Antisense oligomers and methods for inducing immune tolerance and immunosuppression

Abstract

A method and composition for inducing human dendritic cells to a condition of reduced capacity for antigen-specific activation of T cells, and, in mature dendritic cells, increased production of extracellular IL-10 is disclosed. A population of dendritic cells is exposed to a substantially uncharged antisense compound containing 12-40 subunits and a base sequence effective to hybridize to an expression-sensitive region of a preprocessed or processed human CD86 transcript identified, in its processed form, by SEQ ID NO:33, to form a duplex structure between said compound and transcript having a Tm of at least 45° C. Formation of the duplex blocks expression of full-length CD86 in said cells, which in turn leads to reduced capacity for antigen-specific activation of T cells, and, in mature dendritic cells, increased production of extracellular IL-10.

Description

[0001] This application claims priority to U.S. Provisional Application No. 60,538,655, filed Jan. 23, 2004, which is incorporated herein by reference in its entirety.FIELD OF THE INVENTION [0002] The present invention relates to compounds and methods of inducing immunological tolerance using a peptide-antisense conjugate to selectively limit costimulation of naïve T-cells by mature dendritic cells and formation of a cytokine microenvironment that augments tolerized T-cells. REFERENCES [0003] Agrawal, S., et al., Proc Natl Acad Sci USA 87(4):1401-5, (1990). [0004] Akhtar, S., et al., Nucleic Acids Res 19(20):5551-9, (1991). [0005] Anderson, C. M., et al., J Neurochem 73(2):867-73, (1999). [0006] Anderson, K. P., et al., Antimicrob Agents Chemother 40(9):2004-11, (1996). [0007] Bonham, M. A., et al., Nucleic Acids Res 23(7):1197-203, (1995). [0008] Borriello, F., et al., J Immunol 155(12):5490-7, (1995). [0009] Boudvillain, M., et al., Biochemistry 36(10):2925-31, (1997). [0010] Cham...

AI Technical Summary

Benefits of technology

[0035] The invention includes, in one aspect, a method of inducing human dendritic cells to a condition of reduced capacity for antigen-specific activation of T cells, and, in mature dendritic cells, increased production of extracellular IL-10. The method includes exposing a population of human dendritic cells to a substantially uncharged antisense compound containing 12-40 subunits and a base sequence effective to hybridize to an expression-sensitive region of a preprocessed or processed human CD-86 transcript identified, in its processed form, by SEQ ID NO:33, to form, between the compound and transcript, a heteroduplex structure having a Tm of at least 45° C. The heteroduplex formation blocks expression of full-length CD86 in the cells, which in turn, produces inhibition of the expression of full-length CD86 on the surface of dendritic cells, and produces enhanced expression of extracellular IL-10 by mature dendritic cells.

[0037] The compound may be covalently linked, at one compound end, to an arginine-rich peptide effective to enhance uptake of the compound into the dendritic cells. Exemplary arginine-rich peptides are those having the sequences SEQ ID NOS: 1 or 2. Where the dendritic cells include a mixture of immature and mature dendritic cells, the arginine-rich peptide may be an rTAT peptide having the sequence identified by SEQ ID NO: 1. This peptide is effective to achieve a greater level of intracellular uptake of the antisense compound into the mature dendritic cells than is achieved (i) in the immature dendritic cells, or (ii) by exposing the mature dendritic cells to the antisense compound in the absence of the rTAT polypeptide.

[0042] For use in treating an autoimmune condition in a human subject, the compound may be administered to the subject, in an amount effective to reduce the severity of the autoimmune condition. The compound may be administered over an extended period of time, as needed, to control the severity of the autoimmune condition in the subject.

Problems solved by technology

However, allogeneic transplantation involves significant risks and drawbacks, including graft rejection, complications from immunosuppressive therapy and graft-versus host disease which are frequently highly debilitating or lethal.

Although immunosuppressive drugs such as cyclosporine may be used in an attempt to modulate rejection, these immunosuppressive agents have severe side effects and often fail to prevent continued rejection episodes.

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