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ABSTRACT: Granulysin (GNLY) is a novel cytolytic protein lytic against a variety of tumor cells and microbes. The role of GNLY during pregnancy has not been extensively explored. The aim of this study is to examine GNLY expression and distribution in the first trimester pregnancy peripheral blood (PB) and decidua, the ability of decidual and PB natural killer (NK) cells to secrete GNLY spontaneously, and the role of antigen-presenting cells (APC) in the regulation of GNLY expression in decidual NK cells.
GNLY expression was analyzed using cell permeabilization method, flow cytometry, and immunohistochemistry. GNLY secretion by purified NK cells was detected by ELISA method.
GNLY is abundantly expressed at the maternal-fetal interface in the first trimester pregnancy. Decidual T lymphocytes express significantly higher levels of GNLY (58%) then PB T lymphocytes (11%). Over 85% of decidual CD56(+) cells express GNLY and when cultured spontaneously release high quantities of GNLY. Decidual APC participate in the control of GNLY expression in CD56(+) cells.
Abundant expression of GNLY in the decidual immunocompetent cells and the capacity of decidual CD56(+) cells to spontaneously secrete high quantities of GNLY point to important protective and immunomodulatory role that this molecule could play at the maternal-fetal interface.
American Journal Of Reproductive Immunology 05/2011; 66(5):363-72. · 2.17 Impact Factor
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British Journal of Haematology 11/2009; 148(5):812-4. · 4.94 Impact Factor
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ABSTRACT: Immune changes are known to occur in recurrent spontaneous abortion, but it is unclear whether either maternal natural killer (NK) cells or T cells attack fetus-derived trophoblasts. To clarify the immunological causes of spontaneous abortion, we examined the relationship between cytotoxic granule proteins in decidual lymphocytes, such as granulysin, granzyme B, and perforin, and the induction of apoptosis in extravillous trophoblasts (EVTs). The number of granulysin-positive CD56(bright) NK cells increased significantly in the decidua basalis during spontaneous abortion compared with normal pregnancy; however, granzyme B- and perforin-positive cells did not change. Interestingly, the expression of granulysin was also detected in the nuclei of EVTs in spontaneous abortion samples. When IL-2-stimulated CD56(bright) NK cells were cocultured with EVT cells (HTR-8/SV40neo), granulysin was found initially in the cytoplasm and then accumulated in the nuclei of the HTR-8/SV40neo cells. Furthermore, transfected cells expressing a GFP-granulysin fusion protein induced apoptosis in HTR-8/SV40neo cells independently of caspases. Our results suggest that granulysin-positive uterine NK cells attack EVTs; subsequently, the uNK-derived granulysin actively accumulates in the nuclei of EVTs, causing the death of EVTs due to apoptosis. These data support a new apoptosis pathway for trophoblasts via uNK-derived granulysin, suggesting that granulysin is involved in spontaneous abortion.
American Journal Of Pathology 10/2008; 173(3):653-64. · 4.89 Impact Factor
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ABSTRACT: Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is a newly identified monocyte derived cytokine, which has weak apoptosis inducing function against sensitive tumor cell lines in vitro. Also, TWEAK has been reported to have proangiogenic and proinflammatory activities in vivo. However, its functions in pathological situation remain to be elucidated. Here, we analyzed soluble TWEAK in serum of 24 patients with hemophagocytic lymphohistiocytosis (HLH) in combination with interferon-gamma (IFN-gamma) and killer-specific secretory protein of 37 kDa (Ksp37). Soluble TWEAK was not detected in serum of healthy individuals. Soluble TWEAK was markedly elevated in all six primary HLH patients and 12 of 18 secondary HLH patients. Serum IFN-gamma, which is an only known mediator to stimulate TWEAK production in monocyte in vitro, was not elevated despite elevated serum TWEAK in three of six primary HLH patients, although IFN-gamma was markedly elevated in other cases. Ksp37 was only slightly increased in HLH patients. These results indicate that TWEAK may be involved in pathogenesis of HLH and is useful as a clinical marker.
American Journal of Hematology 04/2008; 83(3):222-5. · 4.67 Impact Factor
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International Journal of Hematology 01/2008; 86(5):470-3. · 1.27 Impact Factor
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Susumu Saigusa,
Takashi Ichikura,
Hironori Tsujimoto,
Hidekazu Sugasawa,
Takashi Majima,
Nobuaki Kawarabayashi,
Kentaro Chochi,
Satoshi Ono,
Manabu Kinoshita,
Shuhji Seki, Kazuyuki Ogawa,
Hidetaka Mochizuki
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ABSTRACT: Granulysin is a cytolytic molecule present in human cytotoxic T cells and natural killer cell granules, and plays a key role in the cell-mediated immunity against tumor and infection. However, few studies have estimated serum granulysin concentrations in patients with solid or hematological malignancies.
Peripheral blood samples were taken from patients with gastric carcinoma preoperatively and from healthy volunteers. Serum and tumor tissue granulysin concentrations were measured using a granulysin-specific ELISA kit in order to assess its prognostic value.
Both serum and tumor tissue granulysin concentrations were higher in patients with stage II or III gastric cancer and lower in patients with stage IV disease as compared to healthy controls. The low preoperative granulysin levels were associated with more frequent hepatic and peritoneal metastases, and with a poor outcome of the curative gastrectomy.
Preoperative serum granulysin levels reflect the status of cell-mediated immunity in patients with gastric carcinoma. It has significance as a prognostic determinant following a curative resection.
Journal of Gastroenterology and Hepatology 09/2007; 22(8):1322-7. · 2.87 Impact Factor
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ABSTRACT: Granulysin is a cytolytic granule protein released by natural killer cells and activated cytotoxic T lymphocytes. The influence of exercise training on circulating granulysin concentration is unknown, as is the relationship between granulysin concentration, natural killer cell number and natural killer cell cytotoxicity. We examined changes in plasma granulysin concentration, natural killer cell number and cytotoxicity following acute exercise and different training loads. Fifteen highly trained male cyclists completed a baseline 40-km cycle time trial (TT401) followed by five weeks of normal training and a repeat time trial (TT402). The cyclists then completed four days of high intensity training followed by another time trial (TT403) on day five. Following one final week of normal training cyclists completed another time trial (TT404). Fasting venous blood was collected before and after each time trial to determine granulysin concentration, natural killer cell number and natural killer cell cytotoxicity. Granulysin concentration increased significantly after each time trial (P<0.001). Pre-exercise granulysin concentration for TT403 was significantly lower than pre-exercise concentration for TT401 (-20.3 +/- 7.5%, P<0.026), TT402 (-16.7 +/- 4.3%, P<0.003) and 7T404 (-21 +/- 4.2%, P<0.001). Circulating natural killer cell numbers also increased significantly post-exercise for each time trial (P<0.001), however there was no significant difference across TT40 (P>0.05). Exercise did not significantly alter natural killer cell cytotoxicity on a per cell basis, and there were no significant differences between the four time trials. In conclusion, plasma granulysin concentration increases following moderate duration, strenuous exercise and is decreased in response to a short-term period of intensified training.
Exercise immunology review 01/2007; 13:89-99. · 2.79 Impact Factor
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ABSTRACT: The circulating number of natural killer (NK) cells largely changes after an acute bout of physical exercise. Granulysin is a cytolytic granule protein with a broad range of antimicrobial and tumoricidal activities produced and released by human NK cells and cytolytic T lymphocytes. Since NK cells constitutively produce granulysin, most serum granulysin in healthy humans is derived from NK cells. Serum graulysin levels in the healthy humans may therefore reflect the size of whole-body NK cell population in the body. The aim of this study was to determine the effect of an acute bout of exhaustive exercise on serum granulysin in comparison with the circulating number of NK cells. Six healthy, young male volunteers participated in the study. Each subject underwent both exhaustive exercise and resting sessions in a random order with at least a seven-day interval. Subjects were asked to run to exhaustion on a treadmill with an incremental graded protocol. Blood samples were collected before, immediately after, and 1 hr, 3 hr, 6 hr, 12 hr and 24 hr after exercise. Serum granulysin levels were measured by enzyme-linked immunosorbent assay (ELISA). NK cells were determined by flow cytometry. Exhaustive exercise induced a 4.8-fold increase in peripheral blood NK cells, but no significant change in serum granulysin. Our results support the hypothesis that exhaustive exercise-induced changes in the circulating number of NK cells represent a redistribution of lymphocytes, rather than the change in the size of whole-body NK cell population.
The Tohoku Journal of Experimental Medicine 11/2006; 210(2):117-24. · 1.24 Impact Factor
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ABSTRACT: Epidemiological, clinical and histological data suggest intriguing similarities between preeclampsia and graft-host-rejection. Granulysin, a novel biomarker of overall cellular immunity, is secreted by natural killer cells and cytotoxic T lymphocytes, which are associated with graft-host-rejection. Plasma granulysin was elevated in Japanese preeclamptic women.
50 preeclampsia cases and 50 normotensive controls (USA) were studied. Plasma granulysin at delivery was determined using enzyme immunoassay. Logistic regression procedures were used to estimate odds ratios (OR) and 95% confidence intervals (CI).
Granulysin were elevated in preeclampsia cases compared with controls (3.01+/-0.18 vs. 2.22+/-0.14 ng/mL, p<0.01). After adjusting for age, body-mass-index and race, women with higher granulysin concentrations (> or =1.89 ng/mL) experienced a 2.9-fold (95%CI 1.1-7.8) increased preeclampsia risk compared with women with lower granulysin (<1.89 ng/mL).
These data offer further evidence of a predominant Th1 immune status associated with preeclampsia. Prospective studies are needed to determine whether granulysin is elevated early in pregnancy.
Clinical Biochemistry 11/2006; 39(10):1016-21. · 2.08 Impact Factor
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ABSTRACT: Granulysin is a newly identified CTL/NK cell-related cytotoxic protein, which is secreted in both constitutive and Ca-dependent manner. To evaluate its significance in stem-cell transplantation (SCT), serum granulysin was measured by newly established ELISA method in 26 patients undergoing SCT (21 allogeneic and 5 autologous). In the allogeneic SCT, granulysin was transiently increased in 3 weeks, which was not observed in autologous SCT. In acute GVHD, serum granulysin was markedly increased and correlated with the severity of GVHD. Elevation of granulysin was not necessarily associated with increase of sIL2R or IFN-gamma. In vitro, allospecific T cells released granulysin in an allo-specific manner, and it was correlated with allo-specific cytotoxic activity. These results indicate that increased release of granulysin presents alloreactivity and serum granulysin is useful as a marker of GVHD in SCT.
American Journal of Hematology 06/2006; 81(5):340-8. · 4.67 Impact Factor
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ABSTRACT: Exogenously added granulysin was reported to kill mammalian target cells. The sites of actions and molecular mechanisms of granulysin in target cell killing, however, are presently unclear. We here examine the effects of granulysin with the target HeLa cells transiently expressed with GFP-fused 9 kDa granulysin. Endogenously expressed GFP-fused granulysin was preferentially localized in the nucleus and induced apoptotic cell death accompanying with phosphatidylserine translocation and nuclear condensation in a caspase-independent manner. These results suggest that granulysin enters the nucleus of target cells and induces apoptosis.
Journal of medical and dental sciences 04/2005; 52(1):1-7.
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Kazuyuki Ogawa,
Yasushi Takamori,
Kunou Suzuki,
Masayuki Nagasawa,
Shoichi Takano,
Yoshihito Kasahara,
Yoshiko Nakamura,
Shigemi Kondo,
Kazuo Sugamura,
Masataka Nakamura,
Kinya Nagata
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ABSTRACT: Granulysin is a cytolytic granule protein of natural killer (NK) cells and cytotoxic T lymphocytes (CTL) with a broad range of antimicrobial and tumoricidal activities. Two molecular forms of granulysin, the 15-kDa precursor and 9-kDa mature form, are produced in these cells. In this study, we developed monoclonal antibodies against granulysin and found that the 15-kDa granulysin is spontaneously secreted by peripheral blood NK and T cells via a non-granule exocytotic pathway. When NK cells killed the target cells, the released granulysin levels in culture supernatants significantly increased through the granule exocytosis. The granulysin protein was found in the sera of healthy individuals at an average concentration of 3.7 +/- 3.2 ng/ml (age 0-99 years, n=244). The serum levels of granulysin were transiently highly elevated among patients with acute viral infections. In addition, the serum granulysin levels in patients with severe immunodeficiency treated bycell therapy fluctuated proportionately to the improvement of other immunological parameters. Our results suggest that granulysin is well associated with diverse activities of NK cells and CTL in physiological and pathological settings and could be a useful novel serum marker to evaluate the overall status of host cellular immunity.
European Journal of Immunology 08/2003; 33(7):1925-33. · 5.10 Impact Factor
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ABSTRACT: Maternal cellular immunity is thought to be in a state of tolerance during pregnancy, but the precise mechanism of immunomodulation is not yet known. We investigated a novel serum protein, killer-specific secretory protein of 37 kDa (Ksp37), produced by cytotoxic lymphocytes, during pregnancy.
The level of Ksp37 was determined by enzyme linked immunosorbent assay (ELISA) in the sera of healthy pregnant women. Intracellular Ksp37 expression in mononuclear cells, isolated from peripheral blood and decidua at parturition, was examined with a flow cytometer.
Serum Ksp37 levels significantly increased at late pregnancy, compared with non-pregnant controls and the first trimester of pregnancy. The flow cytometric analysis exhibited that Ksp37 was mainly expressed in CD16+ natural killer (NK) cells in decidua of term placenta.
Serum Ksp37 level was elevated at late gestational period. CD16+ NK cells in decidua seem to be a main maternal source of Ksp37. Innate immunity, with CD16+ NK cells, may play important roles near parturition.
American journal of reproductive immunology (New York, N.Y.: 1989) 08/2002; 48(1):57-62. · 3.05 Impact Factor
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ABSTRACT: Granulysin has been identified as an effector molecule co-localized with perforin in the cytotoxic granules of cytotoxic T lymphocytes and natural killer (NK) cells, and has been reported to kill intracellular pathogens in infected cells in the presence of perforin and to induce a cytotoxic effect against tumor cells. The aim of the present study was to elucidate whether intracellular expression of granulysin and perforin by NK cells might be associated with progression of cancer. Flow cytometric analysis demonstrated high levels of perforin and granulysin expression by CD3(-) CD16(+) cells in healthy controls. In contrast, cancer patients exhibited significantly decreased levels of granulysin expression ( P<0.005), despite having equally high levels of perforin expression in comparison with healthy controls. The tumor-free patients expressed granulysin at levels similar to healthy controls, while the progressive tumor-bearing patients expressed remarkably lower levels of granulysin compared to healthy controls ( P<0.0001). Similarly, patients with an advanced performance status had significantly fewer granulysin-positive NK cells than healthy controls. Meanwhile, a considerable number of the tumor-bearing patients showed a decrease in the number of circulating NK cells, and a correlation between impaired granulysin expression and reduced circulating NK cells was observed. These findings suggest that the tumor-bearing patients with impaired granulysin expression were in an immunosuppressive state. In conclusion, impaired expression of granulysin by NK cells correlates with progression of cancer, and determination of granulysin expression might prove informative for assessing the immunological condition of cancer patients.
Cancer Immunology and Immunotherapy 02/2002; 50(11):604-14. · 3.70 Impact Factor
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Hiroyuki Hirai,
Kazuya Tanaka,
Osamu Yoshie, Kazuyuki Ogawa,
Kazumi Kenmotsu,
Yasushi Takamori,
Michiko Ichimasa,
Kazuo Sugamura,
Masataka Nakamura,
Shoichi Takano,
Kinya Nagata
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ABSTRACT: Prostaglandin (PG)D2, which has long been implicated in allergic diseases, is currently considered to elicit its biological actions through the
DP receptor (DP). Involvement of DP in the formation of allergic asthma was recently demonstrated with DP-deficient mice.
However, proinflammatory functions of PGD2 cannot be explained by DP alone. We show here that a seven-transmembrane receptor, CRTH2, which is preferentially expressed
in T helper type 2 (Th2) cells, eosinophils, and basophils in humans, serves as the novel receptor for PGD2. In response to PGD2, CRTH2 induces intracellular Ca2+ mobilization and chemotaxis in Th2 cells in a Gαi-dependent manner. In addition, CRTH2, but not DP, mediates PGD2-dependent cell migration of blood eosinophils and basophils. Thus, PGD2 is likely involved in multiple aspects of allergic inflammation through its dual receptor systems, DP and CRTH2.
Journal of Experimental Medicine 01/2001; 193(2):255-262. · 13.85 Impact Factor
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ABSTRACT: Cytotoxic lymphocytes such as CTL and NK cells play principal roles in the host defense mechanisms. Monitoring these effector cells in vivo is helpful to understand the immune responses in disorders such as cancer and infectious diseases. In this study, we identified a novel secretory protein, killer-specific secretory protein of 37 kDa (Ksp37), as a Th1-specific protein by a subtractive cloning method between human Th1 and Th2 cells. In peripheral blood leukocytes, Ksp37 expression was limited to Th1-type CD41 T cells, effector CD81 T cells, gd T cells, and CD161 NK cells. Most of these Ksp37-expressing cells coexpressed perforin, indicating that Ksp37 is selectively and commonly expressed in the lymphocytes that have cytotoxic potential. Ksp37 was released at constant rate from both unstimulated and stimulated PBMCs in vitro and also detected in normal human sera. In healthy individuals, serum Ksp37 levels were significantly higher in children (mean 6 SD; 984 6 365 ng/ml for age 0 -9) than in adults (441 6 135 ng/ml for age 20 -99), consistent with reported differences in the absolute counts of blood T and NK cells between children and adults. In patients with infectious mononucleosis, transient elevation of serum Ksp37 levels was observed during the early acute phase of primary EBV infection. These results suggest that Ksp37 may be involved in an essential process of cytotoxic lymphocyte-mediated immunity and that Ksp37 may also have clinical value as a new type of serum indicator for monitoring cytotoxic lymphocytes in vivo. The Journal of Immunology, 2001, 166: 6404 - 6412.
