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Adoptive T-cell therapy improves treatment of canine non-Hodgkin lymphoma post chemotherapy.

O'Connor CM, Sheppard S, Hartline CA, Huls H, Johnson M, Palla SL, Maiti S, Ma W, Davis RE, Craig S, Lee DA, Champlin R, Wilson H, Cooper LJ - Sci Rep (2012)

Bottom Line: Graded doses of autologous T cells were infused after CHOP chemotherapy and persisted for 49 days, homed to tumor, and significantly improved survival.Serum thymidine kinase changes predicted T-cell engraftment, while anti-tumor effects correlated with neutrophil-to-lymphocyte ratios and granzyme B expression in manufactured T cells.The companion canine model has translational implications for human immunotherapy which can be readily exploited since clinical-grade canine and human T cells are propagated using identical approaches.

View Article: PubMed Central - PubMed

Affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

ABSTRACT
Clinical observations reveal that an augmented pace of T-cell recovery after chemotherapy correlates with improved tumor-free survival, suggesting the add-back of T cells after chemotherapy may improve outcomes. To evaluate adoptive immunotherapy treatment for B-lineage non-Hodgkin lymphoma (NHL), we expanded T cells from client-owned canines diagnosed with NHL on artificial antigen presenting cells (aAPC) in the presence of human interleukin (IL)-2 and IL-21. Graded doses of autologous T cells were infused after CHOP chemotherapy and persisted for 49 days, homed to tumor, and significantly improved survival. Serum thymidine kinase changes predicted T-cell engraftment, while anti-tumor effects correlated with neutrophil-to-lymphocyte ratios and granzyme B expression in manufactured T cells. Therefore, chemotherapy can be used to modulate infused T-cell responses to enhance anti-tumor effects. The companion canine model has translational implications for human immunotherapy which can be readily exploited since clinical-grade canine and human T cells are propagated using identical approaches.

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Characterization of T-cell infusion products.(a) The mean inferred cell count for 6 (of the 8) canines infused with T cells that were propagated over 28–35 days on γ-irradiated OKT3-loaded aAPC (CLN4) in presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (b) Percent expression of CD4+ and CD8+ T-cell subsets during propagation on γ-irradiated aAPC in the presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (c) Immunophenotype of the T-cell infusion products (n = 6) at the time of cryopreservation. The horizontal lines describe mean percentage expression. (d) T-cell infusion products were tested for IFN-γ production after stimulation with OKT3-loaded aAPC in the presence of rhIL-2/IL-21 (n = 3). Analysis of multiplexed digital gene profiling of PB-derived canine T cells before and after propagation from healthy subjects (n = 2) and canines with NHL (n = 6) identified mRNA species which were either significantly (p<0.001) (e) up-regulated (> 2 fold change) or (f) down-regulated (< 2 fold change) in both healthy subjects and in at least 5 of 6 canines with NHL. (g) Study design: Enrollment occurred pre-, post, or during treatment with CHOP. Enrolled canines received one course of CHOP, typically administered over 19 weeks, and received one to three infusions of T cells 14 days apart using an intra-patient dose-escalation scheme.
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f3: Characterization of T-cell infusion products.(a) The mean inferred cell count for 6 (of the 8) canines infused with T cells that were propagated over 28–35 days on γ-irradiated OKT3-loaded aAPC (CLN4) in presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (b) Percent expression of CD4+ and CD8+ T-cell subsets during propagation on γ-irradiated aAPC in the presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (c) Immunophenotype of the T-cell infusion products (n = 6) at the time of cryopreservation. The horizontal lines describe mean percentage expression. (d) T-cell infusion products were tested for IFN-γ production after stimulation with OKT3-loaded aAPC in the presence of rhIL-2/IL-21 (n = 3). Analysis of multiplexed digital gene profiling of PB-derived canine T cells before and after propagation from healthy subjects (n = 2) and canines with NHL (n = 6) identified mRNA species which were either significantly (p<0.001) (e) up-regulated (> 2 fold change) or (f) down-regulated (< 2 fold change) in both healthy subjects and in at least 5 of 6 canines with NHL. (g) Study design: Enrollment occurred pre-, post, or during treatment with CHOP. Enrolled canines received one course of CHOP, typically administered over 19 weeks, and received one to three infusions of T cells 14 days apart using an intra-patient dose-escalation scheme.

Mentions: PB-(Supplementary Fig S1 and S2) with NHL recursively stimulated on γ-irradiated CLN4 in the presence of rhIL-2 and rhIL-21 for up to 35 days of culture underwent a mean 109 ± 15 fold expansion (Fig 3A). Seven days after the first aAPC-mediated stimulation the cultures contained T cells that were: 89 ± 6% CD3+, 60 ± 8% CD3+CD8+, and 27 ± 8% CD3+CD4+. By day 14, cultures were predominantly CD3+CD8+ (90 ± 4%) and the mean CD3+CD4+ population had decreased to 8 ± 4% (Fig 3B). T-cell infusion products analyzed for 6 (of the 8) treated dogs at the time of their cryopreservation were on average (Fig 3C): 98 ± 1% CD3+; 88 ± 2% CD3+CD8+; 11 ± 3% CD3+CD4+; 0.2 ± 0.2% CD3negCD21+ (B cells); 73 ± 10% CD3+CCR7+ (central memory); and 90 ± 2% PKH-26+ (fluorescence label to track persistence). To evaluate a T-cell effector function (n = 3) we assessed the IFN-γ production upon cross-linking CD3 with OKT3 loaded on CLN4 (Fig 3D). All the propagated CD3+CD8+ T cells, in contrast to the few numerically expanded CD3+CD4+ T cells, generated IFN-γ (99.1 ± 1% versus 3.7 ± 0.6%). It was noted that of the propagated CD4+ T cells, 8 ± 3% co-expressed CD25 and their reduced production of IFN-γ is consistent with a regulatory function. The selective propagation of CD8+ T cells that could be activated to secrete IFN-γ was exploited for adoptive immunotherapy.


Adoptive T-cell therapy improves treatment of canine non-Hodgkin lymphoma post chemotherapy.

O'Connor CM, Sheppard S, Hartline CA, Huls H, Johnson M, Palla SL, Maiti S, Ma W, Davis RE, Craig S, Lee DA, Champlin R, Wilson H, Cooper LJ - Sci Rep (2012)

Characterization of T-cell infusion products.(a) The mean inferred cell count for 6 (of the 8) canines infused with T cells that were propagated over 28–35 days on γ-irradiated OKT3-loaded aAPC (CLN4) in presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (b) Percent expression of CD4+ and CD8+ T-cell subsets during propagation on γ-irradiated aAPC in the presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (c) Immunophenotype of the T-cell infusion products (n = 6) at the time of cryopreservation. The horizontal lines describe mean percentage expression. (d) T-cell infusion products were tested for IFN-γ production after stimulation with OKT3-loaded aAPC in the presence of rhIL-2/IL-21 (n = 3). Analysis of multiplexed digital gene profiling of PB-derived canine T cells before and after propagation from healthy subjects (n = 2) and canines with NHL (n = 6) identified mRNA species which were either significantly (p<0.001) (e) up-regulated (> 2 fold change) or (f) down-regulated (< 2 fold change) in both healthy subjects and in at least 5 of 6 canines with NHL. (g) Study design: Enrollment occurred pre-, post, or during treatment with CHOP. Enrolled canines received one course of CHOP, typically administered over 19 weeks, and received one to three infusions of T cells 14 days apart using an intra-patient dose-escalation scheme.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3278154&req=5

f3: Characterization of T-cell infusion products.(a) The mean inferred cell count for 6 (of the 8) canines infused with T cells that were propagated over 28–35 days on γ-irradiated OKT3-loaded aAPC (CLN4) in presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (b) Percent expression of CD4+ and CD8+ T-cell subsets during propagation on γ-irradiated aAPC in the presence of rhIL-2/IL-21. Arrows represent days aAPC were added. (c) Immunophenotype of the T-cell infusion products (n = 6) at the time of cryopreservation. The horizontal lines describe mean percentage expression. (d) T-cell infusion products were tested for IFN-γ production after stimulation with OKT3-loaded aAPC in the presence of rhIL-2/IL-21 (n = 3). Analysis of multiplexed digital gene profiling of PB-derived canine T cells before and after propagation from healthy subjects (n = 2) and canines with NHL (n = 6) identified mRNA species which were either significantly (p<0.001) (e) up-regulated (> 2 fold change) or (f) down-regulated (< 2 fold change) in both healthy subjects and in at least 5 of 6 canines with NHL. (g) Study design: Enrollment occurred pre-, post, or during treatment with CHOP. Enrolled canines received one course of CHOP, typically administered over 19 weeks, and received one to three infusions of T cells 14 days apart using an intra-patient dose-escalation scheme.
Mentions: PB-(Supplementary Fig S1 and S2) with NHL recursively stimulated on γ-irradiated CLN4 in the presence of rhIL-2 and rhIL-21 for up to 35 days of culture underwent a mean 109 ± 15 fold expansion (Fig 3A). Seven days after the first aAPC-mediated stimulation the cultures contained T cells that were: 89 ± 6% CD3+, 60 ± 8% CD3+CD8+, and 27 ± 8% CD3+CD4+. By day 14, cultures were predominantly CD3+CD8+ (90 ± 4%) and the mean CD3+CD4+ population had decreased to 8 ± 4% (Fig 3B). T-cell infusion products analyzed for 6 (of the 8) treated dogs at the time of their cryopreservation were on average (Fig 3C): 98 ± 1% CD3+; 88 ± 2% CD3+CD8+; 11 ± 3% CD3+CD4+; 0.2 ± 0.2% CD3negCD21+ (B cells); 73 ± 10% CD3+CCR7+ (central memory); and 90 ± 2% PKH-26+ (fluorescence label to track persistence). To evaluate a T-cell effector function (n = 3) we assessed the IFN-γ production upon cross-linking CD3 with OKT3 loaded on CLN4 (Fig 3D). All the propagated CD3+CD8+ T cells, in contrast to the few numerically expanded CD3+CD4+ T cells, generated IFN-γ (99.1 ± 1% versus 3.7 ± 0.6%). It was noted that of the propagated CD4+ T cells, 8 ± 3% co-expressed CD25 and their reduced production of IFN-γ is consistent with a regulatory function. The selective propagation of CD8+ T cells that could be activated to secrete IFN-γ was exploited for adoptive immunotherapy.

Bottom Line: Graded doses of autologous T cells were infused after CHOP chemotherapy and persisted for 49 days, homed to tumor, and significantly improved survival.Serum thymidine kinase changes predicted T-cell engraftment, while anti-tumor effects correlated with neutrophil-to-lymphocyte ratios and granzyme B expression in manufactured T cells.The companion canine model has translational implications for human immunotherapy which can be readily exploited since clinical-grade canine and human T cells are propagated using identical approaches.

View Article: PubMed Central - PubMed

Affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

ABSTRACT
Clinical observations reveal that an augmented pace of T-cell recovery after chemotherapy correlates with improved tumor-free survival, suggesting the add-back of T cells after chemotherapy may improve outcomes. To evaluate adoptive immunotherapy treatment for B-lineage non-Hodgkin lymphoma (NHL), we expanded T cells from client-owned canines diagnosed with NHL on artificial antigen presenting cells (aAPC) in the presence of human interleukin (IL)-2 and IL-21. Graded doses of autologous T cells were infused after CHOP chemotherapy and persisted for 49 days, homed to tumor, and significantly improved survival. Serum thymidine kinase changes predicted T-cell engraftment, while anti-tumor effects correlated with neutrophil-to-lymphocyte ratios and granzyme B expression in manufactured T cells. Therefore, chemotherapy can be used to modulate infused T-cell responses to enhance anti-tumor effects. The companion canine model has translational implications for human immunotherapy which can be readily exploited since clinical-grade canine and human T cells are propagated using identical approaches.

Show MeSH
Related in: MedlinePlus