Effect ofβ-defensin 2 gene silencing on the inflammatory response induced by Haemophilus influenzae type B combined vaccine immunization in rats
10.3760/cma.j.issn.1000-6680.2016.05.009
- VernacularTitle:β-防御素-2基因沉默对B型流感嗜血杆菌结合疫苗免疫引起的大鼠炎性反应的影响
- Author:
Han LEI
- Publication Type:Journal Article
- Keywords:
Haemophilus influenzae type B vaccine;
β-defensin 2;
Inflammatory factor;
Immunization;
Gene silencing
- From:
Chinese Journal of Infectious Diseases
2016;34(5):292-296
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of β‐defensin 2 (rBD‐2 ) silencing on the inflammatory response induced by Haemophilus influenzae type B vaccine immunization in rats .Methods A total of 144 SPF Sprague Dawley (SD) rats were divided into four groups .Group 1 were immunized with Haemophilus influenzae type B vaccine .Group 2 were immunized with Haemophilus influenzae type B vaccine and administered with lentivirus Lv‐shRNA‐rBD‐2 by intratracheal instillation .Group 3 were administered with the lentivirus Lv‐shRNA‐rBD‐2 by intratracheal instillation .Group 4 were blank controls .Each group was set with 12 rats . The serum was collected at 12 ,24 and 72 hours after immunization ,and tumor necrosis factor (TNF)‐α,interleukine (IL)‐1β and IL‐10 were assessed by enzyme‐linked immuno sorbent assay (ELISA ) .Expression levels of rBD‐2 in lung tissues of rats were measured by western blotting . Results The recombinant virus was successfully produced by a lipofectmaine transfection method .At 12 ,24 ,and 72 h of immunization ,serum levels of TNF‐αof rats in group 1 were (82 .0 ± 8 .0) ,(155 .0 ± 18 .2) ,and (272 .0 ± 32 .5) pg/mL ,respectively ,which were all higher than those in group 4 ([55 .0 ± 6 .2] ,[52 .0 ± 5 .8] ,and [56 .0 ± 4 .8] pg/mL ,respectively ;t=16 .034 ,P=0 .043 ;t=12 .411 ,P=0 .035 ;t=10 .530 ,P=0 .018 ,respectively) .Serum levels of TNF‐αat 12 ,24 ,and 72 h of immunization in group 2 were (66 .0 ± 8 .2) ,(90 .0 ± 12 .6) ,and (108 .0 ± 13 .6) pg/mL ,respectively ,which were all significantly lower than group 1 (t=12 .115 ,P=0 .039 ;t=12 .830 , P=0 .033 ;t=15 .522 ,P=0 .012 ,respectively) .At each time point ,serum levels of IL‐1βand IL‐10 in group 1 were all significantly higher than those in group 4 (IL‐1β:t=18 .032 , P=0 .048 ;t=15 .824 , P=0 .039 ;t=13 .518 , P=0 .021 ,respectively ;IL‐10 :t=15 .410 , P=0 .045 ;t=14 .294 , P=0 .032 ;t=13 .375 ,P=0 .013 ,respectively) .Serum levels of IL‐1βand IL‐10 at each time points in group 2 were all significantly lower than those in group 1 (IL‐1β:t=19 .012 , P=0 .043;t=16 .991 , P=0 .034;t=14 .862 , P=0 .027 ,respectively ;IL‐10 :t=15 .134 , P=0 .048 ;t=15 .264 , P=0 .036 ;t=11 .408 , P=0 .024 ,respectively) .Seventy‐two hours after immunization of SD rats ,the relative content of rBD‐2 protein in lung tissue of rats significantly increased .Protein level in group 1 was significantly higher than that in group 4 (t=10 .582 ,P=0 .035) ,while protein level in group 2 was significantly lower than that in group 1 (t=13 .250 ,P=0 .027) .Conclusions rBD‐2 gene silencing can improve the inflammatory response induced by Haemophilus inf luenzae type B combined vaccine in rats.
