Q3202991 (Q3202991): Difference between revisions
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(Created claim: summary (P836): Objectives: 1) Quantify the frequency and effector capacity of specific cytotoxic T-lymphocytes (TCL) against hepatocarcinoma-associated antigens (AAH) in the peripheral and intrahepatic compartment of patients with hepatocarcinoma (HC), 2) Analyse the phenotype associated with depletion and apoptosis in these cells and its relationship with the expression of cluster miR 17-92, 3) Analyse the effect of in-vitro block with anti-miR cluster miR 17...) |
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Objectives: 1) Quantify the frequency and effector capacity of specific cytotoxic T-lymphocytes (TCL) against hepatocarcinoma-associated antigens (AAH) in the peripheral and intrahepatic compartment of patients with hepatocarcinoma (HC), 2) Analyse the phenotype associated with depletion and apoptosis in these cells and its relationship with the expression of cluster miR 17-92, 3) Analyse the effect of in-vitro block with anti-miR cluster miR 17-92 on the effect of specific LTC-specific AAH. Methodology: In patients with HC HLA-A2+, peripheral blood mononuclear cells (CMSP) and liver (MIHC) are isolated. The frequency of specific LTC-AAH (NY-ESO-1157, MAGE-A3271, Glypican3144) and specific LTC-cytomegalovirus (CMV) (pp65 495-504) [Control group] shall be quantified by marking of CMSP and HMIH with multimeric HLA-A2/epitope complexes and anti-CD8 antibodies. The TAME (Tetramer associated magnetic enrichment) technique will be used to detect cells frequently >10-7 compared to the total CMSP. These cells will analyse the phenotype associated with exhaustion (CD127, PD-1, TRAF1), apoptosis (Mcl-1, Bim) and evaluate the effector capacities (specific antigen proliferation, interferon-? production and cytotoxicity (CD107a)). These analyses will be performed by flow cytometry. By sorting technique specific LTC-AAHs are isolated to obtain their RNA and after retro-transcription to cDNA, the miR 17-92 cluster will be amplified. The expression of the miR 17-92 cluster will be choreolated with the phenotype of exhaustion and apoptosis. In case a positive chorelation is observed between these phenotypes and the miR 17-92 cluster, the miR 17-92 cluster with appropriate anti-miR to restore effector activity of specific LTC-AAHs will be blocked in-vitro. (English) | |||||||||||||||
| Property / summary: Objectives: 1) Quantify the frequency and effector capacity of specific cytotoxic T-lymphocytes (TCL) against hepatocarcinoma-associated antigens (AAH) in the peripheral and intrahepatic compartment of patients with hepatocarcinoma (HC), 2) Analyse the phenotype associated with depletion and apoptosis in these cells and its relationship with the expression of cluster miR 17-92, 3) Analyse the effect of in-vitro block with anti-miR cluster miR 17-92 on the effect of specific LTC-specific AAH. Methodology: In patients with HC HLA-A2+, peripheral blood mononuclear cells (CMSP) and liver (MIHC) are isolated. The frequency of specific LTC-AAH (NY-ESO-1157, MAGE-A3271, Glypican3144) and specific LTC-cytomegalovirus (CMV) (pp65 495-504) [Control group] shall be quantified by marking of CMSP and HMIH with multimeric HLA-A2/epitope complexes and anti-CD8 antibodies. The TAME (Tetramer associated magnetic enrichment) technique will be used to detect cells frequently >10-7 compared to the total CMSP. These cells will analyse the phenotype associated with exhaustion (CD127, PD-1, TRAF1), apoptosis (Mcl-1, Bim) and evaluate the effector capacities (specific antigen proliferation, interferon-? production and cytotoxicity (CD107a)). These analyses will be performed by flow cytometry. By sorting technique specific LTC-AAHs are isolated to obtain their RNA and after retro-transcription to cDNA, the miR 17-92 cluster will be amplified. The expression of the miR 17-92 cluster will be choreolated with the phenotype of exhaustion and apoptosis. In case a positive chorelation is observed between these phenotypes and the miR 17-92 cluster, the miR 17-92 cluster with appropriate anti-miR to restore effector activity of specific LTC-AAHs will be blocked in-vitro. (English) / rank | |||||||||||||||
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| Property / summary: Objectives: 1) Quantify the frequency and effector capacity of specific cytotoxic T-lymphocytes (TCL) against hepatocarcinoma-associated antigens (AAH) in the peripheral and intrahepatic compartment of patients with hepatocarcinoma (HC), 2) Analyse the phenotype associated with depletion and apoptosis in these cells and its relationship with the expression of cluster miR 17-92, 3) Analyse the effect of in-vitro block with anti-miR cluster miR 17-92 on the effect of specific LTC-specific AAH. Methodology: In patients with HC HLA-A2+, peripheral blood mononuclear cells (CMSP) and liver (MIHC) are isolated. The frequency of specific LTC-AAH (NY-ESO-1157, MAGE-A3271, Glypican3144) and specific LTC-cytomegalovirus (CMV) (pp65 495-504) [Control group] shall be quantified by marking of CMSP and HMIH with multimeric HLA-A2/epitope complexes and anti-CD8 antibodies. The TAME (Tetramer associated magnetic enrichment) technique will be used to detect cells frequently >10-7 compared to the total CMSP. These cells will analyse the phenotype associated with exhaustion (CD127, PD-1, TRAF1), apoptosis (Mcl-1, Bim) and evaluate the effector capacities (specific antigen proliferation, interferon-? production and cytotoxicity (CD107a)). These analyses will be performed by flow cytometry. By sorting technique specific LTC-AAHs are isolated to obtain their RNA and after retro-transcription to cDNA, the miR 17-92 cluster will be amplified. The expression of the miR 17-92 cluster will be choreolated with the phenotype of exhaustion and apoptosis. In case a positive chorelation is observed between these phenotypes and the miR 17-92 cluster, the miR 17-92 cluster with appropriate anti-miR to restore effector activity of specific LTC-AAHs will be blocked in-vitro. (English) / qualifier | |||||||||||||||
point in time: 13 October 2021
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Revision as of 00:54, 13 October 2021
Project Q3202991 in Spain
| Language | Label | Description | Also known as |
|---|---|---|---|
| English | No label defined |
Project Q3202991 in Spain |
Statements
44,000.0 Euro
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55,000.0 Euro
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80.0 percent
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1 January 2016
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16 December 2018
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SECRETARIA GENERAL DEL SERVICIO DE SALUD DE CASTILLA-LA MANCHA
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40°44'24.00"N, 2°30'21.35"W
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19130
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Objetivos: 1) Cuantificar la frecuencia y capacidad efectora de linfocitos T citotóxicos (LTC) específicos contra antígenos asociados a hepatocarcinoma (AAH) en el compartimento periférico e intrahepático de pacientes con hepatocarcinoma (HC), 2) Analizar el fenotipo asociado a agotamiento y apoptosis en estas células y su relación con la expresión del cluster miR 17-92, 3) Analizar el efecto del bloqueo in-vitro con anti-miR del cluster miR 17-92 sobre la capacidad efectora de LTC específicos contra AAH. Metodología: En pacientes con HC HLA-A2+ se aislaran células mononucleares de sangre periférica (CMSP) e hígado (CMIH). Se cuantificará la frecuencia de LTC-AAH específicos (NY-ESO-1157, MAGE-A3271, Glypican3144) y LTC-citomegalovirus (CMV) específicos (pp65 495-504) [Grupo control], mediante marcaje de CMSP y CMIH con complejos multiméricos HLA-A2/epítopo y anticuerpos anti-CD8. Se utilizará la técnica TAME ( Tetramer associated magnetic enrichment ) que permite detectar células con frecuencia>10-7 respecto al total de CMSP. En estas células se analizará el fenotipo asociado a agotamiento (CD127, PD-1, TRAF1), apoptosis (Mcl-1, Bim) y se evaluarán las capacidades efectoras (proliferación antígeno específica, producción de interferón-? y citotoxicidad (CD107a)). Estos análisis se realizaran mediante citometría de flujo. Mediante técnica de sorting se aislaran LTC-AAH específicos para obtener su RNA y tras retro-transcripción a cDNA, se amplificará el cluster miR 17-92. Se corelacionará la expresión del cluster miR 17-92 con el fenotipo de agotamiento y apoptosis. En caso de observarse una corelación positiva entre estos fenotipos y el cluster miR 17-92, se bloqueará in-vitro el cluster miR 17-92 con anti-miR apropiados para restaurar la actividad efectora de los LTC-AAH específicos. (Spanish)
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Objectives: 1) Quantify the frequency and effector capacity of specific cytotoxic T-lymphocytes (TCL) against hepatocarcinoma-associated antigens (AAH) in the peripheral and intrahepatic compartment of patients with hepatocarcinoma (HC), 2) Analyse the phenotype associated with depletion and apoptosis in these cells and its relationship with the expression of cluster miR 17-92, 3) Analyse the effect of in-vitro block with anti-miR cluster miR 17-92 on the effect of specific LTC-specific AAH. Methodology: In patients with HC HLA-A2+, peripheral blood mononuclear cells (CMSP) and liver (MIHC) are isolated. The frequency of specific LTC-AAH (NY-ESO-1157, MAGE-A3271, Glypican3144) and specific LTC-cytomegalovirus (CMV) (pp65 495-504) [Control group] shall be quantified by marking of CMSP and HMIH with multimeric HLA-A2/epitope complexes and anti-CD8 antibodies. The TAME (Tetramer associated magnetic enrichment) technique will be used to detect cells frequently >10-7 compared to the total CMSP. These cells will analyse the phenotype associated with exhaustion (CD127, PD-1, TRAF1), apoptosis (Mcl-1, Bim) and evaluate the effector capacities (specific antigen proliferation, interferon-? production and cytotoxicity (CD107a)). These analyses will be performed by flow cytometry. By sorting technique specific LTC-AAHs are isolated to obtain their RNA and after retro-transcription to cDNA, the miR 17-92 cluster will be amplified. The expression of the miR 17-92 cluster will be choreolated with the phenotype of exhaustion and apoptosis. In case a positive chorelation is observed between these phenotypes and the miR 17-92 cluster, the miR 17-92 cluster with appropriate anti-miR to restore effector activity of specific LTC-AAHs will be blocked in-vitro. (English)
13 October 2021
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Guadalajara
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Identifiers
PI15_00074
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