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Systemic T cell-independent tumor immunity after transplantation of universal receptor-modified bone marrow into SCID mice.

Hege KM, Cooke KS, Finer MH, Zsebo KM, Roberts MR - J. Exp. Med. (1996)

Bottom Line: Gene modification of hematopoietic stem cells (HSC) with antigen-specific, chimeric, or "universal" immune receptors (URs) is a novel but untested form of targeted immunotherapy.After transplantation into immunodeficient SCID mice, sustained high level expression of CD4 zeta was observed in circulating myeloid and natural killer cells.These results demonstrate the ability of chimeric receptors bearing zeta-signaling domains to activate non-T cell effector populations in vivo and thereby mediate systemic immunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Cell Biology, Cell Genesys Inc., Foster City, California 94404, USA.

ABSTRACT
Gene modification of hematopoietic stem cells (HSC) with antigen-specific, chimeric, or "universal" immune receptors (URs) is a novel but untested form of targeted immunotherapy. A human immunodeficiency virus (HIV) envelope-specific UR consisting of the extracellular domain of human CD4 linked to the zeta chain of the T cell receptor (CD4 zeta) was introduced ex vivo into murine HSC by retroviral transduction. After transplantation into immunodeficient SCID mice, sustained high level expression of CD4 zeta was observed in circulating myeloid and natural killer cells. CD4 zeta-transplanted mice were protected from challenge with a lethal dose of a disseminated human leukemia expressing HIV envelope. These results demonstrate the ability of chimeric receptors bearing zeta-signaling domains to activate non-T cell effector populations in vivo and thereby mediate systemic immunity.

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Related in: MedlinePlus

Death of CD4ζ-transplanted mice after Raji-env infusion is associated with a loss of gp120 expression in vivo. (a) Flow cytometric analysis.  Raji-env cells sorted from the bone marrow of a CD4ζ-transplanted (CD4ζ sort) and a control mouse (control sort), as well as Raji-p and Raji-env cells  maintained in liquid culture, were incubated with mouse anti-gp120 mAb to detect surface expression of HIV-env or the isotype-negative control, followed by incubation with goat anti–mouse biotin F(ab′)2 and APC (Molecular Probes). (b) Immunoblot analysis. Sorted Raji-env cells (CD4ζ sort and  control sort) and cultured Raji-p and Raji-env cells were lysed and subjected to SDS-PAGE, followed by immunoblotting with anti-gp120 mAb to detect  the presence of the env protein.
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Figure 3: Death of CD4ζ-transplanted mice after Raji-env infusion is associated with a loss of gp120 expression in vivo. (a) Flow cytometric analysis. Raji-env cells sorted from the bone marrow of a CD4ζ-transplanted (CD4ζ sort) and a control mouse (control sort), as well as Raji-p and Raji-env cells maintained in liquid culture, were incubated with mouse anti-gp120 mAb to detect surface expression of HIV-env or the isotype-negative control, followed by incubation with goat anti–mouse biotin F(ab′)2 and APC (Molecular Probes). (b) Immunoblot analysis. Sorted Raji-env cells (CD4ζ sort and control sort) and cultured Raji-p and Raji-env cells were lysed and subjected to SDS-PAGE, followed by immunoblotting with anti-gp120 mAb to detect the presence of the env protein.

Mentions: In the first experiment, it was noted that death was delayed in the two CD4ζ-expressing mice that died after challenge with 105 Raji-env cells (Fig. 2 a). To assay for the maintenance of gp120 expression by Raji-env in vivo, bone marrow was harvested from one of these mice at the time of death, and Raji-env cells were isolated by cell sorting using the human anti-B cell mAb anti–Leu-12 (CD19). Human CD19+ Raji cells constituted 4% of bone marrow leukocytes at the time of death from disseminated leukemia in this mouse. Recovered Raji-env cells were subjected to a sensitive APC staining procedure to detect surface expression of HIV gp120 (Fig. 3 a), as well as immunoblot analysis to detect total protein (Fig. 3 b). HIV gp120 could not be detected in these cells by either technique, suggesting that delayed death of this animal resulted from the outgrowth of Rajienv− revertants. In contrast, Raji-env cells sorted from the bone marrow of a control mouse at the time of death maintained stable expression of HIV gp120 by both flow cytometric and immunoblot analyses (Fig. 3, a and b). In subsequent experiments, death after infusion of 105 Rajienv cells into CD4ζ-transplanted mice was associated with a loss of gp120 expression in three out of three mice analyzed.


Systemic T cell-independent tumor immunity after transplantation of universal receptor-modified bone marrow into SCID mice.

Hege KM, Cooke KS, Finer MH, Zsebo KM, Roberts MR - J. Exp. Med. (1996)

Death of CD4ζ-transplanted mice after Raji-env infusion is associated with a loss of gp120 expression in vivo. (a) Flow cytometric analysis.  Raji-env cells sorted from the bone marrow of a CD4ζ-transplanted (CD4ζ sort) and a control mouse (control sort), as well as Raji-p and Raji-env cells  maintained in liquid culture, were incubated with mouse anti-gp120 mAb to detect surface expression of HIV-env or the isotype-negative control, followed by incubation with goat anti–mouse biotin F(ab′)2 and APC (Molecular Probes). (b) Immunoblot analysis. Sorted Raji-env cells (CD4ζ sort and  control sort) and cultured Raji-p and Raji-env cells were lysed and subjected to SDS-PAGE, followed by immunoblotting with anti-gp120 mAb to detect  the presence of the env protein.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196383&req=5

Figure 3: Death of CD4ζ-transplanted mice after Raji-env infusion is associated with a loss of gp120 expression in vivo. (a) Flow cytometric analysis. Raji-env cells sorted from the bone marrow of a CD4ζ-transplanted (CD4ζ sort) and a control mouse (control sort), as well as Raji-p and Raji-env cells maintained in liquid culture, were incubated with mouse anti-gp120 mAb to detect surface expression of HIV-env or the isotype-negative control, followed by incubation with goat anti–mouse biotin F(ab′)2 and APC (Molecular Probes). (b) Immunoblot analysis. Sorted Raji-env cells (CD4ζ sort and control sort) and cultured Raji-p and Raji-env cells were lysed and subjected to SDS-PAGE, followed by immunoblotting with anti-gp120 mAb to detect the presence of the env protein.
Mentions: In the first experiment, it was noted that death was delayed in the two CD4ζ-expressing mice that died after challenge with 105 Raji-env cells (Fig. 2 a). To assay for the maintenance of gp120 expression by Raji-env in vivo, bone marrow was harvested from one of these mice at the time of death, and Raji-env cells were isolated by cell sorting using the human anti-B cell mAb anti–Leu-12 (CD19). Human CD19+ Raji cells constituted 4% of bone marrow leukocytes at the time of death from disseminated leukemia in this mouse. Recovered Raji-env cells were subjected to a sensitive APC staining procedure to detect surface expression of HIV gp120 (Fig. 3 a), as well as immunoblot analysis to detect total protein (Fig. 3 b). HIV gp120 could not be detected in these cells by either technique, suggesting that delayed death of this animal resulted from the outgrowth of Rajienv− revertants. In contrast, Raji-env cells sorted from the bone marrow of a control mouse at the time of death maintained stable expression of HIV gp120 by both flow cytometric and immunoblot analyses (Fig. 3, a and b). In subsequent experiments, death after infusion of 105 Rajienv cells into CD4ζ-transplanted mice was associated with a loss of gp120 expression in three out of three mice analyzed.

Bottom Line: Gene modification of hematopoietic stem cells (HSC) with antigen-specific, chimeric, or "universal" immune receptors (URs) is a novel but untested form of targeted immunotherapy.After transplantation into immunodeficient SCID mice, sustained high level expression of CD4 zeta was observed in circulating myeloid and natural killer cells.These results demonstrate the ability of chimeric receptors bearing zeta-signaling domains to activate non-T cell effector populations in vivo and thereby mediate systemic immunity.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Cell Biology, Cell Genesys Inc., Foster City, California 94404, USA.

ABSTRACT
Gene modification of hematopoietic stem cells (HSC) with antigen-specific, chimeric, or "universal" immune receptors (URs) is a novel but untested form of targeted immunotherapy. A human immunodeficiency virus (HIV) envelope-specific UR consisting of the extracellular domain of human CD4 linked to the zeta chain of the T cell receptor (CD4 zeta) was introduced ex vivo into murine HSC by retroviral transduction. After transplantation into immunodeficient SCID mice, sustained high level expression of CD4 zeta was observed in circulating myeloid and natural killer cells. CD4 zeta-transplanted mice were protected from challenge with a lethal dose of a disseminated human leukemia expressing HIV envelope. These results demonstrate the ability of chimeric receptors bearing zeta-signaling domains to activate non-T cell effector populations in vivo and thereby mediate systemic immunity.

Show MeSH
Related in: MedlinePlus