Particle-mediated bombardment of DNA sequences into tissue to induce an immune response
First Claim
1. A method of administering a protective immune response-producing protein or peptide to produce a protective immune response in a vertebrate subject by in situ microprojectile bombardment, said method comprising the steps of:
- (a) selecting a target skin tissue residing in a vertebrate subject, the tissue selected from the group consisting of epidermis tissue, dermis tissue, and hypodermis tissue;
(b) providing microprojectiles, the microprojectiles carrying a DNA sequence comprising, in the 5′
to 3′
direction, a regulatory element functional in said target skin tissue and a gene positioned downstream of the regulatory element and under transcriptional control thereof to express said gene, the gene coding for a protective immune response-producing protein or polypeptide, wherein the microprojectiles comprise a material selected from the group consisting of metal, glass, silica, ice, polyethylene, polypropylene, polycarbonate, graphite and diamond;
(c) accelerating the microprojectiles at the subject, with the subject positioned so that the microprojectiles contact the subject'"'"'s epidermis at a speed sufficient to penetrate the epidermis and lodge in said selected target skin tissue cells and deposit the DNA sequence therein to provide transformed skin tissue cells, whereby a protective immune response-producing protein is administered to the vertebrate subject; and
(d) maintaining the transformed skin tissue cells in the subject, wherein a protective immune response is produced to the protein or peptide coded for by said gene in the subject upon expression of said gene in said transformed skin tissue cells, the immune response comprising a response in which B lymphocytes, T lymphocytes, or both B and T lymphocytes are activated by the protein or peptide and travel in circulatory or lymphatic systems of the vertebrate subject to a site removed from the transformed skin tissue cells.
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Abstract
A method of transferring a gene to vertebrate cells is disclosed. The method comprises the steps of: (a) providing microprojectiles, the microprojectiles carrying polynucleic acid sequences, the sequences comprising, in the 5′ to 3′ direction, a regulatory sequence operable in the tissue cells and a gene positioned downstream of the regulatory sequence and under the transcriptional control thereof; and (b) accelerating the microprojectiles at the cells, with the microprojectiles contacting the cells at a speed sufficient to penetrate the cells and deposit the polynucleic acid sequences therein. Preferably, the target cells reside in situ in the animal subject when they are transformed. Preferred target cells are dermis or hypodermis cells, and preferred genes for insertion into the target cells are genes which code for proteins or peptides which produce a physiological response in the animal subject.
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Citations
11 Claims
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1. A method of administering a protective immune response-producing protein or peptide to produce a protective immune response in a vertebrate subject by in situ microprojectile bombardment, said method comprising the steps of:
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(a) selecting a target skin tissue residing in a vertebrate subject, the tissue selected from the group consisting of epidermis tissue, dermis tissue, and hypodermis tissue;
(b) providing microprojectiles, the microprojectiles carrying a DNA sequence comprising, in the 5′
to 3′
direction, a regulatory element functional in said target skin tissue and a gene positioned downstream of the regulatory element and under transcriptional control thereof to express said gene, the gene coding for a protective immune response-producing protein or polypeptide, wherein the microprojectiles comprise a material selected from the group consisting of metal, glass, silica, ice, polyethylene, polypropylene, polycarbonate, graphite and diamond;
(c) accelerating the microprojectiles at the subject, with the subject positioned so that the microprojectiles contact the subject'"'"'s epidermis at a speed sufficient to penetrate the epidermis and lodge in said selected target skin tissue cells and deposit the DNA sequence therein to provide transformed skin tissue cells, whereby a protective immune response-producing protein is administered to the vertebrate subject; and
(d) maintaining the transformed skin tissue cells in the subject, wherein a protective immune response is produced to the protein or peptide coded for by said gene in the subject upon expression of said gene in said transformed skin tissue cells, the immune response comprising a response in which B lymphocytes, T lymphocytes, or both B and T lymphocytes are activated by the protein or peptide and travel in circulatory or lymphatic systems of the vertebrate subject to a site removed from the transformed skin tissue cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method of administering a protective immune response-producing protein or peptide to produce a protective immune response in a vertebrate subject by in situ microprojectile bombardment, said method comprising:
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(a) selecting a target skin tissue residing in a vertebrate subject, the tissue selected from the group consisting of epidermis tissue, dermis tissue, and hypodermis tissue;
(b) providing microprojectiles, the microprojectiles contacting with isolated and purified DNA constructs comprising, in the 5′
to 3′
direction, a regulatory element functional in said target skin tissue and a gene positioned downstream of the regulatory element and under transcriptional control thereof to express said gene, the gene coding for a protective immune response-producing protein or polypeptide, wherein the microprojectiles comprise a material selected from the group consisting of metal, glass, silica, ice, polyethylene, polypropylene, polycarbonate, graphite and diamond;
(c) accelerating the microprojectiles at the subject, with the subject positioned so that the microprojectiles contact the subject'"'"'s epidermis at a speed sufficient to penetrate the epidermis and lodge in said selected target skin tissue cells and deposit the DNA constructs therein to provide transformed skin tissue cells, whereby a protective immune response-producing protein is administered to the vertebrate subject; and
(d) maintaining the transformed skin tissue cells in the subject, wherein a protective immune response is produced to the protein or peptide coded for by said gene in the subject upon expression of said gene in said transformed skin tissue cells, the immune response comprising a response in which B lymphocytes, T lymphocytes, or both B and T lymphocytes are activated by the protein or peptide and travel in circulatory or lymphatic systems of the vertebrate subject to a site removed from the transformed skin tissue cells. - View Dependent Claims (10, 11)
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Specification