administration α-GalCer pulsed DCs.

P6305Activation of invariant natural killer T cells ameliorates doxorubicin-induced cardiomyopathy

2019 ◽  
Vol 40 (Supplement_1) ◽  
Author(s):  
M Sada ◽  
S Matsushima ◽  
S Ikeda ◽  
K Okabe ◽  
A Ishikita ◽  
...  
Keyword(s):  
Natural Killer ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Active Form ◽  
Left Ventricular ◽  
Specific Marker ◽  
Inkt Cells ◽  
Ifn Γ ◽  
Invariant Natural Killer ◽  
Natural Killer T

Abstract Background Invariant natural killer T (iNKT) cells orchestrate tissue inflammation via regulating various cytokine productions, especially strongly upregulating interferon (IFN)-γ. Activation of iNKT cells have been previously reported to exert protective effects against post-infarcted cardiac remodeling and cardiac ischemia/reperfusion injury. However, the role of iNKT cells has not been determined in doxorubicin (DOX)-induced cardiomyopathy. Purpose The purpose of this study was to examine whether the activation of iNKT cells by α-galactosylceramide (αGC), which specifically activates iNKT cells, could affect DOX-induced cardiomyopathy, and if so, to elucidate its downstream target. Methods C57BL/6J mice were received the intraperitoneal injection of either αGC (0.1μg/g, n=11) or vehicle (n=13). After 1 week, these mice were treated with a low dose of DOX (18mg/kg via intravenous 3 injections over 1 week), and were followed during 14 days. Results DOX mice (DOX+vehicle) showed left ventricular (LV) dysfunction and dilatation, which were significantly ameliorated in DOX mice receiving αGC (DOX+αGC) (LV fractional shortening: 27.4±4.31 vs. 31.5±4.62%, p<0.05, LV end-diastolic diameter: 3.70±0.16 vs. 3.32±0.23mm, p<0.05), with no significant changes in arterial pressure, body weight, and food consumption, 14 days after DOX injection. DOX+vehicle demonstrated a significant decrease in myocardial gene expression of Vα14Jα18, a specific marker of iNKT cells, and IFN-γ compared with control mice. Vα14Jα18 expression levels were higher in DOX+αGC than DOX+vehicle by 9.2 folds (p<0.05). Consistent with this change, IFN-γ was higher in DOX+αGC than DOX+vehicle by 4.4 folds (p<0.05), whereas interleukin (IL)-1, IL-4, IL-6, IL-10, IL-17, IL-23, and tumor necrosis factor (TNF)-α were not altered in both groups. Phosphorylation of Akt, its active form, in the heart was significantly increased in DOX+αGC compared with DOX+vehicle by 1.8 folds (p<0.05). Conclusions Activation of iNKT cells by αGC play a protective role against DOX-induced cardiac dysfunction, which was associated with enhancing expression of IFN-γ and activating Akt. Therapies designed to activate iNKT cells might be beneficial to protect the heart from DOX injury.

Activation of invariant natural killer T cells ameliorates doxorubicin-induced cardiotoxicity in mice

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
Y Obata ◽  
N Ishimori ◽  
A Saito ◽  
S Kinugawa ◽  
I Nakano ◽  
...  
Keyword(s):  
Body Weight ◽  
Natural Killer ◽  
Heart Tissue ◽  
M2 Macrophage ◽  
Funding Source ◽  
Inkt Cells ◽  
Arginase 1 ◽  
Relative Ratio ◽  
Invariant Natural Killer ◽  
Natural Killer T

Abstract Objective Doxorubicin (DOX) is one of the most important anticancer agents and widely used to treat cancers but clinical utility of DOX is limited for its dose-dependent cardiotoxicity. The precise mechanism of DOX-induced cardiotoxicity is still not fully understood but it has been reported that cardiac inflammation is involved in the cardiotoxicity. Invariant natural killer T (iNKT) cells, a unique subset of T lymphocytes that recognize glycolipid antigens and secrete a large amount of both Th1 and Th2 cytokines on activation, have been shown to play crucial roles in the regulation of immune responses. However, it remains unclear whether iNKT cells are involved in DOX-induced cardiotoxicity. Methods and results Male C57BL/6J mice were administered DOX (20mg/kg body weight; n=28) or vehicle (Vehicle; n=6). DOX-administered mice were further divided into 2 groups; those treated with α-galactosylceramide (αGC, 0.1μg/g body weight; DOX-αGC; n=14), which specifically activates iNKT cells, or those treated with PBS (DOX-PBS; n=14) by intraperitoneal injections (twice; 4 days before and 3 days after DOX administration).An echocardiography conducted at 14 days after DOX/Vehicle administration revealed that LV fractional shortening was significantly reduced in the DOX-PBS compared to the Vehicle (49.3±0.8% vs. 59.2±1.7%, P&lt;0.05), and this decrease was completely attenuated in the DOX-αGC (57.7±1.3%, P&lt;0.05 vs. DOX-PBS)without affecting LV end-diastolic diameter. Flow cytometric analysis revealed that the ratio of iNKT cells to mononuclear cells infiltrated into the heart tissue was significantly increased in the DOX+αGC compared to the Vehicle and the DOX+PBS (1.00±0.09% vs. 0.54±0.09% and 0.71±0.07%, P&lt;0.05). Immuno-histochemistry revealed that the infiltration number of Iba1+macrophages in the heart tissue was significantly elevated in the DOX+αGC compared to the Vehicle and the DOX+PBS (55.4±3.2 cells/mm2 vs. 21.7±2.0 cells/mm2 and 37.5±5.9 cells/mm2, P&lt;0.05) The ratio of fibrosis area to the heart tissue was markedly higher in the DOX-PBS than in Vehicle (4.3±0.5% vs. 2.2±0.1%, P&lt;0.05), and this increase was completely attenuated in the DOX-αGC (2.8±0.1%, P&lt;0.05 vs.DOX-PBS).Real-time PCR analysis revealed that mRNA expressions of M2 macrophage markers (Arginase 1 and Retnla) and IL-4 were significantly enhanced in the DOX+αGC compared to the DOX+PBS (Arginase 1: 2.5±0.4 vs. 1.6±0.3 [relative ratio to the Vehicle], P=0.08; Retnla: 2.4±0.5 vs. 1.1±0.2 [relative ratio to the Vehicle], P&lt;0.05; IL-4: 1.0±0.3 vs. 8.94±2.8 [relative ratio to the DOX+PBS], P&lt;0.05), while those of M1 macrophage markers (iNOS and MCP-1) did not change among all groups. Conclusions Activation of iNKT cells ameliorates DOX-induced cardiotoxicity in mice via enhanced M2 macrophage polarization with the upregulation of IL-4 and reducing cardiac fibrosis. iNKT cell activation can be a novel preventive strategy against DOX-induced cardiotoxicity. Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): Japan Agency for Medical Research and Development (18lm0203001j0002) and JSPS KAKENHI (18K15834).

scholarly journals Invariant natural killer T cells direct B cell responses to cognate lipid antigen in an IL-21-dependent manner

2011 ◽  
Vol 13 (1) ◽  
pp. 44-50 ◽  
Author(s):  
Irah L King ◽  
Anne Fortier ◽  
Michael Tighe ◽  
John Dibble ◽  
Gerald F M Watts ◽  
...  
Keyword(s):  
T Cells ◽  
B Cell ◽  
Natural Killer ◽  
Natural Killer T Cells ◽  
Dependent Manner ◽  
Cell Responses ◽  
Lipid Antigen ◽  
Killer T Cells ◽  
Invariant Natural Killer ◽  
Natural Killer T

Evaluating The Frequency Of Invariant Natural Killer T (iNKT) Cells In Nasal Polyps

2008 ◽  
Vol 121 (2) ◽  
pp. S263-S263
Author(s):  
M FEREIDOUNI ◽  
R FARIDHOSSEINI ◽  
M MAHMOUDI ◽  
M BAKHSHAEI ◽  
L ALHARTHI
Keyword(s):  
Natural Killer ◽  
Nasal Polyps ◽  
Inkt Cells ◽  
Invariant Natural Killer ◽  
Natural Killer T

scholarly journals Invariant natural killer T cells are functionally impaired in patients with systemic sclerosis

2019 ◽  
Vol 21 (1) ◽  
Author(s):  
Ann-Christin Pecher ◽  
Felix Kettemann ◽  
Elisa Asteriti ◽  
Hannes Schmid ◽  
Silke Duerr-Stoerzer ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Natural Killer ◽  
Immunosuppressive Drugs ◽  
Healthy Controls ◽  
Inkt Cells ◽  
Novel Approach ◽  
Functionally Impaired ◽  
Transfer Strategies ◽  
Invariant Natural Killer ◽  
Natural Killer T

Abstract Background Systemic sclerosis (SSc) is a potentially fatal autoimmune disease that leads to extensive fibrosis of the skin and internal organs. Invariant natural killer T (iNKT) cells are potent immunoregulatory T lymphocytes being able to orchestrate dysregulated immune responses. The purpose of this study was to evaluate numbers and function of iNKT cells in patients with SSc and to analyze their correlation with disease parameters. Methods Human iNKT cells from 88 patients with SSc and 33 healthy controls were analyzed by flow cytometry. Their proliferative capacity and cytokine production were investigated following activation with CD1d ligand α-galactosylceramide (α-GalCer). Results We observed an absolute and relative decrease of iNKT cells in patients with SSc compared with healthy controls. Interestingly, the subtype of SSc, disease severity, or treatment with immunosuppressive drugs did not affect iNKT cell numbers. However, T helper (Th) cell immune polarization was biased towards a Th17 immunophenotype in SSc patients. Moreover, iNKT cells from patients with SSc showed a significantly decreased expansion capacity upon stimulation with α-GalCer. Conclusion iNKT cells are deficient and functionally impaired in patients with SSc. Therefore, adoptive transfer strategies using culture-expanded iNKT cells could be a novel approach to treat SSc patients.

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