Erythrocyte Indices ; Erythrocytes ; Hepacivirus ; Humans ; Liver Diseases ; blood ; virology

OBJECTIVETo investigate the mechanism and the suppression effect of human cytomegalovirus (HCMV) on hematopoietic system.

METHODSSemi-solid culture system was used to observe the effect of HCMV AD169 strain on colony forming unit granulocyte/macrophage (CFU-GM), CFU-erythroid (CFU-E), CFU-multipotent (CFU-Mix) and CFU-megakaryocyte (CFU-MK) growth. The techniques of in situ polymerase chain reaction (IS-PCR) and polymerase chain reaction (PCR) were used to demonstrate the existence of HCMV DNA in the colony cells of cultured CFU-GM, CFU-Mix, CFU-MK and CFU-E, respectively. The immediate early antigen (IEA) mRNA in CFU-MK and late antigen (LA) mRNA in CFU-E were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). HCMV early protein P52 was detected with immunohistochemical technique.

RESULTSHCMV AD169 suppressed the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK in vitro significantly (P < 0.05). The suppression was dose-dependent. HCMV DNA was successfully detected in CFU-GM, CFU-Mix, CFU-MK colony cells from viral infection groups by IS-PCR, and was detected in CFU-E by PCR, while it was negative in blank control or mock control groups. CFU-MK colony cells expressed HCMV IEA mRNA with the size of 340 bp in virus infection groups of 10(3) plague forming unit (PFU), 10(4) PFU and 10(5) PFU, respectively. The HCMV LA mRNA was detected by RT-PCR and was 263 bp long in positive control group of HCMV-infected human embryonic fibroblasts. The expression of HCMV LA mRNA in CFU-E was negative. The early protein P52 of HCMV in 10(4) PFU group was also identified by immunohistochemical staining.

CONCLUSIONHCMV AD169 strains inhibited the differentiation and proliferation of CFU-GM, CFU-E, CFU-Mix and CFU-MK by the infection of the hematopoietic progenitors. HCMV might cause the suppression of hematopoiesis by direct infection, which is thought to be one of the reasons of HCMV infection associated with thrombocytopenia, neutropenia and anemia.

Colony-Forming Units Assay ; Cytomegalovirus ; genetics ; DNA, Viral ; genetics ; Erythrocytes ; virology ; Hematopoietic System ; cytology ; virology ; Humans ; Megakaryocytes ; virology ; Multipotent Stem Cells ; virology ; Polymerase Chain Reaction

Elution time of velogenic, mesogenic and lentogenic strains of Newcastle disease virus was determined. The differences in their elution time were also calculated. Four samples, each of a velogenic strain (VGF2), a mesogenic strain (Komarov) and a lentogenic strain (LaSota) were used for hemagglutination test with 0.6% chicken red blood cells. The time it took for wells of the end hemagglutination points (highest dilution that gave agglutination) to elute was recorded as elution time for each sample. The mean elution time of the three strains of Newcastle disease virus differed significantly (p < 0.05). The velogenic strain gave the highest mean elution time of 118 min, followed by the mesogenic strain with 59 min and the lentogenic strain with 25 min. Based on this result it appears that elution time could form a basis for rough characterization of isolates of Newcastle disease virus into the three major strains.
Animals ; Chickens/blood ; Erythrocytes/*virology ; Hemagglutination Tests ; *Hemagglutination, Viral ; Newcastle disease virus/isolation&purification/*pathogenicity

This study was aimed to evaluate the efficacy of riboflavin photochemical inactivation of virus in red blood cells by using animal models. human cytomegalovirus (HCMV) plus red blood cells were used as indicator, 30 BALA/c mice were divided into the experimental group (n = 10), virus control group (n = 10), visible light control group (n = 5) and red blood cell control group (n = 5). Mice in experimental group were inoculated with red blood cells inactive by the riboflavin photochemical, mice in virus control group was injected with red blood cells without riboflavin photochemical inactivation treatment, and mice in light control group was infused with red blood cells irradiated by visible light, and mice in red blood cells control group was injected with normal red blood cells. The virus was isolated in vitro from mice of various groups, the HCMV UL83 gene was detected by PCR, the PP65 antigen was identified by indirect immunofluorescence. The results indicated that the virus isolation, PCR detection and indirect immunofluorescence identification all showed positive in virus control group and visible light control group, while the results of detection in experimental and red blood cell control groups were negative. It is concluded that riboflavin photochemical viral inactivation of red blood cells is effective.
Animals ; Erythrocytes ; virology ; Humans ; Mice ; Mice, Inbred BALB C ; Models, Animal ; Photochemistry ; Riboflavin ; pharmacology ; Virus Inactivation

BACKGROUND: Validation of hemagglutination inhibition (HI) assays is important for evaluating antibody responses to influenza virus, and selection of erythrocytes for use in these assays is important. This study aimed to determine the correlation between receptor binding specificity and effectiveness of the HI assay for detecting antibody response to pandemic influenza H1N1 (pH1N1) virus. METHODS: Hemagglutination (HA) tests were performed using erythrocytes from 6 species. Subsequently, 8 hemagglutinating units of pH1N1 from each species were titrated by real-time reverse transcription-PCR. To investigate the effect of erythrocyte binding preference on HI antibody titers, comparisons of HI with microneutralization (MN) assays were performed. RESULTS: Goose erythrocytes showed most specific binding with pH1N1, while HA titers using human erythrocytes were comparable to those using turkey erythrocytes. The erythrocyte binding efficiency was shown to have an impact on antibody detection. Comparing MN titers, HI titers using turkey erythrocytes yielded the most accurate results, while those using goose erythrocytes produced the highest geometric mean titer. Human blood group O erythrocytes lacking a specific antibody yielded results most comparable to those obtained using turkey erythrocytes. Further, pre-existing antibody to pH1N1 and different erythrocyte species can distort HI assay results. CONCLUSIONS: HI assay, using turkey and human erythrocytes, yielded the most comparable and applicable results for pH1N1 than those by MN assay, and using goose erythrocytes may lead to overestimated titers. Selection of appropriate erythrocyte species for HI assay allows construction of a more reliable database, which is essential for further investigations and control of virus epidemics.
Adult ; Animals ; Antibodies, Neutralizing/immunology ; Antibodies, Viral/*analysis/immunology ; Chickens ; Erythrocytes/*metabolism ; Female ; Geese ; *Hemagglutination Inhibition Tests ; Horses ; Humans ; Influenza A Virus, H1N1 Subtype/genetics/immunology/*metabolism ; Influenza, Human/epidemiology/immunology/virology ; Male ; Middle Aged ; Neutralization Tests ; Pandemics ; Swine ; Turkeys

OBJECTIVETo research the mechanism of the inhibition effects of BWE on cell attachment of influenza virus by capillary electrophoresis.

METHODThe morphologic difference of red cells after treating with BWE infected by influenza virus was detected with microscope, capillary electrophoresis and HA.

RESULTThe pretreatment of the normal cells with BWE inhibited the attachment of influenza to the cells, while no meaningful inhibition was observed when influenza virus was pretreated before being inoculated to cells.

CONCLUSIONThe results indicate that the inhibition effects of BWE on cell attachment of influenza virus may be an important mechanism of anti-influenza activity of Radix Isatidis Extracts.

Antiviral Agents ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Electrophoresis, Capillary ; Erythrocytes ; ultrastructure ; virology ; Hemagglutination Inhibition Tests ; Humans ; Influenza A virus ; drug effects ; Isatis ; chemistry ; Male ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry