Limits...
Stromal interferon-γ signaling and cross-presentation are required to eliminate antigen-loss variants of B cell lymphomas in mice.

Gerbitz A, Sukumar M, Helm F, Wilke A, Friese C, Fahrenwaldt C, Lehmann FM, Loddenkemper C, Kammertoens T, Mautner J, Schmitt CA, Blankenstein T, Bornkamm GW - PLoS ONE (2012)

Bottom Line: To study mechanisms of T cell-mediated rejection of B cell lymphomas, we developed a murine lymphoma model wherein three potential rejection antigens, human c-MYC, chicken ovalbumin (OVA), and GFP are expressed.Antigen-dependent eradication of lymphoma cells in wild-type animals was dependent on cross-presentation of antigen by cells of the tumor stroma.These findings provide first evidence for an important role of the tumor stroma in T cell-mediated control of hematologic neoplasias and highlight the importance of incorporating stroma-targeting strategies into future immunotherapeutic approaches.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Charité Berlin, Berlin, Germany. armin.gerbitz@uk-erlangen.de

ABSTRACT
To study mechanisms of T cell-mediated rejection of B cell lymphomas, we developed a murine lymphoma model wherein three potential rejection antigens, human c-MYC, chicken ovalbumin (OVA), and GFP are expressed. After transfer into wild-type mice 60-70% of systemically growing lymphomas expressing all three antigens were rejected; lymphomas expressing only human c-MYC protein were not rejected. OVA expressing lymphomas were infiltrated by T cells, showed MHC class I and II upregulation, and lost antigen expression, indicating immune escape. In contrast to wild-type recipients, 80-100% of STAT1-, IFN-γ-, or IFN-γ receptor-deficient recipients died of lymphoma, indicating that host IFN-γ signaling is critical for rejection. Lymphomas arising in IFN-γ- and IFN-γ-receptor-deficient mice had invariably lost antigen expression, suggesting that poor overall survival of these recipients was due to inefficient elimination of antigen-negative lymphoma variants. Antigen-dependent eradication of lymphoma cells in wild-type animals was dependent on cross-presentation of antigen by cells of the tumor stroma. These findings provide first evidence for an important role of the tumor stroma in T cell-mediated control of hematologic neoplasias and highlight the importance of incorporating stroma-targeting strategies into future immunotherapeutic approaches.

Show MeSH

Related in: MedlinePlus

Host IFN-γ and host IFN-γ signaling are required for rejection of 291OVA cells.1×105 291 parental cells (291PC) and retrovirally transduced 291OVA cells were injected s.c. into IFN-γ-deficient (A) and into IFN-γ-receptor- and STAT1-deficient recipient mice (B). Survival (left panels) and corresponding cumulative tumor growth (right panels) were monitored over 100 days. The data are compiled from two independent experiments. (C) Lymphomas developing in IFN-γ-receptor- and STAT1-deficient mice after inoculation of 1×105 291 parental or 291OVA cells were analyzed by immunohistochemistry for infiltration of CD3-positive cells (peroxidase brown staining) and perforin expressing cells (alkaline phosphatase staining) (left panel) in the same fashion as shown in Figure 2C. 10 animals per group were analyzed and Mann Whitney test was used for comparison.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3316708&req=5

pone-0034552-g004: Host IFN-γ and host IFN-γ signaling are required for rejection of 291OVA cells.1×105 291 parental cells (291PC) and retrovirally transduced 291OVA cells were injected s.c. into IFN-γ-deficient (A) and into IFN-γ-receptor- and STAT1-deficient recipient mice (B). Survival (left panels) and corresponding cumulative tumor growth (right panels) were monitored over 100 days. The data are compiled from two independent experiments. (C) Lymphomas developing in IFN-γ-receptor- and STAT1-deficient mice after inoculation of 1×105 291 parental or 291OVA cells were analyzed by immunohistochemistry for infiltration of CD3-positive cells (peroxidase brown staining) and perforin expressing cells (alkaline phosphatase staining) (left panel) in the same fashion as shown in Figure 2C. 10 animals per group were analyzed and Mann Whitney test was used for comparison.

Mentions: In this model lymphoma rejection depends on foreign antigen expression. The marked increase in SIINFEKL-pentamer-positive T cells in mice rejecting the tumors and the elimination of antigen expression in 291OVA cells cocultured with in vivo-primed OT-I cells in vitro point to a T cell-mediated mechanism of rejection. Given the importance of IFN-γ as effector molecule of T cells, we asked whether rejection depends on IFN-γ. 291OVA cells were injected into IFN-γ-deficient recipient animals, and lymphoma growth was monitored over time. As shown in Figure 4A, IFN-γ deficiency in the recipient led to almost complete loss of protection, and 80% of the recipient mice died from outgrowing lymphomas. The kinetics of tumor onset and speed of tumor growth were not markedly different after injection of 291OVA cells and untransduced 291PC cells (Figure 4A, right panel), with the difference that 20% of the IFN-γ-deficient animals receiving 291OVA cells remained tumor-free for at least 100 days. To ask whether IFN-γ acts on the side of the recipient, 291OVA and 291PC lymphoma cells were injected into IFN-γ-receptor- and STAT1-deficient mice lacking essential components of the interferon signaling system. Figure 4B shows that loss of responsiveness to both types of interferons (STAT1−/− recipients) resulted in 100% mortality after inoculation of 291OVA cells. No infiltration of T cells was observed in STAT1−/− recipients that had received 291OVA cells suggesting that recruitment of antigen-specific T cells to the tumor site is severely impaired in STAT1−/− mice (bottom panel, Figure 4C). In recipient mice lacking IFN-γ receptor (IFN-γR−/−) onset of lymphoma growth after inoculation occurred at the same time as in STAT1−/− recipients and lymphomas appeared to grow out slightly faster in IFN-γR−/− recipients. From day 18 on mean tumor diameters were significantly different in IFN-γR−/− and STAT1−/− recipients (p = 0.046–0.013, Mann Whitney test), but this difference did not result in differences in survival. In IFN-γR−/− recipients absence of antigen (291PC) did not result in faster lymphoma growth. T cell infiltration was observed in IFN-γR−/− recipients after inoculation of 291OVA cells in contrast to STAT1−/− recipients (Figure 4C), although to a lesser extent than in wild-type recipients (shown Figure 2C). These results show that both IFN-γ and IFN-γ-signaling are both required in the host to achieve antigen-dependent lymphoma rejection.


Stromal interferon-γ signaling and cross-presentation are required to eliminate antigen-loss variants of B cell lymphomas in mice.

Gerbitz A, Sukumar M, Helm F, Wilke A, Friese C, Fahrenwaldt C, Lehmann FM, Loddenkemper C, Kammertoens T, Mautner J, Schmitt CA, Blankenstein T, Bornkamm GW - PLoS ONE (2012)

Host IFN-γ and host IFN-γ signaling are required for rejection of 291OVA cells.1×105 291 parental cells (291PC) and retrovirally transduced 291OVA cells were injected s.c. into IFN-γ-deficient (A) and into IFN-γ-receptor- and STAT1-deficient recipient mice (B). Survival (left panels) and corresponding cumulative tumor growth (right panels) were monitored over 100 days. The data are compiled from two independent experiments. (C) Lymphomas developing in IFN-γ-receptor- and STAT1-deficient mice after inoculation of 1×105 291 parental or 291OVA cells were analyzed by immunohistochemistry for infiltration of CD3-positive cells (peroxidase brown staining) and perforin expressing cells (alkaline phosphatase staining) (left panel) in the same fashion as shown in Figure 2C. 10 animals per group were analyzed and Mann Whitney test was used for comparison.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3316708&req=5

pone-0034552-g004: Host IFN-γ and host IFN-γ signaling are required for rejection of 291OVA cells.1×105 291 parental cells (291PC) and retrovirally transduced 291OVA cells were injected s.c. into IFN-γ-deficient (A) and into IFN-γ-receptor- and STAT1-deficient recipient mice (B). Survival (left panels) and corresponding cumulative tumor growth (right panels) were monitored over 100 days. The data are compiled from two independent experiments. (C) Lymphomas developing in IFN-γ-receptor- and STAT1-deficient mice after inoculation of 1×105 291 parental or 291OVA cells were analyzed by immunohistochemistry for infiltration of CD3-positive cells (peroxidase brown staining) and perforin expressing cells (alkaline phosphatase staining) (left panel) in the same fashion as shown in Figure 2C. 10 animals per group were analyzed and Mann Whitney test was used for comparison.
Mentions: In this model lymphoma rejection depends on foreign antigen expression. The marked increase in SIINFEKL-pentamer-positive T cells in mice rejecting the tumors and the elimination of antigen expression in 291OVA cells cocultured with in vivo-primed OT-I cells in vitro point to a T cell-mediated mechanism of rejection. Given the importance of IFN-γ as effector molecule of T cells, we asked whether rejection depends on IFN-γ. 291OVA cells were injected into IFN-γ-deficient recipient animals, and lymphoma growth was monitored over time. As shown in Figure 4A, IFN-γ deficiency in the recipient led to almost complete loss of protection, and 80% of the recipient mice died from outgrowing lymphomas. The kinetics of tumor onset and speed of tumor growth were not markedly different after injection of 291OVA cells and untransduced 291PC cells (Figure 4A, right panel), with the difference that 20% of the IFN-γ-deficient animals receiving 291OVA cells remained tumor-free for at least 100 days. To ask whether IFN-γ acts on the side of the recipient, 291OVA and 291PC lymphoma cells were injected into IFN-γ-receptor- and STAT1-deficient mice lacking essential components of the interferon signaling system. Figure 4B shows that loss of responsiveness to both types of interferons (STAT1−/− recipients) resulted in 100% mortality after inoculation of 291OVA cells. No infiltration of T cells was observed in STAT1−/− recipients that had received 291OVA cells suggesting that recruitment of antigen-specific T cells to the tumor site is severely impaired in STAT1−/− mice (bottom panel, Figure 4C). In recipient mice lacking IFN-γ receptor (IFN-γR−/−) onset of lymphoma growth after inoculation occurred at the same time as in STAT1−/− recipients and lymphomas appeared to grow out slightly faster in IFN-γR−/− recipients. From day 18 on mean tumor diameters were significantly different in IFN-γR−/− and STAT1−/− recipients (p = 0.046–0.013, Mann Whitney test), but this difference did not result in differences in survival. In IFN-γR−/− recipients absence of antigen (291PC) did not result in faster lymphoma growth. T cell infiltration was observed in IFN-γR−/− recipients after inoculation of 291OVA cells in contrast to STAT1−/− recipients (Figure 4C), although to a lesser extent than in wild-type recipients (shown Figure 2C). These results show that both IFN-γ and IFN-γ-signaling are both required in the host to achieve antigen-dependent lymphoma rejection.

Bottom Line: To study mechanisms of T cell-mediated rejection of B cell lymphomas, we developed a murine lymphoma model wherein three potential rejection antigens, human c-MYC, chicken ovalbumin (OVA), and GFP are expressed.Antigen-dependent eradication of lymphoma cells in wild-type animals was dependent on cross-presentation of antigen by cells of the tumor stroma.These findings provide first evidence for an important role of the tumor stroma in T cell-mediated control of hematologic neoplasias and highlight the importance of incorporating stroma-targeting strategies into future immunotherapeutic approaches.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Charité Berlin, Berlin, Germany. armin.gerbitz@uk-erlangen.de

ABSTRACT
To study mechanisms of T cell-mediated rejection of B cell lymphomas, we developed a murine lymphoma model wherein three potential rejection antigens, human c-MYC, chicken ovalbumin (OVA), and GFP are expressed. After transfer into wild-type mice 60-70% of systemically growing lymphomas expressing all three antigens were rejected; lymphomas expressing only human c-MYC protein were not rejected. OVA expressing lymphomas were infiltrated by T cells, showed MHC class I and II upregulation, and lost antigen expression, indicating immune escape. In contrast to wild-type recipients, 80-100% of STAT1-, IFN-γ-, or IFN-γ receptor-deficient recipients died of lymphoma, indicating that host IFN-γ signaling is critical for rejection. Lymphomas arising in IFN-γ- and IFN-γ-receptor-deficient mice had invariably lost antigen expression, suggesting that poor overall survival of these recipients was due to inefficient elimination of antigen-negative lymphoma variants. Antigen-dependent eradication of lymphoma cells in wild-type animals was dependent on cross-presentation of antigen by cells of the tumor stroma. These findings provide first evidence for an important role of the tumor stroma in T cell-mediated control of hematologic neoplasias and highlight the importance of incorporating stroma-targeting strategies into future immunotherapeutic approaches.

Show MeSH
Related in: MedlinePlus