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Autologous glioma cell vaccine admixed with interleukin-4 gene transfected fibroblasts in the treatment of patients with malignant gliomas.

Okada H, Lieberman FS, Walter KA, Lunsford LD, Kondziolka DS, Bejjani GK, Hamilton RL, Torres-Trejo A, Kalinski P, Cai Q, Mabold JL, Edington HD, Butterfield LH, Whiteside TL, Potter DM, Schold SC, Pollack IF - J Transl Med (2007)

Bottom Line: Interferon (IFN)-gamma Enzyme-Linked Immuno-SPOT (ELISPOT) assay in another human leukocyte antigen (HLA)-A2+ participant demonstrated systemic T-cell responses against an HLA-A2-restricted glioma-associated antigen (GAA) epitope EphA2883-891.Treatment was well tolerated; however, we were unable to observe detectable IFN-gamma post-vaccine responses or prolonged progression-free survival in these participants.Nevertheless, successful generation of type-1 DCs and preliminary safety in the current study provide a strong rationale for further efforts to develop novel glioma vaccines.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Neurological Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. okadah@upmc.edu

ABSTRACT

Background: The prognosis for malignant gliomas remains dismal. We addressed the safety, feasibility and preliminary clinical activity of the vaccinations using autologous glioma cells and interleukin (IL)-4 gene transfected fibroblasts.

Methods: In University of Pittsburgh Cancer Institute (UPCI) protocol 95-033, adult participants with recurrent glioblastoma multiforme (GBM) or anaplastic astrocytoma (AA) received gross total resection (GTR) of the recurrent tumors, followed by two vaccinations with autologous fibroblasts retrovirally transfected with TFG-IL4-Neo-TK vector admixed with irradiated autologous glioma cells. In UPCI 99-111, adult participants with newly diagnosed GBM or AA, following GTR and radiation therapy, received two intradermal vaccinations with the TFG-IL4-Neo-TK-transfected fibroblasts admixed with type-1 dendritic cells (DC) loaded with autologous tumor lysate. The participants were evaluated for occurrence of adverse events, immune response, and clinical response by radiological imaging.

Results and discussion: In UPCI 95-033, only 2 of 6 participants received the vaccinations. Four other participants were withdrawn from the trial because of tumor progression prior to production of the cellular vaccine. However, both participants who received two vaccinations demonstrated encouraging immunological and clinical responses. Biopsies from the local vaccine sites from one participant displayed IL-4 dose-dependent infiltration of CD4+ as well as CD8+ T cells. Interferon (IFN)-gamma Enzyme-Linked Immuno-SPOT (ELISPOT) assay in another human leukocyte antigen (HLA)-A2+ participant demonstrated systemic T-cell responses against an HLA-A2-restricted glioma-associated antigen (GAA) epitope EphA2883-891. Moreover, both participants demonstrated clinical and radiological improvement with no evidence of allergic encephalitis, although both participants eventually succumbed with the tumor recurrence. In 99-111, 5 of 6 enrolled participants received scheduled vaccinations with no incidence of major adverse events. Monocyte-derived DCs produced high levels of IL-12 p70. Treatment was well tolerated; however, we were unable to observe detectable IFN-gamma post-vaccine responses or prolonged progression-free survival in these participants.

Conclusion: Feasibility challenges inherent in the generation of a patient-specific gene transfection-based vaccine strongly suggests the need for more practical formulations that would allow for the timely administration of vaccines. Nevertheless, successful generation of type-1 DCs and preliminary safety in the current study provide a strong rationale for further efforts to develop novel glioma vaccines.

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Type-1 polarized DCs are efficient producers of IL-12 p70. In UPCI 99-111, on each vaccination day, a small aliquot of DCs (8 × 104) were tested for IL-12 production capability in comparison to DCs that were not stimulated with the type-1 cytokine-mixture (standard DCs). This was done with 24 hr stimulation of DCs (20 × 103per well, duplicates) with CD40L-transfected J558 cells (50 × 103per well). Supernatant was harvested and the production of IL-12 p70 was assayed by specific ELISA. Values indicate averages of duplicate samples. Bars indicate standard errors.
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Figure 1: Type-1 polarized DCs are efficient producers of IL-12 p70. In UPCI 99-111, on each vaccination day, a small aliquot of DCs (8 × 104) were tested for IL-12 production capability in comparison to DCs that were not stimulated with the type-1 cytokine-mixture (standard DCs). This was done with 24 hr stimulation of DCs (20 × 103per well, duplicates) with CD40L-transfected J558 cells (50 × 103per well). Supernatant was harvested and the production of IL-12 p70 was assayed by specific ELISA. Values indicate averages of duplicate samples. Bars indicate standard errors.

Mentions: One innovative aspect of UPCI 99-111 trial was that it employed type-1 polarizing DCs [15]. In 5 patients who received vaccines, we evaluated IL-12 p70 production in response to CD40L stimulation using CD40L-transfected J558 cells (Figure 1). In Case 1, only standard DCs were generated because the protocol was amended after case 1 to employ use of type-1, rather than standard DCs. Type-1 DCs were generated for Cases 3 to 6, and produced high levels of IL-12 p70, ranging 20–116 ng/1 × 106 cells/24 hrs, whereas control standard DCs from the same donors produced only background levels (less than 2 ng/1 × 106 cells/24 hrs) of IL-12 p70. These results demonstrate that type-1 DCs capable of producing high levels of IL-12 were successfully generated in the current trial.


Autologous glioma cell vaccine admixed with interleukin-4 gene transfected fibroblasts in the treatment of patients with malignant gliomas.

Okada H, Lieberman FS, Walter KA, Lunsford LD, Kondziolka DS, Bejjani GK, Hamilton RL, Torres-Trejo A, Kalinski P, Cai Q, Mabold JL, Edington HD, Butterfield LH, Whiteside TL, Potter DM, Schold SC, Pollack IF - J Transl Med (2007)

Type-1 polarized DCs are efficient producers of IL-12 p70. In UPCI 99-111, on each vaccination day, a small aliquot of DCs (8 × 104) were tested for IL-12 production capability in comparison to DCs that were not stimulated with the type-1 cytokine-mixture (standard DCs). This was done with 24 hr stimulation of DCs (20 × 103per well, duplicates) with CD40L-transfected J558 cells (50 × 103per well). Supernatant was harvested and the production of IL-12 p70 was assayed by specific ELISA. Values indicate averages of duplicate samples. Bars indicate standard errors.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2254376&req=5

Figure 1: Type-1 polarized DCs are efficient producers of IL-12 p70. In UPCI 99-111, on each vaccination day, a small aliquot of DCs (8 × 104) were tested for IL-12 production capability in comparison to DCs that were not stimulated with the type-1 cytokine-mixture (standard DCs). This was done with 24 hr stimulation of DCs (20 × 103per well, duplicates) with CD40L-transfected J558 cells (50 × 103per well). Supernatant was harvested and the production of IL-12 p70 was assayed by specific ELISA. Values indicate averages of duplicate samples. Bars indicate standard errors.
Mentions: One innovative aspect of UPCI 99-111 trial was that it employed type-1 polarizing DCs [15]. In 5 patients who received vaccines, we evaluated IL-12 p70 production in response to CD40L stimulation using CD40L-transfected J558 cells (Figure 1). In Case 1, only standard DCs were generated because the protocol was amended after case 1 to employ use of type-1, rather than standard DCs. Type-1 DCs were generated for Cases 3 to 6, and produced high levels of IL-12 p70, ranging 20–116 ng/1 × 106 cells/24 hrs, whereas control standard DCs from the same donors produced only background levels (less than 2 ng/1 × 106 cells/24 hrs) of IL-12 p70. These results demonstrate that type-1 DCs capable of producing high levels of IL-12 were successfully generated in the current trial.

Bottom Line: Interferon (IFN)-gamma Enzyme-Linked Immuno-SPOT (ELISPOT) assay in another human leukocyte antigen (HLA)-A2+ participant demonstrated systemic T-cell responses against an HLA-A2-restricted glioma-associated antigen (GAA) epitope EphA2883-891.Treatment was well tolerated; however, we were unable to observe detectable IFN-gamma post-vaccine responses or prolonged progression-free survival in these participants.Nevertheless, successful generation of type-1 DCs and preliminary safety in the current study provide a strong rationale for further efforts to develop novel glioma vaccines.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Neurological Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA. okadah@upmc.edu

ABSTRACT

Background: The prognosis for malignant gliomas remains dismal. We addressed the safety, feasibility and preliminary clinical activity of the vaccinations using autologous glioma cells and interleukin (IL)-4 gene transfected fibroblasts.

Methods: In University of Pittsburgh Cancer Institute (UPCI) protocol 95-033, adult participants with recurrent glioblastoma multiforme (GBM) or anaplastic astrocytoma (AA) received gross total resection (GTR) of the recurrent tumors, followed by two vaccinations with autologous fibroblasts retrovirally transfected with TFG-IL4-Neo-TK vector admixed with irradiated autologous glioma cells. In UPCI 99-111, adult participants with newly diagnosed GBM or AA, following GTR and radiation therapy, received two intradermal vaccinations with the TFG-IL4-Neo-TK-transfected fibroblasts admixed with type-1 dendritic cells (DC) loaded with autologous tumor lysate. The participants were evaluated for occurrence of adverse events, immune response, and clinical response by radiological imaging.

Results and discussion: In UPCI 95-033, only 2 of 6 participants received the vaccinations. Four other participants were withdrawn from the trial because of tumor progression prior to production of the cellular vaccine. However, both participants who received two vaccinations demonstrated encouraging immunological and clinical responses. Biopsies from the local vaccine sites from one participant displayed IL-4 dose-dependent infiltration of CD4+ as well as CD8+ T cells. Interferon (IFN)-gamma Enzyme-Linked Immuno-SPOT (ELISPOT) assay in another human leukocyte antigen (HLA)-A2+ participant demonstrated systemic T-cell responses against an HLA-A2-restricted glioma-associated antigen (GAA) epitope EphA2883-891. Moreover, both participants demonstrated clinical and radiological improvement with no evidence of allergic encephalitis, although both participants eventually succumbed with the tumor recurrence. In 99-111, 5 of 6 enrolled participants received scheduled vaccinations with no incidence of major adverse events. Monocyte-derived DCs produced high levels of IL-12 p70. Treatment was well tolerated; however, we were unable to observe detectable IFN-gamma post-vaccine responses or prolonged progression-free survival in these participants.

Conclusion: Feasibility challenges inherent in the generation of a patient-specific gene transfection-based vaccine strongly suggests the need for more practical formulations that would allow for the timely administration of vaccines. Nevertheless, successful generation of type-1 DCs and preliminary safety in the current study provide a strong rationale for further efforts to develop novel glioma vaccines.

Show MeSH
Related in: MedlinePlus