Dendritoma vaccination combined with low dose interleukin-2 in metastatic melanoma patients induced immunological and clinical responses

A pilot clinical trial using dendritomas, purified hybrids from the fusion of dendritic/tumor cells combined with a low dose of IL-2, in metastatic melanoma patients was conducted in order to determine its safety and potential immunological and clinical responses. Ten metastatic melanoma patients were enrolled into this study. Dendritoma vaccines were created by fusing dendritic cells stained with green fluorescent dye with irradiated autologous tumor cells stained with red fluorescent dye and purifying the hybrids using immediate fluorescent-activated cell sorting. Initial vaccine was given subcutaneously and followed by IL-2 in serially elevated doses from 3-9 million units/m2 for 5 days. Repeated vaccinations were administered without IL-2, at 3-month intervals for a maximum of 5 times. Immune reactions were measured by the increase of interferon-gamma (IFN-gamma) expressing T cells. Vaccine doses ranged from 250,000 to 1,000,000 dendritomas. There was no grade 2 or higher toxicity directly attributable to the vaccine. All patients experienced toxicity due to IL-2 administration (9-grade 2, 3-grade 3, 1-grade 4). Eight of nine evaluable patients demonstrated immunologic reactions by increased IFN-gamma expressing T cells. One patient developed partial response at 12 weeks after the first vaccine. Nine months later, this patient achieved a complete response. In addition, two patients had stable disease for 9 and 4 months, respectively; one patient had a mixed response. Our findings demonstrated that dendritoma vaccines with a low dose of IL-2 can be safely administered to patients with metastatic melanoma and induce immunological and clinical responses.     

Immunologic and clinical responses after vaccinations with peptide-pulsed dendritic cells in metastatic renal cancer patients

A phase I trial was conducted to evaluate the feasibility, safety, and efficacy of a dendritic cell-based vaccination in patients with metastatic renal cell carcinoma (RCC). Autologous mature dendritic cells derived from peripheral blood monocytes were pulsed with the HLA-A2-binding MUC1 peptides (M1.1 and M1.2). For the activation of CD4(+) T-helper lymphocytes, dendritic cells were further incubated with the PAN-DR-binding peptide PADRE. Dendritic cell vaccinations were done s.c. every 2 weeks for four times and repeated monthly until tumor progression. After five dendritic cell injections, patients additionally received three injections weekly of low-dose interleukin-2 (1 million IE/m(2)). The induction of vaccine-induced T-cell responses was monitored using enzyme-linked immunospot and Cr release assays. Twenty patients were included. The treatment was well tolerated with no severe side effects. In six patients, regression of the metastatic sites was induced after vaccinations with three patients achieving an objective response (one complete response, two partial responses, two mixed responses, and one stable disease). Additional four patients were stable during the treatment for up to 14 months. MUC1 peptide-specific T-cell responses in vivo were detected in the peripheral blood mononuclear cells of the six patients with objective responses. Interestingly, in patients responding to the treatment, T-cell responses to antigens not used for vaccinations, such as adipophilin, telomerase, or oncofetal antigen, could be detected, indicating that epitope spreading might occur. This study shows that MUC1 peptide-pulsed dendritic cells can induce clinical and immunologic responses in patients with metastatic RCC.     

多西他赛对复发转移性乳腺癌免疫功能的影响

目的：探讨多西他赛化疗前、化疗2周期评价疗效时复发转移性乳腺癌（MBC）患者外周血淋巴细胞亚群比率的变化及影响因素。方法应用流式细胞仪检测34例行多西他赛一线化疗的复发MBC患者化疗前、化疗2周期评价疗效时外周血淋巴细胞亚群[CD3+T淋巴细胞、CD3+/CD4+T淋巴细胞、CD3+/CD8+T淋巴细胞、CD3－/CD16+56+自然杀伤细胞（NK）、CD3+/CD16+56+T淋巴细胞、CD19+B淋巴细胞、CD4+/CD25+调节性T细胞（Treg细胞）、CD8+/CD28－T淋巴细胞和CD8+/CD28+T淋巴细胞]比率,并进一步分析患者临床病理因素对于外周血淋巴细胞亚群变化的影响。结果34例患者化疗后CD3+总T淋巴细胞、CD3+/CD4+T淋巴细胞及CD19+B淋巴细胞比率均较化疗前下降（P=0.002、0.044、0.006）,下降平均比率分别为2.2%、4.7%、3.1%。中位年龄﹥54岁的患者CD19+B淋巴细胞下降比率较中位年龄≤54岁的患者小（P=0.031）;中位OS﹥3.6个月的患者CD3+总T淋巴细胞下降比率较中位OS≤33.6个月的患者小（P=0.038）。结论多西他赛化疗后MBC患者外周血总T、B淋巴细胞比率下降,免疫功能降低,而相对保留较好的T淋巴细胞免疫功能的患者可能有更好的生存获益。     

Lymphopenia and clinical outcome of elderly patients treated with sunitinib for metastatic renal cell cancer

ObjectiveLymphopenia is associated with toxicity and outcomes in several cancer types. We assessed the association between pre-treatment lymphopenia, toxicity, and clinical outcomes in elderly patients with metastatic renal cell cancer (mRCC) treated with first-line sunitinib. Prognostic factors in these patients were also evaluated.Patients and MethodsWe reviewed the clinical records of 181 patients with mRCC aged ≥ 70 years treated with first-line sunitinib in 17 Italian Oncology Units from February 2006 to September 2011. Baseline lymphopenia was defined as lymphocyte counts < 1000/μL.ResultsTwenty-nine (16%) patients had a baseline lymphocyte count < 1000/μL (group A) and 152 (84%) patients had a lymphocyte count ≥ 1000/μL (group B). Although no differences between the two groups were reported in terms of overall response rate (P = 0.207), dose reductions (P = 0.740), discontinuation due to adverse events (P = 0.175) or overall incidence of grade 3–4 toxicities (P = 0.112), more patients in the lymphopenia group had grade 3–4 neutropenia (P = 0.017), grade 3–4 thrombocytopenia (P = 0.017) and grade 3–4 diarrhea (P = 0.006). In multivariate analysis, performance status and Heng score were predictors of progression-free survival (P = 0.015 and P = 0.0006, respectively), while performance status, Heng score, and lymphopenia were found to be significantly associated with overall survival (P = 0.007, P < 0.0001 and P = 0.023, respectively).ConclusionsSunitinib appears to be safe and active in elderly patients with lymphopenia. Lymphocyte count is an independent prognostic factor for overall survival in elderly patients with mRCC treated with first-line sunitinib.     

A controlled clinical and immunological evaluation of immunotherapy with Bacillus Calmette--Guerin in patients with metastatic cancer

One hundred thirty adult patients with metastatic solid tumors were studied to evaluate the therapeutic results, toxicity, and immunological effects of three BCG preparations given intradermally by the multiple puncture time technique on either a monthly or weekly schedule: Tice lyophilized BCG, Pasteur lyophilized BCG, and Pasteur fresh-frozen BCG. Patients were randomly assigned in a double-blind manner to receive one of the BCG preparations or saline placebo administered in an identical manner. A battery of immunological tests were performed pretreatment and serially during the investigation. None of the 82 patients with measurable indicator lesions experienced an objective tumor response, nor were there any differences in survival among patients receiving any of the BCG preparations compared to placebo. Toxicity consisted of mild to moderate local cutaneous reactions and liver function abnormalities in BCG-treated patients with no significant differences between BCG preparations. Improvements in each of the assays of immune function performed occurred with roughly equal frequency among patients receiving BCG or placebo. It was not possible to demonstrate any significant clinical or immunological benefit from BCG administration in this patient population with the methodology employed.     

Interferon-alpha plus capecitabine and thalidomide in patients with metastatic renal cell cancer

Thirty patients with progressive metastatic renal cell carcinoma were treated. Eleven patients received 2 or more prior systemic therapies; 2 had a complete response, 7 had a partial response, and 11 had stable disease. The complete responders are off therapy and remain without disease recurrence. The median duration of response was 3.8 months (range 1 - 48+ months). Therapy was well tolerated; predominant toxicities were fatigue, paraesthesias and hand/foot syndrome. The data suggest that the combination of interferon-alpha, thalidomide and capecitabine has anti-tumor activity in previously treated patients with progressive metastatic renal cell carcinoma. A prospective investigation of this combination warranted.     

Soluble immunological parameters and early prognosis of renal cell cancer patients

In local or metastatic cancer, a prognostic tumour marker could be a valuable tool in the selection of different treatments. In renal cell cancer (RCC) no such markers have been available. We therefore evaluated the association between several pretreatment serum markers, tumour classification and short term survival in RCC patients. Serum samples were collected before surgery and three months thereafter from 24 RCC patients. Interleukin-6 (IL-6), IL- 12, soluble IL-2 receptor (sIL-2R) and intercellular adhesion molecule-1 (sICAM-1) were measured in serum samples using specific commercial enzyme immunoassay kits. Serum IL-6, sIL-2R and sICAM-1 levels before nephrectomy were significantly higher in non-local tumours than in local ones (mean IL-6 53 pg/ml versus 6.3 pg/ml, and sICAM-1 443 ng/ml versus 290 ng/ml, sIL-2R 3779 pg/ml versus 1796 pg/ml). In contrast, IL-12 levels were higher in local tumours (148 versus 102 pg/ml) and the levels increased significantly (P < 0.005) after removal of the primary tumour in patients with local disease. All patients with local tumours had normal IL-6 values, while only one with a non-local tumour had IL-6 levels below 10 pg/ml. In addition, IL-6 and sICAM-1 levels before operation were significantly higher in patients with short (less than one year) survival (p=0.007 to IL-6 and p=0.006 to sICAM-1). In contrast, patients with shorter survival had significantly lower IL-12 (p=0.03) levels. Our findings suggest that RCC induces changes in several immunological parameters. These soluble immunological factors, IL-6, IL-12, sIL-2R and sICAM-1, might have a role as prognostic factors in RCC.     

Hybrid cell vaccination for cancer immune therapy: first clinical trial with metastatic melanoma

Hybrid cell vaccination is a new cancer immune therapy approach that aims at recruiting T cell help for the induction of tumour specific cytolytic immunity. The vaccines are generated by fusion of the patients' tumour cells with allogeneic MHC class II bearing cells to combine the tumour's antigenicity with the immunogenicity of allogeneic MHC molecules. Safety and anti-tumour activity of this treatment were assessed in a clinical trial that has yielded one complete and one partial remission, and 5 cases of stable disease among 16 patients with advanced stage metastatic melanoma. As evidenced by histology, the vaccination induced T cell relocation into tumour nodules. Stable disease could be maintained by repeated booster injections for more than 24 months in some patients. The side effects were minor. Occasional occurrences of vitiligo spots after vaccination were indicative of a restricted therapy induced auto-immune reactivity. The results suggest that hybrid cell vaccination is a safe cancer immune therapy potentially effective for induction of acute anti-tumour response as well as long-term maintenance.     

多西他赛解救治疗紫杉醇耐药的转移性乳腺癌的临床研究

目的 评价多西他赛解救治疗紫杉醇耐药复发转移性乳腺癌的临床疗效.方法 回顾性分析60例多西他赛治疗紫杉醇耐药的复发转移性乳腺癌患者的临床资料,对临床疗效及其影响因素等进行统计分析.结果 全组患者中位年龄51岁.雌/孕激素受体阳性率65%,Her-2状态阳性率28.3%.多西他赛解救治疗紫杉醇耐药的复发转移性乳腺癌患者,客观缓解率为18.3%,临床获益率25.0%,中位PFS 4.0个月(3.38～4.62个月);耐药亚组分析结果显示原发耐药组与继发耐药组的客观缓解率及临床获益率无明显统计学差异(P=1.000;P=0.762),但多西他赛治疗紫杉醇耐药复发转移性乳腺癌继发耐药组的无进展生存期(PFS)要优于原发耐药组(2个月vs 6个月,P=0.008).COX多因素分析显示多西他赛PFS与单位体表面积剂量、年龄、病理类型、Her-2状态、雌/孕激素受体表达状态、同侧腋窝淋巴结转移数目、复发转移部位数目等因素均无关,但与耐药情况、无病生存期、解救治疗线数、临床分期(P=0.001;P=0.029;P=0.037;P=0.034)等因素相关.结论 紫杉醇耐药患者接受多西他赛解救治疗仍可获得一定的客观缓解率及临床获益率,二者部分交叉耐药.     

Feasibility study of personalized peptide vaccination for metastatic recurrent triple-negative breast cancer patients

Introduction

Since treatment modalities for metastatic recurrent triple-negative breast cancer (mrTNBC) are limited, a novel treatment approach including immunotherapy is required. We have developed a novel regimen of personalized peptide vaccination (PPV), in which vaccine antigens are individually selected from a pool of different peptide candidates based on the pre-existing host immunity. Herein we conducted a phase II study of PPV for metastatic recurrent breast cancer patients to investigate the feasibility of PPV for mrTNBC.

Methods

Seventy-nine patients with metastatic recurrent breast cancer who had metastases and had failed standard chemotherapy and/or hormonal therapy were enrolled. They were subgrouped as the mrTNBC group (n = 18), the luminal/human epidermal growth factor receptor 2 (HER2)-negative group (n = 41) and the HER2-positive group (n = 18), while the remaining two patients had not been investigated. A maximum of four human leukocyte antigen (HLA)-matched peptides showing higher peptide-specific immunoglobulin G (IgG) responses in pre-vaccination plasma were selected from 31 pooled peptide candidates applicable for the four HLA-IA phenotypes (HLA-A2, -A24, or -A26 types, or HLA-A3 supertypes), and were subcutaneously administered weekly for 6 weeks and bi-weekly thereafter. Measurement of peptide-specific cytotoxic T lymphocyte (CTL) and IgG responses along with other laboratory analyses were conducted before and after vaccination.

Results

No severe adverse events associated with PPV were observed in any of the enrolled patients. Boosting of CTL and/or IgG responses was observed in most of the patients after vaccination, irrespective of the breast cancer subtypes. There were three complete response cases (1 mrTNBC and 2 luminal/HER2-negative types) and six partial response cases (1 mrTNBC and 5 luminal/HER2-negative types). The median progression-free survival time and median overall survival time of mrTNBC patients were 7.5 and 11.1 months, while those of luminal/HER2-negative patients were 12.2 and 26.5 months, and those of HER2-positive patients were 4.5 and 14.9 months, respectively.

Conclusions

PPV could be feasible for mrTNBC patients because of the safety, immune responses, and possible clinical benefits.

Clinical Trial Registration Number

UMIN000001844 (Registration Date: April 5, 2009)     

Aminoglutethimide in patients with metastatic breast cancer

Aminoglutethimide (AG) was administered as palliative therapy in 112 patients with metastatic breast cancer. In 36 patients, the dose level was 1000 mg/day; 76 patients received a dose level of 500 mg/day. Patients with brain or liver metastasis were excluded, as were patients with tumors determined to be negative for estrogen receptors. Objective regression was observed in 35 (31%) patients, with the duration of response ranging from 4 to 36 + months (mean, 12 months; median, 10 months). Response was observed in 11 of 31 (35%) patients with soft tissue metastasis; 16/59 (27%) patients with osseous metastasis; and 8 of 22 (36%) having visceral metastasis. In 93 patients with positive estrogen receptor (ER), 33 responded (35%), whereas in 19 patients with unknown ER status, two responded (11%). Response to previous treatment with tamoxifen (TAM) had occurred in 31 patients; of these, response to AG was noted in 11 (35%). Of 24 patients failing to respond to prior treatment with tamoxifen, four (17%) responded to subsequent therapy with AG. Thirteen patients had previously received combination chemotherapy, and response to AG was noted in two (15%). The side effects observed in this study included skin rash in ten patients, fever in eight, somnolence in three, weakness and dizziness in one, headache in one, insomnia in one, dyspnea in one, and ataxia in one. Treatment had to be discontinued in eight patients, due to the severity of the side effects. As expected, patients receiving AG at the lower dose level of 500 mg/day experienced fewer and less severe side effects than those treated with the higher dose. The response rate in the 1000 mg/day group was 10/36 (28%) and in the 500 mg/day group, it was 25/76 (33%). The lower dosage was better tolerated without apparent compromise in therapeutic efficacy.     

Phase I clinical study applying autologous immunological effector cells transfected with the interleukin-2 gene in patients with metastatic renal cancer, colorectal cancer and lymphoma

Natural killer-like T lymphocytes termed cytokine-induced killer (CIK) cells have been shown to eradicate established tumours in a severe combined immune deficient (SCID) mouse/human lymphoma model. Recently, we demonstrated that CIK cells transfected with cytokine genes possess an improved proliferation rate and a significantly higher cytotoxic activity as compared to non-transfected cells. Here, in a phase I clinical protocol, autologous CIK cells were generated from peripheral blood obtained by leukapheresis in patients with metastatic renal cell carcinoma, colorectal carcinoma and lymphoma. CIK cells were transfected with a plasmid containing the interleukin-2 (IL-2) gene via electroporation. Transfected cells generated IL-2 in the range of 330-1800 pg 10(-6) cells 24 h(-1) with a mean of 836 pg 10(-6) cells 24 h(-1). Ten patients received 1-5 intravenous infusions of IL-2-transfected CIK cells; five infusions with transfected CIK cells were given. In addition, the same patients received five infusions with untransfected CIK cells for control reasons. In three patients, WHO grade 2 fever was observed. Based on polymerase chain reaction of peripheral blood transfected cells could be detected for up to 2 weeks after infusion. There was a significant increase in serum levels of interferon gamma (IFN-gamma), granulocyte-macrophage colony-stimulating factor (GM-CSF) and transforming growth factor beta (TGF-beta) during treatment. Interestingly, there was also an increase in CD3+ lymphocytes in the blood of patients during therapy. In accordance, a partial increase in cytotoxic activity in peripheral blood lymphocytes (PBLs) was documented when patient samples before and after therapy were compared. Concerning clinical outcome, six patients remained in progressive disease, three patients showed no change by treatment, and one patient with lymphoma developed a complete response. In conclusion, we were able to demonstrate that CIK cells transfected with the IL-2 gene can be administered without major side-effects and are promising for future therapeutic trials.     

Impaired primary antibody responses after vaccination against hepatitis B in patients with breast cancer

Summary The response to hepatitis B (HBV) vaccination was studied in 32 breast cancer patients who were HBV antigen- and antibody-negative at the time of diagnosis. Ten of the patients included into the study received radical mastectomy without adjuvant therapy, 15 patients underwent breast-conserving surgery with subsequent irradiation with or without tamoxifen treatment, and 7 patients were treated by modified radical mastectomy plus adjuvant chemotherapy. Eight patients received the first dose of vaccine 3 months after irradiation therapy. Seven patients were first vaccinated 6 months following irradiation therapy. Seven healthy age-matched persons were vaccinated as controls. Antibody response in the patients was significantly delayed. Four weeks after the second vaccination 6 out of 7 controls showed a significant (>10IU/ml) HBs antibody titer, while only 6 out of 32 patients responded. In addition, HBs antibody titers were also significantly lower after the second vaccination even in those patients treated with surgery alone, although they were first vaccinated 3 months after surgery and had no residual tumors. This difference disappeared after the second booster immunization, at which time the frequency of significant antibody titers and the levels of HBs antibody titers were comparable between patients and controls. As the population of patients undergoing treatment for breast cancer is increasing, additional studies are needed to determine optimal immunization regimens in this group of patients.