RESUMO
This review describes clear parameters for designating the correct use of the term Tumor Rejection Antigen [TRA] to define the role of tumor cell constituents which activate adaptive anti-tumor immune reactions in the cancer-bearing host. This is important, especially in defining immunogens which activate the patient's cytotoxic T-cells that are important to immunotherapeutic applications in human cancer treatment. The focus of the review is to correctly delineate the immunogenic properties of 37 kDa oncofetal antigen [OFA], one of only a few true TRAs expressed on human and experimental rodent cancers. The purpose of this review is to provide a background for publication reviewers, journal and text editors, and scientists reporting on TRAs to avoid creating further confusion that has proliferated in the cancer literature to imply traits of so-called tumor-associated antigens that do not qualify as TRAs.
Assuntos
Antígenos de Neoplasias/imunologia , Biomarcadores Tumorais/imunologia , Neoplasias/imunologia , Animais , Humanos , Imunoterapia/métodos , Linfócitos T Citotóxicos/imunologia , Terminologia como AssuntoRESUMO
PURPOSE: We wanted to evaluate feasibility and safety of dendritic cell-based immunotherapy in patients with metastatic renal cell carcinoma (RCC). EXPERIMENTAL DESIGN: Patients with metastatic RCC (n = 35) received vaccinations (i.v. or i.d.) of CD83+ autologous monocyte-derived dendritic cells (moDCs). MoDCs were loaded with lysate of cultured autologous or allogeneic permanent tumor cells (A-498) as well as keyhole limpet hemocyanin as control and helper antigen. Maturation of moDCs was induced by a combination of tumor necrosis factor alpha, interleukin 1 beta, interleukin 6, and prostaglandin E2. RESULTS: Treatment was associated with transient flu-like symptoms. In 2 of 27 evaluable patients, any evidence of disease disappeared (complete response). In both cases, metastatic tissue had been the source of tumor antigen. One patient had an objective partial response. Seven patients had stable disease, the remaining 17 patients had progressive disease. In 11 of 11 patients evaluated, moDCs induced strong immune responses against keyhole limpet hemocyanin. In 5 of 6 patients tested, enhanced immune responses against oncofetal antigen (immature laminin receptor; OFA/LRP) could also be detected. The strongest responses against OFA/LRP were detectable in 2 patients with complete response and partial response, respectively. At the time of submission, mean follow up is 32 months and 8 patients are currently alive. CONCLUSIONS: Our data indicate that moDC-based vaccination is well tolerated and has immunological as well as clinical effects in patients with metastatic RCC. OFA/LRP might be an attractive candidate antigen for DC-based immunotherapy of RCC.
Assuntos
Carcinoma de Células Renais/terapia , Células Dendríticas/metabolismo , Imunoterapia , Neoplasias Renais/terapia , Transplante Autólogo , Adulto , Idoso , Antígenos CD , Carcinoma de Células Renais/patologia , Divisão Celular , Citocinas/biossíntese , Dinoprostona/metabolismo , Feminino , Hemocianinas/metabolismo , Humanos , Imunoglobulinas/biossíntese , Imuno-Histoquímica , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Neoplasias Renais/patologia , Masculino , Glicoproteínas de Membrana/biossíntese , Pessoa de Meia-Idade , Monócitos/metabolismo , Metástase Neoplásica , Resultado do Tratamento , Células Tumorais Cultivadas , Fator de Necrose Tumoral alfa/metabolismo , Antígeno CD83Assuntos
Antígenos de Neoplasias/imunologia , Carcinoma de Células Renais/imunologia , Neoplasias Renais/imunologia , Precursores de Proteínas/imunologia , Receptores de Laminina/imunologia , Carcinoma de Células Renais/terapia , Células Dendríticas/imunologia , Feminino , Humanos , Neoplasias Renais/terapiaRESUMO
The immature laminin receptor (iLR) is a tumor-associated antigen. We analyzed the expression of iLR on malignant B cells of 134 unselected patient samples with CLL and hypothesized that iLR expression would have prognostic significance due to a differential expression pattern. High ILR expression (cut-off value 30%) was correlated with mutated IGVH status (p<0.0001). Patients with high iLR-expression had a significantly longer time to progression (p=0.039). Combination of CD38, ZAP-70, and iLR by flow cytometry can be used to construct a diagnostic score identifying patients with a median progression free survival of 80 months, if no adverse marker is present.
Assuntos
Cadeias Pesadas de Imunoglobulinas/genética , Região Variável de Imunoglobulina/genética , Leucemia Linfocítica Crônica de Células B/diagnóstico , Mutação , Receptores de Laminina/metabolismo , Proteínas Ribossômicas/metabolismo , ADP-Ribosil Ciclase 1/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Western Blotting , Células Cultivadas , Intervalo Livre de Doença , Feminino , Citometria de Fluxo , Humanos , Leucemia Linfocítica Crônica de Células B/genética , Leucemia Linfocítica Crônica de Células B/metabolismo , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Valor Preditivo dos Testes , Prognóstico , Proteína-Tirosina Quinase ZAP-70/metabolismoRESUMO
We describe here for the first time an efficient high yield production method for clinical grade recombinant human Oncofetal Antigen/immature laminin receptor protein (OFA/iLRP). We also demonstrate significant antitumor activity for this protein when administered in liposomal delivery form in a murine model of syngeneic fibrosarcoma. OFA/iLRP is a therapeutically very promising universal tumor antigen that is expressed in all mammalian solid tumors tested so far. We have cloned the human OFA/iLRP cDNA in a bacterial expression plasmid which incorporates a 6x HIS-tag. Large scale cultures of the plasmid transformed Escherichia coli were performed and the crude HIS-tagged OFA/iLRP was isolated as inclusion bodies and solubilized in guanidine chloride. The protein was then purified by successive passage through three column chromatography steps of immobilized metal affinity, anion exchange, and gel filtration. The resulting protein was 94% pure and practically devoid of endotoxin and host cell protein. The purified OFA/iLRP was tested in mice for safety and efficacy in tumor rejection with satisfactory results. This protein will be used for loading onto autologous dendritic cells in an FDA approved phase I/II human cancer vaccine trial in OFA/iLRP-positive breast cancer patients.
Assuntos
Antígenos de Neoplasias/imunologia , Vacinas Anticâncer/imunologia , Fibrossarcoma/imunologia , Receptores de Laminina/imunologia , Proteínas Ribossômicas/imunologia , Sequência de Aminoácidos , Animais , Antígenos de Neoplasias/química , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/isolamento & purificação , Vacinas Anticâncer/efeitos adversos , Vacinas Anticâncer/metabolismo , Linhagem Celular Tumoral/imunologia , Ensaios Clínicos como Assunto , Clonagem Molecular , Modelos Animais de Doenças , Escherichia coli/genética , Feminino , Fibrossarcoma/metabolismo , Guanidina/farmacologia , Histidina/química , Humanos , Corpos de Inclusão/imunologia , Corpos de Inclusão/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Peso Molecular , Plasmídeos/genética , Controle de Qualidade , Receptores de Laminina/química , Receptores de Laminina/genética , Receptores de Laminina/isolamento & purificação , Proteínas Recombinantes/química , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Proteínas Ribossômicas/química , Proteínas Ribossômicas/genética , Proteínas Ribossômicas/isolamento & purificação , Solubilidade , Transformação Bacteriana , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Chronic lymphocytic leukemia (CLL) is characterized by a highly variable clinical course. The role of an autologous tumor-specific immune control contributing to the variable length of survival in CLL is poorly understood. We investigated whether humoral immunity specific for the CLL-associated Ag oncofetal Ag/immature laminin receptor (OFA/iLR) has a prognostic value in CLL. Among sera of 67 untreated patients with CLL, 23 (34.3%) had detectable OFA/iLR Abs that were reactive for at least one specific OFA/iLR epitope. Patients with humoral responses compared with patients with nonreactive sera had a longer progression-free survival (p = 0.029). IgG subclass analyses showed a predominant IgG1 and IgG3 response. OFA/iLR Abs were capable of recognizing and selectively killing OFA/iLR-expressing CLL cells in complement-mediated and Ab-dependent cellular cytotoxicity assays. In the analysis of 11 CLL patients after allogeneic hematopoietic stem cell transplantation, 8 showed high values for OFA/iLR Abs that specifically recognized the extracellular domain of the protein, suggesting a potential role of anti-OFA/iLR-directed immune responses to the graft-vs-leukemia effect in CLL. Our data suggest that spontaneous tumor-specific humoral immune responses against OFA/iLR exist in a significant proportion of CLL patients and that superior progression-free survival in those patients could reflect autologous immune control.
Assuntos
Anticorpos Antineoplásicos/sangue , Formação de Anticorpos , Antígenos de Neoplasias/imunologia , Efeito Enxerto vs Leucemia/imunologia , Imunoglobulina G/sangue , Leucemia Linfocítica Crônica de Células B/sangue , Receptores de Laminina/imunologia , Proteínas Ribossômicas/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antineoplásicos/imunologia , Especificidade de Anticorpos/imunologia , Intervalo Livre de Doença , Feminino , Transplante de Células-Tronco Hematopoéticas , Humanos , Imunoglobulina G/imunologia , Leucemia Linfocítica Crônica de Células B/imunologia , Leucemia Linfocítica Crônica de Células B/mortalidade , Leucemia Linfocítica Crônica de Células B/terapia , Masculino , Pessoa de Meia-Idade , Taxa de Sobrevida , Transplante HomólogoRESUMO
During tumor development in mice and humans, oncofetal Ag/immature laminin receptor (OFA/iLRP)-specific Th1, CTL, and IL-10-secreting T (Ts) cells are induced. The presence of too many Ts or too few effector T cells appears to predict a poor prognosis. We established clones of OFA/iLRP-specific splenic Th1, CTL, and Ts cells from the OFA/iLRP+ MCA1315 fibrosarcoma-bearing BALB/c mice or from BALB/c mice vaccinated with 1 or 10 microg of rOFA/iLRP. The MCA1315 tumor cell-reactive T cell clones were characterized as to surface Ag phenotype, cytokine secretion profile, and specificity for OFA/iLRP presented by syngeneic splenic APC. OFA/iLRP-specific Th1 and Ts clones were established from all mice. OFA/iLRP-specific CTL could be established from all mice except for mice immunized with 10 microg of rOFA/iLRP. Analysis of the proliferation profile of the OFA/iLRP-specific clones to overlapping OFA/iLRP 12-mer peptides that spanned the OFA/iLRP protein sequence defined the epitopes to which the T cell clones responded. There was a similar spatial distribution of the epitopes to which the two types of CD8 T cell clones responded. The nonapeptide epitopes of the Ts clones were located between aa 36 and 147 of OFA/iLRP, while the epitopes of the CTL clones were located between aa 52 and 163. Even though the CTL and Ts epitopes shared part of the protein, all of the CD8 CTL epitopes were distinct and separable from those of CD8 Ts cells.
Assuntos
Antígenos de Neoplasias/análise , Antígenos de Neoplasias/fisiologia , Linfócitos T CD8-Positivos/imunologia , Epitopos de Linfócito T/fisiologia , Ativação Linfocitária/imunologia , Receptores de Laminina/análise , Receptores de Laminina/fisiologia , Linfócitos T Reguladores/imunologia , Transferência Adotiva , Animais , Anticorpos Monoclonais , Antígenos de Neoplasias/administração & dosagem , Linfócitos T CD8-Positivos/metabolismo , Linhagem Celular Tumoral , Células Clonais , Citocinas/biossíntese , Células Dendríticas/imunologia , Células Dendríticas/transplante , Epitopos de Linfócito T/análise , Feminino , Fibrossarcoma/imunologia , Fibrossarcoma/prevenção & controle , Fibrossarcoma/secundário , Inibidores do Crescimento/administração & dosagem , Inibidores do Crescimento/análise , Inibidores do Crescimento/fisiologia , Antígenos H-2/imunologia , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/prevenção & controle , Neoplasias Pulmonares/secundário , Camundongos , Camundongos Endogâmicos BALB C , Receptores de Laminina/administração & dosagem , Baço/citologia , Baço/imunologia , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismo , Linfócitos T Reguladores/metabolismoRESUMO
The oncofetal Ag immature laminin receptor (OFA-iLR) is a potential target molecule for immunotherapeutic studies in several tumor entities, including hematological malignancies. In the present study, we characterize two HLA-A*0201-presented epitopes eliciting strong OFA-iLR peptide-specific human cytotoxic T cell (CTLs) responses in vitro. Both allogeneic HLA-A*0201-matched and autologous CTLs recognized and killed endogenously OFA-iLR-expressing tumor cell lines and primary malignant cells from patients with hemopoietic malignancies in an MHC-restricted fashion but spared nonmalignant hemopoietic cells. Spontaneous OFA-iLR peptide-specific T cell reactivity was detectable in a significant proportion of leukemia patients. Interestingly, in patients with chronic lymphocytic leukemia and multiple myeloma but not in those with acute myeloid leukemia, significant frequencies of OFA peptide-specific CTLs could be detected in an early stage of disease but disappeared in patients with progressive disease. The identification of OFA-iLR-derived peptide epitopes provides a basis for tumor immunological studies and therapeutic vaccination strategies in patients with OFA-iLR-expressing malignancies.
Assuntos
Apresentação de Antígeno , Antígenos de Neoplasias/metabolismo , Epitopos de Linfócito T/metabolismo , Antígenos HLA-A/metabolismo , Leucemia Linfocítica Crônica de Células B/imunologia , Leucemia Mieloide Aguda/imunologia , Mieloma Múltiplo/imunologia , Receptores de Laminina/metabolismo , Adulto , Idoso , Antígenos de Neoplasias/biossíntese , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/imunologia , Testes Imunológicos de Citotoxicidade , Citotoxicidade Imunológica , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Células Dendríticas/patologia , Epitopos de Linfócito T/imunologia , Feminino , Antígenos HLA-A/imunologia , Antígeno HLA-A2 , Humanos , Células K562 , Leucemia Linfocítica Crônica de Células B/metabolismo , Leucemia Linfocítica Crônica de Células B/patologia , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patologia , Masculino , Pessoa de Meia-Idade , Mieloma Múltiplo/metabolismo , Mieloma Múltiplo/patologia , Fragmentos de Peptídeos/imunologia , Fragmentos de Peptídeos/metabolismo , Ligação Proteica/imunologia , Receptores de Laminina/biossíntese , Receptores de Laminina/genética , Receptores de Laminina/imunologia , Proteínas Ribossômicas , Linfócitos T Citotóxicos/imunologia , Linfócitos T Citotóxicos/metabolismo , Linfócitos T Citotóxicos/patologia , TransfecçãoRESUMO
The oncofetal antigen immature laminin receptor protein (OFA-iLRP) is a highly conserved protein that is preferentially expressed in fetal tissues and in many types of cancer, including hematopoietic malignancies, whereas OFA-iLRP is not detectable on healthy differentiated adult cells. To investigate whether OFA-iLRP-specific cytotoxic T lymphocytes (CTLs) are capable of killing OFA-iLRP-expressing hematologic targets, CTLs were generated from healthy HLA-A*0201-positive volunteers by incubating T cells with autologous dendritic cells (DCs) transfected with OFA-iLRP RNA. OFA-iLRP-specific CTLs lysed HLA-A2+ OFA-iLRP+ tumor cells, including several lymphoma and leukemia cell lines, as well as fresh leukemic targets from patients with acute myeloid leukemia (AML) and chronic lymphatic leukemia (CLL), indicating that OFA-iLRP-derived peptides are naturally processed and presented by hematologic tumors. Healthy OFA-iLRP-negative target cells (CD14+ monocytes, activated B cells, DCs, bone marrow cells) were not attacked by OFA-iLRP-specific CTLs. Furthermore, in an established murine B-cell lymphoma model (A20), treatment with syngeneic DCs transfected with OFA-iLRP-coding RNA resulted in powerful antitumor effects in a significant portion of mice. For the first time, these data show that OFA-iLRP can be used as a target for T-cell-based immunotherapeutic strategies against hematologic malignancies.