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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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Recognition of U-87 MG cells and invasive T lymphocytes.Fluorescenceimages of T lymphocytes (green) attached to U-87 MG cells (red) afterinvading through the transwell under (a) PCgel and (b) Matrigel conditions.Nuclei were counterstained with DAPI (blue). The scale bar is 10 μm.(c) Quantification of bound T lymphocytes through either the PCgelor Matrigel to U-87 MG cells (at 60× magnification from 10 differentfields of view on fluorescent images). Data are representative oftwo experiments conducted in duplicate. The statistical differencewith Matrigel was labeled with an asterisk (p <0.05).
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fig7: Recognition of U-87 MG cells and invasive T lymphocytes.Fluorescenceimages of T lymphocytes (green) attached to U-87 MG cells (red) afterinvading through the transwell under (a) PCgel and (b) Matrigel conditions.Nuclei were counterstained with DAPI (blue). The scale bar is 10 μm.(c) Quantification of bound T lymphocytes through either the PCgelor Matrigel to U-87 MG cells (at 60× magnification from 10 differentfields of view on fluorescent images). Data are representative oftwo experiments conducted in duplicate. The statistical differencewith Matrigel was labeled with an asterisk (p <0.05).

Mentions: Once the T lymphocytes invade through the PCgel, they mustremainfunctionally active to specifically target glioblastoma cells. Toinvestigate if T lymphocytes invading through gels specifically bindto tumor cells, 104 U-87 MG cells were seeded on a TCPas the lower chamber of the transwell device and T lymphocytes wereseeded onto transwell plates precoated with the PCgel or Matrigel(as control) the following day. The upper chamber was dissembled fromthe lower chamber 20 h after the addition of T lymphocytes. The interactionbetween the invasive T lymphocytes and U-87 MG cells was imaged usinga fluorescence microscope. The number of T lymphocytes bound to U-87MG cells was quantified by counting on 10 random fields and was reportedin number per field of view. Panels a and b of Figure 7 show the interaction of invasive T lymphocytes with the U-87MG cells after invading through the transwell PCgel and Matrigel,respectively. Green T lymphocytes were observed binding to red U-87MG cells in the PCgel condition, but very few T lymphocytes were observedin the Matrigel condition. Indeed, quantification of bound T lymphocytesrevealed there were significantly more T lymphocytes bound to U-87MG cells in the PCgel transwell invasion condition than in the Matrigeltranswell invasion condition [p = 0.0258 (Figure 7c)].


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)

Recognition of U-87 MG cells and invasive T lymphocytes.Fluorescenceimages of T lymphocytes (green) attached to U-87 MG cells (red) afterinvading through the transwell under (a) PCgel and (b) Matrigel conditions.Nuclei were counterstained with DAPI (blue). The scale bar is 10 μm.(c) Quantification of bound T lymphocytes through either the PCgelor Matrigel to U-87 MG cells (at 60× magnification from 10 differentfields of view on fluorescent images). Data are representative oftwo experiments conducted in duplicate. The statistical differencewith Matrigel was labeled with an asterisk (p <0.05).
© Copyright Policy
Related In: Results  -  Collection

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

fig7: Recognition of U-87 MG cells and invasive T lymphocytes.Fluorescenceimages of T lymphocytes (green) attached to U-87 MG cells (red) afterinvading through the transwell under (a) PCgel and (b) Matrigel conditions.Nuclei were counterstained with DAPI (blue). The scale bar is 10 μm.(c) Quantification of bound T lymphocytes through either the PCgelor Matrigel to U-87 MG cells (at 60× magnification from 10 differentfields of view on fluorescent images). Data are representative oftwo experiments conducted in duplicate. The statistical differencewith Matrigel was labeled with an asterisk (p <0.05).
Mentions: Once the T lymphocytes invade through the PCgel, they mustremainfunctionally active to specifically target glioblastoma cells. Toinvestigate if T lymphocytes invading through gels specifically bindto tumor cells, 104 U-87 MG cells were seeded on a TCPas the lower chamber of the transwell device and T lymphocytes wereseeded onto transwell plates precoated with the PCgel or Matrigel(as control) the following day. The upper chamber was dissembled fromthe lower chamber 20 h after the addition of T lymphocytes. The interactionbetween the invasive T lymphocytes and U-87 MG cells was imaged usinga fluorescence microscope. The number of T lymphocytes bound to U-87MG cells was quantified by counting on 10 random fields and was reportedin number per field of view. Panels a and b of Figure 7 show the interaction of invasive T lymphocytes with the U-87MG cells after invading through the transwell PCgel and Matrigel,respectively. Green T lymphocytes were observed binding to red U-87MG cells in the PCgel condition, but very few T lymphocytes were observedin the Matrigel condition. Indeed, quantification of bound T lymphocytesrevealed there were significantly more T lymphocytes bound to U-87MG cells in the PCgel transwell invasion condition than in the Matrigeltranswell invasion condition [p = 0.0258 (Figure 7c)].

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