RESUMEN
Objectives: Chronic inflammatory demyelinating polyneuropathy (CIDP) is an acquired immune-mediated neuropathy defined by clinical progression for more than 2 months. 16-20% of CIDP patients may present with rapidly progressive weakness that resembles GBS, known as acute-onset CIDP (A-CIDP). However, it is challenging to distinguish from GBS-TRF because of their similar clinical symptom and features. In this case review, we report a patient with A-CIDP with the detection of anti-GM3 and anti-sulfatides antibodies, which rarely have been in A-CIDP and may account for her progressive and recurrent symptoms. Methods: We analyzed existing medical literature and described a clinical case of A-CIDP with antibodies positive. Results: We reported a 56-year-old female presented with bilateral lower extremity weakness and distal numbness. She experienced similar symptoms four times and responded well to the IVIg therapy. Lumbar puncture demonstrated albumin-cytologic dissociation and EDX examination revealed multiple peripheral nerve damage. After ruling out other demyelination diseases, a diagnosis of A-CIDP was made. Discussion: The antiganglioside and anti-sulfatide antibodies are involved in CIDP pathogenesis and can help to distinguish A-CIDP and other variants. To prevent secondary damage, it is important to monitor relapse and remission symptoms along the treatment line. A rare case of A-CIDP is discussed concerning the detection of anti-GM3 and anti-sulfatides antibodies, thus making a retrospective comparison of antibodies in some literature to understand A-CIDP better.
Asunto(s)
Autoanticuerpos , Gangliósido G(M3) , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante , Humanos , Femenino , Persona de Mediana Edad , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/diagnóstico , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/inmunología , Polirradiculoneuropatía Crónica Inflamatoria Desmielinizante/sangre , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Gangliósido G(M3)/inmunología , Gangliósido G(M3)/análogos & derivados , Sulfoglicoesfingolípidos/inmunología , Inmunoglobulinas Intravenosas/uso terapéutico , Diagnóstico DiferencialRESUMEN
Ganglioside GM3 is one of the most common membrane-bound glycosphingolipids. The over-expression of GM3 on tumor cells makes it defined as a tumor-associated carbohydrate antigen (TACA). The specific expression property in cancers, especially in melanoma, make it become an important target to develop anticancer vaccines or immunotherapies. However, in the manner akin to most TACAs, GM3 is an autoantigen facing with problems of low immunogenicity and easily inducing immunotolerance, which means itself only cannot elicit a powerful enough immune response to prevent or treat cancer. With a comparative understanding of the mechanisms that how immune system responses to the carbohydrate vaccines, this review summarizes the studies on the recent efforts to development GM3-based anticancer vaccines.
Asunto(s)
Vacunas contra el Cáncer , Gangliósido G(M3) , Neoplasias , Gangliósido G(M3)/inmunología , Humanos , Vacunas contra el Cáncer/inmunología , Animales , Neoplasias/inmunología , Neoplasias/terapiaRESUMEN
Tumor-associated glycolipids such as NeuGc GM3 are auspicious molecular targets in antineoplastic therapies and vaccine strategies. 14F7 is a monoclonal IgG1 with high clinical potential in cancer immunotherapy as it displays extraordinary specificity for NeuGc GM3, while it does not recognize the very similar, ubiquitous NeuAc GM3. Here we present the 2.3 Å crystal structure of the 14F7 antigen-binding domain (14F7 scFv) in complex with the NeuGc GM3 trisaccharide. Modeling analysis and previous mutagenesis data suggest that 14F7 may also bind to an alternative NeuGc GM3 conformation, not observed in the crystal structure. The most intriguing finding, however, was that a water molecule centrally placed in the complementarity-determining region directly mediates the specificity of 14F7 to NeuGc GM3. This has profound impact on the complexity of engineering in the binding site and provides an excellent example of the importance in understanding the water structure in antibody-antigen interactions.
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Anticuerpos Monoclonales/inmunología , Antineoplásicos/inmunología , Gangliósido G(M3)/inmunología , Agua/química , Anticuerpos Monoclonales/aislamiento & purificación , Especificidad de Anticuerpos , Reacciones Antígeno-Anticuerpo , Gangliósido G(M3)/síntesis química , Gangliósido G(M3)/química , Modelos Moleculares , Estructura MolecularRESUMEN
GM3, a typical tumor-associated carbohydrate antigen, is considered as an important target for cancer vaccine development, but its low immunogenicity limits its application. αGalCer, an iNKT cell agonist, has been employed as an adjuvant via a unique immune mode. Herein, we prepared and investigated two types of antitumor vaccine candidates: (a) self-adjuvanting vaccine GM3-αGalCer by conjugating GM3 with αGalCer and (b) noncovalent vaccine GM3-lipid/αGalCer, in which GM3 is linked with lipid anchor and coassembled with αGalCer. This demonstrated that ßGalCer is an exceptionally optimized lipid anchor, which enables the noncovalent vaccine candidate GM3-ßGalCer/αGalCer to evoke a comparable antibody level to GM3-αGalCer. However, the antibodies induced by GM3-αGalCer are better at recognition B16F10 cancer cells and more effectively activate the complement system. Our study highlights the importance of vaccine constructs utilizing covalent or noncovalent assembly between αGalCer with carbohydrate antigens and choosing an appropriate lipid anchor for use in noncovalent vaccine formulation.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Vacunas contra el Cáncer/farmacología , Gangliósido G(M3)/farmacología , Galactosilceramidas/farmacología , Adyuvantes Inmunológicos/síntesis química , Animales , Vacunas contra el Cáncer/síntesis química , Vacunas contra el Cáncer/inmunología , Secuencia de Carbohidratos , Femenino , Gangliósido G(M3)/análogos & derivados , Gangliósido G(M3)/inmunología , Galactosilceramidas/síntesis química , Galactosilceramidas/inmunología , Humanos , Inmunidad Humoral/efectos de los fármacos , Inmunoglobulina G/inmunología , Liposomas/química , Ratones Endogámicos BALB C , Células T Asesinas Naturales/inmunología , Células THP-1RESUMEN
A 60-year-old woman with a 3-day history of ataxic gait, blurred vision, and upper extremity paresthesia was admitted to our hospital. She presented with severe visual disturbances (finger counting), ophthalmoplegia, neck weakness, and sensory ataxia. Serum anti-GQ1b antibody, anti-GM3 antibody, and anti-GD3 antibody were strongly positive, which might contribute to the pathogenesis. Since we suspected Guillain-Barré syndrome (GBS), intravenous immunoglobulin therapy (IVIg) and high-dose steroid therapy were administered; however, improvements in her visual acuity were minimal. Additional IVIg and high-dose steroid therapy resulted in limited visual acuity improvements. Therapeutic strategies for patients with GBS and refractory optic neuropathy remain controversial.
Asunto(s)
Síndrome de Guillain-Barré/complicaciones , Síndrome de Guillain-Barré/tratamiento farmacológico , Enfermedades del Nervio Óptico/tratamiento farmacológico , Enfermedades del Nervio Óptico/etiología , Autoanticuerpos/sangre , Biomarcadores/sangre , Femenino , Gangliósido G(M3)/inmunología , Gangliósidos/inmunología , Síndrome de Guillain-Barré/diagnóstico , Humanos , Inmunoglobulinas Intravenosas/administración & dosificación , Metilprednisolona/administración & dosificación , Persona de Mediana Edad , Enfermedades del Nervio Óptico/diagnóstico , Quimioterapia por PulsoRESUMEN
The GM3(Neu5Gc) ganglioside represents a tumor-specific antigen that is considered a promising target for cancer immunotherapy. We previously demonstrated that the humanized antibody 14F7hT, specific for this ganglioside, exhibited significant antitumor effects in preclinical hematological tumor models. As this antibody recognizes human tumor tissues from several origins, we addressed its potential effect on different tumor types. The use of cell lines for testing GM3(Neu5Gc)-targeting strategies, in particular for human malignancies, is complicated by the absence in humans of functional cytidine monophospho-N-acetyl-neuraminic acid hydroxylase (CMAH), the enzyme required for Neu5Gc sialic acid biosynthesis. Quantitative flow cytometry revealed the absence of surface GM3(Neu5Gc) in several human but also mouse cell lines, in the last case due to low expression of the enzyme. Hypoxia-induced expression of this ganglioside on human SKOV3 cells was observed upon culture in Neu5Gc-containing medium without evidence for CMAH-independent biosynthesis. However, only transfection of the mouse Cmah gene into human SKOV3 and mouse 3LL cells induced a stable expression of GM3(Neu5Gc) on the cancer cell surface, resulting in effective models to evaluate the antitumor responses by 14F7hT in vitro and in vivo. This antibody exerted antibody-dependent cell-mediated cytotoxicity (ADCC) and in vivo antitumor effects on these Cmah-transfected non-hematological tumors from both mouse and human origin. These results contribute to validate GM3(Neu5Gc) as a relevant target for cancer immunotherapy and reinforces the value of 14F7hT as a novel anti-cancer drug.
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Anticuerpos Monoclonales Humanizados/farmacología , Citotoxicidad Celular Dependiente de Anticuerpos/inmunología , Antineoplásicos/farmacología , Gangliósido G(M3)/inmunología , Oxigenasas de Función Mixta/inmunología , Neoplasias/tratamiento farmacológico , Animales , Antígenos de Neoplasias/inmunología , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Oxigenasas de Función Mixta/química , Neoplasias/inmunología , Neoplasias/patología , Células Tumorales CultivadasAsunto(s)
Anticuerpos/sangre , Gangliósido G(M3)/inmunología , Gangliósidos/inmunología , Síndrome de Miller Fisher/sangre , Síndrome de Miller Fisher/inmunología , Humanos , Inmunoglobulinas Intravenosas/uso terapéutico , Factores Inmunológicos/uso terapéutico , Masculino , Persona de Mediana Edad , Síndrome de Miller Fisher/fisiopatología , Síndrome de Miller Fisher/terapia , Conducción Nerviosa/fisiología , NeuraminidasaRESUMEN
Ganglioside GM3 is strongly related with human tumors, such as lung, brain cancers and melanomas, and more and more evidences have revealed that GM3 possesses powerful effects on cancer development and progression. GM3 is over expressed on several types of cancers, and can be as a tumor-associated carbohydrate antigen, used for immunotherapy of cancers. GM3 can also inhibit tumor cells growth by anti-angiogenesis or motility and so on. Especially, GM3 has effects on the EGFR tyrosine kinase signaling, uPAR-related signaling and glycolipid-enriched microdomains, which are essential for cancer signaling conduction. It is obvious that GM3 will be a promising target for cancer treatment.
Asunto(s)
Gangliósido G(M3)/fisiología , Neoplasias/fisiopatología , Animales , Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/uso terapéutico , Línea Celular Tumoral , Gangliósido G(M3)/inmunología , Humanos , Microdominios de Membrana/fisiología , Neoplasias/terapia , Transducción de Señal/fisiologíaRESUMEN
Numerous molecules have been considered as targets for cancer immunotherapy because of their levels of expression on tumor cells, their putative importance for tumor biology, and relative immunogenicity. In this review we focus on the ganglioside GM3(Neu5Gc), a glycosphingolipid present on the outer side of the plasma membrane of vertebrate cells. The reasons for selecting GM3(Neu5Gc) as a tumor-specific antigen and its use as a target for cancer immunotherapy are discussed, together with the development of antitumor therapies focused on this target by the Center of Molecular Immunology (CIM, Cuba).
Asunto(s)
Anticuerpos Monoclonales/inmunología , Antígenos de Neoplasias/inmunología , Gangliósido G(M3)/inmunología , Neoplasias/inmunología , Animales , Anticuerpos Monoclonales/uso terapéutico , Anticuerpos Monoclonales de Origen Murino , Antígenos de Neoplasias/química , Antígenos de Neoplasias/metabolismo , Secuencia de Carbohidratos , Modelos Animales de Enfermedad , Gangliósido G(M3)/antagonistas & inhibidores , Gangliósido G(M3)/química , Humanos , Inmunoterapia/métodos , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismoRESUMEN
Antitumor strategies based on positive modulation of the immune system currently represent therapeutic options with prominent acceptance for cancer patients' treatment due to its selectivity and higher tolerance compared to chemotherapy. Racotumomab is an anti-idiotype (anti-Id) monoclonal antibody (mAb) directed to NeuGc-containing gangliosides such as NeuGcGM3, a widely reported tumor-specific neoantigen in many human cancers. Racotumomab has been approved in Latin American countries as an active immunotherapy for advanced non-small cell lung cancer (NSCLC) treatment. In this work, we evaluated the induction of Ab-dependent cell-mediated cytotoxicity (ADCC) in NSCLC patients included in a phase III clinical trial, in response to vaccination with racotumomab. The development of anti-NeuGcGM3 antibodies (Abs) in serum samples of immunized patients was first evaluated using the NeuGcGM3-expressing X63 cells, showing that racotumomab vaccination developed antigen-specific Abs that are able to recognize NeuGcGM3 expressed in tumor cell membranes. ADCC response against NeuGcGM3-expressing X63 (target) was observed in racotumomab-treated- but not in control group patients. When target cells were depleted of gangliosides by treatment with a glucosylceramide synthase inhibitor, we observed a significant reduction of the ADCC activity developed by sera from racotumomab-vaccinated patients, suggesting a target-specific response. Our data demonstrate that anti-NeuGcGM3 Abs induced by racotumomab vaccination are able to mediate an antigen-specific ADCC response against tumor cells in NSCLC patients.
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Anticuerpos Monoclonales/farmacología , Citotoxicidad Celular Dependiente de Anticuerpos , Carcinoma de Pulmón de Células no Pequeñas/terapia , Gangliósido G(M3)/análogos & derivados , Inmunoterapia Activa , Neoplasias Pulmonares/terapia , Anticuerpos Monoclonales de Origen Murino , Apoptosis , Carcinoma de Pulmón de Células no Pequeñas/inmunología , Carcinoma de Pulmón de Células no Pequeñas/patología , Gangliósido G(M3)/inmunología , Humanos , Neoplasias Pulmonares/inmunología , Neoplasias Pulmonares/patología , Células Tumorales CultivadasAsunto(s)
Parálisis Bulbar Progresiva/complicaciones , Gangliósido G(M3)/inmunología , Síndrome de Guillain-Barré , Inmunoglobulina G/sangre , Plexo Braquial/fisiopatología , Médula Cervical/fisiopatología , Femenino , Síndrome de Guillain-Barré/sangre , Síndrome de Guillain-Barré/patología , Síndrome de Guillain-Barré/fisiopatología , Humanos , Persona de Mediana Edad , Orofaringe/fisiopatologíaRESUMEN
A murine monoclonal antibody (MAb-1) specific for GM3 has been generated by immunizing ß3Gn-T5 knockout mice with purified GM3 ganglioside. The binding specificity of MAb-1 (IgG3 subclass) was established by an enzyme-linked immunosorbent assay (ELISA) and FACS and the antibody showed high binding specificity with GM3. Cell viability assay showed that MAb-1 significantly suppressed cell growth. Immunohistochemistry analysis revealed that MAb-1 was strongly expressed in human ovarian cancer tissues, whereas it was hardly expressed in normal tissues. Finally, antibody-dependent cellular cytotoxicity (ADCC) activities were determined by measuring lactate dehydrogenase (LDH) releasing assay and the results showed high ADCC activities in two representative ovarian cancer cell lines (OVHM and ID8). All of these data indicate that MAb-1 may be potentially used as a therapeutic antibody against ovarian cancers in clinical trials.
Asunto(s)
Anticuerpos Monoclonales de Origen Murino/inmunología , Gangliósido G(M3)/análogos & derivados , Inmunoglobulina G/inmunología , Neoplasias Ováricas/inmunología , Animales , Anticuerpos Monoclonales de Origen Murino/genética , Especificidad de Anticuerpos , Línea Celular Tumoral , Pruebas Inmunológicas de Citotoxicidad , Femenino , Gangliósido G(M3)/inmunología , Humanos , Inmunoglobulina G/genética , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, ß-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins.
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Diferenciación Celular/inmunología , Glicoesfingolípidos/química , Macrófagos/química , Monocitos/química , Polisacáridos/química , Conformación de Carbohidratos , Secuencia de Carbohidratos , Línea Celular , Fucosiltransferasas/genética , Fucosiltransferasas/inmunología , Gangliósido G(M3)/química , Gangliósido G(M3)/inmunología , Regulación de la Expresión Génica , Glicoesfingolípidos/inmunología , Glicosilación , Glicosiltransferasas/genética , Glicosiltransferasas/inmunología , Humanos , Macrófagos/citología , Macrófagos/inmunología , Manosa/química , Manosa/inmunología , Monocitos/citología , Monocitos/inmunología , N-Acetilglucosaminiltransferasas/genética , N-Acetilglucosaminiltransferasas/inmunología , Neuraminidasa/genética , Neuraminidasa/inmunología , Polisacáridos/inmunología , Ácidos Siálicos/química , Ácidos Siálicos/inmunología , Sialiltransferasas/genética , Sialiltransferasas/inmunologíaRESUMEN
The expression of N-glycolyl-monosialodihexosyl-ganglioside (NGcGM3) in humans is restricted to cancer cells; therefore, it is a tumor antigen. There are measurable quantities of circulating anti-NGcGM3 antibodies (aNGcGM3 Abs) in human serum. Interestingly, some people have circulating Ag-specific immunoglobulins G (IgGs) that are capable of complement mediated cytotoxicity against NGcGM3 positive cells, which is relevant for tumor surveillance. In light of the chemical nature of Ag, we postulated it as a candidate ligand for CD1d. Furthermore, we hypothesize that the immune mechanism involved in the generation of these Abs entails cross talk between B lymphocytes (Bc) and invariant natural killer T cells (iNKT). Combining cellular techniques, such as flow cytometry and biochemical assays, we demonstrated that CD1d binds to NGcGM3 and that human Bc present NGcGM3 in a CD1d context according to two alternative strategies. We also showed that paraformaldehyde treatment of cells expressing CD1d affects the presentation. Finally, by co-culturing primary human Bc with iNKT and measuring Ki-67 expression, we detected a reproducible increment in the proliferation of the iNKT population when Ag was on the medium. Our findings identify a novel, endogenous, human CD1d ligand, which is sufficiently competent to stimulate iNKT. We postulate that CD1d-restricted Bc presentation of NGcGM3 drives effective iNKT activation, an immunological mechanism that has not been previously described for humans, which may contribute to understanding aNGcGM3 occurrence.
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Antígenos CD1d/inmunología , Linfocitos B/inmunología , Comunicación Celular/inmunología , Gangliósido G(M3)/inmunología , Células T Asesinas Naturales/inmunología , Adulto , Presentación de Antígeno/inmunología , Antígenos CD1d/metabolismo , Linfocitos B/metabolismo , Línea Celular , Proliferación Celular , Células Cultivadas , Técnicas de Cocultivo , Citometría de Flujo , Gangliósido G(M3)/metabolismo , Humanos , Ligandos , Activación de Linfocitos/inmunología , Células T Asesinas Naturales/metabolismo , Tonsila Palatina/citología , Unión Proteica/inmunologíaRESUMEN
INTRODUCTION: Racotumomab (originally known as 1E10 mAb) is an anti-idiotype murine IgG1 directed to membrane glycoconjugates expressed in aggressive solid tumors. It was developed as a mirror image of the idiotype of another antibody against N-glycolyl-containing molecules, such as the NeuGcGM3 ganglioside. After a successful phase II/III study, racotumomab formulated in alum was conditionally approved in Latin American countries as maintenance therapy for advanced non-small cell lung cancer. AREAS COVERED: This review analyzes the biology of the target antigen, summarizes preclinical studies and discusses clinical trials in adults and the pediatric experience with racotumomab. EXPERT OPINION: Proper patient selection and combination with chemotherapy, radiotherapy or checkpoint inhibitors appear to be critical issues to maximize the effects of racotumomab vaccination in lung cancer. In a recent phase I clinical trial in children with relapsed or resistant neuroectodermal malignancies, racotumomab was well tolerated and immunogenic, and its evaluation as immunotherapy for high-risk neuroblastoma is warranted.
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Anticuerpos Monoclonales/uso terapéutico , Vacunas contra el Cáncer/uso terapéutico , Carcinoma de Pulmón de Células no Pequeñas/tratamiento farmacológico , Neoplasias Pulmonares/tratamiento farmacológico , Animales , Anticuerpos Antiidiotipos/uso terapéutico , Anticuerpos Monoclonales de Origen Murino , Niño , Gangliósido G(M3)/análogos & derivados , Gangliósido G(M3)/inmunología , HumanosRESUMEN
Membrane-wrapped nanoparticles represent a versatile platform for utilizing specific lipid-receptor interactions, such as siallyllactose-mediated binding of the ganglioside GM3 to Siglec1 (CD169), for targeting purposes. The membrane wrap around the nanoparticles not only serves as a matrix to incorporate GM3 as targeting moiety for antigen-presenting cells but also offers unique opportunities for constructing a biomimetic surface from lipids with potentially protein-repellent properties. We characterize nonspecific protein adsorption (corona formation) to membrane-wrapped nanoparticles with core diameters of approximately 35 and 80 nm and its effect on the GM3-mediated targeting efficacy as a function of surface charge through combined in vitro and in vivo studies. The stability and fate of the membrane wrap around the nanoparticles in a simulated biological fluid and after uptake in CD169-expressing antigen-presenting cells is experimentally tested. Finally, we demonstrate in hock immunization studies in mice that GM3-decorated membrane-wrapped nanoparticles achieve a selective enrichment in the peripheral regions of popliteal lymph nodes that contain high concentrations of CD169-expressing antigen-presenting cells.
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Células Dendríticas/efectos de los fármacos , Gangliósido G(M3)/administración & dosificación , Liposomas/administración & dosificación , Ganglios Linfáticos/inmunología , Nanopartículas/administración & dosificación , 1,2-Dipalmitoilfosfatidilcolina/análogos & derivados , 1,2-Dipalmitoilfosfatidilcolina/química , Animales , Materiales Biomiméticos/química , Línea Celular , Células Dendríticas/citología , Células Dendríticas/inmunología , Gangliósido G(M3)/química , Gangliósido G(M3)/inmunología , Expresión Génica , Células HeLa , Humanos , Inmunización , Liposomas/química , Liposomas/inmunología , Ganglios Linfáticos/citología , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Ratones , Nanopartículas/química , Fosfatidilserinas/química , Cultivo Primario de Células , Lectina 1 Similar a Ig de Unión al Ácido Siálico/genética , Lectina 1 Similar a Ig de Unión al Ácido Siálico/inmunología , Propiedades de Superficie , Virión/química , Virión/inmunologíaRESUMEN
Following the mass vaccinations against pandemic influenza A/H1N1 virus in 2009, a sudden increase in juvenile onset narcolepsy with cataplexy (NC) was detected in several European countries where AS03-adjuvanted Pandemrix vaccine had been used. NC is a chronic neurological disorder characterized by excessive daytime sleepiness and cataplexy. In human NC, the hypocretin-producing neurons in the hypothalamus or the hypocretin signaling pathway are destroyed by an autoimmune reaction. Both genetic (e.g. HLA-DQB1*0602) and environmental risk factors (e.g. Pandemrix) contribute to the disease development, but the underlying and the mediating immunological mechanisms are largely unknown. Influenza virus hemagglutinin is known to bind gangliosides, which serve as host cell virus receptors. Anti-ganglioside antibodies have previously been linked to various neurological disorders, like the Guillain-Barré syndrome which may develop after infection or vaccination. Because of these links we screened sera of NC patients and controls for IgG anti-ganglioside antibodies against 11 human brain gangliosides (GM1, GM2, GM3, GM4, GD1a, GD1b, GD2, GD3, GT1a, GT1b, GQ1b) and a sulfatide by using a line blot assay. Samples from 173 children and adolescents were analyzed: 48 with Pandemrix-associated NC, 20 with NC without Pandemrix association, 57 Pandemrix-vaccinated and 48 unvaccinated healthy children. We found that patients with Pandemrix-associated NC had more frequently (14.6%) anti-GM3 antibodies than vaccinated healthy controls (3.5%) (P = 0.047). Anti-GM3 antibodies were significantly associated with HLA-DQB1*0602 (P = 0.016) both in vaccinated NC patients and controls. In general, anti-ganglioside antibodies were more frequent in vaccinated (18.1%) than in unvaccinated (7.3%) individuals (P = 0.035). Our data suggest that autoimmunity against GM3 is a feature of Pandemrix-associated NC and that autoantibodies against gangliosides were induced by Pandemrix vaccination.
Asunto(s)
Autoanticuerpos/inmunología , Gangliósido G(M3)/inmunología , Cadenas beta de HLA-DQ/inmunología , Vacunas contra la Influenza/efectos adversos , Narcolepsia/etiología , Adolescente , Niño , Preescolar , Europa (Continente)/epidemiología , Humanos , Subtipo H1N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza/inmunología , Gripe Humana/prevención & control , Vacunación Masiva/efectos adversos , Narcolepsia/inmunologíaRESUMEN
While not naturally expressed in normal human tissues, N-glycolylated (NeuGc) gangliosides are overexpressed in several tumors and have immunosuppressive capacity, which contributes to cancer progression. Naturally occurring antibodies against NeuGcGM3 exist in healthy donors that specifically recognize and kill tumor cells expressing the antigen by complement-dependent and -independent mechanisms, the latter resembling an oncotic necrosis-type of cell death. Both the levels of anti-NeuGcGM3 antibodies in the sera of healthy donors and the percentage of donors with these natural antibodies decrease with age. Our work has shown that anti-NeuGcGM3 antibodies are not detected in the sera of non-small cell lung cancer (NSCLC) patients, compared to age- and sex-matched healthy donors, which have anti-NeuGcGM3. Interestingly, the level of serum total IgM, but not IgG, was significantly lower in cancer patients than in healthy donors. Screening of immortalized mouse splenic and peritoneal-derived hybridomas showed that peritoneal B-1 cells secrete anti-NeuGcGM3 with tumor cytotoxic capacity. Defects in the natural surveillance against tumor antigens could increase the risk of elderly donors developing cancer and affect the capacity of cancer patients to effectively fight against tumor cells.
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Autoanticuerpos/inmunología , Subgrupos de Linfocitos B/inmunología , Gangliósido G(M3)/análogos & derivados , Hibridomas/inmunología , Vigilancia Inmunológica/inmunología , Animales , Autoanticuerpos/sangre , Subgrupos de Linfocitos B/metabolismo , Línea Celular Tumoral , Perros , Femenino , Gangliósido G(M3)/sangre , Gangliósido G(M3)/inmunología , Caballos , Humanos , Hibridomas/metabolismo , Ratones , Ratones Endogámicos BALB CRESUMEN
The ganglioside GM3(Neu5Gc) has gained increasing attention as therapeutic target because of its selective expression in various human tumours, such as melanoma, breast and lung cancer. 14F7 is a mouse IgG1 with specific reactivity to GM3(Neu5Gc)-positive tumours. The therapeutic activity of 14F7 has also been demonstrated in vivo, through its repetitive passive administration in tumour-bearing animals. In this work we used an alternative strategy to deliver recombinant 14F7 in vivo and analysed the therapeutic efficacy of this approach. We engineered a recombinant adeno-associated vector to direct the expression of secretable recombinant 14F7 in BALB/c animals. A single administration of the rAAV induced efficient production and secretion of the antibody in the bloodstream, with an expression level reaching plateau at â¼3 weeks after injection and persisting for almost a year. Strikingly, upon challenge with GM3(Neu5Gc)-positive X63-AG8.653 myeloma cells, tumour development was significantly delayed in animals treated with rAAV-14F7 with respect to animals treated with a control rAAV codifying for an irrelevant antibody. Finally, no significant differences in survival proportion were detected in animals injected with rAAV-14F7 or treated by standard administration of repetitive doses of purified monoclonal antibody 14F7.
Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Anticuerpos Monoclonales/inmunología , Gangliósido G(M3)/inmunología , Secuencia de Aminoácidos , Animales , Dependovirus/genética , Dependovirus/metabolismo , Células HEK293 , Humanos , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/inmunología , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Neoplasias Experimentales/inmunología , Neoplasias Experimentales/terapia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
Gangliosides are sialic acid-bearing glycosphingolipids expressed on all mammalian cell membranes, and participate in several cellular processes. During malignant transformation their expression changes, both at the quantitative and qualitative levels. Of particular interest is the overexpression by tumor cells of Neu5Gc-gangliosides, which are absent, or detected in trace amounts, in human normal cells. The GM3(Neu5Gc) ganglioside in particular has been detected in many human tumors, and it is considered one of the few tumor specific antigen. We previously demonstrated that a humanized antibody specific for this molecule, named 14F7hT, retained the binding and cytotoxic properties of the mouse antibody. In this work, we confirm that 14F7hT exerts a non-apoptotic cell death mechanism in vitro and shows its potent in vivo antitumor activity on a solid mouse myeloma model. Also, we demonstrate, in contrast to the murine counterpart, the capacity of this antibody to induce antibody-dependent cell-mediated cytotoxicity using human effector cells, which increases its potential for the treatment of GM3(Neu5Gc)-expressing human tumors.