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Thermoreversible poly(ethylene glycol)-g-chitosan hydrogel as a therapeutic T lymphocyte depot for localized glioblastoma immunotherapy.

Tsao CT, Kievit FM, Ravanpay A, Erickson AE, Jensen MC, Ellenbogen RG, Zhang M - Biomacromolecules (2014)

Bottom Line: Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors.Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C.Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Washington , Seattle, Washington 98195, United States.

ABSTRACT
The outcome for glioblastoma patients remains dismal for its invariably recrudesces within 2 cm of the resection cavity. Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors. Here, we report the development of a biodegradable hydrogel containing therapeutic T lymphocytes for localized delivery to glioblastoma cells for brain tumor immunotherapy. Thermoreversible poly(ethylene glycol)-g-chitosan hydrogels (PCgels) were optimized for steady T lymphocyte release. Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C. T lymphocyte invasion through the PCgel and subsequent cytotoxicity to glioblastoma were assessed in vitro. The PCgel was shown to be cellular compatible with T lymphocytes, and the T lymphocytes retain their anti-glioblastoma activity after being encapsulated in the PCgel. T lymphocytes in the PCgel were shown to be more effective in killing glioblastoma than those in the Matrigel control. This may be attributed to the optimal pore size of the PCgel allowing better invasion of T lymphocytes. Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

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Related in: MedlinePlus

Evaluationof U-87 MG cell death by the live/dead flow cytometryassay. (a and b) Flow cytometry histograms of the gated U-87 MG cellpopulation (RFP+) assessed for the percentage of dead cells (BV 450-A+)after (a) invasive T lymphocyte through PCgel or Matrigel and (b)H2O2 and heat as positive controls. Data forthe untreated U-87 MG cells are labeled as U87. (c) Quantificationof the percentage of dead cells based on BV 450-A positivity. Dataare representative of triplicate samples. The statistical differencewith untreated tumor cells (U87) is labeled with an asterisk (p < 0.05).
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fig8: Evaluationof U-87 MG cell death by the live/dead flow cytometryassay. (a and b) Flow cytometry histograms of the gated U-87 MG cellpopulation (RFP+) assessed for the percentage of dead cells (BV 450-A+)after (a) invasive T lymphocyte through PCgel or Matrigel and (b)H2O2 and heat as positive controls. Data forthe untreated U-87 MG cells are labeled as U87. (c) Quantificationof the percentage of dead cells based on BV 450-A positivity. Dataare representative of triplicate samples. The statistical differencewith untreated tumor cells (U87) is labeled with an asterisk (p < 0.05).

Mentions: Figure 8a shows that there were more deadU-87 MG cells (BV 450-A+) under the PCgel transwell invasion conditionthan in the Matrigel. Figure 8b shows thatthere were a large number of dead U-87 MG cells killed by heat orH2O2. Moreover, quantitative analysis of theflow cytometry data (Figure 8c) shows thatthe frequency of U-87 MG death was significantly higher under thePCgel transwell invasion condition than for untreated cells (p = 0.0003) and under the Matrigel transwell invasion condition(p = 0.0057). This confirms the T lymphocytes thatinvaded through the PCgel and bound to U-87 MG cells were active andcould induce a therapeutic response. Therefore, our study shows theutility of the PCgel as a depot for release of T lymphocytes and aconvenient and valuable in vitro model for studyingclinically relevant improvements to glioblastoma immunotherapy.


Thermoreversible poly(ethylene glycol)-g-chitosan hydrogel as a therapeutic T lymphocyte depot for localized glioblastoma immunotherapy.

Tsao CT, Kievit FM, Ravanpay A, Erickson AE, Jensen MC, Ellenbogen RG, Zhang M - Biomacromolecules (2014)

Evaluationof U-87 MG cell death by the live/dead flow cytometryassay. (a and b) Flow cytometry histograms of the gated U-87 MG cellpopulation (RFP+) assessed for the percentage of dead cells (BV 450-A+)after (a) invasive T lymphocyte through PCgel or Matrigel and (b)H2O2 and heat as positive controls. Data forthe untreated U-87 MG cells are labeled as U87. (c) Quantificationof the percentage of dead cells based on BV 450-A positivity. Dataare representative of triplicate samples. The statistical differencewith untreated tumor cells (U87) is labeled with an asterisk (p < 0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215871&req=5

fig8: Evaluationof U-87 MG cell death by the live/dead flow cytometryassay. (a and b) Flow cytometry histograms of the gated U-87 MG cellpopulation (RFP+) assessed for the percentage of dead cells (BV 450-A+)after (a) invasive T lymphocyte through PCgel or Matrigel and (b)H2O2 and heat as positive controls. Data forthe untreated U-87 MG cells are labeled as U87. (c) Quantificationof the percentage of dead cells based on BV 450-A positivity. Dataare representative of triplicate samples. The statistical differencewith untreated tumor cells (U87) is labeled with an asterisk (p < 0.05).
Mentions: Figure 8a shows that there were more deadU-87 MG cells (BV 450-A+) under the PCgel transwell invasion conditionthan in the Matrigel. Figure 8b shows thatthere were a large number of dead U-87 MG cells killed by heat orH2O2. Moreover, quantitative analysis of theflow cytometry data (Figure 8c) shows thatthe frequency of U-87 MG death was significantly higher under thePCgel transwell invasion condition than for untreated cells (p = 0.0003) and under the Matrigel transwell invasion condition(p = 0.0057). This confirms the T lymphocytes thatinvaded through the PCgel and bound to U-87 MG cells were active andcould induce a therapeutic response. Therefore, our study shows theutility of the PCgel as a depot for release of T lymphocytes and aconvenient and valuable in vitro model for studyingclinically relevant improvements to glioblastoma immunotherapy.

Bottom Line: Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors.Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C.Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

View Article: PubMed Central - PubMed

Affiliation: Department of Materials Science and Engineering, University of Washington , Seattle, Washington 98195, United States.

ABSTRACT
The outcome for glioblastoma patients remains dismal for its invariably recrudesces within 2 cm of the resection cavity. Local immunotherapy has the potential to eradicate the residual infiltrative component of these tumors. Here, we report the development of a biodegradable hydrogel containing therapeutic T lymphocytes for localized delivery to glioblastoma cells for brain tumor immunotherapy. Thermoreversible poly(ethylene glycol)-g-chitosan hydrogels (PCgels) were optimized for steady T lymphocyte release. Nuclear magnetic resonance spectroscopy confirmed the chemical structure of poly(ethylene glycol)-g-chitosan, and rheological studies revealed that the sol-to-gel transition of the PCgel occurred around ≥32 °C. T lymphocyte invasion through the PCgel and subsequent cytotoxicity to glioblastoma were assessed in vitro. The PCgel was shown to be cellular compatible with T lymphocytes, and the T lymphocytes retain their anti-glioblastoma activity after being encapsulated in the PCgel. T lymphocytes in the PCgel were shown to be more effective in killing glioblastoma than those in the Matrigel control. This may be attributed to the optimal pore size of the PCgel allowing better invasion of T lymphocytes. Our study suggests that this unique PCgel depot may offer a viable approach for localized immunotherapy for glioblastoma.

Show MeSH
Related in: MedlinePlus