ABSTRACT
Objective:To build the DNA vaccine encoding hantavirus glycoprotein fused with lysosome-associated membrane protein (LAMP) and assess its immune response.Methods:BALB/c mice were immuned with the previous experimental expressing purified recombinant plasmids pVAX-Gn,pVAX-Gc,pVAX-LAMP/Gn,pVAX-LAMP/Gc,and inactivated vaccine.Indirect ELISA and neutralization test was used to detect specific antibody and neutralizing antibody in the serum of mice.The mice were tested to detect the protective effects in vivo.Results:Indirect ELISA results showed that the pVAX-LAMP/Gc group was the highest,followed by pVAX-Gc,pVAX-LAMP/Gn,and PVAX-Gn,and inactivated vaccine group.In neutralization test,there were significantly higher serum antibodies in LAMP group than those in conventional DNA group,which were higher than the inactivated vaccine group.The mice immuned had good protective effect in vivo without the specific antigen of hantavirus in vivo.Conclusion:Chimeric DNA vaccines induced higher levels of antibody in BALB/c mice and produced better protective efficacy,which illustrate the targeting strategy is expected to be an effective way to improve the DNA vaccine efficacy.
ABSTRACT
<p><b>UNLABELLED</b>Dendritic cells (DC) are very potent antigen-presenting cells (APC) with a unique ability to activate naive T cells to induce the differentiation of TH1/TH2. Monocytes can develop into DC in the presence of different cytokines such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4. DCs are thought to play a key role in the initiation and maintenance of T cell immunity to inhaled antigens. While the density of DC within the bronchial mucosa is increased in asthma, there is little information currently available concerning the effects of DC in asthmatic children.</p><p><b>OBJECTIVE</b>To investigate the role of dendritic cells in the pathogenesis of acute attack of asthma in children.</p><p><b>METHODS</b>Thomas' method was adopted. The adherent precursors of DC were isolated from peripheral blood of asthmatic children in acute attack stage and healthy controls. The adherent cells were induced with granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-alpha) to DC in vitro. The expression of the surface molecules CD80, CD86, HLA-DR etc. on the DC was examined by fluorescent activated cell sorter (FACS). And the ability to secret IL-10, IL-12 and their potentials to stimulate the proliferative reaction of DC inductive self T- lymphocyte were observed.</p><p><b>RESULTS</b>The results showed that in asthmatic children's acute attack stage, self T- lymphocyte proliferative reaction induced by DC was remarkably increased compared with normal control subjects (P < 0.01). At the same time, the asthmatic children in acute attack stage had remarkably decreased the ability to secret IL-10 compared with normal control subjects (P < 0.01), while the ability to secret IL-12 remarkably decreased compared with normal control subjects (P < 0.01); meanwhile, the HLA-DR and co-stimulating factor CD86(B(7-2)) expressed by DCs remarkably increased in the asthmatic children in acute attack stage compared with normal control subjects (P < 0.01).</p><p><b>CONCLUSION</b>DC possibly plays a vital role in the immunological mechanism of asthma by means of inducing the differentiation of TH1/TH2, that is DC may be the key factor in initiating the airway allergic reaction and the possible mechanism may involve interleukins (especially IL-10 and IL-12, etc.) secreted by DCs.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Male , Antigens, CD , Metabolism , Asthma , Metabolism , Dendritic Cells , Cell Biology , Metabolism , Flow Cytometry , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , HLA-DR Antigens , Metabolism , Interleukin-10 , Bodily Secretions , Interleukin-12 , Bodily Secretions , Interleukin-4 , PharmacologyABSTRACT
Objective IL-2,6 production by PBMNC (peripheral blood mononucleat cells) were ineaeured with an attempt to illustrate the immune function of children with bronchopneumonia.Methods The biological activities of IL-2,6 were measured by CTLL and CESS cell lines respectively In addition, mitogen-stimulated blastogenic transformation (MSBT) was also measured. The results were analysed statistically. Patients or Other Farticipants T enty-eight cases of children with bronshopneumonia, male, 16,female: 12. Age: 8 months to 7 years, average 3. 9 years. 15 normol controls, male: 8, female 7.Age: 2 to 6 years, average 3 .5 years.Interventions 5～6ml of venous blood was obtaincd, followed PBMNC were centrifugalized by. FicollHypaque. Wash three:times with RPMI-1640 medium. Resuspend cell at 5 ?106/ml. Add 1% PHA and culture for 48h. Horvest supernatant and store at - 20℃.Measurement and Main Rcsults The levels of IL-2 in children with bronchopneumonia and normal children were (CPM) 9 538 + 3 363, 16 152 ? 3 411 (t test, P