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Multimeric immunotoxins
| Details |
Inventors: Vallera, Daniel A.; Blazar, Bruce R.;
Assignee: Regents of the University of Minnesota (Minneapolis, MN)
Primary Examiner: Huff; Sheela
Assistant Examiner: Helms; Larry R.
Attorney, Agent or Firm: Fish & Richardson P.C.
The invention features fusion protein monomers, multimeric immunotoxic proteins, nucleic acids encoding fusion protein monomers, vectors containing the nucleic acids, and cells containing the vectors. Also encompassed by the invention are methods of killing pathogenic cells and making multimeric immunotoxic proteins. |
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DETAILED DESCRIPTION The invention derives from the finding that treatment of animals undergoing lethal GVHD with a homodimeric immunotoxin abrogated the GVHD.
Treatment with this immunotoxin allowed for a therapeutic window in that the dose of the dimeric immunotoxin required for abrogation of GVHD was more than four-fold lower than the toxic dose.
The invention includes monomeric fusion proteins useful for the production of multimeric immunotoxic proteins, multimeric immunotoxic proteins, nucleic acid molecules encoding fusion protein monomers, vectors containing the nucleic acid molecules, and cells containing the vectors.
Also included in the invention are in vitro and in vivo methods of killing a target cell involving delivery of a multimeric immunotoxic protein to the surface of a target cell and methods of producing both a fusion protein monomer of the invention and the multimeric immunotoxic proteins of the invention.
More specifically, the invention features a fusion protein molecule containing a toxic domain, a targeting domain, and at least one heterologous coupling moiety.
In this fusion protein, cysteine residues forming disulfide bonds are cysteine residues native to the toxic domain or the targeting domain.
Other aspects of the invention are: (a) an isolated nucleic acid molecule containing a nucleic acid sequence encoding the above fusion protein; (b) a vector containing the nucleic acid molecule of (a), e.
g.
, a vector in which transcriptional regulatory elements (TRE) are operably linked to the nucleic acid sequence; (c) a cell containing the vector of (b); and (d) a method of making a fusion protein in which the cell of (c) is cultured and the fusion protein is extracted from the culture, i.
e.
, either from the culture medium or from the cells.
The invention also provides a multimeric immunotoxic protein containing at least two of the above fusion protein molecules, each fusion protein molecule being joined by at least one of the heterologous coupling moieties to one or more of the other fusion protein molecules
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