Hepacivirus ; isolation & purification ; Humans ; Leukocytes, Mononuclear ; virology ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Viremia ; virology

Adult ; Epstein-Barr Virus Infections ; complications ; Humans ; Leukocytes, Mononuclear ; virology ; Nasopharyngeal Neoplasms ; pathology ; virology ; Viral Matrix Proteins ; genetics

BACKGROUNDTo reveal the characteristics of genotype and phenotype of HIV strains in blood and some tissues of AIDS patients.

METHODSThe virus was isolated from peripheral blood mononuclear cell (PBMC),cerebrospinal fluid (CSF)and lymph nodes of 3 AIDS patients by coculture with PBMC stimulated by PHA for 72 hours from uninfected donor. The cytopathic effect of the HIV isolates was determined in cultured MT2 cell line. The env gene sequences form proviral DNA were analyzed by GCG software.

RESULTSIn one patient,there were differences between the strains from blood and different tissues both in genotype and phenotype. The biological phenotypes of two strains from CSF were non syncytium (NSI) type, their env sequences were similar to standard CNS tropic strain (SF162).

CONCLUSIONSThe viral heterogeneity exists in different body compartments within an infected individual. The neurotropic isolate which is similar to international standard strain exists in some AIDS patients in China.

Acquired Immunodeficiency Syndrome ; virology ; Adult ; Coculture Techniques ; Female ; Genetic Heterogeneity ; Genotype ; HIV ; isolation & purification ; Humans ; Leukocytes, Mononuclear ; virology ; Lymph Nodes ; virology ; Male ; Phenotype

BACKGROUNDTo study the significance of the expression of HCV core protein in PBMC of patients with chronic hepatitis C and to evaluate the relationship between HCV core protein expression and clinical states.

METHODSIdentification of HCV protein antigen (Ag) in PBMC of 66 hepatitis C patients by immunohistochemical method and clinical status of the patients with HCV protein positive expression were investigated. In 27 out off 66 patients the HCV RNA and HCV Ag in PBMC were detected with RT-PCR and immunohistochemical method.

RESULTSThe HCV Ag (core+NS3) was identified in PBMC in 51 out of 66 patients (77.27%). It was also demonstrated that HCV core protein in nucleus showed strong expression and NS3 protein in cytoplasm showed weak expression. The expression of core protein in nucleus of PBMC were 35.29% in advanced chronic hepatitis patients, which was significantly higher than that from moderate cases (5.88%).

CONCLUSIONSThis study suggested that the expression of PMBC-HCV core protein may be related to the clinical state of the patients. The nuclear expression of HCV core protein in PBMC of patients with hepatitis C may be related to the persistence and activity of the chronic hepatitis C virus and play an important role in the pathogenesis of cirrhrosis and hepatocellular carcinoma.

Hepatitis C Antigens ; blood ; Hepatitis C, Chronic ; virology ; Humans ; Immunohistochemistry ; Leukocytes, Mononuclear ; metabolism ; RNA, Viral ; blood ; Viral Core Proteins ; blood

This study was aimed to explore the method for induction and expansion of EB virus specific cytotoxic T lymphocytes (EBV-CTL) in vitro, and to detect their killing effect. Peripheral blood mononuclear cells (PBMNC) were collected from 6 EBV seropositive healthy donors, and EBV-transformed B lymphoblastoid cells (BLCL)were used as the antigen-presenting cells and antigen stimulant which was irradiated by 40 Gy (60)Co irradiator. The autologous PBMNC and irradiated BLCL were cultured to induce and expand the EBV-CTL, and the immunophenotype was identified by the flow cytometry. The killing effect of the EBV-CTL against the autologous BLCL (autoBLCL), the autologous PHA cultured B lymphoblastoid cells( PHA-BLCL), the allogeneic BLCL (alloBLCL) and the K562 cells were measured with LDH release assay under different effector-to-target ratio. The results showed that the 6 cell lines of EBV-CTL were induced and expanded from the EBV seropositive healthy donors, the overall increase in cell numbers varied from 18.6 to 55.0 times. After 10 stimulations, the specific killing efficiency of the EBV-CTL for the autoBLCL were 59.4%, 43.2% and 29.0% under the effector-to-target ratio of 20: 1, 10: 1 and 5: 1. The nonspecific killing efficiency for the PHA-blast, alloBLCL and K562 cells were 7.1%, 9.4% and 10.3% (P < 0.05) under the 20: 1 ratio; 6.6%, 8.3% and 8.1% (P < 0.05) under 10: 1; 5.4%, 7.3% and 6.3% (P < 0.05) under 5: 1, respectively. It is concluded that the EBV-CTL can be successfully induced and expanded ex vivo for specific killing of HLA matched BLCL and may become a potential treatment for EBV related post-transplant lymphoproliferative disorders.
B-Lymphocytes ; immunology ; Cell Line, Transformed ; Herpesvirus 4, Human ; immunology ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; immunology ; virology ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; virology

OBJECTIVETo investigate the potential transmissibility of porcine endogenous retrovirus (PERV) from a newly-developed porcine hepatocyte bioartificial liver (BAL) system prior to human clinical trial by using a live canine model.

METHODSFive normal beagles were treated with the new BAL support system for six hours. Samples of plasma from the BAL system and whole blood from the beagles were collected at regular intervals over the six month study period. DNA and RNA were isolated from both the peripheral blood mononuclear cells (PBMCs) and plasma for evaluation by polymerase chain reaction (PCR) and reverse transcription (RT)-PCR, respectively, to detect PERV and the Sus scrofa cytochrome B normalization standard. In addition, RT activity and the in vitro infectivity of the plasma were detected in HEK293 cells.

RESULTSAll five beagles remained in stable physical health throughout the treatment and survived until the end of the study. PERV RNA-positivity and RT activity were only detected in the plasma samples from the 3rd BAL treatment cycle. All other samples, including PBMCs and plasma, were negative for PERV RNA, PERV DNA, and RT activity. In addition, none of the sera samples showed in vitro infectivity.

CONCLUSIONApplication of our BAL system does not lead to PERV transmission.

Animals ; Cell Line ; Dogs ; Endogenous Retroviruses ; HEK293 Cells ; Hepatocytes ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Liver, Artificial ; adverse effects ; Models, Animal ; Swine

OBJECTIVETo investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C to IFN treatment.

METHODSThe HCV RNAs in PBMC were detected at the end of treatment, and 24 week and 1 year follow up after end treatment, in 16 patients who acquired complete response to IFN in 12 weeks of 24 weeks therapy.

RESULTS9 patients were HCV RNA positive in their PBMC at the end of treatment, the serum HCV RNA of 8 turned positive after 24 weeks and 1 year follow-up. In 7 patients with negative HCV RNA in PBMC, only two patients relapsed in serum HCV RNA after 1 year follow-up, and others remained viral response after 3.5 years.

CONCLUSIONHCV RNA in PBMC at the end of treatment was a predictor of the durable response to antiviral therapy in chronic hepatitis C.

Adult ; Female ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Leukocytes, Mononuclear ; virology ; Male ; Middle Aged ; RNA, Viral ; blood ; Recombinant Proteins

Adolescent ; Adult ; Aged ; Child ; DNA, Circular ; blood ; DNA, Viral ; blood ; Female ; Hepatitis B, Chronic ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; Middle Aged

OBJECTIVEIn order to investigate the epidemics of borna disease virus (BDV) in Ningxia and its vicinal regions.

METHODSp24 fragment of BDV from: (1) peripheral blood mononuclear cells (PBMC) and cerebrospinal fluid mononuclear cells (CSFMC) from 52 patients with viral encephalitis (VE) and 32 healthy donors, (2) peripheral blood mononuclear cells (PBMC) from 53 patients with depressive disorder (DD) and from 360 sheep, were examined by nested reverse transcriptase polymerase chain reaction(PCR) with fluorescence quantitative PCR. Gene sequence and amino acid sequence were analysed for positive product and the molecular epidemiologic characteristics by drawing phylogenetic trees.

RESULTSThe positive rate of BDV p24 in CSFMC from VE (11.54%) and in PBMC from DD 11.32% was significantly higher than that in healthy donors (0%) (P < 0.05). The phylogenetic trees indicating the genetic relationship of the p24 fragment of BDV in both sheep and VE, DD in China and was similar to the nucleotide sequence of H1766 strain in Germany.

CONCLUSIONData indicated that the BDV infection was possibly existing in VE, DD patients and health sheep in Ningxia and its vicinal regions with confined locality which called for further study.

Animals ; Borna Disease ; epidemiology ; genetics ; Borna disease virus ; genetics ; isolation & purification ; China ; epidemiology ; Depressive Disorder ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Molecular Epidemiology ; Phylogeny ; Polymerase Chain Reaction ; Sheep

Adult ; Female ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Transcriptome ; Young Adult