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Rapid induction of tumor-specific type 1 T helper cells in metastatic melanoma patients by vaccination with mature, cryopreserved, peptide-loaded monocyte-derived dendritic cells.

Schuler-Thurner B, Schultz ES, Berger TG, Weinlich G, Ebner S, Woerl P, Bender A, Feuerstein B, Fritsch PO, Romani N, Schuler G - J. Exp. Med. (2002)

Bottom Line: The induction of strong, ex vivo detectable type 1 Th cell responses has not been reported to date.We demonstrate now that the subcutaneous injection of cryopreserved, mature, antigen-loaded, monocyte-derived dendritic cells (DCs) rapidly induces unequivocal Th1 responses (ex vivo detectable IFN-gamma-producing effectors as well as proliferating precursors) both to the control antigen KLH and to major histocompatibility complex (MHC) class II-restricted tumor peptides (melanoma-antigen [Mage]-3.DP4 and Mage-3.DR13) in the majority of 16 evaluable patients with metastatic melanoma.These Th1 cells recognized not only peptides, but also DCs loaded with Mage-3 protein, and in case of Mage-3DP4-specific Th1 cells IFN-gamma was released even after direct recognition of viable, Mage-3-expressing HLA-DP4+ melanoma cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Hospital of Erlangen, D-91052 Erlangen, Germany.

ABSTRACT
There is consensus that an optimized cancer vaccine will have to induce not only CD8+ cytotoxic but also CD4+ T helper (Th) cells, particularly interferon (IFN)-gamma-producing, type 1 Th cells. The induction of strong, ex vivo detectable type 1 Th cell responses has not been reported to date. We demonstrate now that the subcutaneous injection of cryopreserved, mature, antigen-loaded, monocyte-derived dendritic cells (DCs) rapidly induces unequivocal Th1 responses (ex vivo detectable IFN-gamma-producing effectors as well as proliferating precursors) both to the control antigen KLH and to major histocompatibility complex (MHC) class II-restricted tumor peptides (melanoma-antigen [Mage]-3.DP4 and Mage-3.DR13) in the majority of 16 evaluable patients with metastatic melanoma. These Th1 cells recognized not only peptides, but also DCs loaded with Mage-3 protein, and in case of Mage-3DP4-specific Th1 cells IFN-gamma was released even after direct recognition of viable, Mage-3-expressing HLA-DP4+ melanoma cells. The capacity of DCs to rapidly induce Th1 cells should be valuable to evaluate whether Th1 cells are instrumental in targeting human cancer and chronic infections.

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Related in: MedlinePlus

Induction of tumor-specific recall Th responses by DC immunization pre- and post therapy (i.e., 4 wk after vaccination no. 5) PBMCs were thawed, CD8 depleted, pulsed with gp100 DR4, tyrosinase DR4, Mage-3.DP4, and Mage-3.DR13 peptides and proliferation measured on day 5. In parallel cultures SEA (staphylococcal enterotoxin A) was added as a control both for the functioning of CD4+ T cells and an enhanced background reactivity, and proliferation measured on day 3. The results shown represent the average cpm 3[H]thymidine incorporation) and SEM of triplicate cultures. An increased proliferative response to Mage-3 peptide is seen in 12/16 patients but is ambiguous in patients 08 and 28 due to an increased background postvaccination.
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fig5: Induction of tumor-specific recall Th responses by DC immunization pre- and post therapy (i.e., 4 wk after vaccination no. 5) PBMCs were thawed, CD8 depleted, pulsed with gp100 DR4, tyrosinase DR4, Mage-3.DP4, and Mage-3.DR13 peptides and proliferation measured on day 5. In parallel cultures SEA (staphylococcal enterotoxin A) was added as a control both for the functioning of CD4+ T cells and an enhanced background reactivity, and proliferation measured on day 3. The results shown represent the average cpm 3[H]thymidine incorporation) and SEM of triplicate cultures. An increased proliferative response to Mage-3 peptide is seen in 12/16 patients but is ambiguous in patients 08 and 28 due to an increased background postvaccination.

Mentions: Induction of tumor-specific recall Th1 responses by DC immunization Pre and posttherapy (i.e., 4 wk after vaccination no. 5) CD4+ T cells from patients 12 (a) and 15 (b) were stimulated once in vitro with autologous, peptide-loaded mature DCs (10 μg/ml). On day 8 T cells were stimulated either with peptide alone (Mage-3.DP-4 or Mage-3.DR13 peptide) or with peptide-loaded mature DCs and IFN-γ spot-forming cells were measured by the Elispot technique. The results shown represent the average and SD of triplicate cocultures. Note that Mage-3.DP4-specific and Mage-3.DR13-specific IFN-γ spot-producing CD4+ Th1 cells are clearly expanded after DC vaccination in patients 12 and 15, respectively which fits to the results of the respective proliferative assays (Fig. 5). Note also that (auto-) reactivity to peptide-unloaded DCs (generated in patient 12 from post, and in patient 15 from prevaccination PBMCs) is not increased, but rather decreased upon DC vaccination and induction of specific immunity.


Rapid induction of tumor-specific type 1 T helper cells in metastatic melanoma patients by vaccination with mature, cryopreserved, peptide-loaded monocyte-derived dendritic cells.

Schuler-Thurner B, Schultz ES, Berger TG, Weinlich G, Ebner S, Woerl P, Bender A, Feuerstein B, Fritsch PO, Romani N, Schuler G - J. Exp. Med. (2002)

Induction of tumor-specific recall Th responses by DC immunization pre- and post therapy (i.e., 4 wk after vaccination no. 5) PBMCs were thawed, CD8 depleted, pulsed with gp100 DR4, tyrosinase DR4, Mage-3.DP4, and Mage-3.DR13 peptides and proliferation measured on day 5. In parallel cultures SEA (staphylococcal enterotoxin A) was added as a control both for the functioning of CD4+ T cells and an enhanced background reactivity, and proliferation measured on day 3. The results shown represent the average cpm 3[H]thymidine incorporation) and SEM of triplicate cultures. An increased proliferative response to Mage-3 peptide is seen in 12/16 patients but is ambiguous in patients 08 and 28 due to an increased background postvaccination.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193752&req=5

fig5: Induction of tumor-specific recall Th responses by DC immunization pre- and post therapy (i.e., 4 wk after vaccination no. 5) PBMCs were thawed, CD8 depleted, pulsed with gp100 DR4, tyrosinase DR4, Mage-3.DP4, and Mage-3.DR13 peptides and proliferation measured on day 5. In parallel cultures SEA (staphylococcal enterotoxin A) was added as a control both for the functioning of CD4+ T cells and an enhanced background reactivity, and proliferation measured on day 3. The results shown represent the average cpm 3[H]thymidine incorporation) and SEM of triplicate cultures. An increased proliferative response to Mage-3 peptide is seen in 12/16 patients but is ambiguous in patients 08 and 28 due to an increased background postvaccination.
Mentions: Induction of tumor-specific recall Th1 responses by DC immunization Pre and posttherapy (i.e., 4 wk after vaccination no. 5) CD4+ T cells from patients 12 (a) and 15 (b) were stimulated once in vitro with autologous, peptide-loaded mature DCs (10 μg/ml). On day 8 T cells were stimulated either with peptide alone (Mage-3.DP-4 or Mage-3.DR13 peptide) or with peptide-loaded mature DCs and IFN-γ spot-forming cells were measured by the Elispot technique. The results shown represent the average and SD of triplicate cocultures. Note that Mage-3.DP4-specific and Mage-3.DR13-specific IFN-γ spot-producing CD4+ Th1 cells are clearly expanded after DC vaccination in patients 12 and 15, respectively which fits to the results of the respective proliferative assays (Fig. 5). Note also that (auto-) reactivity to peptide-unloaded DCs (generated in patient 12 from post, and in patient 15 from prevaccination PBMCs) is not increased, but rather decreased upon DC vaccination and induction of specific immunity.

Bottom Line: The induction of strong, ex vivo detectable type 1 Th cell responses has not been reported to date.We demonstrate now that the subcutaneous injection of cryopreserved, mature, antigen-loaded, monocyte-derived dendritic cells (DCs) rapidly induces unequivocal Th1 responses (ex vivo detectable IFN-gamma-producing effectors as well as proliferating precursors) both to the control antigen KLH and to major histocompatibility complex (MHC) class II-restricted tumor peptides (melanoma-antigen [Mage]-3.DP4 and Mage-3.DR13) in the majority of 16 evaluable patients with metastatic melanoma.These Th1 cells recognized not only peptides, but also DCs loaded with Mage-3 protein, and in case of Mage-3DP4-specific Th1 cells IFN-gamma was released even after direct recognition of viable, Mage-3-expressing HLA-DP4+ melanoma cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Hospital of Erlangen, D-91052 Erlangen, Germany.

ABSTRACT
There is consensus that an optimized cancer vaccine will have to induce not only CD8+ cytotoxic but also CD4+ T helper (Th) cells, particularly interferon (IFN)-gamma-producing, type 1 Th cells. The induction of strong, ex vivo detectable type 1 Th cell responses has not been reported to date. We demonstrate now that the subcutaneous injection of cryopreserved, mature, antigen-loaded, monocyte-derived dendritic cells (DCs) rapidly induces unequivocal Th1 responses (ex vivo detectable IFN-gamma-producing effectors as well as proliferating precursors) both to the control antigen KLH and to major histocompatibility complex (MHC) class II-restricted tumor peptides (melanoma-antigen [Mage]-3.DP4 and Mage-3.DR13) in the majority of 16 evaluable patients with metastatic melanoma. These Th1 cells recognized not only peptides, but also DCs loaded with Mage-3 protein, and in case of Mage-3DP4-specific Th1 cells IFN-gamma was released even after direct recognition of viable, Mage-3-expressing HLA-DP4+ melanoma cells. The capacity of DCs to rapidly induce Th1 cells should be valuable to evaluate whether Th1 cells are instrumental in targeting human cancer and chronic infections.

Show MeSH
Related in: MedlinePlus