Use of human skin substitutes expressing exogenous IL-12 to treat a wound bed
The present invention relates generally to compositions for treating patients that have skin cancer or have recently had skin cancers removed. More specifically, the present invention provides human skin substitutes engineered to express exogenous IL-12 and compositions and methods for making human skin substitutes engineered to express exogenous IL-12. In addition, the present invention provides methods for treatment of sites on a patient where skin cancers have been removed with human skin substitutes engineered to express exogenous IL-12.
1. A method of treating a wound bed resulting from removal of a skin cancer in a patient, comprising:
a) providing a human skin substitute comprising cells expressing exogenous Interleukin-12, wherein the cells are selected from the group consisting of primary keratinocytes, keratinocyte precursors, transdifferentiated keratinocytes, and immortalized keratinocytes, wherein said skin substitute comprises a stratified squamous epithelium comprising said cells expressing exogenous Interleukin-12 supported on a dermal equivalent; and
b) placing said human skin substitute on said wound bed resulting from removal of a skin cancer in said patient such that the dermal equivalent is in contact with the wound bed and separates the wound bed from the cells expressing exogenous Interleukin-12.
2. The method of claim 1 , wherein said skin cancer is selected from the group consisting of melanoma and basal cell carcinoma.
3. The method of claim 1 , wherein said human skin substitute inhibits the spread of said skin cancer.
4. The method of claim 1 , wherein said human skin substitute produces amounts of bioactive IL-12 sufficient for inducing the proliferation of PHA-stimulated lymphoblasts.
5. The method of claim 1 , wherein said exogenous IL-12 is a fusion of the p35 and p40 IL-12 subunits.
6. The method of claim 5 , wherein said fusion protein has the amino acid sequence of SEQ ID NO:3.
7. The method of claim 6 , wherein said human skin substitute comprises near diploid immortalized keratinocytes (NIKS) cells.
8. A method of treating a wound bed resulting from removal of a skin cancer in a patient, comprising:
a) providing a human skin substitute comprising cells expressing exogenous Interleukin-12, wherein the cells are selected from the group consisting of primary keratinocytes, keratinocyte precursors, transdifferentiated keratinocytes, and immortalized keratinocytes, wherein said skin substitute comprises a stratified squamous epithelium comprising said cells expressing exogenous Interleukin-12 supported on a dermal equivalent; and
b) placing said human skin substitute on said wound bed resulting from removal of a skin cancer in said patient such that the dermal equivalent is in direct contact with the wound bed and separates the wound bed from the cells expressing exogenous Interleukin-12;
wherein the human skin substitute suppresses growth of cancer cells in the wound bed.
9. A method of treating a wound bed resulting from removal of a skin cancer in a patient, comprising:
a) providing a human skin substitute comprising cells expressing exogenous Interleukin-12, wherein the cells are selected from the group consisting of primary keratinocytes, keratinocyte precursors, transdifferentiated keratinocytes, and immortalized keratinocytes, wherein said skin substitute comprises a stratified squamous epithelium comprising said cells expressing exogenous Interleukin-12 supported on a dermal equivalent; and
b) placing said human skin substitute on said wound bed resulting from 9 removal of a skin cancer in said patient such that the dermal equivalent is in direct contact with the wound bed and separates the wound bed from the cells expressing exogenous Interleukin-12;
wherein the human skin substitute stimulates an immune response against residual cancer cells in the wound bed.