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Objective To evaluate the effects of fluticasone propionate (FP)on Foxp3 expression in CD4+T cells, to explore the possible mechanisms of childhood asthma. Methods Thirty asthmatic children, 15 with inhaled FP and 15 without inhaled FP, and 16 healthy children were recruited. Peripheral blood mononuclear cells (PBMC)labeled for CD4 and intracellular Foxp3 were analyzed using flow eytometry. The levels of IL-2 and IL-6 in serum and supernatant before and after stimulation by Phytohemagglutination (PHA)were measured by ELISA. The expression of phosphorylated signal transducer and activator of transcription 5 (STAT5)in PBMC was detected by Western-blot.Results Compared with healthy control, the percentage of CD4+ Foxp3+ cells in PBMC in asthmatic children without inhaled FP was significantly decreased. After inhaled FP and in remission stage, the percentage of CD4+ Foxp3+ cells in asthmatic children was up regulated with a decreased serum IL-6 level and an increased phosphorylated STATS expression. Conclusions Decreased Foxp3 protein expression in peripheral CD4+ T regulatory cells (Treg)is characterized in childhood asthma. Inhaled glucoeorticoid therapy of childhood asthma might be attributed to its ability of increasing Foxp3 expression by upregulation of phosphorylated STAT5 to balance the T cell response.
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Objective To investigate the effects of deoxyribozyme(DZ) against respiratory syncytial virus(RSV) infection in BALB/c mice and nude mice. Methods RSV infected BALB/c mice and nude mice were nasally dripped with DZ. Pulmonary viral titers were detected by plaque forming experiment,and viral mRNA expression was assayed by RT-PCR. Leukocytes and the subgroup cells in bronchoalveolar lavage fluid (BALF) were counted, cytokines of TNF-α, IL-12, IFN-γand IL-10 in BALF were assayed by ELISA. Pulmonary histopathology was examined to realize the inflammation of airway. Results Pulmonary titers of 0.2 mg, 0.4 mg and 0.8 mg DZ treated BALB/c mice were lg(3.65 ±0.12) PFU/g lung,lg( 3.25 ± 0.10) PFU/g lung and lg( 3.03 ±0.08 ) PFU/g lung, decreased as compared with that of infected control BALB/c mice lg(4.35 ± 0.11 ) PFU/g lung ( P<0.05 ). Meanwhile viral titers of 0.2 mg,0.4 mg and 0.8 mg DZ treated nude mice were lg(4.82 ±0.15) PFU/g lung, lg(4.47 ±0.12) PFU/g lung and lg(4.21 ±0.11 ) PFU/g lung, declined dramatically as compared with that of infected control nude mice lg(6.23 ± 0.15) PFU/g lung( P<0.01 ). 0.2 mg, 0.4 mg and 0.8 mg DZ reduced BALB/c mice pulmonary viral mRNA expression by 30.51% ,47.38% ( P<0.05 ) and 53.97% ( P<0.01 ) and nude mice by 36.59% (P ＜0.05 ), 48.72%, 59.78% ( P<0.01 ) respectively as compared with their infected control groups. In 0.4 mg DZ treated BALB/c mice and nude mice, total numbers of leukocytes in BALF were decreased dramatically and pulmonary histology was significantly improved compared with their infected controls( P<0.05 ). And the treatment of 0.4 mg DZ reduced productions of TNF-α, IL-12 and IFN-γin BALF of RSV infected nude mice ( P<0.05 ). Conclusion DZ effectively inhibits viral replication and reduces airway inflammation in RSV infected BALB/c mice and nude mice, and the effects in nude mice are more significant. DZ is a potential therapeutic agent against RSV infection in vivo.
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Human metapneumovirus [hMPV] is a respiratory pathogen responsible for disease and subsequent hospitalizations in young children around the world. The disease pathology, including how viral load correlates with respiratory disease severity, remains unclear. This study investigated the correlation between viral load and clinical characteristics of hMPV infections. Nasopharyngeal aspirate [NPA] samples collected from 18 infants hospitalized for lower respiratory tract infections [LRTIs] in winter were tested for hMPV by reverse transcriptase polymerase chain reaction [RT-PCR] and real-time RT-PCR. Their NPA samples were collected every-other-day to monitor changes in hMPV viral load during hospitalization. Also all these 18 patients were monitored to characterize clinically their illness. hMPV load was not correlated with infection severity [P=0.5, 0.9, 0.5]. In contrast the log[10] of hMPV viral load was significantly different between those lasted for 6-11 days and those for less than 5 days [P=0.01], also the significant difference was shown between those of 6-11 days duration and those of more than 11 days [P=0.006], but there was no significant difference between those lasted for less than 5 days and those for more than 11 days [P=0.4]. Additionally, high hMPV viral shedding occurred between 6 and 11 days. hMPV load was significantly correlated with the course of illness. The association between Hmpv viral load and the course of disease suggested that hMPV is an important pathogen in lower respiratory tract infection in children. But hMPV did not always lead to more sever respiratory illness
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Humans , Male , Female , Paramyxoviridae Infections , Viral Load , Child , Reverse Transcriptase Polymerase Chain Reaction , Respiratory Tract Infections , NasopharynxABSTRACT
Objective To investigate the antiviral effects of antisense phosphorothioate oligodeoxynucleotide (ASODN)in 9HTEO infected with influenza A virus (IFAV) in vitro. Methods The ASODN which complemented to genomic PB1, M2, NS, PB2, HA mRNA of IFAV was used to investigate antiviral effection in vitro. The cytopathic effect (CPE) was observed, and the cell survival rates were measured by MTF assay, plaque assay, RT-PCR, Western blot and Immunofluorescence were performed to test anti-viral efficiency of PB1, M2, NS in cells mRNA and protein level. Results 9HTEO cells infected with IFAV almost all died when the multipicity of infection (MOI) is aboved after 5 days cell culture. The ASODN could increase the cell survival rates. The IFAV PB1, M2, NS significantly reduced CPE of IFAV infected 9HTEO cells, reduced the viral replication of IFAV in the cells (P <0.05). Conclusion The ASODN which targeted the mRNA of IFAV gene showed a significant and specific anti-IFAV effect both in mRNA and protein level in cells culture system. The study indicates that the PB1, M2 and NS mRNA may play an important role in regulating IFAV replication, and ASODN may have inhibitory activity on IFAV replication. The results established the basis for further study on new drugs against IFAV infection include the highly pathogenic H5N1 influenza virus.
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Objective To explore the effects of BCG and Poly I:C co-vaccination on the development of spleen T cell subsets of neonatal BALB/c mice. Methods Neonatal BALB/c mice were inoculated with BCG and/or Poly I:C intraperitoneally within 2-3 d after birth. Four weeks later, spleen cells of mice were isolated and the percentage of CD3+ CD8+ IFN-γ+,CD3+ CD8-IFN-γ+,CD3+ CD8+ IL-4+,CD3+ CD8- IL-4+,CD4+ Foxp3+ T cells,which represent Tc1,TH1,Tc2,TH2,Treg cells,respectively,were tested by flow cytometry at single cell level,and the ratios of TH 1/TH 2 and Tc1/Tc2 were calculated. Results The percentages of TH1 and Tc1 cells of BCG-vaccinated mice,Poly I:C-vaccinated mice and BCG plus Poly I:C-vaccinated mice were significantly higher than that of control mice(P＜0.05 or P＜0.01),and there was no difference among the three vaccinated group. The ratios of TH1/TH2 and total IFN-γ/IL-4 of the three vaccinated groups were higher than that of control group,but not the ratio of Tc1/Tc2. The TH1/TH2 ratio of BCG plus Poly I:C-vaccinated group was higher than that of BCG-vaccinated group(P＜0.05).The percentages of Trge cells showed no difference among the four groups(P＞0.05). Conclusion BCG and Poly I:C co-vaccination can significantly increase the number of Tc1 and TH 1 cells and TH 1/TH2 ratio in spleen cells. BCG and Poly I:C vaccination may have a synergistic effect on TH 1/TH2 ratio of spleen cells in neonataI mice. The percentage of CD4+ Foxp3+ T cells among four groups showed no significant difference.
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Objective To compare respiratory syncytial virus(RSV)infection and inflammatory responses between immunocompetent BALB/c mice and immanodeficient nude mice.Methods At various time points after BSV infection of BALB/c mice and nude mice,pulmonary viral titers were assayed.Leukocvtes in bronchoalveolar lavage fluid(BALF)and pulmonary histology were identified.F4/80+cells and CD49b+cells in lung tissue were examined by immunohistochemistry,and the cytokines of TNF-α,IL-12,IFN-r and IL-10 in BALF were assayed by ELISA.Results RSV titers in infected BALB/c mice and nude mice peaked on the 3rd day postinoculation,and nude mice had higher-level and more durative viral replication than BALB/c mice.RSV infection induced more severe pulmonary histopathology and larger number of leukocytes in airway in nude mice than in BALB/c mice.RSV infection enhanced more pulmonary F4/80+macrophages,CD49b+ NK cells in both mice.Furthermore infected nude mice had larger amount of pulmonary macrophages and NK cells than infected BALB/c mice.RSV infected BALB/c mice secreted more TNF-α,IL-12,IFN-r and IL-10 as compared with control BALB/c mice,and infected nude mice had hisher level of TNF-α.IL-12 and IL-10 than infected BALB/c mice.Conclusion Nude mice are good model for severe and pemistent RSV infection in immunocomprised hosts.The inflammation induced by RSV infection is not parallel with the immune response of T cells,and macrophages and NK cells are potent immunocytes and inflammatory cells in RSV infection especially when T lymphocytes are absent.
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Objective To investigate the role of Huang-qi in balance of TH1/TH2 in asthma on dendritic cells level. Methods DCs from human peripheral blood mononuclear cells(PBMC) were induced by rhGM-CSF and rhIL-4, and then were identified. The level of IL-12 and IL-10 produced by DCs were de-tected by ELISA assay. After autoreactive T cells, mRNA of T-bet and GATA-3 was measured by RT-PCR. Simultaneously, IL-4 and IFN-γ were determined by flow cytometer. Results After 7 days culture, IL-12 was significantly decreased in asthma group compared to control group (P ＜ 0.01), whereas IL-10 on the opposite. At the 7th day of co-culture with T cells derived from floating cells, the IFN-γ/and T-bet mRNA level in asthma group were significantly decreased than that in control group, whereas IL-4, GATA-3 mRNA level, the ratio of IL-4/IFN-γ and GATA-3/T-bet were apparently increased in asthma group than that in control group(P＜0.01). After Huang-qi treatment, the IFN-γ/and T-bet mRNA level were significantly in-creased, whereas the ratio of IL-4/IFN-γ and GATA-3/T-bet, and the IL-10 level were apparently de-creased, but the level of IL-12, IL-4 and GATA-3 mRNA were not changed significantly. Conclusion DCs in asthma regulated the balance of TH1/TH2 by means of secreting decreased IL-12 and increased IL-10, that made TH2 playing a dominance role which is the key factor in initiating asthma. Huang-qi regulated DCs through decreasing the level of IL-10, and thus decreased the ability of inhibiting the differentiation of TH1 from TH0, that is also inhibiting the differentiation of TH2 from TH0 directly.
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Objective To investigate the role of interleukin-1 beta (IL-1?) in glomerulosclerosis secondary to adriamycin (ADR)-induced nephropathy in rats, and the effect of simvastatin on the expression of IL-1?. Methods Male Sprague-Dawley rats were randomly divided into normal control, ADR-induced nephropathy (model), and simvastatin-treated ADR nephropathy (treatment) groups. After ADR-induced nephropathy establishment in model and treatment groups, and eleven weeks of intragastrical administer of normal saline in normal control and model groups and of simvastatin in treatment group, the expression of IL-1? was detected by immunohistochemistry, RT-PCR and ELISA. Histopathological change of renal tissues was observed under light microscope, and glomerulosclerosis index (GSI) was also evaluated. Results Higher expression of IL-1? in kidney and GSI, as well as more severe loss of renal function were observed in model group than those in control group (all P
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Objective:To explore the role of T lymphocytes activation co-stimulation pathway in asthma pathogenesis and the ability of therapy asthma with Anti-CD86mAb.Methods:The blood samples were taken from 28 asthma children( including 18 male and 10 female, age 1 year-8.08 years) and 15 normal children( including 7 male and 8 female, age 3.25 years-10 years).ELISA was used to detect the levels of IL-4、IL-13、IFN-? in culture supernatants of PBMC stimulated with PHA and treated with mouse anti human CD86mAb. Results:①When treated control PBMC with anti-CD86mAb, the level of IL-4 in control group(13.30?4.66 pg/ml) was lower than that of mouse IgG control group (15.20?5.22 pg/ml,P
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0.5).IgG、IgA and IgM of controlgroup and observation group in 10～13 years old are similar to that in 5～9 years old group. The IgG_1of control group and observation group in 5～9 years old are 5.501?0.976 and 9.715?3.746g/L respe-etively (t=5.046, P0.05),IgG_3 are0.517?0.167 and 0.828?0.578g/L respectively (t=2.132, P0.05).The IgG_2 Levels of observation groupis higher than that of control group in 10～13 years old(P