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ObjectiveTo explore effect of modified Wuhutang on airway inflammation and expression of mucin （Muc） 5AC， signal transducer and activator of transcription 3 （STAT3）， nuclear factor kappa B （NF-κB）， and NOD-like receptor thermal protein domain associated protein 3 （NLRP3） in respiratory syncytial virus （RSV）-infected asthmatic mice. MethodSeventy male BALB/c mice of 6-8 weeks old were randomized into normal control （CON）， asthma （ovalbumin， OVA）， RSV infection-induced asthma （OVA+RSV）， high-， medium-， and low-dose （4.08， 2.04， 1.02 g·kg-1·d-1， respectively） modified Wuhutang， and dexamethasone （Dxms， 0.1 g·kg-1d-1） groups （n=10）. The model of asthma was established by sensitization and atomization inhalation with OVA. The RSV infection-induced asthma model was established by three consecutive RSV nasal infusions （1.0 × 106 PFU·mL-1， 50 μL）. Wuhutang was administrated by gavage， and Dxms by intraperitoneal injection. The CON group was given the same amount of normal saline by gavage. The mice were anesthetized with 2.5% pentobarbital sodium 24 h after the last administration， and then the lung tissue was stained by hematoxylin-eosin （HE） and Van Gieson （VG） for observation of airway inflammation. The immunohistochemical assay was employed to detect the expression of Muc5AC. Western blot was employed to determine the protein levels of phosphorylated （p）-STAT3， STAT3， p-NF-κB， NF-κB， and NLRP3. ResultCompared with the CON group， the OVA group presented airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition. The OVA+RSV group showed severer airway inflammatory cell infiltration and tissue hyperemia and edema than the OVA group. Compared with the OVA+RSV group， modified Wuhutang alleviated the airway inflammatory cell infiltration， tissue hyperemia and edema， and collagen fiber deposition， and the high-dose group had the best performance. Compared with the CON group， the OVA group and the OVA+RSV group showed increased expression level of Muc5AC （P<0.01）. Compared with the OVA+RSV group， modified Wuhutang reduced the expression level of Muc5AC， and the reduction was significant in the high-dose group （P<0.05）. Compared with the high-dose modified Wuhutang group， Dxms lowered the expression level of Muc5AC （P<0.05）. Compared with the CON group， the OVA and OVA+RSV groups showed up-regulated protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.05， P<0.01）. Compared with the OVA+RSV group， modified Wuhutang down-regulated the protein levels of p-STAT3， p-NF-κB， and NLRP3 （P<0.01）. Compared with the high-dose modified Wuhutang group， the Dxms group showed up-regulated levels of p-STAT3， p-NF-κB proteins （P<0.01）. ConclusionModified Wuhutang can reduce airway inflammation and down-regulate the expression of Muc5AC， p-STAT3， p-NF-κB， and NLRP3 in RSV-infected asthmatic mice， which suggests that Wuhutang reduces airway inflammation in RSV-infected asthma by regulating the STAT3/NF-κB signaling pathway.
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@#Abstract：In recent years，the outbreak and prevalence of respiratory infectious diseases in the world seriously endanger human health，among which the respiratory infectious diseases caused by viral infection account for a large proportion. The use of vaccines and common antiviral drugs is an effective way to fight viral infection，but there are also problems such as lag and drug resistance. Monoclonal antibodies against respiratory viral infections provide a new strategy for clinical treatment. This paper reviews the development of monoclonal antibody against respiratory virus and its application in respiratory viral infectious diseases. Keywords：Respiratory viral infectious diseases；Respiratory syncytial virus（RSV）；Influenza virus（IFV）；Coronavirus （CoV）；Monoclonal antibody
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Objective To identify the role of IL-17A during respiratory syncytial virus (RSV) in-fection in a mouse model. Methods Female wild-type C57BL/6 mice and IL-17A knockout ( IL-17A-/-) mice at the age of 6 to 8 weeks were both randomly divided into two groups:control and RSV groups. Mice in the control groups were given the supernatant of Hep-2 cell culture, while those in the RSV groups were treated with RSV A2 through intranasal administration. Leukocytes in bronchoalveolar lavage fluid ( BALF) samples were counted. Left lung tissues were stained with hematoxylin and eosin ( HE ) to evaluate his-topathological scores. Airway hyperresponsiveness ( AHR) was measured by whole-body plethysmography. The concentrations of IFN-γ were determined with ELISA. RSV titers were measured by plaque assay. To assess the effects of IL-17A on IFN-γproduction and its role in RSV infection, IL-17A-/- mice were treated with exogenous recombinant murine IFN-γ or IL-17A, while wild-type mice were given IFN-γ neutralizing antibody intervention. Results The counts of inflammatory cells and neutrophils in BALF, lung tissue his-topathological scores, AHR, IFN-γlevels and virus titers of the wild-type group were higher than those of the IL-17A-/-group after RSV infection. IFN-γlevels, inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly increased in RSV-infected IL-17A-/- mice after the intervention of recombinant murine IL-17A or IFN-γ. RSV titers were much higher in the recombinant murine IL-17A-trea-ted group, but not affected by the recombinant murine IFN-γ intervention. Inflammatory cell counts in BALF, AHR and lung tissue histopathological scores were significantly decreased in RSV-infected wild-type mice following IFN-γ neutralizing antibody treatment, but no significant changes were found in RSV titers. Conclusions IL-17A might be involved in the pathogenesis of pulmonary diseases during RSV infection through promoting IFN-γ production and inhibiting viral clearance in mice.
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Extract of Nauclea latifolia (NL) root bark collected from the Nigerian flora was examined for anti-RSV activity. Preliminary data showed anti-RSV activities with IC50 =75.62 µg/ml when tested against the recombinant strain rgRSV expressing the green fluorescent protein. Corresponding assays for the cytotoxic effect of the extract against utilized cell lines gave TC50 = 333.82 µg/ml. Further screening of against the circulating RSV A2 strain established their promising anti-RSV utility. Time of additional studies for the elucidation of the possible mechanism of action gave 74.38, 69.42, and 71.90% reduction of RSV plaque forming units at the respective 0, 2, and 4 hours post-infection addition times.
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Objective Objective to establish a method for identification of respiratory syncytial virus (RSV) A and B subtypes for clinical research and application.Methods Using fluorescent quantitative polymerase chain reaction (FQ-PCR) screened 50 cases of throat swab that RSV were positive in hospitalized children from 2015 to 2017.The genotyping was performed according to the nucleotide sequence of G protein coding gene,and a single tube double nested PCR primers was designed for it.A and B subgroup by sequencing to conduct comparative analysis with nucleotide sequence in the Genebank.The results were analyzed by chi-square test.Results In the 50 cases of throat swab RSV positive children,respiratory syncytial virus A and B subtype and mixed infection rates were 82.00％,14.00％ and 4.00％,respectively.The difference was statistically significant (x2=81.06,P＜0.01).The RSV fractal results were consistent with the gene sequencing results.Conclusion The eastern part of shenzhen was dominated by respiratory syncytial virus A subtype epidemic and mixed infection.Single tube double nested polymerase chain reaction (PCR) and gene sequencing technique is suitable for the identification of A and B subtypes of RSV.It is characterized by high sensitivity,specificity and high accuracy.
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Objective:To investigate the changes of thymic lymphocyte procyanidin(TSLP)and interleukin-4(IL-4),interleukin-10(IL-10)and interleukin-13(IL-13)in the lung tissue of young rats infected with respiratory syncytial virus,and to investigate the changes of TSLP and Th2 Correlation.Methods:18 Wistar rats were randomly divided into RSV infection group and normal group,9 rats in each group.Application of RSV virus droplets nasal modeling,saline diarrhea as a control.The lung tissue of rats was taken for pathological observation;the load of syncytial virus at different time points and the expression of TSLP in lung tissue were detected by Real-time quantitative polymerase chain reaction(q-PCR);at the same time,the expression of TSLP at the protein level was detected by Western blot.The levels of cytokines IL-4,IL-10 and IL-13 in serum were measured by enzyme-linked immunosorbent assay(ELISA).And the relationship between TSLP and IL-4,IL-10 and IL-13 was analyzed by Spearman correlation.Results:Compared with the normal control group,the rats in the model group showed symptoms such as poor mental state,rough hair,decreased activity and shortness of breath after intranasal infection,and the symptoms were aggravated.Pathological observation showed that the alveolar wall was thickened in the model group,and a large number of lymphocytes,plasma cells,eosinophil infiltration.q-PCR was used to detect the syncytial virus load in model group,and reached the peak at about 5 days,then decreased gradually.RSV infection increased the secretion of TSLP in rat respiratory tract.q-PCR and Western blot showed that TSLP concentration in model group was higher than that in control group(P<0.05).The levels of IL-4,IL-10 and IL-13 in the serum of the model group were higher than those in the control group(P<0.05).Spearman correlation analysis showed that TSLP was positively correlated with IL-4 and IL-10 expression in RSV model group,and there was no correlation between TSLP and IL-13 expression.Conclusion:RSV infection in rat lung tissue can induce increased expression of TSLP,while promoting the occurrence of Th2 type of inflammatory response and the expression of related cytokines,in order to further study of RSV infection caused by bronchiolitis and asthma and other respiratory diseases,the mechanism of action and clinical treatment laid the foundation.
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Objective To study effect of traditional Chinese medicine chrysanthemum decoction on RSV infection after A549 cells chemotactic factor.Methods Subjects from The Second Hospital Affilated To Suzhou University in recent days, 64 cases of patients diagnosed with RSV effected pneumonia by clinical examination, RSV infection A549 cell model was established and the infected cells were cultured, chrysanthemums with TCM decoction and conventional antiviral observed its effect on RSV infection cell survival, compared two groups of cell SAP was determined by ELISA kit and the expression of RANTEs and chemokines MCP-1.Results Chrysanthemum, according to the results of in vitro cell culture studies traditional Chinese medicine decoction had the effect of directly inhibit the growth of virus replication, and there were some positive effects RSV infected A549 cells, MCP-1 and RANTEs expression or release significantly increased ( P<0.05 ) .Chrysanthemum Chinese medicine decoction and antiviral drugs after the intervention, the expression of MCP-1 and RANTES reduced significantly (P<0.05), but overall volume was still significantly higher than the normal. Conclusion Chrysanthemum Chinese medicine decoction can inhibit RSV infected A549 cells release chemokine RANTEs and MCP-1 to some extent, and then reduce the airway inflammation reaction of children.
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Objective To investigate the epidemiological features of respiratory syncytial virus (RSV) A and B in hospitalized children with acute respiratory tract infections(ARTIs) and to analyze the genetic characteristics of G protein gene of RSV in Chongqing area, especially for BA strains. Methods Nasopharyngeal secretions collected from 508 hospitalized children with ARTls from April, 2008 to March, 2009 were screened for RSV using RT-PCR. Full length G protein gene was amplified by RT-PCR from 10 RSV subtype A and 29 RSV subtype B strains. Results Out of the total 508 specimens, 126 (24.8%) were revealed positive for RSV. RSV subtype A and B viruses accounted for 34.1% and 63.5% of the total positive specimens, respectively. The remaining 2.4% of the specimens were positive for both subtype A and B. At the nucleotide level, identities between the 10 subtype A virus G genes and that of the prototype strain A2 were 91.4% -92.0%, indicating genotype GA2. Identities of the 29 subtype B virus G genes and that of the CH18537 strain were 92.0%-93.0%. Nineteen out of 29 RSV subtype B isolates contained highly repeated 60 nucleotides insertion in G protein gene, namely BA strain. As compared to the prototypes, the RSV G protein gene included nucleotide deletion, insertion, substitutions, especially in the carboxy-terminal third of the G gene. Condnsion RSV has been the major cause of acute respiratory tract infections in children in Chongqing area. Subtype B strains, especially BA strains, were predominant during the study peried. Whether the predominated circulation of BA strain is resulted from enhanced attachment function of G protein remains unknown.
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PURPOSE: Respiratory syncytial virus (RSV) is one of the main pathogens causing lower respiratory infections (LRI) in young children, usually of limited severity. However, in congenital heart disease (CHD) patients, one of the high-risk groups for RSV infection, RSV can cause serious illnesses and fatal results. To elucidate the effects of RSV infection in CHD patients, we observed RSV infection cases among CHD patients and non-CHD patients. METHODS: On admission of 343 LRI patients over 3 years, 77 cases of RSV infection were detected by the RSV antigen rapid test of nasopharyngeal secretion. We compared RSV infection cases among groups of CHD and non-CHD patients. RESULTS: During the winter season, RSV caused 20-50% of LRI admissions in children. In patients with completely repaired simple left to right (L-R) shunt diseases such as ventricular septal defect, atrial septal defect, and patent ductus arteriosus, RSV infections required short admission days similar to non-CHD patients. In patients with repaired CHD other than simple L-R shunt CHD, for whom some significant hemodynamic problems remained, RSV infection required long admission days with severe clinical course. In children with unrepaired CHD, RSV infection mostly occurred in early infant age, with long admission days. RSV infections within a month after cardiac surgery also required long admission days and severe clinical course. CONCLUSION: To avoid the tragedic outcome of severe RSV infection in the CHD patients, efforts to find the subgroups of CHD patients at high risk to RSV infection are needed, and effective preventive treatment should be applied.
Subject(s)
Child , Humans , Infant , Antibodies, Monoclonal, Humanized , Bronchiolitis , Ductus Arteriosus, Patent , Heart , Heart Diseases , Heart Septal Defects, Atrial , Heart Septal Defects, Ventricular , Hemodynamics , Pneumonia , Respiratory Syncytial Viruses , Respiratory Tract Infections , Seasons , Thoracic Surgery , PalivizumabABSTRACT
Objective To detect the antiviral effect and its antiviral mechanism of the sulfated polysaccharide SP2 isolated from the brown alga Sargassum patens C. Ag. against RSV.Methods The antiviral activity,cytotoxicity and its possible antiviral mechanism were assayed by cytopathogenic effect (CPE) inhibition assay,MTT assay and plaque reduction assay. Reduction assay when it was added during virus adsorption. It did not show any antiviral activity against influenza A and B viruses and parainfluenza type 3 virus up to the concentration of treatment of cells with SP2 did not protect the cells from RSV infection. This polysaccharide addition assay showed that when SP2 was present only during virus adsorption phase at 37℃, it could significantly inhibit RSV-induced plaque formation. Conclusions SP2 might be a potent substance for treating RSV infection. The mode of action of SP2 might depend on inhibiting RSV attachment to host cells.
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Objective: To study the effect of Jinxin Oral Liqiud medicated serum on RS virus virus (RSV) infected cell apoptosis and the regulation gene as Bcl-2, Bax on the different point in early time. Methods: Cell culture, serum pharmacology and Annexin V/PI technique combining with flow cytometry were used to detect cell apoptosis, Bcl-2 and Bax expression. Results: The total apoptosis rates of human embryonic lung fibroblast in JOL group in early time of 12h and 24h were significantly higher than that in RSV infected group (P
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PURPOSE: Recent studies have demonstrated and suggested that Interleukin (IL) -10 and IL-11 are implicated in the pathophysiology of RSV infection and may act in the regulation of inflammatory response. We measured IL-10 and IL-11 in nasal secretions of infants with acute RSV bronchiolitis to investigate if there is any difference in the production of these anti-inflammatory cytokines between atopic and non-atopic subjects. METHODS: We measured IL-10, IL-11 in nasal secretions of 44 infants (20 were atopic) with acute RSV bronchiolitis. The nasal secretion samples were obtained from patients on admission and stored immediately at -70degrees C until analysis. Atopy was defined as having at least one positive skin prick test to common allergens, a positive history of atopic dermatitis or age-matched, high serum IgE level. RESULTS: IL-10 and IL-11 increased significantly in nasal secretion of infants with acute RSV bronchiolitis. Both IL-10 and IL-11 were significantly lower in atopic patients than in non-atopic patients. There was no significant relation between the severity of symptoms and IL-10 or IL-11 levels. CONCLUSION: Our study showed that both IL-10 and IL-11 increased in nasal secretion during acute RSV bronchiolitis, and the levels were significantly lower in atopic patients than in non-atopic patients. It suggests that the airway inflammation induced by RSV may be different between atopic and non-atopic patients and this may be associated with lower induction of these anti-inflammatory cytokines in atopic patients.
Subject(s)
Humans , Infant , Allergens , Bronchiolitis , Cytokines , Dermatitis, Atopic , Immunoglobulin E , Inflammation , Interleukin-10 , Interleukin-11 , Interleukins , Respiratory Syncytial Viruses , SkinABSTRACT
5#. The hybridoma cell lines grew well after continuous culture for more than three months or after being stored in liquid nitrogen for six months, and the titers of secreting McAbs were stable.Conclusion:The McAbs against respiratory syncytial virus are obtained. It lay a foundation for early diagnosis and further study of RSV.
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BACKGROUND: Respiratory syncytial virus (RSV) is the single most common cause of lower respiratory tract infections in infants and young children. RSV can be classified into two major groups, A and B with subgroups based on their reactivity with monoclonal antibodies. There were no reports on the subgroups of RSV isolates in Korea. The purpose of this study is to identify RSV isolates from patients with lower respiratory tract infections to subgroup level and to examine clinical characteristics of subgroup infections. METHODS: RSV infection was diagnosed by viral culture of nasopharyngeal aspirates in patients with lower respiratory infection. Forty two RSV isolates over four successive outbreaks (94/95, November 1994-January 1995; 95/96, Nov. 95-Jan. 96; 96/97, Nov. 96-Jan. 97; 97/98, Nov. 97-Jan. 98) were subgrouped by indirect immunofluorescence with subgroup-specific monoclonal antibodies. Clinical characteristics of subgroup infections were evaluated by review of medical records. RESULTS: Twenty eight (67%) isolates were identified as group A and 14 (37%) strains as group B. Group A isolates of the 94/95, 95/96, and 96/97 seasons were subgroup A/4, and those of 97/98 season were subgroup A/2. Group B isolates were all identified as subgroup B/2. There was no statistically significant difference in clinical characteristics according to the subgroup infections. CONCLUSIONS: This study show that RSV subgroup A/4, A/2 and B/2 isolated over recent four successive epidemic seasons in Seoul. There was no significant difference in clinical characteristics or severity according to the subgroup infections.
Subject(s)
Child , Humans , Infant , Antibodies, Monoclonal , Disease Outbreaks , Fluorescent Antibody Technique, Indirect , Korea , Medical Records , Respiratory Syncytial Viruses , Respiratory Tract Infections , Seasons , SeoulABSTRACT
PURPOSE: Respiratory syncytial virus (RSV) is the most common cause of bronchiolitis in infants and also responsible for the production of asthma syndrome which can persist after the acute infection. This study evaluated the role of chemokine RANTES in airway inflammation in RSV bronchiolitis. METHODS: RANTES in nasal secretion examined in 20 infants with RSV bronchiolitis (RSV group) and 22 infants with non-RSV bronchiolitis (non-RSV group) and 10 age-matched controls. Eosinophil cationic protein (ECP) was determined to evaluate the RANTES-induced eosinophil activation. The severity of disease in RSV group was investigated by the presence of hypoxemia and the duration of wheezing. RESULTS: RANTES in RSV group (131+/-77 pg/mL) was significantly higher than those in non-RSV group (55+/-41 pg/mL) and controls (16+/-13 pg/mL) (P=0.0002, P= 0.0001, respectively). ECP in RSV group (49+/-57 ng/mL) was significantly higher than in controls (6.4+/-12.3 ng/mL) (P<0.05). RANTES and ECP concentrations showed significant correlation in both RSV and non-RSV groups (r=0.59, P<0.01 and r=0.64, P<0.01, respectively). Concentration of RANTES in RSV group was not significantly higher in infants with severe symptoms than in those with mild symptoms. CONCLUSION: RANTES, an effective eosinophil chemoattractant and activator, may play a role in the pathogenesis of airway inflammation and later airway hyperreactivity induced by RSV bronchiolitis in infants.