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Route of immunization with peptide-pulsed dendritic cells controls the distribution of memory and effector T cells in lymphoid tissues and determines the pattern of regional tumor control.

Mullins DW, Sheasley SL, Ream RM, Bullock TN, Fu YX, Engelhard VH - J. Exp. Med. (2003)

Bottom Line: In contrast, i.v. immunization-induced memory was restricted to spleen and failed to impart protective immunity against subcutaneously growing tumors.Using peripheral LN-ablated mice, these LNs were shown to be essential for control of subcutaneously growing tumors but not lung metastases; in contrast, using immunized asplenic mice, we found that the spleen is necessary and sufficient for control of lung tumors, but unnecessary for control of subcutaneously growing tumors.These data demonstrate the existence of a previously undescribed population of splenic-resident memory CD8 T cells that are essential for the control of lung metastases.

View Article: PubMed Central - PubMed

Affiliation: Carter Immunology Center, University of Virginia, Box 801386, Charlottesville, VA 22908, USA.

ABSTRACT
We have established that the route of immunization with peptide-pulsed, activated DC leads to memory CD8+ T cells with distinct distributions in lymphoid tissues, which determines the ability to control tumors growing in different body sites. Both intravenous (i.v.) and subcutaneous (s.c.) immunization induced memory T cells in spleen and control of metastatic-like lung tumors. s.c. immunization also induced memory T cells in lymph nodes (LNs), imparting protection against subcutaneously growing tumors. In contrast, i.v. immunization-induced memory was restricted to spleen and failed to impart protective immunity against subcutaneously growing tumors. Memory cell distribution and tumor control were both linked to injection route-dependent localization of DCs in lymphoid compartments. Using peripheral LN-ablated mice, these LNs were shown to be essential for control of subcutaneously growing tumors but not lung metastases; in contrast, using immunized asplenic mice, we found that the spleen is necessary and sufficient for control of lung tumors, but unnecessary for control of subcutaneously growing tumors. These data demonstrate the existence of a previously undescribed population of splenic-resident memory CD8 T cells that are essential for the control of lung metastases. Thus, regional immunity based on memory T cell residence patterns is an important factor in DC-based tumor immunotherapy.

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s.c., but not i.v., immunization with peptide-pulsed DCs controls s.c. melanoma outgrowth. Mice were immunized once with unpulsed or 1 μM peptide-pulsed DCs, and B16-AAD melanoma was injected s.c. 21 d later. (A) Kinetics of tumor outgrowth after i.v. or s.c. immunization with 105 DCs pulsed with either Tyr369Y or a peptide corresponding to residues 57–66 of the influenza A M1 protein. Data are the mean of five separate animals ± SD; representative data from one of six similar experiments are shown. Additional representative experiments are shown in Fig. S1. (B) Kaplan-Meier survival curves after B16-AAD challenge in mice immunized i.v. or s.c. with 105 DCs pulsed with Tyr369Y- or M1 flu peptide-pulsed DCs, scored as days from challenge until tumor diameter exceeds 300 mm2. Survival was significantly increased in s.c. immunized animals compared with untreated (P < 0.001). Data are for groups of five animals; representative data from one of six similar experiments are shown. (C) Day 21 s.c. tumor size in mice immunized i.v. or s.c. with 103 to 5 × 105 DCs pulsed with Tyr369Y (1 μM). P-values indicate statistical significance compared with unimmunized control (black bar) by two-sample Student's t test.
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fig1: s.c., but not i.v., immunization with peptide-pulsed DCs controls s.c. melanoma outgrowth. Mice were immunized once with unpulsed or 1 μM peptide-pulsed DCs, and B16-AAD melanoma was injected s.c. 21 d later. (A) Kinetics of tumor outgrowth after i.v. or s.c. immunization with 105 DCs pulsed with either Tyr369Y or a peptide corresponding to residues 57–66 of the influenza A M1 protein. Data are the mean of five separate animals ± SD; representative data from one of six similar experiments are shown. Additional representative experiments are shown in Fig. S1. (B) Kaplan-Meier survival curves after B16-AAD challenge in mice immunized i.v. or s.c. with 105 DCs pulsed with Tyr369Y- or M1 flu peptide-pulsed DCs, scored as days from challenge until tumor diameter exceeds 300 mm2. Survival was significantly increased in s.c. immunized animals compared with untreated (P < 0.001). Data are for groups of five animals; representative data from one of six similar experiments are shown. (C) Day 21 s.c. tumor size in mice immunized i.v. or s.c. with 103 to 5 × 105 DCs pulsed with Tyr369Y (1 μM). P-values indicate statistical significance compared with unimmunized control (black bar) by two-sample Student's t test.

Mentions: Mice expressing a recombinant class I MHC molecule (AAD) were immunized with CD40L-activated and tyrosinase peptide (Tyr369Y)–pulsed DCs by either the s.c. or i.v. route. 3 wk later, mice were challenged with a s.c. bolus of B16-AAD melanoma cells. In six experiments involving five animals in each group (representative experiments shown in Fig. 1 A and Fig. S1, available at http://www.jem.org/cgi/content/full/jem.20021348/DC1), measurable outgrowth was delayed by 8.7 ± 1.5 d in mice immunized s.c. with Tyr369Y-pulsed DCs, compared with unimmunized littermates (P < 0.005). Survival, defined as the time point when tumors achieved or exceeded 300 mm2, was also significantly (P < 0.001) enhanced by a mean of 22.3 ± 2.4 d in the same experiments (one representative experiment is shown in Fig. 1 B). Surprisingly, tumor outgrowth was not delayed in animals immunized i.v. with the same DCs and no survival advantage was conferred, although this immunization has been shown previously to induce a significant antigen-specific CD8+ T cell response in spleen (20). s.c. immunization with unpulsed DCs or DCs pulsed with influenza M1 peptide did not delay tumor outgrowth or enhance survival, establishing that a tumor antigen-specific immune response was required. In these experiments, 105 DCs were injected; however, no control of outgrowth of subcutaneously growing tumors was observed in mice immunized i.v. with numbers of Tyr369Y-pulsed DCs ranging between 103 and 5 × 105 per animal (Fig. 1 C). In contrast, as few as 104 DCs injected by the s.c. route were sufficient for maximal control (Fig. 1 C). These data demonstrate that the immunization with DCs via the i.v. route fails to induce an immune response capable of controlling subcutaneously growing melanomas.


Route of immunization with peptide-pulsed dendritic cells controls the distribution of memory and effector T cells in lymphoid tissues and determines the pattern of regional tumor control.

Mullins DW, Sheasley SL, Ream RM, Bullock TN, Fu YX, Engelhard VH - J. Exp. Med. (2003)

s.c., but not i.v., immunization with peptide-pulsed DCs controls s.c. melanoma outgrowth. Mice were immunized once with unpulsed or 1 μM peptide-pulsed DCs, and B16-AAD melanoma was injected s.c. 21 d later. (A) Kinetics of tumor outgrowth after i.v. or s.c. immunization with 105 DCs pulsed with either Tyr369Y or a peptide corresponding to residues 57–66 of the influenza A M1 protein. Data are the mean of five separate animals ± SD; representative data from one of six similar experiments are shown. Additional representative experiments are shown in Fig. S1. (B) Kaplan-Meier survival curves after B16-AAD challenge in mice immunized i.v. or s.c. with 105 DCs pulsed with Tyr369Y- or M1 flu peptide-pulsed DCs, scored as days from challenge until tumor diameter exceeds 300 mm2. Survival was significantly increased in s.c. immunized animals compared with untreated (P < 0.001). Data are for groups of five animals; representative data from one of six similar experiments are shown. (C) Day 21 s.c. tumor size in mice immunized i.v. or s.c. with 103 to 5 × 105 DCs pulsed with Tyr369Y (1 μM). P-values indicate statistical significance compared with unimmunized control (black bar) by two-sample Student's t test.
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fig1: s.c., but not i.v., immunization with peptide-pulsed DCs controls s.c. melanoma outgrowth. Mice were immunized once with unpulsed or 1 μM peptide-pulsed DCs, and B16-AAD melanoma was injected s.c. 21 d later. (A) Kinetics of tumor outgrowth after i.v. or s.c. immunization with 105 DCs pulsed with either Tyr369Y or a peptide corresponding to residues 57–66 of the influenza A M1 protein. Data are the mean of five separate animals ± SD; representative data from one of six similar experiments are shown. Additional representative experiments are shown in Fig. S1. (B) Kaplan-Meier survival curves after B16-AAD challenge in mice immunized i.v. or s.c. with 105 DCs pulsed with Tyr369Y- or M1 flu peptide-pulsed DCs, scored as days from challenge until tumor diameter exceeds 300 mm2. Survival was significantly increased in s.c. immunized animals compared with untreated (P < 0.001). Data are for groups of five animals; representative data from one of six similar experiments are shown. (C) Day 21 s.c. tumor size in mice immunized i.v. or s.c. with 103 to 5 × 105 DCs pulsed with Tyr369Y (1 μM). P-values indicate statistical significance compared with unimmunized control (black bar) by two-sample Student's t test.
Mentions: Mice expressing a recombinant class I MHC molecule (AAD) were immunized with CD40L-activated and tyrosinase peptide (Tyr369Y)–pulsed DCs by either the s.c. or i.v. route. 3 wk later, mice were challenged with a s.c. bolus of B16-AAD melanoma cells. In six experiments involving five animals in each group (representative experiments shown in Fig. 1 A and Fig. S1, available at http://www.jem.org/cgi/content/full/jem.20021348/DC1), measurable outgrowth was delayed by 8.7 ± 1.5 d in mice immunized s.c. with Tyr369Y-pulsed DCs, compared with unimmunized littermates (P < 0.005). Survival, defined as the time point when tumors achieved or exceeded 300 mm2, was also significantly (P < 0.001) enhanced by a mean of 22.3 ± 2.4 d in the same experiments (one representative experiment is shown in Fig. 1 B). Surprisingly, tumor outgrowth was not delayed in animals immunized i.v. with the same DCs and no survival advantage was conferred, although this immunization has been shown previously to induce a significant antigen-specific CD8+ T cell response in spleen (20). s.c. immunization with unpulsed DCs or DCs pulsed with influenza M1 peptide did not delay tumor outgrowth or enhance survival, establishing that a tumor antigen-specific immune response was required. In these experiments, 105 DCs were injected; however, no control of outgrowth of subcutaneously growing tumors was observed in mice immunized i.v. with numbers of Tyr369Y-pulsed DCs ranging between 103 and 5 × 105 per animal (Fig. 1 C). In contrast, as few as 104 DCs injected by the s.c. route were sufficient for maximal control (Fig. 1 C). These data demonstrate that the immunization with DCs via the i.v. route fails to induce an immune response capable of controlling subcutaneously growing melanomas.

Bottom Line: In contrast, i.v. immunization-induced memory was restricted to spleen and failed to impart protective immunity against subcutaneously growing tumors.Using peripheral LN-ablated mice, these LNs were shown to be essential for control of subcutaneously growing tumors but not lung metastases; in contrast, using immunized asplenic mice, we found that the spleen is necessary and sufficient for control of lung tumors, but unnecessary for control of subcutaneously growing tumors.These data demonstrate the existence of a previously undescribed population of splenic-resident memory CD8 T cells that are essential for the control of lung metastases.

View Article: PubMed Central - PubMed

Affiliation: Carter Immunology Center, University of Virginia, Box 801386, Charlottesville, VA 22908, USA.

ABSTRACT
We have established that the route of immunization with peptide-pulsed, activated DC leads to memory CD8+ T cells with distinct distributions in lymphoid tissues, which determines the ability to control tumors growing in different body sites. Both intravenous (i.v.) and subcutaneous (s.c.) immunization induced memory T cells in spleen and control of metastatic-like lung tumors. s.c. immunization also induced memory T cells in lymph nodes (LNs), imparting protection against subcutaneously growing tumors. In contrast, i.v. immunization-induced memory was restricted to spleen and failed to impart protective immunity against subcutaneously growing tumors. Memory cell distribution and tumor control were both linked to injection route-dependent localization of DCs in lymphoid compartments. Using peripheral LN-ablated mice, these LNs were shown to be essential for control of subcutaneously growing tumors but not lung metastases; in contrast, using immunized asplenic mice, we found that the spleen is necessary and sufficient for control of lung tumors, but unnecessary for control of subcutaneously growing tumors. These data demonstrate the existence of a previously undescribed population of splenic-resident memory CD8 T cells that are essential for the control of lung metastases. Thus, regional immunity based on memory T cell residence patterns is an important factor in DC-based tumor immunotherapy.

Show MeSH
Related in: MedlinePlus