Objective:The high heterogeneity of hepatocellular carcinoma(HCC)renders traditional therapies unable to effectively activate the patient's immune system to combat tumors.Patients with advanced HCC and T cell func...Objective:The high heterogeneity of hepatocellular carcinoma(HCC)renders traditional therapies unable to effectively activate the patient's immune system to combat tumors.Patients with advanced HCC and T cell functional deficiencies may benefit more from cellular immunotherapy,especially tumor neoepitope-targeted T cell receptor(TCR)-T cells.Neoepitopes with strong immunogenicity provide precise targets for HCC,further enhancing the efficacy of cellular immunotherapy.Methods:A scalable workflow for identifying neoepitopes from 7 HLA-A*02:01-restricted patients with HCC was established based on whole exome sequencing and bioinformatics analyses,followed by identification of neoepitope-specific TCRs through tetramerbased screening and single-cell TCR cloning technology,which were further validated in the JC4 cell model.The cytotoxicity of CD8+TCR-T cells was evaluated in neoepitope-positive tumor cell lines or NCG mice.Results:Ten specific neoepitopes were identified,among which neoepitope B and T lymphocyte attenuatorP267L[BTLAP267L(SLNHSVIGL)]exhibited advantageous properties as a potential tumor target.Three TCRs(85-3,126-5,and 52-3)were confirmed to specifically recognize the neoepitope BTLAP267L,while no cross-recognition of irrelevant or wild-type epitopes was observed.Activated BTLAP267L-specific CD8+TCR-T cells released extensive perforin,granzyme B,IFN-γ,and TNF-αin vitro,thereby inducing strong cytotoxic effects against BTLAP267L-positive T2 or HCC cell lines.BTLAP267L-specific CD8+TCR-T cells mediated robust tumor regression due to long-lasting survival and released perforin without causing significant cytotoxic effects on normal organs in murine experiments.Conclusions:This preclinical study demonstrated the beneficial effects of neoepitope BTLAP267L-specific TCR-T cell immunotherapy,unlocking a novel strategy for personalized precision therapy in HCC.展开更多
Objective:Neoantigens arising from gene mutations in tumors can induce specific immune responses,and neoantigen-based immunotherapies have been tested in clinical trials.Here,we characterized the efficacy of altered n...Objective:Neoantigens arising from gene mutations in tumors can induce specific immune responses,and neoantigen-based immunotherapies have been tested in clinical trials.Here,we characterized the efficacy of altered neoepitopes in improving immunogenicity against gastric cancer.Methods:Raw data of whole-exome sequencing derived from a patient with gastric cancer were analyzed using bioinformatics methods to identify neoepitopes.Neoepitopes were modified by P1Y(the first amino acid was replaced by tyrosine)and P2L(the second amino acid was replaced by leucine).T2 binding and stability assays were used to detect the affinities between the neoepitopes and the HLA molecules,as well as the stabilities of complexes.Dendritic cells(DCs)presented with neoepitopes stimulated naïve CD8+T cells to induce specific cytotoxic T lymphocytes.ELISA and carboxyfluorescein succinimidyl ester were used to detect IFN-γand TNF-αlevels,and T cell proliferation.Perforin was detected by flow cytometry.The cytotoxicity of T cells was determined using the lactate dehydrogenase assay.Results:Bioinformatics analysis,T2 binding,and stability assays indicated that residue substitution increased the affinity between neoepitopes and HLA molecules,as well as the stabilities of complexes.DCs presented with altered neoepitopes stimulated CD8+T cells to release more IFN-γand had a greater effect on promoting proliferation than wild-type neoepitopes.CD8+T cells stimulated with altered neoepitopes killed more wild-type neoepitope-pulsed T2 cells than those stimulated with wild-type neoepitopes,by secreting more IFN-γ,TNF-α,and perforin.Conclusions:Altered neoepitopes exhibited greater immunogenicity than wild-type neoepitopes.Residue substitution could be used as a new strategy for immunotherapy to target neoantigens.展开更多
Objective： The aim of this study was to evaluate the efficacy of transcatheter arterial chemoembolization （TACE） combined with a Chinese compound preparation of ganfule on advanced hepatocellular carcinoma （HCC）....Objective： The aim of this study was to evaluate the efficacy of transcatheter arterial chemoembolization （TACE） combined with a Chinese compound preparation of ganfule on advanced hepatocellular carcinoma （HCC）. Methods： The study population consisted of 132 advanced HCC patients with Child-pugh NB. Tumor in all patients was involved with main trunk of portal vein and/or inferior vena cava, or local lymph node metastasis, or distant metastasis. TACE combined with ganfule were performed in 65 patients with advanced HCC （interventional treatment group）, 67 patients were treated with traditional Chinese herbal drug alone （Chinese herb group）. The prime end point was overall survival （OS）, and prognostic factors were analyzed. Results： The median OS was 205 days [95% confidence interval （CI）, 155-255 days] in interventional treatment group and 127 days （95% CI, 70-184 days） in Chinese herb group （P 〈 0.05）. The 6-month, 1-year, and 2-year OS rates were 58.9%, 29.1%, 7.7% in interventional treatment group, and 33.3%, 12.3%, 1.8% in Chinese herb group, respectively. The portal vein thrombosis, ECOG performance status （PS） were independent prognostic factors for OS. Conclusion： Ttranscatheter arterial chemoembolization combined with a Chinese compound preparation of ganfule could greatly prolong the OS of advanced HCC patients. The portal vein thrombosis and ECOG PS were independent prognostic factors for OS.展开更多
Dear Editor,Cytokine-induced killer(CIK)cells have been recognized as a new type of anti-tumor effector cells.CIK cells are a mixture of T lymphocytes.Among them,CD3+/CD56+T cells,which are rare in uncultured peripher...Dear Editor,Cytokine-induced killer(CIK)cells have been recognized as a new type of anti-tumor effector cells.CIK cells are a mixture of T lymphocytes.Among them,CD3+/CD56+T cells,which are rare in uncultured peripheral blood,are the main effector cells.CIK cells can proliferate rapidly in vitro,with stronger antitumor activity,broader target tumor spectrum,and lower adverse effect than other reported antitumor effector cells.1 Their ease of production in vitro and antitumor potential have made them suitable candidates for cell therapy regimens in solid and hematopoietic tumor treatments.1,2 Our previous retrospective study showed that the median progression-free survival(PFS)and overall survival(OS)in untreated,advanced non-small-cell lung cancer(NSCLC)patients who received CIK cell immunotherapy plus chemotherapy(13 and 24 months,respectively)were significantly longer than in those who received chemotherapy alone(6 and 10 months,respectively).2 But so far,there is no prospective,multicenter clinical study in lung cancer.Based on our previous study,we designed this randomized,multicenter,open-label trial to further evaluate the clinical efficacy of CIK cell immunotherapy plus chemotherapy in patients with advanced squamous NSCLC(ClinicalTrials.gov number,NCT01631357).展开更多
基金supported by grants from the Joint Funds for the Innovation of Science and Technology of Fujian Province(Grant No.2018Y9108 and 2024Y9607)the Natural Science Foundation of Fujian Province(Grant No.2023J011247)+1 种基金the Science and Technology Project of Fujian Province(Grant No.2021Y0054)the Fujian Provincial Health Technology Project(Grant No.2020GGA018)。
文摘Objective:The high heterogeneity of hepatocellular carcinoma(HCC)renders traditional therapies unable to effectively activate the patient's immune system to combat tumors.Patients with advanced HCC and T cell functional deficiencies may benefit more from cellular immunotherapy,especially tumor neoepitope-targeted T cell receptor(TCR)-T cells.Neoepitopes with strong immunogenicity provide precise targets for HCC,further enhancing the efficacy of cellular immunotherapy.Methods:A scalable workflow for identifying neoepitopes from 7 HLA-A*02:01-restricted patients with HCC was established based on whole exome sequencing and bioinformatics analyses,followed by identification of neoepitope-specific TCRs through tetramerbased screening and single-cell TCR cloning technology,which were further validated in the JC4 cell model.The cytotoxicity of CD8+TCR-T cells was evaluated in neoepitope-positive tumor cell lines or NCG mice.Results:Ten specific neoepitopes were identified,among which neoepitope B and T lymphocyte attenuatorP267L[BTLAP267L(SLNHSVIGL)]exhibited advantageous properties as a potential tumor target.Three TCRs(85-3,126-5,and 52-3)were confirmed to specifically recognize the neoepitope BTLAP267L,while no cross-recognition of irrelevant or wild-type epitopes was observed.Activated BTLAP267L-specific CD8+TCR-T cells released extensive perforin,granzyme B,IFN-γ,and TNF-αin vitro,thereby inducing strong cytotoxic effects against BTLAP267L-positive T2 or HCC cell lines.BTLAP267L-specific CD8+TCR-T cells mediated robust tumor regression due to long-lasting survival and released perforin without causing significant cytotoxic effects on normal organs in murine experiments.Conclusions:This preclinical study demonstrated the beneficial effects of neoepitope BTLAP267L-specific TCR-T cell immunotherapy,unlocking a novel strategy for personalized precision therapy in HCC.
基金This work was supported by grants from the Science and Technology Project in Fujian Province of China(Grant No.2018I0004)Joint Funds for the innovation of Science and Technology,Fujian Province of China(Grant No.2018Y9108).
文摘Objective:Neoantigens arising from gene mutations in tumors can induce specific immune responses,and neoantigen-based immunotherapies have been tested in clinical trials.Here,we characterized the efficacy of altered neoepitopes in improving immunogenicity against gastric cancer.Methods:Raw data of whole-exome sequencing derived from a patient with gastric cancer were analyzed using bioinformatics methods to identify neoepitopes.Neoepitopes were modified by P1Y(the first amino acid was replaced by tyrosine)and P2L(the second amino acid was replaced by leucine).T2 binding and stability assays were used to detect the affinities between the neoepitopes and the HLA molecules,as well as the stabilities of complexes.Dendritic cells(DCs)presented with neoepitopes stimulated naïve CD8+T cells to induce specific cytotoxic T lymphocytes.ELISA and carboxyfluorescein succinimidyl ester were used to detect IFN-γand TNF-αlevels,and T cell proliferation.Perforin was detected by flow cytometry.The cytotoxicity of T cells was determined using the lactate dehydrogenase assay.Results:Bioinformatics analysis,T2 binding,and stability assays indicated that residue substitution increased the affinity between neoepitopes and HLA molecules,as well as the stabilities of complexes.DCs presented with altered neoepitopes stimulated CD8+T cells to release more IFN-γand had a greater effect on promoting proliferation than wild-type neoepitopes.CD8+T cells stimulated with altered neoepitopes killed more wild-type neoepitope-pulsed T2 cells than those stimulated with wild-type neoepitopes,by secreting more IFN-γ,TNF-α,and perforin.Conclusions:Altered neoepitopes exhibited greater immunogenicity than wild-type neoepitopes.Residue substitution could be used as a new strategy for immunotherapy to target neoantigens.
基金Supported by a grant from the Research Foundation of Fujian Provincial Health Department(No.2010-7)
文摘Objective： The aim of this study was to evaluate the efficacy of transcatheter arterial chemoembolization （TACE） combined with a Chinese compound preparation of ganfule on advanced hepatocellular carcinoma （HCC）. Methods： The study population consisted of 132 advanced HCC patients with Child-pugh NB. Tumor in all patients was involved with main trunk of portal vein and/or inferior vena cava, or local lymph node metastasis, or distant metastasis. TACE combined with ganfule were performed in 65 patients with advanced HCC （interventional treatment group）, 67 patients were treated with traditional Chinese herbal drug alone （Chinese herb group）. The prime end point was overall survival （OS）, and prognostic factors were analyzed. Results： The median OS was 205 days [95% confidence interval （CI）, 155-255 days] in interventional treatment group and 127 days （95% CI, 70-184 days） in Chinese herb group （P 〈 0.05）. The 6-month, 1-year, and 2-year OS rates were 58.9%, 29.1%, 7.7% in interventional treatment group, and 33.3%, 12.3%, 1.8% in Chinese herb group, respectively. The portal vein thrombosis, ECOG performance status （PS） were independent prognostic factors for OS. Conclusion： Ttranscatheter arterial chemoembolization combined with a Chinese compound preparation of ganfule could greatly prolong the OS of advanced HCC patients. The portal vein thrombosis and ECOG PS were independent prognostic factors for OS.
基金supported by the National Key Technologies R&D Program of China grant Awards No.2015BAI12B12(to X.R.)and 2018YFC1313400(to J.X.)the National Natural Science Foundation of China grants Awards No.81572913 and 81872487(to L.L.).
文摘Dear Editor,Cytokine-induced killer(CIK)cells have been recognized as a new type of anti-tumor effector cells.CIK cells are a mixture of T lymphocytes.Among them,CD3+/CD56+T cells,which are rare in uncultured peripheral blood,are the main effector cells.CIK cells can proliferate rapidly in vitro,with stronger antitumor activity,broader target tumor spectrum,and lower adverse effect than other reported antitumor effector cells.1 Their ease of production in vitro and antitumor potential have made them suitable candidates for cell therapy regimens in solid and hematopoietic tumor treatments.1,2 Our previous retrospective study showed that the median progression-free survival(PFS)and overall survival(OS)in untreated,advanced non-small-cell lung cancer(NSCLC)patients who received CIK cell immunotherapy plus chemotherapy(13 and 24 months,respectively)were significantly longer than in those who received chemotherapy alone(6 and 10 months,respectively).2 But so far,there is no prospective,multicenter clinical study in lung cancer.Based on our previous study,we designed this randomized,multicenter,open-label trial to further evaluate the clinical efficacy of CIK cell immunotherapy plus chemotherapy in patients with advanced squamous NSCLC(ClinicalTrials.gov number,NCT01631357).
