The Suppressed Induction of Human Mature Cytotoxic T Lymphocytes Caused by Asbestos Is Not due to Interleukin-2 Insufficiency

We previously reported that exposure to chrysotile B (CB) asbestos suppressed the induction of mature cytotoxic T lymphocytes (CTLs) during mixed lymphocyte reaction assays (MLRs) with a decrease in the proliferation of immature CTLs. However, the mechanism responsible for the effect of asbestos fibers on the differentiation of CTLs remains unclear. Since interleukin-2 (IL-2) is a regulator of T lymphocyte proliferation, we examined the effect of IL-2 addition on suppressed CTL differentiation in CB-exposed cultures using flow cytometry (FCM). When IL-2 was added at 1 ng/mL on the second day of MLRs, the asbestos-caused decreases in the proliferation and percentages of CD25+and CD45RO+cells in CD8+lymphocytes were not recovered by IL-2 addition, although the decrease in percentage of granzyme B+cells was partially recovered. CD8+lymphocytes from the IL-2-treated culture with asbestos showed the same degree of cytotoxicity as those in cultures without IL-2 or asbestos. These findings indicate that IL-2 insufficiency is not the main cause for the suppressed induction of CTLs by asbestos exposure, although they suggest a potential for the improvement of such suppressed CTL functions. Secretory factors other than IL-2 in addition to membrane-bound stimulatory molecules may play a role in asbestos-caused suppressed CTL differentiation.

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Effect of Asbestos Exposure on Differentiation of Cytotoxic T Lymphocytes in Mixed Lymphocyte Reaction of Human Peripheral Blood Mononuclear Cells

American Journal of Respiratory Cell and Molecular Biology ◽

10.1165/rcmb.2012-0134oc ◽

2013 ◽

Vol 49 (1) ◽

pp. 28-36 ◽

Cited By ~ 26

Author(s):

Naoko Kumagai-Takei ◽

Yasumitsu Nishimura ◽

Megumi Maeda ◽

Hiroaki Hayashi ◽

Hidenori Matsuzaki ◽

...

Keyword(s):

T Lymphocytes ◽

Cytotoxic T Lymphocytes ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mixed Lymphocyte Reaction ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Asbestos Exposure ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

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Effect of IL-15 addition on asbestos-induced suppression of human cytotoxic T lymphocyte induction

Environmental Health and Preventive Medicine ◽

10.1186/s12199-021-00967-9 ◽

2021 ◽

Vol 26 (1) ◽

Author(s):

Naoko Kumagai-Takei ◽

Yasumitsu Nishimura ◽

Hidenori Matsuzaki ◽

Suni Lee ◽

Kei Yoshitome ◽

...

Keyword(s):

T Cells ◽

T Lymphocyte ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Granzyme B ◽

Interleukin 2 ◽

Functional Markers ◽

Asbestos Fibers ◽

T Lymphocyte Proliferation ◽

Ctl Induction

Abstract Background Asbestos fibers possess tumorigenicity and are thought to cause mesothelioma. We have previously reported that exposure to asbestos fibers causes a reduction in antitumor immunity. Asbestos exposure in the mixed lymphocyte reaction (MLR) showed suppressed induction of cytotoxic T lymphocytes (CTLs), accompanied by a decrease in proliferation of CD8+ T cells. Recently, we reported that asbestos-induced suppression of CTL induction is not due to insufficient levels of interleukin-2 (IL-2). In this study, we continue to investigate the mechanism responsible for the effect of asbestos fibers on the differentiation of CTLs and focus on interleukin-15 (IL-15) which is known to be a regulator of T lymphocyte proliferation. Methods For MLR, human peripheral blood mononuclear cells (PBMCs) were cultured with irradiated allogenic PBMCs upon exposure to chrysotile B asbestos at 5 μg/ml for 7 days. After 2 days of culture, IL-15 was added at 1 ng/ml. After 7 days of MLR, PBMCs were collected and analyzed for phenotypic and functional markers of CD8+ T cells with fluorescence-labeled anti-CD3, anti-CD8, anti-CD45RA, anti-CD45RO, anti-CD25, and anti-granzyme B antibodies using flow cytometry. To examine the effect of IL-15 on the expression level of intracellular granzyme B in proliferating and non-proliferating CD8+ lymphocytes, PBMCs were stained using carboxyfluorescein diacetate succinimidyl ester (CFSE) and then washed and used for the MLR. Results IL-15 addition partially reversed the decrease in CD3+CD8+ cell numbers and facilitated complete recovery of granzyme B+ cell percentages. IL-15 completely reversed the asbestos-induced decrease in percentage of granzyme B+ cells in both non-proliferating CFSE-positive and proliferating CFSE-negative CD8+ cells. The asbestos-induced decrease in the percentage of CD25+ and CD45RO+ cells in CD8+ lymphocytes was not reversed by IL-15. Conclusion These findings indicate that CTLs induced upon exposure to asbestos possess dysfunctional machinery that can be partly compensated by IL-15 supplementation, and that IL-15 is more effective in the recovery of proliferation and granzyme B levels from asbestos-induced suppression of CTL induction compared with IL-2.

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Faculty Opinions recommendation of Serglycin-deficient cytotoxic T lymphocytes display defective secretory granule maturation and granzyme B storage.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1027289.336049 ◽

2005 ◽

Author(s):

Eckhard Podack

Keyword(s):

T Lymphocytes ◽

Secretory Granule ◽

Cytotoxic T Lymphocytes ◽

Granzyme B ◽

Granule Maturation

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CD27 Expression Promotes Long-Term Survival of Functional Effector–Memory CD8+Cytotoxic T Lymphocytes in HIV-infected Patients

Journal of Experimental Medicine ◽

10.1084/jem.20040717 ◽

2004 ◽

Vol 200 (11) ◽

pp. 1407-1417 ◽

Cited By ~ 92

Author(s):

Adrian F. Ochsenbein ◽

Stanley R. Riddell ◽

Michele Brown ◽

Lawrence Corey ◽

Gabriela M. Baerlocher ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Cytotoxic T Lymphocytes ◽

Cd8 T Cells ◽

Interleukin 2 ◽

Tumor Necrosis Factor Receptor ◽

Cell Receptor ◽

Effector Memory ◽

Term Survival ◽

Functional Consequences

Human immunodeficiency virus (HIV)-specific CD8+ T cells persist in high frequencies in HIV-infected patients despite impaired CD4+ T helper response to the virus, but, unlike other differentiated effector cytotoxic T lymphocytes, most continue to express the tumor necrosis factor receptor family member CD27. Because the ligand for CD27 (CD70) is also overexpressed in HIV-infected hosts, we examined the nature of expression and potential functional consequences of CD27 expression on HIV-specific CD8+ T cells. Analysis of CD27+ and CD27− T cells derived from the same HIV-specific clone revealed that retention of CD27 did not interfere with acquisition of effector functions, and that after T cell receptor stimulation, CD27+ cells that concurrently were triggered via CD27 exhibited more resistance to apoptosis, interleukin 2 production, and proliferation than CD27− T cells. After transfer back into an HIV-infected patient, autologous HIV-specific CD27− T cells rapidly disappeared, but CD27+ T cells derived from the same clone persisted at high frequency. Our findings suggest that the CD27–CD70 interaction in HIV infection may provide CD27+ CD8+ T cells with a survival advantage and compensate for limiting or absent CD4+ T help to maintain the CD8 response.

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

Blood ◽

10.1182/blood-2003-06-2156 ◽

2004 ◽

Vol 103 (10) ◽

pp. 3845-3853 ◽

Cited By ~ 36

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.

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

Endocrinology ◽

10.1210/en.2005-0996 ◽

2006 ◽

Vol 147 (3) ◽

pp. 1419-1426 ◽

Cited By ~ 36

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.

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