This study was to establish a model to explore anti- RSV effect of different administration method of Chinese medicine realgar on respiratory syncytial virus type A (RSV-A) replication in Hep-2 cells. Using high-energy ball milling with distilled water to prepare realgar nanoparticles,the concentration of nanometer realgar was tested by molybdenum blue staining method and the size of realgar nanoparticles was tested on Nano Series. Cell culture with ribavirin as a positive control was applied to observe the effect of anti-respiratory syncytial virus type A replication through prevention, treatment or direct inactivation of three different drug administration methods. Realgar nano-particles was found to be a potential inhibitor of RSV-A in a concentration-dependent manner with the median toxic concentration(TC50) of 0.649 microg/mL in Hep-2 cell culture. The median inhibition concentration (IC50) was 0.20 microg/mL when drug was added before virus infection. The IC50 was 0.13 microg/mL when drug was added after virus infection,and it was 0.16 microg/mL when the drug was mixed with virus and added. The therapeutic index (TI) was 3.18, 4.99 and 4.11, respectively. The results showed realgar nanoparticles could inhibit the replication of the RSV and inactivate the RSV in vitro.
Arsenicals ; pharmacology ; Nanoparticles ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Sulfides ; pharmacology ; Virus Replication ; drug effects

OBJECTIVETo investigate the core functional region of antimicrobial peptide LL-37, which inhibites RSV replication and could be developed for theraputic aplication.

METHODSA panel of 6 partial LL-37 peptides (referred to as P1 to P6) was synthesized according to LL-37 amino acide sequence. Hep-2 cells were infected with RSV, treated with LL-37 or partial peptides respectively. Cells were collected after 24 hours incubation at 37 degrees C, CO2 5%. Total RNA was obtained from the cells. Expression level of RSV N gene was quantified by real-time PCR. Meanwhile enzyme-linked immunosorbent assay (ELISA) was used to quantify the chemokines RANTES, IL-8, MCP1 in the supernatants of Hep-2 cultures after 24 h incubation with or without LL-37 and partial peptide P6.

RESULTSN-terminal partial LL-37 peptide (corresponding to residues 1-12 of LL-37) had no significant effects on RSV replication (P > 0.05). In contrast, C-terminal (corresponding to residues 13-37) and a panel of 4 overlapping 22-mer partial peptides (from the peptide incorporating aa 13-34 through that spanning aa 16-37) showed significant inhibitory effect on RSV replication to some extent (P < 0.05 or P < 0.01). LL-37 induced significant expression of chemokine RANTES, IL-8 and MCP-1 in Hep-2 cells. In contrast, partial peptide P6 had no significant effect on expression of the chemokines in Hep-2 cells.

CONCLUSIONThe LL-37 C-terminal 22-mer partial peptide P6 was putative core functional region for inhibition of RSV replication. The partial peptide didn't induce significant expression of chemokine RANTES, IL-8 and MCP-1.

Cathelicidins ; pharmacology ; Cell Line ; Cytokines ; genetics ; immunology ; Humans ; Respiratory Syncytial Virus Infections ; genetics ; immunology ; virology ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Virus Replication ; drug effects

OBJECTIVETo evaluate possible inactivating effect of recombined decoction in on mumps virus.

METHODSBy adopting tissue cell culturing technology, a group of viruses including the mumps virus, herpes simplex virus (type I, II), rubella virus, cytomegalovirus (CMV), herpes zoster virus, influenza virus, parainfluenza virus, adeno viruses, respiratory syneytial virus (RSV) were cultured. The cells infected with the viruses were treated with the decoction.

RESULTSThe decoction showed remarkable inhibitory and killing effects on the mumps virus while had no obvious inhibitory and killing effects on host's cells, herpes simplex virus (type I, II), rubella virus, cytomegalovirus (CMV), herpes zoster virus, influenza virus, parainfluenza virus, adeno viruses, respiratory syneytial virus (RSV).

CONCLUSIONSThe decoction had obvious inhibitory and killing effects on mumps virus during single layer cells culture.

Cells, Cultured ; Cytomegalovirus ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Mumps virus ; drug effects ; Respiratory Syncytial Viruses ; drug effects ; Respirovirus ; drug effects ; Rubella virus ; drug effects ; Simplexvirus ; drug effects

OBJECTIVETo search the effect of PPARgamma agonists for infection of RSV in vitro.

METHODSThe CPE of Hep-2 and A549 cells induced by RSV infection were observed. The effects of 15d-PGJ2 and rosiglitazone on change of CPE of A549 cells induced by RSV infection for 48 h were observed, too. MTT assay was used to detect the rate of viral suppression, and the protective effects of 15d-PGJ2 and rosiglitazone on A549 cells induced by RSV infection for 48 h.

RESULTSA549 cells interfered by 15d-PGJ2 (5 -25 micromol/L) and rosiglitazone (10-50 micromol/L) did not show obvious CPE, MTT assay also showed that the survival rate of A549 cells induced by RSV infection with PPARgamma agonists added, was significantly higher than that of RSV infection without PPARgamma agonists added, the difference was statistically significant (P < 0.01), but comparision between the two drugs showed no statistical significance. The optimal concentrations of 15d-PGJ2 and rosiglitazone were 5 micromol/L and 10 micromol/L respectively.

CONCLUSIONSPPARgamma agonist can reduce the CPE of A549 cells after RSV infection and improve the survival rate of A549 cells. PPARgamma agonist can counteract the infection of RSV in A549 cells.

Cells, Cultured ; Humans ; PPAR gamma ; agonists ; Prostaglandin D2 ; analogs & derivatives ; pharmacology ; Respiratory Syncytial Viruses ; drug effects ; Thiazolidinediones ; pharmacology

OBJECTIVETo explore the antivirus function in vitro of earthworm coelomic fluid (ECF) by researching its effect on inhibiting respiratory syncytial virus (RSV).

METHODSBy the method of Hep-2 cell culture and using ribavirin as a positive control, the anti-RSV effect of ECF was investigated by observing cytopathic effect (CPE) with MTT colorimetric assay.

RESULTSIn Hep-2 cells, the CC50 of ECF and ribavirin were 3.11 mg/ml and 1.35 mg/ml separately. In the experiment of ECF directly killing RSV, the IC50 of ECF was 184.1 microg/ml, SI was 16.87; In the experiment of ECF preventing RSV invasion, no antiviral function of ECF within the experimental concentration range was observed; In the experiment of ECF inhibiting RSV replication, the IC50 of ECF was 1555. 8 microg/ml, SI was 1.99.

CONCLUSIONECF couldn't prevent virus from invading into host cell, but showed direct killing-virus function and inhibition of the virus replication.

Animals ; Antiviral Agents ; chemistry ; pharmacology ; Cell Line ; Humans ; Inhibitory Concentration 50 ; Oligochaeta ; chemistry ; Respiratory Syncytial Viruses ; drug effects ; Virus Replication ; drug effects

OBJECTIVETo study the effect on anti-respiratory syncytial virus of an active compound (AP3) from a Chinese medicinal herb-Herba patriniae in vitro.

METHODSActive component of herba patriniae (AP3) was extracted and its anti-respiratory syncytial virus (RSV) effect was tested. A water soluble substance (AP3) was isolated from a Chinese herb Herba patriniae, by hot water extraction, ethol precipitation and gel permeation column chromatography. The cytotoxicity of AP3 was tested by adding it to HeLa cells directly. Its effect against RSV was estimated by CPEI assay while ribavirin was used as positive control.

RESULTSChemical test showed that the nature of substance AP3 was polysaccharide. The median cytotoxic concentration (TC(50)) of AP3 was 11.45 mg/ml by morphological observation and the median effective concentration (50% effective concentration, EC(50)) of it against replication of the long strain of RSV in HeLa cells was 0.0986 mg/ml. The Therapeutic index (TI = TC(50)/EC(50)) of AP3 was 116.12, much higher than the TI of herba patriniae (AP1) (TI = 59.26) and ribavirin (TI = 53.45). Moreover, AP3 gave a dose-dependent response in inhibiting RSV. In the assay, the effect of AP3 against RSV growth was also tested. In addition, the effect of AP3 on virus growth, AP3 inhibited replication of RSV in HeLa cells, when added at 0 h, 2 h, 4 h after virus infection, were also tested.

CONCLUSIONThis study suggested that the AP3 exerted an obvious inhibitory effect to RSV in HeLa cell culture. This study furnished a reliable evidence for development of a new antiviral drug.

Antiviral Agents ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; HeLa Cells ; Humans ; Respiratory Syncytial Viruses ; drug effects ; Time Factors

OBJECTIVE: To explore the mechanisms for an increase in susceptibility of asthma induced by respiratory syncytial virus (RSV), to observe the expression of interleukin-8 (IL-8) in human bronchial epithelial cells (HBECs) after RSV infection and to invesigate the regulatory effect of IL-8 on Th17/Treg differentiation.
 METHODS: HBECs were divided into a control group and a RSV infected group. The RSVE-infected model of HBECs was established and examined. The expression of IL-8 mRNA was detected by real-time PCR, and the levels of IL-8 were measured by ELISA. Peripheral blood lymphocytes in healthy people were extracted and divided into a control group and an IL-8 treatment group. Based on concentration of IL-8 in RSV-infected HBECs, lymphocytes were treated by a matched concentration of human recombinant IL-8 for 24 h. The distribution of Th17 and Treg subsets in lymphocytes were examined by flow cytometry.
 RESULTS: The RSV-infected HBECs model was successfully established. The infected HBECs were still able to split and passage. The RSV could be detected in every passage in the infected cells. Virus particles indicated by bright yellow green fluorescence were seen under fluorescence microscope. Edema of mitochondrias, expansion of endoplasmic reticulum, fissure around nucleus and intracellular virus particles were all observed under electron microscope. The expression IL-8 mRNA were significantly enhanced in the RSV-infected group, and the level of IL-8 in the RSV-infected group was higher than that in the control group (P<0.05). After IL-8 treatment for 24 h, the ratio of Th17 subsets in lymphocytes were dramatically increased compared to the control group (P<0.05), but there was no difference in the ratio of Treg subsets between the 2 groups (P>0.05).
 CONCLUSION Over-secretion of IL-8 by the RSV-infected HBECs may promote the differentiation of Th17 subsets and maintain the Th17/Tred imbalance.
Cell Differentiation ; Cells, Cultured ; Epithelial Cells ; drug effects ; virology ; Flow Cytometry ; Humans ; Interleukin-8 ; immunology ; pharmacology ; Real-Time Polymerase Chain Reaction ; Recombinant Proteins ; pharmacology ; Respiratory Syncytial Virus Infections ; immunology ; Respiratory Syncytial Viruses ; T-Lymphocytes, Regulatory ; cytology ; Th17 Cells ; cytology

OBJECTIVETo understand the clinical characteristics and outcome associated with respiratory syncytial virus (RSV) infection in hematopoietic stem cell transplantation (HSCT) recipients with primary immunodeficiencies (PIDs).

METHODNasopharyngeal aspirate samples were collected consecutively before and after HSCT from 9 recipients from Apr. 2009 to Sep. 2010 and analyzed for the presence of RSV using real-time polymerase chain reaction assay. To further verify the presence of the virus, positive samples for PCR were isolated for RSV. RSV G gene was amplified, sequenced and used for phylogenetic analysis.

RESULTThe presence of RSV was detected in 3 out of 9 children. The viral replication in all the patients was prolonged for months. All the 3 patients with RSV infection were treated with intravenous immune globulin (IVIG) and one was treated with antiviral medication. All patients survived and achieved successful immune reconstitution.

CONCLUSIONThis study indicates that the HSCT recipients with PID are at increased risk for RSV infection. RSV can shed for months after the initial infection and the patients recover with the course of immune reconstitution.

Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunologic Deficiency Syndromes ; surgery ; virology ; Infant ; Prognosis ; Respiratory Syncytial Virus Infections ; diagnosis ; drug therapy ; Respiratory Syncytial Viruses ; genetics ; isolation & purification ; physiology ; Virus Replication ; Virus Shedding

OBJECTIVEDNAzyme/Deoxyribozyme is another novel molecular biological tool following the ribozyme. DNAzyme consists of a 15-nucleotide (nt) internal loop as its catalytic domain and two flanking substrate-recognition domains of 7 to 8 nt each which is complementary to substrate. The RNA substrate is cleaved at a particular phosphodiester located between an unpaired purine and a paired pyrimidine residue. DNAzyme has been applied in fields such as viral infectious disease, tumor, cardiovascular disease and genetic disease. But there is no report about using DNAzyme for anti-respiratory syncytial virus purpose. To observe the inhibitory effects of RNA-cleaving DNAzymes on respiratory syncytial virus (RSV) replication in cultured cells.

METHODSAnti-RSV RNA-cleaving DNAzyme DZ604 was designed to target the RSV genome at the start of the NS2 gene in an effort to inhibit the RNA replication. Microscope and electron microscope were used to observe the effects of anti-RSV genomic RNA DNAzyme on cytopathogenic effect (CPE) and ultrastructural change of 9HTE cell induced by RSV infection. Viral plaque forming reduction assay and MTT assay were used to detect the anti-RSV activity and protective function for RSV infected 9HTE cells of DNAzyme.

RESULTSAnti-RSV genomic RNA DNAzyme (DZ604) significantly improved CPE of RSV-infected 9HTE cells. The time to appearance of CPE and of total CPE was delayed by using DZ604 in a dose-dependent manner. At a 5 micro mol/L concentration of DZ604, CPE of 9HTE cells induced by RSV infection at 10 and 1 multiplicity of infection (MOI) was not improved. At smaller MOI (0.1, 0.01, 0.001, 0.0001) of RSV infection, CPE of 9HTE cells was significantly lightened by DZ604 at the same concentration. DZ604 also significantly improved ultrastructural change of 9HTE cells at early stage of RSV infection. Reduction in RSV yield was 85.56% and 8.33% at concentrations of 5 micro mol/L and 0.25 micro mol/L of DZ604. DZ604 inhibited RSV yield in a dose-dependent manner (P < 0.05). Non-specific DNAzyme did not have anti-RSV activity (P > 0.05).

CONCLUSIONAnti-RSV genomic RNA DNAzyme designed and synthesized in our laboratory was capable of inhibiting respiratory syncytial virus replication specifically in cultured cells. Our data indicated that DNAzymes could be useful for the prevention against respiratory syncytial virus infection.

Animals ; Cell Line ; Cercopithecus aethiops ; DNA, Catalytic ; pharmacology ; Dose-Response Relationship, Drug ; Humans ; Microscopy, Electron ; Respiratory Mucosa ; cytology ; ultrastructure ; virology ; Respiratory Syncytial Viruses ; drug effects ; genetics ; physiology ; Vero Cells ; Virus Replication ; drug effects

OBJECTIVETo study the effective part of solution prescription of Zhidanhuayu (ZDHY) against respiratory syncytial virus (RSV) in vitro.

METHODSObserve the pathology of RSV to Hep-2 under the condition of different concentrations and each effective part of ZDHY.

RESULTSThe concentration limit causing celluar toxicity of ZDHY is 5.5 mg/ml. The ZDHY failed to block the absorption of RSV to Hep-2 within this concentration, and consequently the cell fell into the full pathological changes. During the concentration of 2.75-5.50 mg/ml, the ZDHY directly destroyed virus array,meanwhile, the infected cells that treated by the medicine kept healthy also.

CONCLUSIONZDHY could not defend the infection of RSV, but is able to destroy the RSV directly and inhibit the RSV inhabiting in the cell.

Animals ; Antiviral Agents ; adverse effects ; therapeutic use ; Cell Line ; Cells, Cultured ; Cercopithecus aethiops ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Respiratory Syncytial Virus Infections ; drug therapy ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Vero Cells ; drug effects ; physiology ; Virus Replication ; drug effects