[Construction of recombinant eukaryotic expression plasmid of murine chemokine MIP-1alpha and coimmunization with HSV-II DNA vaccine].
OBJECTIVE
To construct the recombinant eukaryotic expression plasmid of murine macrophage inflammatory protein-1alpha (MIP-1alpha) and investigate the effect of MIP-1alpha as an adjuvant on immune response induced by herpes simplex virus type II glycoprotein D (HSV-II gD) DNA vaccine.


METHODS
Using total RNA from RAW264.7 cells stimulated with LPS, the whole code sequence of murine MIP-1alpha was amplified by RT-PCR and inserted into pcDNA3 at Hind III/Xba I restriction sites. The recombinant eukaryotic expression plasmid Pm was transiently expressed in COS-7 cells and its specificity was demonstrated by RT-PCR and Boyden chemotaxis chamber assay. BALB/c mice were immunized with gD DNA vaccine and/or MIP-1alpha, and the effect of MIP-1alpha on gD DNA vaccine was evaluated by detecting anti-HSV-II antibody, antigen-specific lymphoproliferative responses, and examining survival rates after mice were challenged intravaginally with HSV-II.


RESULTS
The recombinant eukaryotic expression plasmid of murine MIP-1alpha was constructed, and it was revealed that immune responses of HSV-II gD DNA vaccine were enhanced by coimmunization with MIP-1alpha.


CONCLUSIONS
The murine MIP-1alpha can be used as an adjuvant of HSV-II gD DNA vaccine.
Shao
Wen-ai
WA
Department of Microbiology and Immunology, Shantou University Medical College, Shantou 515031, China. swa001@126.com
Li
Xiao-mian
XM
D000818:Animals; D019556:COS Cells; D054405:Chemokine CCL3 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002522:Chlorocebus aethiops; D004279:DNA, Viral / Q000276:immunology*; D005260:Female; D015870:Gene Expression; D006568:Herpesvirus 1, Cercopithecine / Q000235:genetics*; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*
D054405:Chemokine CCL3; D004279:DNA, Viral