Melanoma
Conditions
Keywords
Melanoma, Stage II-IV, Immunotherapy, Vaccination, HLA class I and II tumor-specific peptides, IMP321, Montanide ISA-51
Brief summary
The purpose of this study is to determine whether vaccination with tumor antigenic peptides and both IMP321/LAG-3 and Montanide adjuvants can induce an immune response in melanoma patients and to assess the safety and tolerability of this vaccination. Tumor responses following this vaccination will also be documented.
Detailed description
The primary objective of this study is: * to evaluate melanoma antigen specific immune response induced by this vaccination with tumor antigenic peptides derived from MAGE-A3 (Melanoma Antigen family A3) (MHCI: MAGE-A3.A2 and MHCII: MAGE-A3.DP4), NY-ESO-1 (New York Esophageal squamous cell carcinoma antigen-1), Melan A (analog ELA and native EAA) and NA-17A with IMP321 (ImmuFact)/ LAG-3Ig (Lymphocyte activation gene-3 immunoglobulin-like domains) as adjuvant/immunostimulant, formulated with Montanide ISA-51. * to assess the safety and tolerability of this vaccination The secondary objective is to document tumor responses in patients following this vaccination.
Interventions
BIOLOGICAL2 vaccine injections in 1 limb
Participants receive the vaccine separated in 2 syringes with syringe 1 containing NA-17, MAGE-3.A2 and NY-ESO-1 peptides with IMP321/LAG3 ± Montanide, and syringe 2 containing Melan-A and MAGE-A3-DP4 peptides with IMP321/LAG-3 ± Montanide. The content of each syringe is injected s.c. in the same limb at about 5 cm distance from each other.
BIOLOGICAL2 vaccine injections in distinct limbs
Participants receive the vaccine separated in 2 syringes with syringe 1 containing NA-17, MAGE-3.A2 and NY-ESO-1 peptides with IMP321/LAG3 ± Montanide, and syringe 2 containing Melan-A and MAGE-A3-DP4 peptides with IMP321/LAG-3 ± Montanide.The content of each syringe is injected s.c. in different limbs.
Sponsors
Immutep S.A.S.
Centre Hospitalier Universitaire Vaudois
Study design
Allocation
NON_RANDOMIZED
Intervention model
PARALLEL
Primary purpose
TREATMENT
Masking
NONE
Intervention model description
open label, non comparative study in patients suffering from stage II, III or IV melanoma
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
No
Inclusion criteria
* Histologically confirmed stage II, III or IV melanoma patients. * Tumor expression of Melan-A. * Human leukocyte antigen-A2 (HLA-A2) positive. * Expected survival of at list 3 months. * Karnofsky scale performance status of 70 % or more. * Age ≥ 18 years. * Able to give a written informed consent. * The following laboratory results: Hemoglobin ≥ 100g/L, Neutrophil count ≥ 1.5 x 109/L, Lymphocyte count ≥ 0.5 x 109/L, Platelet count ≥ 100 x 109/L, Serum creatinine ≤ 2 mg/dL (0.18mmol/L), Serum bilirubin ≤ 2mg/dL (0.034mmol/L), Granulocyte count \> 2.5x109/L, Aspartate Amino Transférase (ASAT), Alanine Amino Transferase (ALAT) \< 2.5 x upper limit of normal, Activated Partial Thromboplastin Time (aPTT) within the normal ranges ±25%, Thromplastin (TP) ≥ 80%
Exclusion criteria
* Clinically significant heart disease. * Serious illness, eg. serious infections requiring antibiotics, uncontrolled peptic ulcer, or central nervous system disorders. * History of immunodeficiency disease or autoimmune disease. * Metastatic disease to the central nervous system, unless treated and stable. * Known HIV positivity. * Known seropositivity for hepatitis B surface antigen. * Concomitant treatment with steroids, antihistamine drugs. Topical or inhalation steroids are permitted. * Participation in any other clinical trial involving another investigational agent within 4 weeks prior to enrollment. * Pregnancy or lactation. * Women of childbearing potential not using a medically acceptable means of contraception. * Psychiatric or addictive disorders that may compromise the ability to give informed consent. * Lack of availability of the patient for immunological and clinical follow-up assessment. * Coagulation or bleeding disorders. * Kidney dysfunction with creatinine \> 2 X the upper limit of the normal value. * Reported strong (allergic) reactions to previous vaccination.
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | MAGE A3.DP4 specific CD4+ T-cells producing TNF-α were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3). | The activation of peptide-specific CD4+ T cells was analyzed in vitro before and after vaccination by ICS. From each patient, total CD4+ T-cells were stimulated in the presence of peptide MAGE-A3.DP4 LP (MAGE-A3243-258 peptide presented by autologous cells). After 10 days, cell cultures were challenged for 4h with the peptide or left unchallenged. Specific CD4+ T cells responses were identified via detection of TNF-α producing cells. |
| Ex Vivo Frequency of Melan-A Specific CD8+T Cells | Melan-A specific CD8+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3). | Cellular immunity was evaluated through the activation and the expansion of Melan-A-specific CD8+ cytotoxic T lymphocytes. Their frequency was measured in the peripheral blood mononuclear cells (PBMC) directly ex vivo (i.e. without prior in vitro expansion) by multicolor flow cytometry with Melan-A ELA tetramers. The fold change for each time point compared to baseline was calculated as: Melan-A-specific CD8+ T cell frequency at the time point/ Melan-A-specific CD8+ T cell frequency at baseline. Significant T cell response is defined by at least 2-fold change of Melan-A-specific CD8+ T cell frequency as compared to pre-immunotherapy. |
| In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | MAGE A3.DP4 specific CD4+ T cells producing IFN-γ were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3). | The activation of peptide-specific CD4+ T cells was analyzed in vitro before and after vaccination by Intracellular Cytokine Staining (ICS). From each patient, total CD4+ T-cells were stimulated in the presence of peptide MAGE-A3.DP4 LP (MAGE-A3243-258 peptide presented by autologous cells). After 10 days, cell cultures were challenged for 4h with the peptide or left unchallenged. Specific CD4+ T cells responses were identified via detection of IFN-γ producing cells. |
| In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | In vitro stimulated NY-EYO-1 specific CD8+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3). | After 12 days of in vitro stimulation with NY-ESO-1 peptide, CD8+ T cells were analyzed by flow cytometry using tetramer staining. The fold change for each time point compared to baseline was calculated as: NY-ESO-1-specific CD8+ T cell frequency at the time point/ NY-ESO-1-specific CD8+ T cell frequency at baseline. |
| In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | MAGE A3.DP4 specific CD4+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40) and Follow-up (6 to 18 months after the end of Cycle 3). | Frequencies of specific MAGE-A3.DP4-specific CD4+ T cells were quantified by flow cytometry using class II tetramers after 10 days of in vitro stimulation. The fold change for each time point compared to baseline was calculated as: MAGE-A3.DP4-specific CD4+ T cell frequency at the time point/ MAGE-A3.DP4-specific CD4+ T cell frequency at baseline. |
| In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | In vitro stimulated Melan-A specific CD8+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40) and Follow-up (6 to 18 months after the end of Cycle 3). | After 12 days of in vitro stimulation with Melan-A peptides, CD8+ T cells were analyzed by flow cytometry using tetramer staining. The fold change for each time point compared to baseline was calculated as: Melan-A-specific CD8+ T cell frequency at the time point/ Melan-A-specific CD8+ T cell frequency at baseline. Significant T cell response is defined by at least 2-fold change of Melan-A-specific CD8+ T cell frequency as compared to pre-immunotherapy. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Tumor Response | Change from baseline in tumor response at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25) and end of Cycle 3 (Week 40) | The assessment of the baseline disease status was performed, using CT (Computed Tomography) scan or PET (Positron Emission Tomography)/ CT scan, at screening visit or within 8 weeks preceding the screening visit. Imagery examinations occurred after the end of each vaccination cycle. The tumor response was assessed according to the classification World Health Organization (WHO) 1979 and defined as: * No evidence of disease (NED), * Stable disease (SD): change in size of all measurable lesions (the sum of the products of the greatest and perpendicular parameters), of less than a 25% increase or 25% decrease from baseline for at least 4 weeks, without appearance of new lesions or progression of any lesion. * Progressive disease (PD): appearance of new tumors, or increase in size of any measurable tumor by at least 25% of the sum of the product of the greatest and perpendicular diameter. |
Countries
Switzerland
Participant flow
Recruitment details
16 patients were selected between June 15, 2010 and January 24, 2013. Among them 10 patients were included in the group 1 and 6 in the group 2. Since January 2013, 11 patients have been prescreened but could not be included in the study. On April 15, 2014, the decision had to taken to abort the study (16 patients included, while 28 were planned).
Pre-assignment details
An unexplained high percentage, i.e. about 90% of the 11 prescreened patients were HLA-A2 negative. We assume that this was due to the increasing ethnic diversity of patients. The consequence of the time delay was that some of the peptides expired definitively.
Participants by arm
| Arm | Count |
|---|---|
| Group 1: 2 Vaccine Injections in 1 Limb 9 patients initially planned: patients received peptides with IMP321/LAG-3Ig and Montanide in 2 injections sites at 5 cm distance from each other 2 vaccine injections in same limb (vaccine 1 : NY-ESO-1, MAGE-3.A2, NA-17 peptides + IMP321 + Montanide)(vaccine 2 : Melan-A, MAGE-A3-DP4 peptides + IMP321 + Montanide)
2 vaccine injections in 1 limb: Participants receive the vaccine separated in 2 syringes with syringe 1 containing NA-17, MAGE-3.A2 and NY-ESO-1 peptides with IMP321/LAG3 ± Montanide, and syringe 2 containing Melan-A and MAGE-A3-DP4 peptides with IMP321/LAG-3 ± Montanide. The content of each syringe is injected s.c. in the same limb at about 5 cm distance from each other. | 10 |
| Group 2: 2 Injections in Different Limbs 9 patients initially planned: patients received the same vaccine in 2 syringes injected each in a distinct limb 2 vaccine injections in different limbs (vaccine 1: NY-ESO-1, MAGE-3.A2, NA-17 peptides + IMP321 + Montanide) (vaccine 2: Melan-A, MAGE-A3-DP4 peptides + IMP321 + Montanide)
2 vaccine injections in different limbs: Participants receive the vaccine separated in 2 syringes with syringe 1 containing NA-17, MAGE-3.A2 and NY-ESO-1 peptides with IMP321/LAG3 ± Montanide, and syringe 2 containing Melan-A and MAGE-A3-DP4 peptides with IMP321/LAG-3 ± Montanide.The content of each syringe is injected s.c. in different limbs. | 6 |
| Group 3: 2 Injections in Different Limbs 9 patients initially planned: due to premature trial termination, no patients could be enrolled in this last group. 2 vaccine injections in different limbs; patients of this group should have received the same vaccine but without the MHC class II peptide (MAGE-A3 LP) in 2 syringes injected each in a distinct limb.
2 vaccine injections without MHC class II peptide in different limbs: Participants receive the vaccine separated in 2 syringes with syringe 1 containing NA-17, MAGE-3.A2 and NY-ESO-1 peptides with IMP321/LAG3 ± Montanide, and syringe 2 containing Melan-A peptide with IMP321/LAG-3 ± Montanide. | 0 |
| Total | 16 |
Baseline characteristics
| Characteristic | Total | Group 1: 2 Vaccine Injections in 1 Limb | Group 2: 2 Injections in Different Limbs | Group 3: 2 Injections in Different Limbs |
|---|---|---|---|---|
| Age, Categorical <=18 years | 0 Participants | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical >=65 years | 6 Participants | 5 Participants | 1 Participants | 0 Participants |
| Age, Categorical Between 18 and 65 years | 10 Participants | 5 Participants | 5 Participants | 0 Participants |
| Age, Continuous | 62.7 years | 65.3 years | 50.2 years | — |
| Region of Enrollment Switzerland | 16 participants | 10 participants | 6 participants | — |
| Sex: Female, Male Female | 8 Participants | 3 Participants | 5 Participants | — |
| Sex: Female, Male Male | 8 Participants | 7 Participants | 1 Participants | — |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk | EG002 affected / at risk |
|---|---|---|---|
| deaths Total, all-cause mortality | 0 / 10 | 0 / 6 | 0 / 0 |
| other Total, other adverse events | 10 / 10 | 6 / 6 | 0 / 0 |
| serious Total, serious adverse events | 1 / 10 | 0 / 6 | 0 / 0 |
Outcome results
Primary
Ex Vivo Frequency of Melan-A Specific CD8+T Cells
Cellular immunity was evaluated through the activation and the expansion of Melan-A-specific CD8+ cytotoxic T lymphocytes. Their frequency was measured in the peripheral blood mononuclear cells (PBMC) directly ex vivo (i.e. without prior in vitro expansion) by multicolor flow cytometry with Melan-A ELA tetramers. The fold change for each time point compared to baseline was calculated as: Melan-A-specific CD8+ T cell frequency at the time point/ Melan-A-specific CD8+ T cell frequency at baseline. Significant T cell response is defined by at least 2-fold change of Melan-A-specific CD8+ T cell frequency as compared to pre-immunotherapy.
Time frame: Melan-A specific CD8+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3).
Population: No patients were included in the group 3
| Arm | Measure | Group | Value (MEDIAN) | Dispersion |
|---|---|---|---|---|
| Group 1 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | End cycle 2 | 1.09 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 18.21 |
| Group 1 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | Follow up | 1.00 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 2.14 |
| Group 1 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | End cycle 3 | 0.91 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 5.77 |
| Group 1 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | Max | 1.21 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 18.21 |
| Group 1 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | End cycle 1 | 1.21 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 0.47 |
| Group 2 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | Max | 1.83 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 2.14 |
| Group 2 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | End cycle 1 | 1.25 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 1.44 |
| Group 2 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | End cycle 2 | 1.67 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 2.14 |
| Group 2 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | End cycle 3 | 1.83 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 1.79 |
| Group 2 | Ex Vivo Frequency of Melan-A Specific CD8+T Cells | Follow up | 0.28 Fold change of % Melan-A CD8+T ex vivo | Standard Deviation 0.39 |
Primary
In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells
Frequencies of specific MAGE-A3.DP4-specific CD4+ T cells were quantified by flow cytometry using class II tetramers after 10 days of in vitro stimulation. The fold change for each time point compared to baseline was calculated as: MAGE-A3.DP4-specific CD4+ T cell frequency at the time point/ MAGE-A3.DP4-specific CD4+ T cell frequency at baseline.
Time frame: MAGE A3.DP4 specific CD4+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40) and Follow-up (6 to 18 months after the end of Cycle 3).
Population: No patients were included in group 3
| Arm | Measure | Group | Value (MEDIAN) | Dispersion |
|---|---|---|---|---|
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | End of cycle 2 | 0.88 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 1.79 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | Follow up | 0.63 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 2.63 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | End of cycle 3 | 1.57 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 1.64 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | Max | 1.96 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 2.67 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | End cycle 1 | 0.19 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 2.02 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | Max | 1.28 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 1.69 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | End cycle 1 | 0.22 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 0.17 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | End of cycle 2 | 1.97 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 1.77 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | End of cycle 3 | 0.66 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 0.61 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells | Follow up | 0.62 Fold change of % MageA3.DP4 CD4+T | Standard Deviation 0.32 |
Primary
In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ)
The activation of peptide-specific CD4+ T cells was analyzed in vitro before and after vaccination by Intracellular Cytokine Staining (ICS). From each patient, total CD4+ T-cells were stimulated in the presence of peptide MAGE-A3.DP4 LP (MAGE-A3243-258 peptide presented by autologous cells). After 10 days, cell cultures were challenged for 4h with the peptide or left unchallenged. Specific CD4+ T cells responses were identified via detection of IFN-γ producing cells.
Time frame: MAGE A3.DP4 specific CD4+ T cells producing IFN-γ were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3).
Population: No patients were included in the group 3
| Arm | Measure | Group | Value (MEDIAN) | Dispersion |
|---|---|---|---|---|
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | End of cycle 1 | 0.47 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 1.05 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | End of cycle 3 | 2.00 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 1.74 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | End of cycle 2 | 1.11 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 1.06 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | Follow up | 1.46 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 1.97 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | Baseline | 0 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 0 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | Follow up | 0.77 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 0.91 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | Baseline | 0 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 0 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | End of cycle 1 | 1.89 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 2.23 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | End of cycle 2 | 1.63 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 1.6 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Interferon-gamma (IFN-γ) | End of cycle 3 | 0.29 % MAGE-A3.DP4 specific CD4+T IFN-γ | Standard Deviation 0.91 |
Primary
In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α)
The activation of peptide-specific CD4+ T cells was analyzed in vitro before and after vaccination by ICS. From each patient, total CD4+ T-cells were stimulated in the presence of peptide MAGE-A3.DP4 LP (MAGE-A3243-258 peptide presented by autologous cells). After 10 days, cell cultures were challenged for 4h with the peptide or left unchallenged. Specific CD4+ T cells responses were identified via detection of TNF-α producing cells.
Time frame: MAGE A3.DP4 specific CD4+ T-cells producing TNF-α were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3).
Population: No patients were included in the group 3
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | End of cycle 1 | 0.70 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 2.33 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | End of cycle 3 | 1.63 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 3.9 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | End of cycle 2 | 1.05 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 2.74 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | Follow up | 2.04 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 1.57 |
| Group 1 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | Baseline | 0.01 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 0.01 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | Follow up | 3.63 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 3.06 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | Baseline | 0 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 0 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | End of cycle 1 | 3.39 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 2.35 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | End of cycle 2 | 2.31 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 2.04 |
| Group 2 | In Vitro Frequency of MAGE A3.DP4-specific CD4+ T Cells Producing Tumor Necrosis Factor-alpha (TNF-α) | End of cycle 3 | 0.60 % MAGE A3.DP4 specific CD4+T TNFα | Standard Deviation 0.8 |
Primary
In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation
After 12 days of in vitro stimulation with Melan-A peptides, CD8+ T cells were analyzed by flow cytometry using tetramer staining. The fold change for each time point compared to baseline was calculated as: Melan-A-specific CD8+ T cell frequency at the time point/ Melan-A-specific CD8+ T cell frequency at baseline. Significant T cell response is defined by at least 2-fold change of Melan-A-specific CD8+ T cell frequency as compared to pre-immunotherapy.
Time frame: In vitro stimulated Melan-A specific CD8+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40) and Follow-up (6 to 18 months after the end of Cycle 3).
Population: No patients were included in group 3
| Arm | Measure | Group | Value (MEDIAN) | Dispersion |
|---|---|---|---|---|
| Group 1 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | End cycle 2 | 1.10 Fold change of % Melan-A CD8+T cells | Standard Deviation 4.61 |
| Group 1 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | Follow up | 2.83 Fold change of % Melan-A CD8+T cells | Standard Deviation 16.73 |
| Group 1 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | End cycle 3 | 0.69 Fold change of % Melan-A CD8+T cells | Standard Deviation 9.57 |
| Group 1 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | Max | 2.83 Fold change of % Melan-A CD8+T cells | Standard Deviation 16.73 |
| Group 1 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | End cycle 1 | 0.97 Fold change of % Melan-A CD8+T cells | Standard Deviation 3.75 |
| Group 2 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | Max | 5.33 Fold change of % Melan-A CD8+T cells | Standard Deviation 44.83 |
| Group 2 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | End cycle 1 | 2.60 Fold change of % Melan-A CD8+T cells | Standard Deviation 1.84 |
| Group 2 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | End cycle 2 | 5.33 Fold change of % Melan-A CD8+T cells | Standard Deviation 11.02 |
| Group 2 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | End cycle 3 | 2.84 Fold change of % Melan-A CD8+T cells | Standard Deviation 44.83 |
| Group 2 | In Vitro Frequency of Melan-A-specific CD8+T Cells After Stimulation | Follow up | 0 Fold change of % Melan-A CD8+T cells | Standard Deviation 0 |
Primary
In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells
After 12 days of in vitro stimulation with NY-ESO-1 peptide, CD8+ T cells were analyzed by flow cytometry using tetramer staining. The fold change for each time point compared to baseline was calculated as: NY-ESO-1-specific CD8+ T cell frequency at the time point/ NY-ESO-1-specific CD8+ T cell frequency at baseline.
Time frame: In vitro stimulated NY-EYO-1 specific CD8+ T cells were measured in PBMC collected at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25), end of Cycle 3 (Week 40), and Follow-up (6 to 18 months after the end of Cycle 3).
Population: No patients were included in the group 3
| Arm | Measure | Group | Value (MEDIAN) | Dispersion |
|---|---|---|---|---|
| Group 1 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | End cycle 3 | 0.005 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.23 |
| Group 1 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | Follow up | 0.02 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.64 |
| Group 1 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | End cycle 2 | 0 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.3 |
| Group 1 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | Max | 0.02 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.64 |
| Group 1 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | End cycle 1 | 0.01 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.45 |
| Group 2 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | Max | 0.06 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.95 |
| Group 2 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | End cycle 1 | 0 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0 |
| Group 2 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | End cycle 2 | 0 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.2 |
| Group 2 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | Follow up | 0 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0 |
| Group 2 | In Vitro Frequency of NY-ESO-1-specific CD8+ T Cells | End cycle 3 | 0.06 Fold change of % NY-ESO-1 CD8+T cells | Standard Deviation 0.95 |
Secondary
Tumor Response
The assessment of the baseline disease status was performed, using CT (Computed Tomography) scan or PET (Positron Emission Tomography)/ CT scan, at screening visit or within 8 weeks preceding the screening visit. Imagery examinations occurred after the end of each vaccination cycle. The tumor response was assessed according to the classification World Health Organization (WHO) 1979 and defined as: * No evidence of disease (NED), * Stable disease (SD): change in size of all measurable lesions (the sum of the products of the greatest and perpendicular parameters), of less than a 25% increase or 25% decrease from baseline for at least 4 weeks, without appearance of new lesions or progression of any lesion. * Progressive disease (PD): appearance of new tumors, or increase in size of any measurable tumor by at least 25% of the sum of the product of the greatest and perpendicular diameter.
Time frame: Change from baseline in tumor response at the end of Cycle 1 (Week 7), end of Cycle 2 (Week 25) and end of Cycle 3 (Week 40)
Population: No patients were included in the group 3
| Arm | Measure | Group | Category | Value (COUNT_OF_PARTICIPANTS) |
|---|---|---|---|---|
| Group 1 | Tumor Response | Disease status at cycle 2 end | NED | 8 Participants |
| Group 1 | Tumor Response | Disease status at cycle 2 end | PD | 2 Participants |
| Group 1 | Tumor Response | Disease status at cycle 1 end | SD | 0 Participants |
| Group 1 | Tumor Response | Disease status at cycle 3 end | NED | 8 Participants |
| Group 1 | Tumor Response | Disease status at cycle 1 end | PD | 3 Participants |
| Group 1 | Tumor Response | Disease status at cycle 3 end | SD | 0 Participants |
| Group 1 | Tumor Response | Disease status at cycle 2 end | SD | 0 Participants |
| Group 1 | Tumor Response | Disease status at cycle 3 end | PD | 2 Participants |
| Group 1 | Tumor Response | Disease status at cycle 1 end | NED | 7 Participants |
| Group 2 | Tumor Response | Disease status at cycle 3 end | PD | 0 Participants |
| Group 2 | Tumor Response | Disease status at cycle 1 end | NED | 5 Participants |
| Group 2 | Tumor Response | Disease status at cycle 1 end | SD | 0 Participants |
| Group 2 | Tumor Response | Disease status at cycle 1 end | PD | 1 Participants |
| Group 2 | Tumor Response | Disease status at cycle 2 end | NED | 4 Participants |
| Group 2 | Tumor Response | Disease status at cycle 2 end | SD | 0 Participants |
| Group 2 | Tumor Response | Disease status at cycle 2 end | PD | 1 Participants |
| Group 2 | Tumor Response | Disease status at cycle 3 end | NED | 4 Participants |
| Group 2 | Tumor Response | Disease status at cycle 3 end | SD | 0 Participants |