Towards a novel vaccine against human cytomegalovirus based on a chimeric Ad5F35 adenovirus vector expressing the immunodominant antigenic domain 1 epitope.
BACKGROUND
Antibodies induced from glycoprotein B (gB) by antigenic domain (AD)-1 demonstrate broad neutralizing activity across different human cytomegalovirus (HCMV) types. This study aimed to prepare a novel HCMV vaccine using the modified adenoviral vector Ad5F35 to direct the expression of the conserved HCMV epitope AD-1 and to determine its transfer and expression in peripheral blood mononuclear cells (PBMCs).


METHODS
AD-1 genes were amplified from AD169 HCMV strain and cloned into the Ad5F35 vector. Ad5F35-AD-1 virus vaccine was prepared by packaging Ad5F35-AD-1 into HEK293 cells. RT-PCR and fluorescence detection were used to detect the expression of AD-1 in HEK293 cells. PBMCs were stimulated in vitro with Ad5F35-AD-1 virus vaccine. The AD-1 expression in PBMCs was determined with immunocytochemistry and cell viability was measured to observe the possible adverse effects of AD-1 on PBMCs.


RESULTS
We constructed an Ad5F35-AD-1 vector and transferred it into HEK293 cells to prepare the Ad5F35-AD-1 virus vaccine successfully. The AD-1 gene was proved to be expressed in HEK293 cells. In vitro stimulation of PBMCs with Ad5F35-AD-1 showed the highly efficient expression of AD-1 and low cytopathic activity in PBMCs.


CONCLUSIONS
The novel vaccine Ad5F35-AD-1 is a promising candidate for clinical trials and may be of utility in prime-boost strategies for HCMV prevention and control.
Department of Hematology, Qilu Hospital, Shandong University, Jinan, PR China.
Ping
P
Zhao
Daoxin
D
Ma
Shuxin
S
Yan
Na
N
Shao
Jingru
J
Zhang
Zheng
Z
Bi
Jianjian
J
Dai
Min
M
Ji
Chunyan
C
Ji
D000256:Adenoviridae / Q000235:genetics; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology; D003586:Cytomegalovirus Infections / Q000276:immunology* / Q000517:prevention & control; D022482:Cytomegalovirus Vaccines / Q000276:immunology* / Q000627:therapeutic use; D004274:DNA, Recombinant / Q000378:metabolism; D005822:Genetic Vectors / Q000378:metabolism; D006801:Humans; D016056:Immunodominant Epitopes / Q000096:biosynthesis / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D017434:Protein Structure, Tertiary; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D022482:Cytomegalovirus Vaccines; D004274:DNA, Recombinant; D016056:Immunodominant Epitopes; D014759:Viral Envelope Proteins; C040495:glycoprotein B, Simplexvirus