Human autologous in vitro models of glioma immunogene therapy using B7-2, GM-CSF, and IL12

Ian F Parney, Maxine A. Farr-Jones, Kevin Kane, Lung Ji Chang, Kenneth C. Petruk

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Background: Cancer immunogene therapy is based on vaccination with radiated, autologous tumor cells transduced with immunostimulatory genes. To help determine an optimal glioma immunogene therapy strategy, we stimulated lymphocytes with autologous human glioma cells transduced with B7-2 (CD86), granulocyte-macrophage colony - stimulating factor (GM-CSF), and/or interleukin-12 (IL12). Methods: A human glioma-derived cell culture (Ed147.BT) was transduced with B7-2, GM-CSF, and/or IL12 using retroviral vectors. Autologous peripheral blood mononuclear cells (PBMC) were co-cultured with irradiated gene-transduced tumor alone or a combination of radiated wild type and gene-transduced cells. Peripheral blood mononuclear cells proliferation was determined by serial cell counts. Peripheral blood mononuclear cells phenotype was assessed by flow cytometry for CD4, CD8, and CD16. Anti-tumor cytotoxicity was determined by chromium-51 (51Cr) release assay. Results: Peripheral blood mononuclear cells cell numbers all decreased during primary stimulation but tumor cells expressing B7-2 or GM-CSF consistently caused secondary proliferation. Tumors expressing B7-2 and GM-CSF or B7-2, GM-CSF, and IL12 consistently increased PBMC CD8+ (cytotoxic T) and CD16+ (natural killer) percentages. Interestingly, anti-tumor cytotoxicity only exceeded that of PBMC stimulated with wild type tumor alone when peripheral blood mononuclear cells were stimulated with both wild type tumor and B7-2/GM-CSF- (but not IL12) transduced cells. Conclusions: PBMC proliferation and phenotype is altered as expected by exposure to immunostimulatory gene- transduced tumor. However, transduced tumor cells alone do not stimulate greater anti-tumor cytotoxicity than wild type tumor. Only B7-2/GM-CSF-transduced cells combined with wild type produced increased cytotoxicity. This may reflect selection of tumor subclones with limited antigenic spectra during retrovirus-mediated gene transfer.

Original languageEnglish (US)
Pages (from-to)267-275
Number of pages9
JournalCanadian Journal of Neurological Sciences
Volume29
Issue number3
StatePublished - Aug 2002
Externally publishedYes

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Interleukin-12
Granulocyte-Macrophage Colony-Stimulating Factor
Glioma
Blood Cells
Neoplasms
Therapeutics
Genes
In Vitro Techniques
Cell Count
Cell Proliferation
Phenotype
Chromium
Retroviridae
Flow Cytometry
Vaccination
Cell Culture Techniques

ASJC Scopus subject areas

  • Clinical Neurology
  • Neuroscience(all)

Cite this

Human autologous in vitro models of glioma immunogene therapy using B7-2, GM-CSF, and IL12. / Parney, Ian F; Farr-Jones, Maxine A.; Kane, Kevin; Chang, Lung Ji; Petruk, Kenneth C.

In: Canadian Journal of Neurological Sciences, Vol. 29, No. 3, 08.2002, p. 267-275.

Research output: Contribution to journalArticle

Parney, Ian F ; Farr-Jones, Maxine A. ; Kane, Kevin ; Chang, Lung Ji ; Petruk, Kenneth C. / Human autologous in vitro models of glioma immunogene therapy using B7-2, GM-CSF, and IL12. In: Canadian Journal of Neurological Sciences. 2002 ; Vol. 29, No. 3. pp. 267-275.
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abstract = "Background: Cancer immunogene therapy is based on vaccination with radiated, autologous tumor cells transduced with immunostimulatory genes. To help determine an optimal glioma immunogene therapy strategy, we stimulated lymphocytes with autologous human glioma cells transduced with B7-2 (CD86), granulocyte-macrophage colony - stimulating factor (GM-CSF), and/or interleukin-12 (IL12). Methods: A human glioma-derived cell culture (Ed147.BT) was transduced with B7-2, GM-CSF, and/or IL12 using retroviral vectors. Autologous peripheral blood mononuclear cells (PBMC) were co-cultured with irradiated gene-transduced tumor alone or a combination of radiated wild type and gene-transduced cells. Peripheral blood mononuclear cells proliferation was determined by serial cell counts. Peripheral blood mononuclear cells phenotype was assessed by flow cytometry for CD4, CD8, and CD16. Anti-tumor cytotoxicity was determined by chromium-51 (51Cr) release assay. Results: Peripheral blood mononuclear cells cell numbers all decreased during primary stimulation but tumor cells expressing B7-2 or GM-CSF consistently caused secondary proliferation. Tumors expressing B7-2 and GM-CSF or B7-2, GM-CSF, and IL12 consistently increased PBMC CD8+ (cytotoxic T) and CD16+ (natural killer) percentages. Interestingly, anti-tumor cytotoxicity only exceeded that of PBMC stimulated with wild type tumor alone when peripheral blood mononuclear cells were stimulated with both wild type tumor and B7-2/GM-CSF- (but not IL12) transduced cells. Conclusions: PBMC proliferation and phenotype is altered as expected by exposure to immunostimulatory gene- transduced tumor. However, transduced tumor cells alone do not stimulate greater anti-tumor cytotoxicity than wild type tumor. Only B7-2/GM-CSF-transduced cells combined with wild type produced increased cytotoxicity. This may reflect selection of tumor subclones with limited antigenic spectra during retrovirus-mediated gene transfer.",
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