ABSTRACT
Objective:To investigate the impact of dexamethasone on the balance of natural regulatory T cells(nTreg) and typeⅠ regulatory T cell(Tr1) in vitro.Methods:Peripheral blood lymphocytes(PBMCs) recruited from healthy donor,divided into dexam-ethasone stimulus group and negative control group.After 3 days treatment,cells were stained.The expression of CD25,CD127, Lymphocyte-activation 3 (LAG-3) and Forkhead box P3 (Foxp3) and frequency of Treg and Tr1 were analyzed by flow cytometry.Results:Compared to the control group,after stimulation with dexamethasone for 3 days,the percentage of CD4+T cells increased in a dose-dependent manner.The expression of CD25 and Foxp3 in CD4+T cells decreased significantly (P=0.006, P<0.000 1),while CD127 and LAG-3 expression increased significantly in CD4+T cells (P<0.000 1,P=0.011).Dexamethasone treatment significantly enhance the frequency of Tr1(P=0.051),reduce the frequency of Treg (P<0.001),and the ratio of Tr1/Treg also increased(P=0.044).Conclusion:Short-term treatment with dexamethasone in vitro change the balance of natural regulatory and type Ⅰ regulatory T cells.
ABSTRACT
<p><b>OBJECTIVE</b>To construct human metapneumovirus (hMPV) DNA vaccines and evaluate the cellular and humoral immune response in mice.</p><p><b>METHODS</b>Fusion protein FdeltaTM (without transmembrane domain) gene and M gene of hMPV were amplified from cDNA by PCR, then DNA vaccines pcDNA3.1His-FdeltaTM and pcDNA3.1His-M were constructed to verify the expression of F and M protein by Western blotting and indirect immunofluorescent assay (IFA) respectively. Serum IgG and spleen cell CTL were detected with ELISA and ELISPOT assay after the BALB/c mice were immunized intramuscularly with the vaccines.</p><p><b>RESULTS</b>The candidate DNA vaccines could express FdeltaTM and M protein as detected with Western blotting and IFA. The IgG antibody titers of mice was 1:44 when immunized with pcDNA3.1His-FdeltaTM, but could increase to 1:64 when co-immunized with pcDNA3.1His-M. ELISPOT assay demonstrated that IFN-gamma-secreting effector T cells reached 42 +/- 8.9 in co-immunization group, higher than single vaccine pcDNA3.1His-FdeltaTM group (32 +/- 7.4).</p><p><b>CONCLUSION</b>DNA vaccine pcDNA3.1His-FdeltaTM could induce specific cellular and humoral immune responses, and the immune response could increase when co-immunization with pcDNA3.1His-M was carried out.</p>
Subject(s)
Animals , Female , Humans , Mice , Antibodies, Viral , Blood , Immunization , Metapneumovirus , Genetics , Allergy and Immunology , Mice, Inbred BALB C , Paramyxoviridae Infections , Allergy and Immunology , Virology , Vaccines, DNA , Genetics , Allergy and Immunology , Viral Proteins , Genetics , Allergy and Immunology , Viral Vaccines , Genetics , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To understand the genotypes of human metapneumovirus (hMPV) and the genetic character of hMPV attachment protein G sequence in Hunan, China.</p><p><b>METHODS</b>232 nasopharyngeal aspirates (NPA) samples from hospitalized children with acute respiratory infections were collected from Hunan, China in 2005. HMPV was detected. The full length of G glycoprotein genes were amplified and sequenced. Bioinformatics soft-wares were employed to analyze the sequences.</p><p><b>RESULTS</b>17/232 (7.3%) were showed hMPV positive. And co-infection rate with other viruses is 35%. The diagnoses of these hMPV positive cases are pneumonia, bronchiolitis and bronchopneumonia. Phylogenetic analysis for G genes from 13 hMPVs revealed the existence of four major subgroups: A1, A2, B1, B2 in Hunan, China in 2005. There are four types of sequence lengths of hMPV G glycoprotein, which are 711, 675, 660, 696nt. It is different in potential N-linked glycosylation sites and number of cysteine residues among these hMPVs of Hunan, China and Beijing, China. Also it is different from those in Japan and North America.</p><p><b>CONCLUSION</b>The investigation of hMPV from Hunan, China in 2005 revealed the high speed of genetic variation and the marked character of geographic epidemic differences.</p>
Subject(s)
Child , Humans , Amino Acid Sequence , China , Epidemiology , Genotype , Glycoproteins , Classification , Genetics , Metapneumovirus , Classification , Genetics , Molecular Sequence Data , Phylogeny , RNA, Viral , Genetics , Respiratory Syncytial Virus Infections , Epidemiology , Virology , Sequence Homology, Amino Acid , Viral Proteins , Classification , GeneticsABSTRACT
The full-length genome of one human bocavirus (HBoV) and the VP1 sequences of nine HBoV were amplified from patients' samples by PCR, cloned into pGEM-T vector separately, and sequenced. In this study, the one full length gemome and nine VP1 sequences of HBoV were aligened with 14 sequences of Parvoviruses which were canonical exemplars in Parvovirinae. Phylogenetic analysis showed that HBoV capsid sequences positioned closely to B19 parvovirus, although they positioned far in phylogenetic tree based on full length genome. Many similarities were found between HBoV and B19 in capsid by alignment on secondary structural elements. Because both B19 and HBoV are the only Parvoviruses that infect mankind, so study on HBoV may be used for reference to B19 which had been studied for about 30 years. By analysis of mutational sites, HBoV capsid protein showed a highly conserved secondary structural elements, but highly active in VP1-U, leading end of VP2 and insertions between the strands of the betaG-H. This cued that HBoV inclined to immune evasion and infectant adaptive faculty.