OBJECTIVETo obtain the recombinant Eotaxin N Terminal Mutant (Met/Eotaxin) adenovirus, and to study its role in allergic diseases.

METHODSA 400 bp DNA fragment of Eotaxin was cloned from human peripheral blood monocyte (PBMC), and the Met/Eotaxin DNA fragment was obtained by NH2-terminal mutant. Then, this DNA fragment was inserted into AdEasy transfer vector -pAdtrack-CMV plasmid. The plasmid pAdtrack-CMV-Met/Eotaxin was then linearized with Pme I and co-transformed into the E coli strain BJ5183 together with pAdEasy-1, the viral DNA plasmid. The recombinant Met/Eotaxin adenovirus vector was then transfected into 293 cells to produce and amplify viral particles. The allergic rhinitis in mice was induced by sensitization and challenging with ovalbumin (OVA). Fourty mice were equally divided into allergic rhinitis group (AR group) and Ad-Met/Eotaxin group. 2 ml Ad-Met/Eotaxin with titer of 5 x 10(9) pfu/ml was intraperitoneal injected in each mice in Ad-Met/Eotaxin group 30 min before each challenge. The symptom and the morphological change of nasal mucosa were compared in those two groups. The enhanced green fluorescence protein (EGFP) expression corresponding to the efficiency of transfection in the Ad-Met/Eotaxin group was observed under fluorescence microscope.

RESULTSThe results of sequencing and PCR showed that the Met/Eotaxin gene recombinant replication-deficient adenovirus was successfully constructed. The adenovirus marker of EGFP could be observed in a higher intensity after Ad-Met/Eotaxin intraperitoneal injection. Ad-Met/Eotaxin adenovirus significantly alleviated the symptoms and nasal histological changes of allergic rhinitis.

CONCLUSIONAd-Met/Eotaxin could be transfected efficiently with high titer in nasal mucosa of mice, and could alleviate the symptoms of allergic rhinitis in mice.