OBJECTIVETo investigate the telomerase activity and the telomere length in CD4(+), CD8(+) and CD19(+) lymphocytes from patients with systemic lupus erythematosus (SLE).

METHODSTelomerase activity of CD4(+), CD8(+) and CD19(+) cells from patients with SLE and normal controls was examined by telomeric repeats amplification protocol. Telomere length in those cells was measured by Southern blot method.

RESULTCD4(+), CD8(+) and CD19(+) cells in patients with SLE showed high telomerase activity, but only telomerase activity of CD19(+) cells was significantly higher than that in controls. The length of telomere in CD4(+) and CD8(+) cells was significantly shorter than that in controls, but not in CD19(+) cells.

CONCLUSIONHigher telomerase activity in CD19(+) cells and shortened telomere length in CD4(+) and CD8(+) cells of patients with SLE may be associated with pathogenesis of SLE.

Adolescent ; Adult ; Antigens, CD19 ; immunology ; CD4-Positive T-Lymphocytes ; enzymology ; CD8-Positive T-Lymphocytes ; enzymology ; Female ; Humans ; Lupus Erythematosus, Systemic ; enzymology ; immunology ; Lymphocytes ; enzymology ; Male ; Middle Aged ; Telomerase ; metabolism ; Telomere ; genetics

Cysteine and aspartic proteases possess high elastolytic activity and might contribute to the degradation of the abdominal aortic aneurysm (AAA) wall. The aim of this study was to analyze, in detail, the proteases (cathepsins B, D, K, L and S, and inhibitor cystatin C) found in human AAA and healthy aortic tissue samples. The vessel walls from AAA patients (n=36) and nonaneurysmal aortae (n=10) were retrieved using conventional surgical repair and autopsy methods. Serum samples from the same AAA patients and 10 healthy volunteers were also collected. Quantitative expression analyses were performed at the mRNA level using real-time reverse transcriptase-PCR (RT-PCR). Furthermore, analyses at the protein level included western blot and immunoprecipitation analyses. Cellular sources of cysteine/aspartic proteases and cystatin C were identified by immunohistochemistry (IHC). All cysteine/aspartic proteases and cystatin C were detected in the AAA and control samples. Using quantitative RT-PCR, a significant increase in expression was observed for cathepsins B (P=0.021) and L (P=0.018), compared with the controls. Cathepsin B and cystatin C were also detected in the serum of AAA patients. Using IHC, smooth muscle cells (SMCs) and macrophages were positive for all of the tested cathepsins, as well as cystatin C; in addition, the lymphocytes were mainly positive for cathepsin B, followed by cathepsins D and S. All cysteine/aspartic proteases analyzed in our study were detected in the AAA and healthy aorta. The highest expression was found in macrophages and SMCs. Consequently, cysteine/aspartic proteases might play a substantial role in AAA.
Aged ; Aorta/enzymology ; Aortic Aneurysm, Abdominal/*enzymology ; Aspartic Acid Proteases/genetics/*metabolism ; Case-Control Studies ; Cathepsins/genetics/metabolism ; Cysteine Proteases/genetics/*metabolism ; Humans ; Lymphocytes/enzymology ; Macrophages/enzymology ; Middle Aged ; Myocytes, Smooth Muscle/enzymology ; RNA, Messenger/genetics/metabolism

OBJECTIVETo study the effects of benzene and selenium on telomerase in mouse lymphocytes in vivo and evaluate telomerase activity as an early marker of benzene effects on lymphocytes.

METHODSMale Kunming mice were divided randomly into 8 groups, including negative control group, reagent control group, 100 mg/kg benzene group, 200 mg/kg benzene group, 400 mg/kg benzene group, 200 mg/kg benzene + 0.75 mg/kg selenium group, 200 mg/kg benzene + 1.50 mg/kg selenium group and 200 mg/kg benzene + 3.00 mg/kg selenium group, 5 mice in each group. The mice in different groups were treated with different methods, once daily for 5 days. After 48 hours of the final exposure, lymphocytes were separated and the telomerase activities were detected with TRAPELISA.

RESULTSCompared with negative and reagent control groups, the telomerase activity was increased after treatment with different dose of benzene and at the dose of 100 mg/kg benzene group it was significantly increased (P < 0.01). At the dose of 200 mg/kg benzene + 0.75 mg/kg selenium group, it was significantly increased (P < 0.01). Compared with the counterpart treated with 200 mg/kg benzene group, the expression of telomerase was increased at the different concentrations after treatment with benzene combined with selenium and it was significantly increased at the dose of 200 mg/kg benzene + 0.75 mg/kg selenium group (P < 0.05).

CONCLUSIONIncreased telomerase activity in lymphocytes stimulated by benzene at different concentrations indicates activation and proliferation of these lymphocytes of mice in vivo. Telomerase activity is probably a sensitive early marker of lymphocyte proliferation by benzene. Selenium can upregulate the telomerase activity.

Animals ; Benzene ; pharmacology ; Cells, Cultured ; Lymphocytes ; drug effects ; enzymology ; Male ; Mice ; Selenium ; pharmacology ; Telomerase ; metabolism

OBJECTIVE: To explore the expression of perforin and granzyme-B in peripheral blood lymphocyte (PBL) in patients with prostate cancer (PCa) and the clinical significance. METHODS: The expressions of perforin and granzyme-B in PBL were detected by fluorescence quantitative reverse transcription polymerase chain reaction. The results of perforin and granzyme-B expression were compared among patients with PCa (n=60), patients with BPH (benign prostatic hyperplasia, n=40) and healthy controls (n=20). RESULTS: Th e expressions of perforin and granzyme-B in patients with PCa were significantly lower than that in patients with BPH or that in the healthy controls (P<0.05), respectively. Furthermore, in PCa patients with low pathological grade, the expressions of perforin and granzyme-B in PBL was statistically higher than that in patients with high pathological grade (P<0.05). The expressions of perforin and granzyme-B in PCa patients at high clinical stage was statistically lower than that in PCa patients at low clinical stage (P<0.05). CONCLUSION The results of this study suggest that development and progression of PCa might be associated with poor immune status of patients.
Case-Control Studies ; Granzymes ; metabolism ; Humans ; Lymphocytes ; enzymology ; Male ; Perforin ; metabolism ; Prostatic Hyperplasia ; Prostatic Neoplasms ; immunology

Several studies have shown the mechanisms and importance of immune responses against Toxoplasma gondii infection and the notable role of cholinesterases in inflammatory reactions. However, the association between those factors has not yet been investigated. Therefore, the aim of this study was to evaluate the acetylcholinesterase (AChE) activity in blood and lymphocytes and the activity of butyrylcholinesterase (BChE) in serum of rats experimentally infected with T. gondii during the acute phase of infection. For that, an in vivo study was performed with evaluations of AChE and BChE activities on days 5 and 10 post-infection (PI). The activity of AChE in blood was increased on day 5 PI, while in lymphocytes its activity was enhanced on days 5 and 10 PI (P<0.05). No significant difference was observed between groups regarding to the activity of BChE in serum. A positive (P<0.01) correlation was observed between AChE activity and number of lymphocytes. The role of AChE as an inflammatory marker is well known in different pathologies; thus, our results lead to the hypothesis that AChE has an important role in modulation of early immune responses against T. gondii infection.
Acetylcholinesterase/blood/*metabolism ; Animals ; Butyrylcholinesterase/blood/*metabolism ; Humans ; Lymphocytes/enzymology/parasitology ; Male ; Rats ; Rats, Wistar ; Toxoplasma/*physiology ; Toxoplasmosis/*enzymology/genetics/parasitology

OBJECTIVEAcute idiopathic thrombocytopenic purpura (AITP) is a common autoimmune disease in children. Thrombocytes decrease extremely in serious patients, its pathogenesis involves abnormal activation of T lymphocytes and T cell-dependent production of autoantibody. The aim of the present study was to investigate changes of protein kinase C (PKC) activity in peripheral blood T lymphocytes in children with AITP and the relationships between PKC activity and T lymphocytes activation and thrombocytopenia.

METHODSPeripheral blood specimens were collected from children with acute ITP (n = 35) and healthy children (n = 30), and T lymphocytes were isolated and purified by using T cells Segregation Enrichment Column. PKC activity was detected by using PepTag Assay, a non-radioactive detection method. The reaction mixture, in a final volume of 25 microl, consisted of 5 microl reaction buffer, PepTag C1 5 microl (0.4 microg/microl), PKC activator solution (DG) 5 microl, peptide protection solution 1 microl and sample 9 microl. Phosphorylation reaction was allowed to continue for 30 minutes, then 25 microl reaction mixture was subjected to electrophoresis on a 0.8% agarose gel at 100 V for about 20 minutes. After electrophoresis, the PepTag C1 peptides which were phosphorylated and non-phosphorylated were separated, phosphorylated PepTag C1 peptide with negative electricity migrated toward the anode (+), but nonphosphorylated PepTag C1 peptide with positive electricity migrated toward cathode (-), the gel was photographed. Electrophoresis bands on anode represented PKC activity and were analyzed quantitatively. FasL, which is T cell activation marker, was determined by flow cytometer and platelet was counted by cell counting meter.

RESULTSCompared with healthy children, children with AITP had significantly higher PKC total activity [(0.97 +/- 0.21) nmol/(min.ml) vs. (0.55 +/- 0.13) nmol/(min.ml), (P < 0.05)]. Expression of FasL on T cell subpopulation in children with AITP was significantly higher [Th FasL: (32.7 +/- 3.4) vs. (14.7 +/- 4.2); Tc FasL: (17.3 +/- 9.7) vs. (11.6 +/- 8.5)%, (P < 0.05)]. Besides, relationships between the changes of PKC activity, Th FasL and Tc FasL had positive correlation (r(1) = 0.68, r(2) = 0.53, P < 0.05). However, PKC activity and platelet count had a significantly negative correlation (r = -0.75, P < 0.05).

CONCLUSIONIncreased PKC activity was seen in children with AITP, which can cause damage to thrombocytes and reduction of thrombocytes. PKC signal transduction pathway might play an important role in the immunopathogenesis of AITP.

Acute Disease ; Case-Control Studies ; Cell Separation ; Child ; Electrophoresis ; Flow Cytometry ; Humans ; Lymphocyte Activation ; Platelet Count ; Protein Kinase C ; metabolism ; Purpura, Thrombocytopenic, Idiopathic ; enzymology ; immunology ; T-Lymphocytes ; enzymology

Thymidine phosphorylase (TP) has shown to be up-regulated in several cancers and to play a role in angiogenesis and invasion. Most studies regarding TP have focused on cancer cells. Recently, evidences suggest that TP in cancer-infiltrating inflammatory cells (CIICs) also affect the cancer cell behavior. To evaluate the significance of TP expression of CIICs in gastric cancer, we assessed TP expression of cancer cells and CIICs separately using immunohistochemical assay on 116 paraffin-embedded tissue samples from stomach cancer patients and investigated their clinical significance. When subjects were divided into 4 groups according to the TP expression: cancer/matrix (+/+), C/M (+/-), C/M (-/+), and C/M (-/-), intratumoral microvessel density scores were higher in the C/M (+/-) group than in the C/M (-/-) group (p=0.02). For lymph node metastasis and survival, there were no significant differences among the 4 groups. However, there were significant differences in survival (p=0.035) and LN metastasis (p=0.023) between the two groups divided by TP expression of CIICs alone irrespective of TP expression of cancer cells. Taken together, this study suggested the TP expression in CIICs could affect lymph node metastasis and patients' survival in gastric cancer.
Adult ; Aged ; Female ; Humans ; Immunohistochemistry ; Inflammation/*enzymology/pathology ; Kaplan-Meiers Estimate ; Lymphatic Metastasis ; Lymphocytes, Tumor-Infiltrating/*enzymology/pathology ; Male ; Microcirculation/pathology ; Middle Aged ; Neovascularization, Pathologic ; Prognosis ; Stomach Neoplasms/blood supply/*enzymology/mortality/pathology ; Thymidine Phosphorylase/*metabolism

BACKGROUND/AIMS: Infectious mononucleosis (IM) is the clinical presentation of primary infection with Epstein-Barr virus. Although the literature contains a massive amount of information on IM, most of this is related specifically to only children or adults separately. In order to distinguish any differences between preschool children and youth patients, we retrospectively analyzed their demographic and clinical features. METHODS: Records of patients hospitalized from December 2001 to September 2011 with a diagnosis of IM were retrieved from Peking University First Hospital, which is a tertiary teaching hospital in Beijing. The demographic data and clinical characteristics were collected. RESULTS: IM was diagnosed in 287 patients during this 10-year period, with incidence peaks among preschool children (< or =7 years old, 130/287, 45.3%) and youth patients (>15 and <24 years old, 101/287, 35.2%). Although the complaints at admission did not differ between these two patient groups, the incidence of clinical signs (tonsillopharyngitis, lymphadenopathy, hepatomegaly, and edema of the eyelids) was much higher in preschool children. The incidence of liver lesion and percentage of atypical lymphocytes were significantly higher in the youth group (P<0.001), and the average hospital stay was longer in this group. Pneumonia was the most common complication, and there was no case of mortality. CONCLUSIONS: The incidence of IM peaks among preschool children and youth patients in Beijing, China. The levels of liver enzymes and atypical lymphocytes increase with age.
Adolescent ; Adult ; Alanine Transaminase/blood ; Aspartate Aminotransferases/blood ; Child ; Child, Preschool ; Demography ; Fever/etiology ; Humans ; Incidence ; Infant ; Infectious Mononucleosis/*diagnosis/enzymology/epidemiology/pathology ; Liver/*enzymology ; Lymphocytes/cytology/*immunology/metabolism ; Pharyngitis/etiology ; Retrospective Studies ; Young Adult ; gamma-Glutamyltransferase/blood

The enteric protozoan parasite Entamoeba histolytica is the causative agent of human amebiasis. During infection, adherence of E. histolytica through Gal/GalNAc lectin on the surface of the amoeba can induce caspase-3-dependent or -independent host cell death. Phosphorylinositol 3-kinase (PI3K) and protein kinase C (PKC) in E. histolytica play an important function in the adhesion, killing, or phagocytosis of target cells. In this study, we examined the role of amoebic PI3K and PKC in amoeba-induced apoptotic cell death in Jurkat T cells. When Jurkat T cells were incubated with E. histolytica trophozoites, phosphatidylserine (PS) externalization and DNA fragmentation in Jurkat cells were markedly increased compared to those of cells incubated with medium alone. However, when amoebae were pretreated with a PI3K inhibitor, wortmannin before being incubated with E. histolytica, E. histolytica-induced PS externalization and DNA fragmentation in Jurkat cells were significantly reduced compared to results for amoebae pretreated with DMSO. In addition, pretreatment of amoebae with a PKC inhibitor, staurosporine strongly inhibited Jurkat T cell death. However, E. histolytica-induced cleavage of caspase-3, -6, and -7 were not inhibited by pretreatment of amoebae with wortmannin or staurosporin. In addition, we found that amoebic PI3K and PKC have an important role on amoeba adhesion to host compartment. These results suggest that amebic PI3K and PKC activation may play an important role in caspase-independent cell death in Entamoeba-induced apoptosis.
*Apoptosis ; Caspases/metabolism ; Entamoeba histolytica/*enzymology/*growth & development ; Humans ; Hydrolysis ; Jurkat Cells ; Phosphatidylinositol 3-Kinases/*metabolism ; Protein Kinase C/*metabolism ; T-Lymphocytes/*parasitology/*physiology

This study was aimed to investigate the mechanism of indoleamine 2, 3-dioxygenase (IDO) activity in acute myeloid leukemia cells contributing to tumor immune escape. Myeloid leukemia cells were isolated from bone marrow of 23 patients with acute myeloid leukemia (AML) and IDO expression was detected by immunochemistry and RT-PCR methods. Then mixed lymphocyte reaction (MLR) of one way was carried out, leukemia cells were used as stimulating cells and T-lymphocytes were used as reactive cells in culture with or without 1-MT. T-lymphocyte proliferation rate was determined by MTT assay and IDO activity in supernatant of MLR was detected by high-performance liquid chromatography (HPLC). The results showed that IDO expression was found in 17 out of 23 cases of acute myeloid leukemia cells; IDO enzyme activity in leukemia cells inhibited T-lymphocyte proliferation in MLR cultures. It is concluded that IDO activity expressing in leukemia cells can suppress T-lymphocyte proliferation responses, which may be contributing to tumor immune escape.
Cell Proliferation ; Humans ; Immune Tolerance ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism ; Leukemia, Myeloid, Acute ; enzymology ; immunology ; pathology ; T-Lymphocytes ; cytology ; immunology ; Tumor Cells, Cultured ; Tumor Escape ; immunology