Objective:To investigate the distribution of γδT17,Th17 and Tc17 cells in the lung of mice severely infected by influenza A(H1N1)pdm09 virus and the relationship between these cells with lung immunopathalogical injury.Methods:Intranasal infection was used to establish mouse model of severe H1N1 infection.Flow cytometry assay was used to detect the proportion and number of γδT17 cells,Th17 cells and Tc17 cells in the lung.The concentrations of interleukin-17A(IL-17A),interleukin-1β(IL-1β)and interleukin-23(IL-23) in the bronchoalveolar lavage fluid and serum were assayed by enzyme-linked immunosorbent assay and Lu-minex assay.Results:①The model of mice severely infected by influenza A(H1N1)pdm09 virus was established successfully.②The ratio of γδT cells,but not CD4+T and CD8+T cells in total lymphocytes of the lung of infected mice significantly increased compared with uninfected control mice at the third day post infection(DPI)(P<0.01).③The proportion and number of γδT17 cells,Th17 cells and Tc17 cells in total γδT cells,Th cells and Tc cells in the lung of infected mice were significant higher than that in uninfected control mice at the first DPI,respectively.However,the absolute number of γδT17 cells was far more than Th17 and Tc17 cells(P<0.05);④The concentration of IL-17A in BALF increased significantly after infection(P<0.05),and the concentration of IL-17A in serum increased significantly at the third DPI(P<0.05).The concentrations of both IL-1β and IL-23 in BALF probably participating in the activation of γδT17 cells increased significantly after infection compared with uninfected control mice.Conclusion:The γδT17 cells could be activated and secreted IL-17A via γδTCR non-depended pathway and involved in inflammatory pathological injury of lung at the early stage of severe H1N1 infection.

OBJECTIVETo analyze the effect of small interfering RNA (siRNA) targeting mouse epididymis-specific colipase-like (meClps) gene on mouse sperm mobility.

METHODSThe eukaryotic expression vector pDsRed2.0-C1-meClps was constructed and transfected into NIH-3T3 cells, and the protein expression was detected with anti-meClps serum. Three interfering sequences targeting meClps (RNAi-251, 224 and 286) were inserted into lentiviral vectors pRNAT-U6.2/lenti, which were co-transfected with pDsRed2.0-C1-meClps into NIH-3T3 cells. The RNA interfering efficiency was confirmed by semi-quantitative PCR and Western blotting. The lentivirus, packed with the lentiviral vector with the highest interfering efficiency, was injected into the caput tissues of mouse epididymis, and its effect on sperm mobility of the cauda epididymis was evaluated.

RESULTSAll the 3 lentiviral RNAi vectors targeting meClps could inhibit the mRNA and protein expressions of meClps, among which pRNAT-U6.2/lenti-RNAi-251 had the highest interfering efficiency. The lentivirus packed with pRNAT-U6.2/lenti-RNAi-251 significantly reduced the path velocity of cauda sperm after injection into the caput epididymis of the mice (P<0.05).

CONCLUSIONKnock-down meClps expression by lentiviral-mediated RNA interference can lower sperm mobility of mice.

Animals ; Epididymis ; Gene Targeting ; Genetic Vectors ; Lentivirus ; Male ; Mice ; NIH 3T3 Cells ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; Sperm Motility ; Spermatozoa ; Transfection

Humans ; Aged ; Lung Diseases ; Pneumonia/drug therapy* ; Adrenal Cortex Hormones/therapeutic use* ; Pulmonary Disease, Chronic Obstructive/drug therapy* ; Administration, Inhalation ; Community-Acquired Infections/drug therapy*