The Study Of Ebola Virus Envelope Glycoprotein Enhancing The Immunogenicity Of HIV-1 Virus-like Particles

Objective:Ebola virus envelope glycoprotein enhancing the immunogenicity of HIV-1 virus-like particles.Methods:1.Transfection:(1)Obtain the desired VLP by PEI transfection with the plasmids HIVgp120,EbovGp,EbovGp ? M,? 8.2 and Gluc:HIVgp-VLP?EbovGp-VLP?EbovGp?M-VLP?HIVgp/EboGp-VLP and HIVgp/EboGp?M-VLP;(2)Check the res?lting of transfection is successf?l:Western Blot(WB)detection the proteins in VLP:HIVgp120 protein,HIV P24 protein,EboGp protein and EboGp?M protein;The amount of HIV P24 in the VLP was detected by Enzyme-link Immunosorbent Assay(ELISA).2.Cell study:VLP of 100ng/100?lPBS:HIVgp-VLP,EbovGp-VLP,EbovGp ? M-VLP,HIVgp/EboGp-VLP and HIVgp/EboGp ? M-VLP infect C8166 cells,THP1 cells,Monocyte Derived Macrophages(MDMs)and Monocyte Derived Dendritic cells,(MDDCs),detecting the Gaussia Luciferase Assay(Gluc)of four cell supernatants after 3 days of infection,to compare the amount of different VLPs targeted into the target cells.3.Animal study:(1)HIVgp(M)group animal study:Nine Balb/c female mice age 6 weeks were randomly divided into three group:PBS,HIVgp(M)and HIVgp(M)/EboGp.On the 0th,14th and 49th day,the VLP 100ng/100?l PBS was injected into the neck of each group mice,On the 56th day,take the mice's leg saphenous vein blood 200?l,detection the anti-HIVgp 120 antibody and anti-HIV P24 antibody in serum by ELISA.(2)HIVgp(T)group animal study:Twelve Balb/c female mice age 6 weeks were randomly divided into four group:PBS,HIVgp(T)-VLP,HIVgp(T)/EboGp-VLP and HIVgp(T)/Eb oGp?M-VLP.On the 0th,14th and 49th day,the VLP 100ng/100?l PBS was injected into the neck of each group mice,On the 56th day,take the mice's leg saphenous vein blood 200?l,detection the anti-HIVgp120 antibody and anti-HIV P24 antibody in serum by ELISA.Results:1.Transfected VLP:HIVgp-VLP,EbovGp-VLP,EbovGpAM-VLP,HIVgp/EboGp-VLP and HIVgp/EboGpAM-VLP.HIVgp120 protein(120KD),HIV P24 protein(24KD),EboGp protein(75KD)and EboGpAM(40KD)protein can be seen in WB.2.Cell study:(1)EboGp-VLP,HIVgp-VLP and HIVgp/EboGp-VLP respectively infect C8166 cells,The Gluc value of HIVgp(T)-VLP group was higher than other groups(P<0.05).Infected MDMs and MDDCs,the Gluc value of EboGp-VLP group and HIVgp(M)/EboGp-VLP group was higher than that of HIVgp(M)-VLP group(P<0.05).(2)HIVgp-VLP,EboGp-VLP,HIVgp/EboGp-VLP,EboGp?M-VLP,and HIVgp/EboGp?M-VLP infect THP1 cells,MDMs,and MDDCs respectively.The Gluc of HIVgp/EboGpAM-VLP was higher than other groups(P<0.05).3.Animal study:(1)HIVgp(M)group:The anti-HIVgp120 antibody and anti-HIV P24 antibody in female Balb/c mice immunized with HIVgp(M)/EboGp-VLP were significantly higher than other groups(P<0.01);(2)HIVgp(T)group:The anti-HIVgp 120 antibody and anti-HIV P24 antibody in female Balb/c mice immunized with HIVgp(T)/EboGp-VLP were significantly higher than other groups(P<0.01);HIVgp(T)/EboGp-VLP compared with HIVgp(M)/EboGp-VLP showed no significant difference in antibody(P>0.05).Conclusion:(1)Both EbovGp and EbovGp?M can promote VLP targeting into C8166 cells,THP1 cells,MDMs and MDDCs,EbovGpAM is more potent than EbovGp;(2)HIVgp(M)/EboGp-VLP targeting macrophages,HIVgp(T)/EboGp-VLP targeting T cell both induce higher anti-HIV antibody levels in female Balb/c mice.