Transgenically produced non-secreted proteins
The invention provides a method of making and secreting a non-secreted protein. The method includes expressing the protein from a nucleic acid construct which includes: (a) a mammary epithelial specific promoter; (b) a milk protein specific signal sequence which can direct the secretion of a protein; (c) optionally, a sequence which encodes a sufficient portion of the amino terminal coding region of a secreted protein to allow secretion in the milk of a transgenic mammal, of the non-secreted protein; and (d) a sequence which encodes a non-secreted protein, wherein elements (a), (b), optionally (c), and (d) are preferably operatively linked in the order recited. Both glutamic acid decarboxylase (GAD) and myelin basic protein (MBP), which are cytoplasmic proteins, have been produced by the methods of the present invention. The invention also provides methods for treating diabetes and multiple sclerosis using proteins produced by the methods of the present invention.
1 - 25 . (canceled)
26 . A method of inducing tolerance in a subject to an antigen, comprising:
providing a tolerogen expressed in a transgenic mammal which comprises the antigen; and
administering the tolerogen to the subject in an amount sufficient to induce tolerance to the antigen.
27 . The method of claim 26 , wherein the tolerogen is administered orally to the subject in the milk of a transgenic mammal.
28 . The method of claim 26 , wherein the tolerogen is an inactive protein.
29 . The method of claim 26 , wherein the tolerogen is a non-secreted protein.
30 . The method of claim 26 , wherein the tolerogen is a protein antigen fused to all or part of a secreted protein.
31 . The method of claim 26 , wherein the antigen is an antigen which is characteristic of an autoimmune disorder selected from the group consisting of diabetes, lupus, multiple sclerosis and rheumatoid arthritis.
32 . The method of claim 26 , wherein the subject is at risk of developing, or has, an autoimmune disorder selected from the group consisting of diabetes, lupus, multiple sclerosis and rheumatoid arthritis.
33 . A method of treating insulin-dependent diabetes mellitus (IDDM) in a subject, comprising administering to the subject therapeutically effective amount of a transgenically produced tolerogen which comprises glutamic acid decarboxylase, or an effective amount of a fusion protein which comprises glutamic acid decarboxylase.
34 . The method of claim 33 , wherein the subject is orally administered milk from a transgenic mammal which expresses the transgenically produced tolerogen which comprises glutamic acid decarboxylase, or an effective amount of a fusion protein which comprises glutamic acid decarboxylase.
35 . The method of claim 33 , wherein the transgenically produced tolarogen is in inactive form.