Plasmid Construction of tgfβ1 and 1l10 in pVAX1

Mouse cytokines TGFβ and IL10 were used as immunomodulators and were expressed by host cells under the control of the CMV promoter (4–6). The mammalian expression vector pVAX1 (Invitrogen, CA) was used to be consistent with the Food and Drug Administration (FDA) guidelines. For the construction of recombinant mouse TGFβ and mouse IL10 into pVAX1, PCR amplification of mouse tgfβ1 and il10 (from Origen, Table 1 ) sequences using the primers TGFb-HindII-For/TGFb-XhoI-Rev and mIL10-HindII-For/mIL10-XhoI-Rev was undertaken ( Table 2 ). During the digestion with HindII/XhoI, the products were subcloned into HindII/XhoI that was digested by pVAX1 to produce pMH 524 and pMH 525 ( Table 1 ).