Immune response after adenoviral gene transfer in syngeneic heart transplants: effects of anti-CD4 monoclonal antibody therapy.
BACKGROUND
E1-deleted adenoviral vectors are frequently used for in vivo gene therapy. However, gene expression after adenovirus-(ad) mediated gene transfer is known to be transient due to the generation of an immune response against virus-infected cells. In this study, we asked whether an anti-CD4 mAb (RIB 5/2) treatment may improve the gene transfer into rat cardiac grafts.


METHODS
We injected recombinant ad-constructs encoding for Escherichia coli beta-gal into syngeneic rat heart transplants via the proximal aorta. One-half of the recipients of genetically modified grafts received the anti-CD4 mAb RIB 5/2, whereas the other half received no monoclonal antibody treatment. Genetically unmodified isografts without any treatment of the recipients were used as additional controls. At different time points hearts were harvested and analyzed for reporter gene expression, intragraft cellular infiltration, and cytokine gene expression (quantitative "real time" reverse transcriptase polymerase chain reaction). Serum samples were analyzed for anti-ad-Ig using enzyme-linked-immunosorbent-assay.


RESULTS
In control animals the beta-gal reporter gene expression slowly increased until day 7 and then declined. The immunohistological and reverse transcriptase polymerase chain reaction intragraft analyses revealed a strong inflammatory response (cellular infiltration, cytokine expression) in ad-transfected grafts that may explain the delayed expression and fast down-regulation of the transgene. Treatment with RIB 5/2 mAb resulted in a faster and prolonged reporter gene expression, reduced graft infiltration, reduced anti-ad-Ig titers and less interferon-gamma up-regulation.


CONCLUSIONS
Our results indicate that modulation of the anti-ad immune response using a nondepleting anti-CD4 mAb may increase the efficiency of ad-vectors for gene therapy in the transplant setting.
Institute of Medical Biochemistry and Molecular Biology, University Rostock, Department of Nephrology and Internal Care, Charité, Germany.
G
G
Schröder
K
K
Risch
H
H
Nizze
J
J
Kolls
P
P
Reinke
J
J
Brock
M
M
Lehmann
H D
HD
Volk
T
T
Ritter
D000256:Adenoviridae / Q000235:genetics* / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000627:therapeutic use*; D000914:Antibodies, Viral / Q000097:blood; D015704:CD4 Antigens / Q000276:immunology*; D018014:Gene Transfer Techniques; D016027:Heart Transplantation; D007371:Interferon-gamma / Q000235:genetics; D008297:Male; D051379:Mice; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011917:Rats, Inbred Lew; D019076:Transgenes
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015704:CD4 Antigens; D012333:RNA, Messenger; D007371:Interferon-gamma