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Objective@#To investigate the levels of NK cells and their relevant cytokines (IL-10, TGF-β and IFN-γ) in patients with primary immune thrombocytopenia (ITP) .@*Methods@#All samples were obtained from 42 patients (22 newly diagnosed and 20 in remission) and 20 healthy volunteers. The levels of IL-10 and IFN-γ in blood serum were detected by enzyme-linked immunosorbent assay (ELISA) . The percentage of CD3- CD56+ NK cell, CD3- CD56bright CD16- NK cell, CD3- CD56dim CD16+ NK cell in peripheral blood lymphocyte were detected by flow cytometry. The NK cells were isolated by immunomagnetic microbeads. The mRNA expression levels of IL-10, TGF-β, and IFN-γ in NK cells were detected by real-time fluorescent quantitative PCR. Correlation between the above measured results was analyzed.@*Results@#① The blood serum level of IFN-γ in newly diagnosed ITP patients [ (653.0±221.6) ng/L] was higher than that in remission ITP patients [ (484.4±219.5) ng/L] and healthy control [ (390.9±253.5) ng/L] (P=0.022, P=0.001) . The blood serum level of IL-10 in newly diagnosed ITP patients was lower than that in healthy control [ (52.09±26.66) ng/L vs (79.44±38.43) ng/L, P=0.007]. ②The percentage of NK cell in newly diagnosed and remission ITP patients [ (9.53±3.93) %, (9.03±3.78) %] were significantly lower than that in healthy control [ (13.72±7.42) %] (P=0.013, P=0.007) . The ratio of CD3- CD56bright CD16- NK cell/total NK cells in newly diagnosed ITP patients was higher than that in healthy control [ (6.85±4.43) % vs (4.05±2.81) %, P=0.032]. The ratio of CD3-CD56dim CD16- NK cell/total NK cells in newly diagnosed ITP patients was lower than that in healthy control [ (93.14±4.43) % vs (95.94±2.81) %, P=0.032]. ③ There was no significant difference in the mRNA expression level of IFN-γ in NK cells of ITP patients and healthy control (all P>0.05) . The mRNA expression levels of IL-10 and TGF-β in NK cells in newly diagnosed ITP patients were significantly higher than that in healthy control (1.82±1.32 vs 1.02±1.03, P=0.023; 2.80±2.31 vs 1.46±1.37, P=0.028) . The ratio of CD3-CD56bright CD16- NK cell/total NK cells was positively correlated with the mRNA expression levels of IL-10, TGF-β in NK cells (r=0.424, P=0.001; r=0.432, P<0.001) .@*Conclusion@#NK cells may compensate for the deficiency of the number by enhancing the secretion of negative regulation cytokines, acting as "protective" roles in the disease.
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Objective To investigate the effects of hydrocortisone (HC) on proliferation and killing activity of NK cells against pancreatic cancer SW1990 cells in vitro.Methods Peripheral blood mononuclear cells of healthy people were isolated and cultured with NK cells medium containing IL-1S.When the purity of NK cells reached above 70％,different concentrations of HC (10-6,10-5,10-4,10-3 μmol/L) were added and co-cultured with NK cells for 7 days.And NK cells without HC were used as control.CD3-CD56 + NK cell numbers of each group were countered by trypan blue staining.Perforin,granzyme B and IFN-γ expression of CD3-CD56+ + NK cells were verified by flow cytometry.NK cells and SW1990 cells were co-cultured with a 20∶1effector to target ratio,then the cytotoxic activity of NK cells against SW1990 cells were analyzed by CCK-8 kit.Results After treatment with different concentration of HC for 7 days,NK cells purity of each group reached 70.72％ ～ 76.39％,and it was not significantly different with that in control group [(72.61 ± 3.76) ％].The proliferation folds of NK cells treated with 10-6,10-5,10-4,10-3 μmol/L HC were (9.13 ± 0.94),(9.67 ±1.51),(10.33±1.07),(8.40±1.47) times,respectively,while it was (4.23 ±0.82) times in control group (all P ＜0.01).The killing effects of NK cells on SW1990 cells were (58.58 ± 4.89) ％,(62.27 ± 5.63) ％,(64.02 ± 5.79) ％,(63.88 ± 3.61) ％,which were higher than that in control group [(57.46 ± 5.11) ％],moreover,the difference between NK cells of 10-4 μmol/L HC treatment group and control group was statistically significant(P ＜ 0.05).The expressions of perforins of 10-4,10-3 μmol/L HC treatment group were (96.71 ± 3.04) ％,(97.56 ± 2.18) ％,which were significantly higher than that in control group [(92.40 ±3.53)％,P ＜0.05 or 0.01].The expression of granzyme B in 10-5 μmol/L HC treatment group was (78.23 ±2.94)％,which were significantly higher than that in control group [(73.68 ±3.52) ％,P ＜0.05].The expressions of IFN-γ in 10-5,10-4,10-3 μmol/L HC treatment group were (96.61 ±2.04)％,(97.58 ± 2.17)％,(98.00 ± 1.77)％,which were significantly higher than that in control group [(92.44 ± 2.74)％,P＜0.01].Conclusions HC can promote IL-15 activated NK cells proliferation and enhance NK cells mediated killing activity against SW1990 cells with proper concentration,and up-regulation of perforin,granzyme B and IFN-γ expression may be the main mechanisms.
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Objective To detect the serum level of interleukin (IL)-1 1, lymphocyte subsets and NK cells in patients with idiopathic thrombocytopenic purpura (ITP), and explore the related factors in the pathogenesis of ITP. Methods The serum level of IL-11, lymphocyte subsets and NK cells were detected by double antibody sandwich enzyme linked immunosorbent assay (ELISA) and flow cytometry in 50 ITP patients (ITP group) and 30 controls (control group). Results The platelet in ITP group [ (30.21 ± 19.40) ×109/L] was lower than that in control group [ (207.21 ± 31.55 ) × 109/L] obviously (P ＜ 0.05 ); the serum level of IL-11 in ITP group [(255.72 ± 163.43) ng/L] was significantly higher than that in control group [ (40.60 ± 5.57 ) ng/L ] (P ＜ 0.05 ). The correlation analysis indicated that the blood serum levels of IL- 11 had negative relationship with the platelet (r = -0.557 ,P ＜ 0.05). The percentage of CD3+ and CD4+ T lymphocyte percentage, CD4+/CD8+ in ITP group were lower and the percentage of CD8+ T lymphocyte was higher than those in control group obviously (P ＜ 0.05 ). The percentage of CD3- CD(16+56) +NK cell in ITP group was lower than that in control group (P ＜ 0.05). Conclusion IL-11, lymphocyte subgroup and NK cell change correlate with ITP morbidity closely, and the IL-11 level and the platelet possibly have the negative feedback control action.
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Objective To compare the effects of different anesthesia on immunologic function during perioperative radical mastectomy in patients having undergone preoperative chemotherapy. Methods Sixty ASA Ⅰ or Ⅱ patients, aged 28-64 yr, weighing 55-70 kg, scheduled for radical mastectomy, were studied. Thirty patients received preoperative chemical therapy for 3-4 weeks were randomly divided into CP group and CS group ( n = 15 each); thirty patients received no chemical therapy were also randomly divided into NCP group and NCS group (n=15 each).Propofol at a rate of 4-6 mg·kg-1·h-1 was infused in group CP and CS and sevoflurane with the end-tidal concentration of 1.5%-2.5% inhaled in group NPC and NCS to maintain anesthesia. Peripheral venous blood samples were taken before chemical therapy, before anesthesia, immediately after operation and at 72 h after operation for determination of the levels of T lymphocyte subsets and NK cells and plasma CK19 mRNA expression (by RT-PCR). CD4+/CD8+ ratio and the micrometastatic rate of the tumor cells were calculated. Results The CD3+ , CD8+ and NK cell level were significantly lower before anesthesia, immediately after operation and at 72 h after operation than before chemical therapy ( P < 0.05). The CD8+ immediately after operation and NK cell level at 72 h after operation in group CP, and CD4+ and NK cell levels immediately and at 72 h after operation in group CS were significantly lower than those before anesthesia ( P < 0.05). CD4+ and NK cell levels immediately and at 72 h after operation were significantly lower in group CS than in group CP, and in group NCS than in group NCP (P < 0.05). The CD4+/CD8+ ratio was significantly lower immediately after operation in group CS than in group CP ( P < 0.05) .There was no significant difference in the micrometastatic rate of the tumor cells among all groups (P > 0.05). Conclusion Sevoflurane combined anesthesia has a stronger inhibitory effect on immunologic function in radical mastectomy patients with preoperative chemotherapy than propofol combined anesthesia.
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Objective To identify the clinical and pathological features of aggressive natural killercell leukemia (ANKL). Methods 9 cases of ANKL fulfilling the criteria defined by the World Health Organization classification were retrospectively analyzed with literature review, Results Systemic symptoms, hepatomegaly, splenomegaly, lymphadenopathy were frequently observed. Liver dysfunction, neutropenia, anemia and thrombocytopenia were often seen during the course of the disease. Most of the bone marrow shows focal or subtle infiltration by the neoplastic cells. The immunophenotype of cells was characteristic for CD+56, sCD-36, and variable expression of CD2, CD7, CD8 and CD11b. T-cell receptor (TCR) genes rearrangement were in germline configuration. Median survival time was 9 weeks. Conclusion ANKL is an entity of mature cytotoxic NK-cell neoplasms with distinct phenotype and disease presentations. The disease often has a fulminant course with a poor response to chemotherapy and a short survival time. Patients achieving CR showed significantly longer survival time, but the remission did not translate into cure of the disease.
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To evaluate the frequency of bone marrow involvement by nasal-type NK/T cell lymphoma, we retrospectively studied biopsy specimens from 40 patients by EBV in situ hybridization (ISH). Three patients had marrow involvement at initial diagnosis (7.5%). In one patient (1/40, 2.5%), the disease in bone marrow was recognized by routine morphological assessment, while two other patients had minimal involvement of lymphoma cells which was recognized only by EBV in situ hybridization (2/40, 5%). Two patients had a disseminated disease at diagnosis and died 6 days and 214 days after diagnosis. One patient had diffuse colonic lesion and died 82 days later. In conclusion, marrow involvement in nasal NK/T cell lymphoma is infrequent at initial diagnosis, and EBV ISH is a useful technique for identifying the minor subgroup of patients which have easily overlooked neoplastic involvement.
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Biopsy , Bone Marrow/pathology , Epstein-Barr Virus Infections/complications , Herpesvirus 4, Human/genetics , Killer Cells, Natural/pathology , Lymphoma, T-Cell/complications , Nasal Cavity/pathology , Prognosis , RNA, Viral/analysis , Retrospective StudiesABSTRACT
To evaluate the induction of preneoplastic hepatic foci in relation to natural killer cell (NK) activity, we sequentially analyzed glutathione S-transferase placental form positive (GST-P+) hepatocytes and NK activity during diethylnitrosamine (DEN) and phenobarbital (PB)-induced hepatocarcinogenesis in Sprague-Dawley rats. Previous studies have shown that NK activity can modulate the carcinogenic process induced by chemical carcinogens. Newborn females were initially given a single intraperitoneal injection of 15 mg DEN/kg and three weeks later, they were treated with 500 ppm phenobarbital (PB). From week 3, PB was administered in drinking water for 9 weeks. Interim and terminal sacrifices were performed at weeks 12, 15 and 30. GST-P+ hepatocytes increased with age in DEN-treated rats, especially in the population of more than two GST-P+ hepatocytes. The NK activity of DEN-treated rats did not significantly differ from that of control rats until week 12, but it progressively decreased from week 15 to 30. These results indicate that changes of NK activity inversely correlated with the induction of preneoplastic hepatic foci. This strong correlation of decreased NK activity with enhanced induction of GST-P+ foci suggests that NK activity is important in the early progression of hepatocarcinogenesis in rats.
Subject(s)
Female , Rats , Animals , Body Weight , Carcinogens/pharmacology , Diethylnitrosamine/pharmacology , Glutathione Transferase/metabolism , Killer Cells, Natural/immunology , Liver/enzymology , Liver/cytology , Liver Neoplasms/physiopathology , Organ Size , Placenta , Rats, Sprague-DawleyABSTRACT
The level of interferon (IFN) in serum and homogenized cardiac tissue, and the activity of natural killer cells in Coxsackievirus B3 induced myocarditis in BALB/c mice were determined. NK cell cytolytic activity and IFN titers peaked on day 3~ 7 postinoculation (p.i.) and then declined. Virus titers in heart tissue reached a maximum on day 7 p.i. and then declined. These results suggest that the increase of NK cell activity and IFN titers provides some protection against Cox B, induced myocarditis by limiting virus replication in heart tissue.