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Treatment of experimental autoimmune encephalomyelitis with genetically modified memory T cells.

Mathisen PM, Yu M, Johnson JM, Drazba JA, Tuohy VK - J. Exp. Med. (1997)

Bottom Line: Isolated T cell clones demonstrated antigen-inducible expression of transgene IL-10 and expressed cell surface markers consistent with the phenotype of normal memory T cells.Semiquantitative immunocytochemistry showed a significant correlation between decreased demyelination and treatment with the transfected T cells.Taken together, these data indicate the autoreactive T cells can be genetically designed to produce therapeutic factors in an antigen-inducible manner resulting in a decreased severity of clinical and histological autoimmune demyelinating disease.

View Article: PubMed Central - PubMed

Affiliation: The Cleveland Clinic Foundation, Research Institute, Department of Immunology, FFb-1, Cleveland, Ohio 44195, USA.

ABSTRACT
The migratory properties of memory T cells provide a model vector system for site-specific delivery of therapeutic transgene factors to autoimmune inflammatory lesions. Lymph node cells from (SWRxSJL)F1 mice immunized with the p139-151 determinant of myelin proteolipid protein (PLP) were transfected with a DNA construct that placed the anti-inflammatory cytokine interleukin-10 (IL-10) cDNA under control of an antigen-inducible IL-2 promoter region. Isolated T cell clones demonstrated antigen-inducible expression of transgene IL-10 and expressed cell surface markers consistent with the phenotype of normal memory T cells. Upon adoptive transfer, transfected T cell clones were able to inhibit onset of experimental autoimmune encephalomyelitis (EAE) and to treat EAE animals therapeutically after onset of neurologic signs. Semiquantitative immunocytochemistry showed a significant correlation between decreased demyelination and treatment with the transfected T cells. Taken together, these data indicate the autoreactive T cells can be genetically designed to produce therapeutic factors in an antigen-inducible manner resulting in a decreased severity of clinical and histological autoimmune demyelinating disease.

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Related in: MedlinePlus

Detection of transgene IL-10 mRNA in T10.11 clone cells.  An RNase protection assay (RPA) was used for distinguishing endogenous and transgene mRNA. (a) The DNA construct used to generate the  RNA probe for measuring IL-10 expression. The RNA probe is represented by an arrow and protected RNase digestion products are shown.  (b) RNase-digestion products after hybridization to 32P-labeled RNA probe.  Lane 1, yeast RNA; lane 2, RNA from rested T10.11 clone cell 11 d after  activation with PLP 139–151; lane 3, RNA from activated T10.11 clone  cells 48 h after activation with PLP 139–151; lane 4, spleen RNA.
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Figure 2: Detection of transgene IL-10 mRNA in T10.11 clone cells. An RNase protection assay (RPA) was used for distinguishing endogenous and transgene mRNA. (a) The DNA construct used to generate the RNA probe for measuring IL-10 expression. The RNA probe is represented by an arrow and protected RNase digestion products are shown. (b) RNase-digestion products after hybridization to 32P-labeled RNA probe. Lane 1, yeast RNA; lane 2, RNA from rested T10.11 clone cell 11 d after activation with PLP 139–151; lane 3, RNA from activated T10.11 clone cells 48 h after activation with PLP 139–151; lane 4, spleen RNA.

Mentions: An RPA was used to differentiate between IL-2Prom→ IL-10cDNA transgene and endogenous IL-10 gene expression. RNA from resting and antigen-activated T10.11 cells was hybridized to an RNA probe prepared from the 5′ end of the IL-10 cDNA that incorporated transcribed-transgene vector sequences (Fig. 2 a). Two RNA transcripts were protected from activated T10.11 cells, indicating that both endogenous and transgene IL-10 expression were induced after stimulation with antigen (Fig. 2 b). Transgene expression represented 40% of the total IL-10 mRNA, and increased 7.5-fold after activation with PLP 139–151 compared with a 6.1-fold antigen-induced increase in endogenous IL-10 mRNA. Thus, antigen-inducible expression of transgene IL-10 mRNA occurred concomitant with endogenous IL-10 gene expression.


Treatment of experimental autoimmune encephalomyelitis with genetically modified memory T cells.

Mathisen PM, Yu M, Johnson JM, Drazba JA, Tuohy VK - J. Exp. Med. (1997)

Detection of transgene IL-10 mRNA in T10.11 clone cells.  An RNase protection assay (RPA) was used for distinguishing endogenous and transgene mRNA. (a) The DNA construct used to generate the  RNA probe for measuring IL-10 expression. The RNA probe is represented by an arrow and protected RNase digestion products are shown.  (b) RNase-digestion products after hybridization to 32P-labeled RNA probe.  Lane 1, yeast RNA; lane 2, RNA from rested T10.11 clone cell 11 d after  activation with PLP 139–151; lane 3, RNA from activated T10.11 clone  cells 48 h after activation with PLP 139–151; lane 4, spleen RNA.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2198970&req=5

Figure 2: Detection of transgene IL-10 mRNA in T10.11 clone cells. An RNase protection assay (RPA) was used for distinguishing endogenous and transgene mRNA. (a) The DNA construct used to generate the RNA probe for measuring IL-10 expression. The RNA probe is represented by an arrow and protected RNase digestion products are shown. (b) RNase-digestion products after hybridization to 32P-labeled RNA probe. Lane 1, yeast RNA; lane 2, RNA from rested T10.11 clone cell 11 d after activation with PLP 139–151; lane 3, RNA from activated T10.11 clone cells 48 h after activation with PLP 139–151; lane 4, spleen RNA.
Mentions: An RPA was used to differentiate between IL-2Prom→ IL-10cDNA transgene and endogenous IL-10 gene expression. RNA from resting and antigen-activated T10.11 cells was hybridized to an RNA probe prepared from the 5′ end of the IL-10 cDNA that incorporated transcribed-transgene vector sequences (Fig. 2 a). Two RNA transcripts were protected from activated T10.11 cells, indicating that both endogenous and transgene IL-10 expression were induced after stimulation with antigen (Fig. 2 b). Transgene expression represented 40% of the total IL-10 mRNA, and increased 7.5-fold after activation with PLP 139–151 compared with a 6.1-fold antigen-induced increase in endogenous IL-10 mRNA. Thus, antigen-inducible expression of transgene IL-10 mRNA occurred concomitant with endogenous IL-10 gene expression.

Bottom Line: Isolated T cell clones demonstrated antigen-inducible expression of transgene IL-10 and expressed cell surface markers consistent with the phenotype of normal memory T cells.Semiquantitative immunocytochemistry showed a significant correlation between decreased demyelination and treatment with the transfected T cells.Taken together, these data indicate the autoreactive T cells can be genetically designed to produce therapeutic factors in an antigen-inducible manner resulting in a decreased severity of clinical and histological autoimmune demyelinating disease.

View Article: PubMed Central - PubMed

Affiliation: The Cleveland Clinic Foundation, Research Institute, Department of Immunology, FFb-1, Cleveland, Ohio 44195, USA.

ABSTRACT
The migratory properties of memory T cells provide a model vector system for site-specific delivery of therapeutic transgene factors to autoimmune inflammatory lesions. Lymph node cells from (SWRxSJL)F1 mice immunized with the p139-151 determinant of myelin proteolipid protein (PLP) were transfected with a DNA construct that placed the anti-inflammatory cytokine interleukin-10 (IL-10) cDNA under control of an antigen-inducible IL-2 promoter region. Isolated T cell clones demonstrated antigen-inducible expression of transgene IL-10 and expressed cell surface markers consistent with the phenotype of normal memory T cells. Upon adoptive transfer, transfected T cell clones were able to inhibit onset of experimental autoimmune encephalomyelitis (EAE) and to treat EAE animals therapeutically after onset of neurologic signs. Semiquantitative immunocytochemistry showed a significant correlation between decreased demyelination and treatment with the transfected T cells. Taken together, these data indicate the autoreactive T cells can be genetically designed to produce therapeutic factors in an antigen-inducible manner resulting in a decreased severity of clinical and histological autoimmune demyelinating disease.

Show MeSH
Related in: MedlinePlus