scholarly journals G6PD functions as a metabolic checkpoint to regulate granzyme B expression in tumor-specific cytotoxic T lymphocytes

2022 ◽  
Vol 10 (1) ◽  
pp. e003543
Author(s):  
Chunwan Lu ◽  
Dafeng Yang ◽  
John D Klement ◽  
Yolonda L Colson ◽  
Nicholas H Oberlies ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Immune Evasion ◽  
Aerobic Glycolysis ◽  
Granzyme B ◽  
Pentose Phosphate ◽  
Expression Level ◽  
Tumor Immune Evasion ◽  
Specific Ctls ◽  
Tumor Bearing

BackgroundGranzyme B is a key effector of cytotoxic T lymphocytes (CTLs), and its expression level positively correlates with the response of patients with mesothelioma to immune checkpoint inhibitor immunotherapy. Whether metabolic pathways regulate Gzmb expression in CTLs is incompletely understood.MethodsA tumor-specific CTL and tumor coculture model and a tumor-bearing mouse model were used to determine the role of glucose-6-phosphate dehydrogenase (G6PD) in CTL function and tumor immune evasion. A link between granzyme B expression and patient survival was analyzed in human patients with epithelioid mesothelioma.ResultsMesothelioma cells alone are sufficient to activate tumor-specific CTLs and to enhance aerobic glycolysis to induce a PD-1hi Gzmblo CTL phenotype. However, inhibition of lactate dehydrogenase A, the key enzyme of the aerobic glycolysis pathway, has no significant effect on tumor-induced CTL activation. Tumor cells induce H3K9me3 deposition at the promoter of G6pd, the gene that encodes the rate-limiting enzyme G6PD in the pentose phosphate pathway, to downregulate G6pd expression in tumor-specific CTLs. G6PD activation increases acetyl-coenzyme A (CoA) production to increase H3K9ac deposition at the Gzmb promoter and to increase Gzmb expression in tumor-specific CTLs converting them from a Gzmblo to a Gzmbhi phenotype, thus increasing CTL tumor lytic activity. Activation of G6PD increases Gzmb+ tumor-specific CTLs and suppresses tumor growth in tumor-bearing mice. Consistent with these findings, GZMB expression level was found to correlate with increased survival in patients with epithelioid mesothelioma.ConclusionG6PD is a metabolic checkpoint in tumor-activated CTLs. The H3K9me3/G6PD/acetyl-CoA/H3K9ac/Gzmb pathway is particularly important in CTL activation and immune evasion in epithelioid mesothelioma.

Induction of Multiple Myeloma-Specific Cytotoxic T Lymphocytes Using HLA-A2.1-Specific CD19 and CD20 Peptides.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2477-2477
Author(s):  
Jooeun Bae ◽  
Jeff A. Martinson ◽  
Hans G. Klingemann ◽  
Steven Treon ◽  
Kenneth C. Anderson ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
T Lymphocytes ◽  
Cytotoxic Activity ◽  
Cell Lines ◽  
Cytotoxic T Lymphocytes ◽  
Myeloma Cell ◽  
Myeloma Cells ◽  
Specific Ctls ◽  
Ifn Γ ◽  
Hla A2.1

Abstract We have identified novel CD19 and CD20 antigen-derived HLA-A2.1-specific immunogenic peptides, CD19150–158 (KLMSPKLYV) and CD20188–196 (SLFLGILSV), for generating cytotoxic T lymphocytes (CTLs) against malignant B-cell diseases. Initial testing showed that the CTLs displayed antigen-specific and HLA-A2.1-restriced cytotoxic activity against both Burkitt’s lymphoma and chronic lymphoid leukemia cell lines. The observed cytotoxic activity of the CTLs was shown to be specific to the CD19150–158 or the CD20188–196 peptides. Additionally, the CTLs displayed a distinct phenotype (majority CD69+/CD45RO+) along with a significant (p<0.05) increase in cell proliferation and IFN-γ release following re-stimulation with HLA-A2.1+/CD19+/CD20+ tumor cell lines. Based on emerging information that clonogenic myeloma cells express CD19 and/or CD20, we evaluated the activity of the CD19 and CD20 peptide specific-CTLs against several multiple myeloma cell lines. Five of 10 myeloma cell lines evaluated were HLA-A2.1-positive and expressed both CD19 and CD20 antigens. CD19 peptide specific-CTLs generated from normal donors were able to specifically lyse CD19+/HLA-A2.1+ MM cell lines (30% lysis; 10:1 E:T ratio) but did not lyse CD19−/HLA-A2.1+ or CD19+/HLA-A2.1− cell lines. Similarly, the CD20-specific CTLs generated from normal donors lysed CD20+/HLA-A2.1+ MM cell lines (25% lysis; 10:1 E:T ratio), in a manner restricted to HLA-A2.1 and specific to antigens. We next showed IFN-γ production by the CTLs after exposure to CD19+/HLA-A2.1+ or CD20+/HLA-A2.1+ MM cells. Moreover, we have demonstrated the ability to expand CD20-CTLs under serum-free culture conditions while maintaining their cytotoxic activity (28–49%). In ongoing studies, we are evaluating the ability of CD19- and CD20-specific CTLs to eliminate clonogenic myeloma cells in vitro and in vivo in a SCID mouse model of myeloma. These preclinical studies strongly suggest that immunogenic CD19 and CD20 peptide-based vaccines represent a promising immunotherapeutic approach in myeloma.

Target Epitopes of Human T Lymphotropic Virus 1 Recognized by Class I MHC-Restricted Cytotoxic T Lymphocytes in HAM/TSP Patients and Infected Patients with Autoimmune Disorders.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2660-2660
Author(s):  
Tomohiro Kozako ◽  
Masaki Akimoto ◽  
Yohann White ◽  
Shingo Toji ◽  
Kakushi Matsushita ◽  
...  
Keyword(s):  
T Cell ◽  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Spastic Paraparesis ◽  
T Cell Leukemia ◽  
Cell Leukemia ◽  
Class I Mhc ◽  
Collagen Diseases ◽  
Specific Ctls ◽  
Significant Difference

Abstract Abstract 2660 Poster Board II-636 Human T-cell leukemia virus-1 (HTLV-1) causes adult T cell leukemia/lymphoma (ATL) and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) after long-term infection. A variety of collagen diseases such as Sjögren's syndrome and systemic lupus erythematosus has been reported in HTLV-1-infected individuals, although the precise relationship between these disorders and HTLV-1 infection remains unknown. We have previously reported the decreased frequency and function of HTLV-1 Tax-specific cytotoxic T lymphocytes (CTLs) in ATL patients due to insufficient cytolytic effector molecules compared with asymptomatic carriers (ACs) (ASH annual meeting 2007). However, there is no report on the repertoire of HTLV-1-specific CTLs in HAM/TSP patients or carriers with collagen diseases (CCs), both of which are characterized by abnormal immune states. In order to characterize HTLV-1-specific CTLs in ACs, HAM/TSP patients and CCs, we examined the frequency and diversity of HTLV-1-specific CTLs using 16 distinct HTLV-1 Tax and Env HLA-A*0201 and HLA-A*2402 tetramers. There was no statistically significant difference in the proportion of patients having HTLV-1 Tax specific CTLs among ACs, HAM/TSP and CC patients, but a greater proportion of patients with HAM/TSP or collagen disease had HTLV-1 Env-specific CTLs compared with ACs (Env175-183, Env239-246, Env442-450 for HLA-A*0201 and Env11-19, Env21-29, Env153-161 for HLA-A*2402) (Table 1). Within CD8+ lymphocyte subsets, the percentage of cells binding either Tax11-19/HLA-A*0201 or Tax 301-309/HLA-A*2402 tetramer (Figure 1), was significantly higher for HAM/TSP patients compared to ACs, being lowest among CCs. Additionally, the number of epitope repertoires found on HTLV-1 Env-specific CD8+ cells in ACs was considerably lower than for HAM/TSP patients and CCs (ACs, 1/6; HAM/TSP, 4/6; CCs, 6/6). This study demonstrates the relatively greater importance of CTLs recognizing HTLV-1 envelope tetramers compared to HTLV-1 Tax tetramers, thereby suggesting that the diversity, frequency and repertoire of HTLV-1-specific CTL clones, especially Env-CTLs, may be related to the hyperimmune response in HAM/TSP and carriers with collagen diseases. Disclosures: No relevant conflicts of interest to declare.

The granzyme B–serglycin complex from cytotoxic granules requires dynamin for endocytosis

Blood ◽  
2004 ◽  
Vol 103 (10) ◽  
pp. 3845-3853 ◽  
Author(s):  
Kirstin Veugelers ◽  
Bruce Motyka ◽  
Christine Frantz ◽  
Irene Shostak ◽  
Tracy Sawchuk ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Granzyme B ◽  
Endocytic Pathway ◽  
Target Cells ◽  
Ongoing Debate ◽  
Effector Molecules ◽  
Key Factor ◽  
Dependent Pathway ◽  
Endocytic Pathways

Abstract Cytotoxic T lymphocytes and natural killer cells destroy target cells via the directed exocytosis of lytic effector molecules such as perforin and granzymes. The mechanism by which these proteins enter targets is uncertain. There is ongoing debate over whether the most important endocytic mechanism is nonspecific or is dependent on the cation-independent mannose 6-phosphate receptor. This study tested whether granzyme B endocytosis is facilitated by dynamin, a key factor in many endocytic pathways. Uptake of and killing by the purified granzyme B molecule occurred by both dynamin-dependent and -independent mechanisms. However most importantly, serglycin-bound granzyme B in high-molecular-weight degranulate material from cytotoxic T lymphocytes predominantly followed a dynamin-dependent pathway to kill target cells. Similarly, killing by live cytotoxic T lymphocytes was attenuated by a defect in the dynamin endocytic pathway, and in particular, the pathways characteristically activated by granzyme B were affected. We therefore propose a model where degranulated serglycin-bound granzymes require dynamin for uptake.

scholarly journals Estrogen Induction of the Granzyme B Inhibitor, Proteinase Inhibitor 9, Protects Cells against Apoptosis Mediated by Cytotoxic T Lymphocytes and Natural Killer Cells

Endocrinology ◽  
2006 ◽  
Vol 147 (3) ◽  
pp. 1419-1426 ◽  
Author(s):  
Xinguo Jiang ◽  
Brent A. Orr ◽  
David M. Kranz ◽  
David J. Shapiro
Keyword(s):  
T Lymphocytes ◽  
Natural Killer ◽  
Cytotoxic T Lymphocytes ◽  
Proteinase Inhibitor ◽  
Nk Cell ◽  
Granzyme B ◽  
Liver Cells ◽  
Target Cells ◽  
Proteinase Inhibitor 9 ◽  
Estrogen Induction

Exposure to estrogens is associated with an increased risk of developing breast, cervical, and liver cancer. Estrogens strongly induce the human granzyme B inhibitor, proteinase inhibitor 9 (PI-9). Because cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells use the granzyme pathway to induce apoptosis of target cells, we tested the ability of activated CTLs and the human NK cell line, YT cells, to lyse human liver cells. Estrogen induction of PI-9 protected the liver cells against CTL and NK cell-mediated, granzyme-dependent, apoptosis. Knockdown of PI-9 by RNA interference blocked the protective effect of estrogen. This work demonstrates that estrogens can act on target cells to control their destruction by immune system cells and shows that induction of PI-9 expression can inhibit both CTL and NK cell-mediated apoptosis. Estrogen induction of PI-9 may reduce the ability of cytolytic lymphocytes-mediated immune surveillance to destroy newly transformed cells, possibly providing a novel mechanism for an estrogen-mediated increase in tumor incidence.

Increased Numbers of Granzyme-B-Expressing Cytotoxic T-Lymphocytes in the Small Intestine of HIV-Infected Patients

1996 ◽  
Vol 12 (3) ◽  
pp. 276-281 ◽  
Author(s):  
Fransje Snijders ◽  
Peter C. Wever ◽  
Sven A. Danner ◽  
C. Erik Hack ◽  
Fiebo J. W. ten Kate ◽  
...  
Keyword(s):  
Small Intestine ◽  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Granzyme B

Presence of survivin-specific cytotoxic T-lymphocytes (CTLs) in pancreatic cancer patients, and specific killing of human pancreatic carcinoma cells in vitro by survivin-specific CTLs

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 3070-3070
Author(s):  
D. Z. Chang ◽  
K. Zhu ◽  
S. Kopetz ◽  
K. Voo ◽  
Y. Li ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
T Lymphocytes ◽  
Pancreatic Carcinoma ◽  
Cell Lines ◽  
Cytotoxic T Lymphocytes ◽  
Carcinoma Cell ◽  
Specific Ctls ◽  
Carcinoma Cell Lines ◽  
Ifn Γ

3070 Background: The apoptosis inhibitor protein Survivin is an attractive target for a cancer vaccine because of its differential overexpression by most human tumors, its importance for tumor survival, and its demonstrated immunogenicity. To examine Survivin’s potential as a vaccine target for pancreatic cancer (PC), we evaluated the presence of Survivin-specific cytotoxic T-lymphocytes (CTLs) in PC patients and assessed the specific killing of HLA-A*0201+ human pancreatic carcinoma cell lines in vitro by Survivin-specific CTL. Methods: Mononuclear cells from the peripheral blood (PBMC) of PC patients and normal donors and Survivin peptides were used to generate Survivin-specific CTLs in vitro. The presence of Survivin-specific CTLs was evaluated by IFN-γ ELISPOT analysis. Recognition of human PC cell lines (e.g., Panc-1 and CF-Pac-1) was analyzed by 51Cr release assays. Results: Multiple CTL lines were generated from different donors against a number of Survivin 9-mer and 10-mer peptides, including Sur5–14, Sur95–104, and Sur96–104 and altered forms of these peptides with enhanced HLA-A*0201 binding. Sur5–14 (TLPPAWQPFL) and new modified mSur95–104 (ELMLGEFLKL) peptides could generate specific CTL from the PBMC of most donors, and the resulting CTL showed cytotoxicity against Survivin peptide-pulsed Panc-1 and CF-Pac-1 in an HLA-A2-dependent manner. We found for the first time that mSur95–104 could elicit CTL activity that cross-reacted with wild-type peptide and 9-mer Sur96–104 in IFN-γ ELISPOT assays. In our preliminary analysis, we detected Survivin peptide-specific CTLs in four of five PC patients for Sur5–14 and three of five for mSur95–104. All five PC patients had a positive response to a pool of A2 peptides as positive controls, indicating their immune competence. Conclusions: CTLs specific for human Survivin peptide Sur5–14 and the modified peptides mSur95–104 were detected in majority of the PC patients tested. Further, Survivin-specific CTLs killed pancreatic carcinoma cell lines in vitro. Our results suggest that these two peptides are potential vaccines for the treatment of PC. No significant financial relationships to disclose.

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