Expression of a tumor-reactive antibody-interleukin 2 fusion protein after in vivo particle-mediated gene delivery.
We have used a particle-mediated gene transfer method to analyze the posttransfection expression pattern of an antibody-cytokine fusion protein (FP) in vivo. The FP, denoted CC49-IL2, consists of a single-chain antibody containing the antigen recognition domain from the murine monoclonal antibody CC49 (recognizing the tumor-associated antigen TAG-72), a human IgG1 constant heavy chain, and human interleukin-2 (IL-2). This FP can bind to TAG-72-expressing tumor cells and exhibits IL-2 activity. To induce systemic levels of this FP in vivo, we have transferred the FP gene into murine epidermal cells by direct delivery of DNA-coated gold particles using a transcutaneous "gene gun." After the pericutaneous delivery of the FP gene via gold particles, production of the exogenous FP was detected at the epidermal target site. The FP produced in vivo at the site of gene delivery has cytokine activity and antigen recognition capabilities similar to those present in CC49-IL2 FP purified from hybridoma culture supernatants in vitro. FP was also detectable in the serum from test animals treated with particle-mediated gene transfer. Time course experiments indicated that serum levels of FP reached a peak level within 8 hours after DNA delivery, whereas the epidermal target tissue levels continued to increase for 24 hours before plateauing. Our results indicate that exogenous protein levels consistent with immunotherapeutic effects of the FP can be readily achieved at the skin tissue site of gene delivery, with the potential for achieving therapeutic levels systemically.
Nicolet
C M
CM
University of Wisconsin Comprehensive Cancer Center, Madison 53792, USA.
Burkholder
J K
JK
Gan
J
J
Culp
J
J
Kashmiri
S V
SV
Schlom
J
J
Yang
N S
NS
Sondel
P M
PM
D000818:Animals; D000912:Antibodies, Neoplasm / Q000235:genetics*; D004817:Epidermis / Q000187:drug effects; D018014:Gene Transfer Techniques; D006801:Humans; D007376:Interleukin-2 / Q000097:blood / Q000235:genetics*; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000097:blood / Q000235:genetics; D012867:Skin / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors
D000912:Antibodies, Neoplasm; C091202:B72.3 antibody; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins