Cyclophosphamide diminishes inflammation and prolongs transgene expression following delivery of adenoviral vectors to mouse liver and lung.

Immune responses to adenovirus-mediated gene transfer contribute to the problems of transient recombinant gene expression, inflammation, and difficulties with vector readministration. Activation of CD4+ T cells is required for full realization of effector function of both CD8+ T cells (i.e., cytotoxic T cells) and B cells (i.e., neutralizing antibody). We evaluate in this study the effectiveness of a short course of high-dose cyclophosphamide to block immune responses in mice administered vector into lung and liver of C57BL/6 mice. Administration of cyclophosphamide with vector directed to liver blocked activation and mobilization of both CD4+ and CD8+ T cells. As a result, transgene expression was prolonged, inflammation was reduced, and, at the higher doses of cyclophosphamide, formation of neutralizing antibody was prevented and the vector was successfully readministered. Similar studies in the lung demonstrated an effective blockade of T and B cell responses. In contrast to the liver, where it was easier to stabilize transgene expression than to prevent neutralizing antibody, cyclophosphamide prevented the formation of neutralizing antibodies at all doses in the lung, whereas stabilization of transgene expression was only achieved at the highest dose. These experiments begin to define the parameters by which cyclophosphamide could be used as an adjunct in gene therapy.

[1]  J. Wilson,et al.  Transient immune blockade prevents formation of neutralizing antibody to recombinant adenovirus and allows repeated gene transfer to mouse liver. , 1996, Gene therapy.

[2]  J. Wilson,et al.  Gene therapy for cystic fibrosis: challenges and future directions. , 1995, The Journal of clinical investigation.

[3]  Ronald G. Crystal,et al.  Transfer of Genes to Humans: Early Lessons and Obstacles to Success , 1995, Science.

[4]  K. Weinberg,et al.  Engraftment of gene–modified umbilical cord blood cells in neonates with adenosine deaminase deficiency , 1995, Nature Medicine.

[5]  A. Gown,et al.  Long–term hepatic adenovirus–mediated gene expression in mice following CTLA4Ig administration , 1995, Nature Genetics.

[6]  M. Kay,et al.  Gene therapy for hemophilia B: host immunosuppression prolongs the therapeutic effect of adenovirus-mediated factor IX expression. , 1995, Human gene therapy.

[7]  Z. Xiang,et al.  Upregulation of class I major histocompatibility complex antigens by interferon gamma is necessary for T-cell-mediated elimination of recombinant adenovirus-infected hepatocytes in vivo. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[8]  E. Smit,et al.  Oral therapy for small cell lung cancer. , 1995, Lung cancer.

[9]  J. Lynch,et al.  Immunosuppressive treatment of the pulmonary manifestations of progressive systemic sclerosis. , 1995, Current opinion in rheumatology.

[10]  J. Whitsett,et al.  Persistence of replication-deficient adenovirus-mediated gene transfer in lungs of immune-deficient (nu/nu) mice. , 1995, Human gene therapy.

[11]  M. Kay,et al.  Strain related variations in adenovirally mediated transgene expression from mouse hepatocytes in vivo: comparisons between immunocompetent and immunodeficient inbred strains. , 1995, Gene therapy.

[12]  N. Sarvetnick,et al.  Cellular and humoral immune responses to adenoviral vectors containing factor IX gene: tolerization of factor IX and vector antigens allows for long-term expression. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[13]  D. Waxman,et al.  Intratumoral activation and enhanced chemotherapeutic effect of oxazaphosphorines following cytochrome P-450 gene transfer: development of a combined chemotherapy/cancer gene therapy strategy. , 1995, Cancer research.

[14]  P. Schellekens,et al.  Immunosuppressive drugs in clinical medicine. , 1994, The Netherlands journal of medicine.

[15]  J. Wilson,et al.  Ablation of E2A in recombinant adenoviruses improves transgene persistence and decreases inflammatory response in mouse liver. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[16]  E. Furth,et al.  Cellular immunity to viral antigens limits E1-deleted adenoviruses for gene therapy. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[17]  B. Trapnell,et al.  Adenovirus-mediated gene transfer for cystic fibrosis: quantitative evaluation of repeated in vivo vector administration to the lung. , 1994, Gene therapy.

[18]  S. Wessler,et al.  Mobile inverted-repeat elements of the Tourist family are associated with the genes of many cereal grasses. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[19]  G. F. Weber,et al.  Differential activation of cyclophosphamide and ifosphamide by cytochromes P-450 2B and 3A in human liver microsomes. , 1993, Cancer research.

[20]  M. Kay,et al.  Assessment of recombinant adenoviral vectors for hepatic gene therapy. , 1993, Human gene therapy.

[21]  G. F. Weber,et al.  Activation of the anti-cancer drug ifosphamide by rat liver microsomal P450 enzymes. , 1993, Biochemical pharmacology.

[22]  M. Perricaudet,et al.  In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium , 1992, Cell.

[23]  M. Perricaudet,et al.  Evaluation of the transfer and expression in mice of an enzyme-encoding gene using a human adenovirus vector. , 1990, Human gene therapy.

[24]  D. Waxman,et al.  Oxidative metabolism of cyclophosphamide: identification of the hepatic monooxygenase catalysts of drug activation. , 1989, Cancer research.