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Anti-Tumor Effects after Adoptive Transfer of IL-12 Transposon-Modified Murine Splenocytes in the OT-I-Melanoma Mouse Model.

Galvan DL, O'Neil RT, Foster AE, Huye L, Bear A, Rooney CM, Wilson MH - PLoS ONE (2015)

Bottom Line: Adoptive transfer of gene modified T cells provides possible immunotherapy for patients with cancers refractory to other treatments.We have previously used the non-viral piggyBac transposon system to gene modify human T cells for potential immunotherapy.We next gene-modified OT-I cells to express mIL-12.

View Article: PubMed Central - PubMed

Affiliation: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas, United States of America.

ABSTRACT
Adoptive transfer of gene modified T cells provides possible immunotherapy for patients with cancers refractory to other treatments. We have previously used the non-viral piggyBac transposon system to gene modify human T cells for potential immunotherapy. However, these previous studies utilized adoptive transfer of modified human T cells to target cancer xenografts in highly immunodeficient (NOD-SCID) mice that do not recapitulate an intact immune system. Currently, only viral vectors have shown efficacy in permanently gene-modifying mouse T cells for immunotherapy applications. Therefore, we sought to determine if piggyBac could effectively gene modify mouse T cells to target cancer cells in a mouse cancer model. We first demonstrated that we could gene modify cells to express murine interleukin-12 (p35/p40 mIL-12), a transgene with proven efficacy in melanoma immunotherapy. The OT-I melanoma mouse model provides a well-established T cell mediated immune response to ovalbumin (OVA) positive B16 melanoma cells. B16/OVA melanoma cells were implanted in wild type C57Bl6 mice. Mouse splenocytes were isolated from C57Bl6 OT-I mice and were gene modified using piggyBac to express luciferase. Adoptive transfer of luciferase-modified OT-I splenocytes demonstrated homing to B16/OVA melanoma tumors in vivo. We next gene-modified OT-I cells to express mIL-12. Adoptive transfer of mIL-12-modified mouse OT-I splenocytes delayed B16/OVA melanoma tumor growth in vivo compared to control OT-I splenocytes and improved mouse survival. Our results demonstrate that the piggyBac transposon system can be used to gene modify splenocytes and mouse T cells for evaluating adoptive immunotherapy strategies in immunocompetent mouse tumor models that may more directly mimic immunotherapy applications in humans.

No MeSH data available.


Related in: MedlinePlus

IL-12 piggyBac-modified mouse splenocytes exhibit anti-tumor activity in vivo.A, 5 X 105 B16/OVA cells were transplanted into the flank of C57Bl6 mice.OT-I splenocytes modified with pT-mIL12 and pCMV-m7pB (compared to pCMV-m7pB alone) were adoptively transferred on day 0 and day 8 and tumor growth was monitored in vivo via caliper measurement of tumor diameter. *, p<0.05 using the student’s T test on the given day of comparison. OT-I splenocytes modified with pT-mIL12 slowed tumor growth in vivo. B, Adoptive transfer of piggyBac modified OT-1 splenocytes also improved mouse survival in the B16 melanoma model. The Mantel-Cox test exhibited a statistically different survival between the two groups, N = 10.
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pone.0140744.g006: IL-12 piggyBac-modified mouse splenocytes exhibit anti-tumor activity in vivo.A, 5 X 105 B16/OVA cells were transplanted into the flank of C57Bl6 mice.OT-I splenocytes modified with pT-mIL12 and pCMV-m7pB (compared to pCMV-m7pB alone) were adoptively transferred on day 0 and day 8 and tumor growth was monitored in vivo via caliper measurement of tumor diameter. *, p<0.05 using the student’s T test on the given day of comparison. OT-I splenocytes modified with pT-mIL12 slowed tumor growth in vivo. B, Adoptive transfer of piggyBac modified OT-1 splenocytes also improved mouse survival in the B16 melanoma model. The Mantel-Cox test exhibited a statistically different survival between the two groups, N = 10.

Mentions: We next gene-modified mouse splenocytes with pT-IL12-Thy1.1 to evaluate the effect of mIL-12 on tumor growth in vivo. We implanted 5 X 105 B16/OVA cells into the flank of 5 gray irradiated C57Bl6 mice. IL-12-modified OT-I splenocytes were adoptively transferred on day 0 and day 8 and tumor growth was monitored in vivo. Untreated, i.e. no adoptive transfer of OT-I splenocytes, mice exhibited rapid tumor growth (Fig 6A). Adoptive transfer of piggyBac alone modified splenocytes slowed tumor growth as expected given the antigen specificity of OT-I T cells directed towards the melanoma cells expressing the OVA antigen. Splenocytes modified with pT-mIL-12 slowed tumor growth in vivo even further when compared to OT-I cells without IL-12 (Fig 6A). Adoptive transfer of piggyBac-mIL-12 modified splenocytes also improved mouse survival in the B16 melanoma model (Fig 6B). Therefore, piggyBac modified mouse splenocytes expressing mIL-12 were capable of anti-tumor activity in an in vivo melanoma tumor model and had improved anti-tumor activity compared to splenocytes containing antigen specific T cells alone not expressing mIL-12.


Anti-Tumor Effects after Adoptive Transfer of IL-12 Transposon-Modified Murine Splenocytes in the OT-I-Melanoma Mouse Model.

Galvan DL, O'Neil RT, Foster AE, Huye L, Bear A, Rooney CM, Wilson MH - PLoS ONE (2015)

IL-12 piggyBac-modified mouse splenocytes exhibit anti-tumor activity in vivo.A, 5 X 105 B16/OVA cells were transplanted into the flank of C57Bl6 mice.OT-I splenocytes modified with pT-mIL12 and pCMV-m7pB (compared to pCMV-m7pB alone) were adoptively transferred on day 0 and day 8 and tumor growth was monitored in vivo via caliper measurement of tumor diameter. *, p<0.05 using the student’s T test on the given day of comparison. OT-I splenocytes modified with pT-mIL12 slowed tumor growth in vivo. B, Adoptive transfer of piggyBac modified OT-1 splenocytes also improved mouse survival in the B16 melanoma model. The Mantel-Cox test exhibited a statistically different survival between the two groups, N = 10.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4608718&req=5

pone.0140744.g006: IL-12 piggyBac-modified mouse splenocytes exhibit anti-tumor activity in vivo.A, 5 X 105 B16/OVA cells were transplanted into the flank of C57Bl6 mice.OT-I splenocytes modified with pT-mIL12 and pCMV-m7pB (compared to pCMV-m7pB alone) were adoptively transferred on day 0 and day 8 and tumor growth was monitored in vivo via caliper measurement of tumor diameter. *, p<0.05 using the student’s T test on the given day of comparison. OT-I splenocytes modified with pT-mIL12 slowed tumor growth in vivo. B, Adoptive transfer of piggyBac modified OT-1 splenocytes also improved mouse survival in the B16 melanoma model. The Mantel-Cox test exhibited a statistically different survival between the two groups, N = 10.
Mentions: We next gene-modified mouse splenocytes with pT-IL12-Thy1.1 to evaluate the effect of mIL-12 on tumor growth in vivo. We implanted 5 X 105 B16/OVA cells into the flank of 5 gray irradiated C57Bl6 mice. IL-12-modified OT-I splenocytes were adoptively transferred on day 0 and day 8 and tumor growth was monitored in vivo. Untreated, i.e. no adoptive transfer of OT-I splenocytes, mice exhibited rapid tumor growth (Fig 6A). Adoptive transfer of piggyBac alone modified splenocytes slowed tumor growth as expected given the antigen specificity of OT-I T cells directed towards the melanoma cells expressing the OVA antigen. Splenocytes modified with pT-mIL-12 slowed tumor growth in vivo even further when compared to OT-I cells without IL-12 (Fig 6A). Adoptive transfer of piggyBac-mIL-12 modified splenocytes also improved mouse survival in the B16 melanoma model (Fig 6B). Therefore, piggyBac modified mouse splenocytes expressing mIL-12 were capable of anti-tumor activity in an in vivo melanoma tumor model and had improved anti-tumor activity compared to splenocytes containing antigen specific T cells alone not expressing mIL-12.

Bottom Line: Adoptive transfer of gene modified T cells provides possible immunotherapy for patients with cancers refractory to other treatments.We have previously used the non-viral piggyBac transposon system to gene modify human T cells for potential immunotherapy.We next gene-modified OT-I cells to express mIL-12.

View Article: PubMed Central - PubMed

Affiliation: Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas, United States of America.

ABSTRACT
Adoptive transfer of gene modified T cells provides possible immunotherapy for patients with cancers refractory to other treatments. We have previously used the non-viral piggyBac transposon system to gene modify human T cells for potential immunotherapy. However, these previous studies utilized adoptive transfer of modified human T cells to target cancer xenografts in highly immunodeficient (NOD-SCID) mice that do not recapitulate an intact immune system. Currently, only viral vectors have shown efficacy in permanently gene-modifying mouse T cells for immunotherapy applications. Therefore, we sought to determine if piggyBac could effectively gene modify mouse T cells to target cancer cells in a mouse cancer model. We first demonstrated that we could gene modify cells to express murine interleukin-12 (p35/p40 mIL-12), a transgene with proven efficacy in melanoma immunotherapy. The OT-I melanoma mouse model provides a well-established T cell mediated immune response to ovalbumin (OVA) positive B16 melanoma cells. B16/OVA melanoma cells were implanted in wild type C57Bl6 mice. Mouse splenocytes were isolated from C57Bl6 OT-I mice and were gene modified using piggyBac to express luciferase. Adoptive transfer of luciferase-modified OT-I splenocytes demonstrated homing to B16/OVA melanoma tumors in vivo. We next gene-modified OT-I cells to express mIL-12. Adoptive transfer of mIL-12-modified mouse OT-I splenocytes delayed B16/OVA melanoma tumor growth in vivo compared to control OT-I splenocytes and improved mouse survival. Our results demonstrate that the piggyBac transposon system can be used to gene modify splenocytes and mouse T cells for evaluating adoptive immunotherapy strategies in immunocompetent mouse tumor models that may more directly mimic immunotherapy applications in humans.

No MeSH data available.


Related in: MedlinePlus