Dendritic cell immunotherapy in disseminated parathyroid carcinoma

Abstract

Arthur Bradwell and Timothy Harvey (Jan 30, p 370)1Bradwell AR Harvey TC Control of hypercalcaemia of parathyroid carcinoma by immunisation.Lancet. 1999; 353: 370-373Summary Full Text Full Text PDF PubMed Scopus (92) Google Scholar report on tumour immunotherapy with parathyroid hormone (PTH) and PTH peptides with complete Freund's adjuvant in a patient with parathyroid carcinoma.We describe a new tumour therapy in a 50-year-old woman with disseminated parathyroid carcinoma by use of tumour lysate-pulsed dendritic cells. Several studies in mice and human beings have shown the potent capacity of dendritic cells to induce specific antitumour immunity.2Nestle FO Alijagic S Gilliet M Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells.Nat Med. 1998; 4: 328-332Crossref PubMed Scopus (2680) Google Scholar, 3Hsu FJ Benike C Fagnoni F Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells.Nat Med. 1996; 2: 52-58Crossref PubMed Scopus (1694) Google Scholar Mature dendritic cells were generated from peripheral blood monocytes in the presence of granulocyte-macrophage colony stimulating factor (800 U/mL, day 1–7), interleukin 4 (500 U/mL, day 1–6), and tumour necrosis factor α (1000 U/mL; day 7 only).4Sallusto F Lanzavecchia A Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha.J Exp Med. 1994; 179: 1109-1111Crossref PubMed Scopus (4475) Google Scholar After loading with antigen (100 μg tumour lysate per 1×105 dendritic cells/mL medium for 4 h at 37°C), 5×106 dendritic cells were delivered every week by subcutaneous and intralymphatical injections into regional lymph nodes. While vaccinations (six cycles) were continued, serum calcium increased from 2·50 mmol/L to 2·99 mmol/L and total serum PTH from 202 pg/mL to 523 pg/mL. We therefore decided to introduce keyhole limpet haemocyanin (KLH), which is known to be a CD4-lymphocyte helper antigen and immunological tracer molecule. After four cycles of combined immunisation with tumour lysate and KLH, tumour-specific proliferation of the patients' peripheral blood mononuclear cells (PBMC) was observed. After treatment, PBMC exhibited strong, dose-dependent invitro reactivity (stimulation index 1·8–5·7) compared with the PBMC response before immunotherapy (1·0–1·1, compared with ovalbumin; figure).To investigate immune response in vivo, we used delayed-type hypersensitivity reaction as a convenient way to detect antigen-specific immunity. Skin tests were done with KLH (5μg) and tumour lysate (5 μg), respectively. Before therapy, neither KLH nor tumour lysate induced any visible reactivity. 1 week after the first combined treatment, the patient had a strong delayed-type hypersensitivity response towards KLH. While combined vaccinations (four cycles) were continued, an erythema and induration (18 mm diameter) was also observed against tumour lysate, indicating the efficient generation of tumour lysate-specific memory T cells. In addition, serum PTH decreased from 523 pg/mL to 401 pg/mL in the last 4 weeks.Our results and observations from other scientists indicate that dendritic cells can be pulsed with tumour protein and administered repeatedly without any significant side-effects. Small numbers of antigen-pulsed dendritic cells are effective in stimulating an invitro and in-vivo immune response after several treatments. It is unknown whether this vaccination strategy will induce sufficient clinical response with tumour eradication. Nevertheless, the use of tumour lysate-pulsed dendritic cells may represent a promising new approach to induce a potent immune response against weak antigens such as tumour proteins. Arthur Bradwell and Timothy Harvey (Jan 30, p 370)1Bradwell AR Harvey TC Control of hypercalcaemia of parathyroid carcinoma by immunisation.Lancet. 1999; 353: 370-373Summary Full Text Full Text PDF PubMed Scopus (92) Google Scholar report on tumour immunotherapy with parathyroid hormone (PTH) and PTH peptides with complete Freund's adjuvant in a patient with parathyroid carcinoma. We describe a new tumour therapy in a 50-year-old woman with disseminated parathyroid carcinoma by use of tumour lysate-pulsed dendritic cells. Several studies in mice and human beings have shown the potent capacity of dendritic cells to induce specific antitumour immunity.2Nestle FO Alijagic S Gilliet M Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells.Nat Med. 1998; 4: 328-332Crossref PubMed Scopus (2680) Google Scholar, 3Hsu FJ Benike C Fagnoni F Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells.Nat Med. 1996; 2: 52-58Crossref PubMed Scopus (1694) Google Scholar Mature dendritic cells were generated from peripheral blood monocytes in the presence of granulocyte-macrophage colony stimulating factor (800 U/mL, day 1–7), interleukin 4 (500 U/mL, day 1–6), and tumour necrosis factor α (1000 U/mL; day 7 only).4Sallusto F Lanzavecchia A Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha.J Exp Med. 1994; 179: 1109-1111Crossref PubMed Scopus (4475) Google Scholar After loading with antigen (100 μg tumour lysate per 1×105 dendritic cells/mL medium for 4 h at 37°C), 5×106 dendritic cells were delivered every week by subcutaneous and intralymphatical injections into regional lymph nodes. While vaccinations (six cycles) were continued, serum calcium increased from 2·50 mmol/L to 2·99 mmol/L and total serum PTH from 202 pg/mL to 523 pg/mL. We therefore decided to introduce keyhole limpet haemocyanin (KLH), which is known to be a CD4-lymphocyte helper antigen and immunological tracer molecule. After four cycles of combined immunisation with tumour lysate and KLH, tumour-specific proliferation of the patients' peripheral blood mononuclear cells (PBMC) was observed. After treatment, PBMC exhibited strong, dose-dependent invitro reactivity (stimulation index 1·8–5·7) compared with the PBMC response before immunotherapy (1·0–1·1, compared with ovalbumin; figure). To investigate immune response in vivo, we used delayed-type hypersensitivity reaction as a convenient way to detect antigen-specific immunity. Skin tests were done with KLH (5μg) and tumour lysate (5 μg), respectively. Before therapy, neither KLH nor tumour lysate induced any visible reactivity. 1 week after the first combined treatment, the patient had a strong delayed-type hypersensitivity response towards KLH. While combined vaccinations (four cycles) were continued, an erythema and induration (18 mm diameter) was also observed against tumour lysate, indicating the efficient generation of tumour lysate-specific memory T cells. In addition, serum PTH decreased from 523 pg/mL to 401 pg/mL in the last 4 weeks. Our results and observations from other scientists indicate that dendritic cells can be pulsed with tumour protein and administered repeatedly without any significant side-effects. Small numbers of antigen-pulsed dendritic cells are effective in stimulating an invitro and in-vivo immune response after several treatments. It is unknown whether this vaccination strategy will induce sufficient clinical response with tumour eradication. Nevertheless, the use of tumour lysate-pulsed dendritic cells may represent a promising new approach to induce a potent immune response against weak antigens such as tumour proteins.