AIM:To investigate the prognostic role of invariant natural killer T(iNKT) cells and antibody-dependent cell-mediated cytotoxicity(ADCC) in wild type KRAS metastatic colorectal cancer(mC RC) patients treated with cetu...AIM:To investigate the prognostic role of invariant natural killer T(iNKT) cells and antibody-dependent cell-mediated cytotoxicity(ADCC) in wild type KRAS metastatic colorectal cancer(mC RC) patients treated with cetuximab.METHODS: Forty-one KRAS wt mC RC patients,treated with cetuximab and irinotecan-based chemotherapy in Ⅱ and Ⅲ lines were analyzed. Genotyping of single nucleotide polymorphism(SNP)s in the FCGR2A,FCGR3A and in the 3' untranslated regions of KRAS and mutational analysis for KRAS,BRAF and NRAS genes was determined either by sequencing or allelic discrimination assays. Enriched NK cells were obtained from lymphoprepperipheral blood mononuclear cell and iN KT cells were defined by co-expression of CD3,TCRVα24,TCRVβ11. ADCC was evaluated as ex vivo NK-dependent activity,measuring lactate dehydrogenase release.RESULTS: At basal,mCRC patients performing ADCC activity above the median level(71%) showed an improved overall survival(OS) compared to patients with ADCC below(median 16 vs 8 mo;P=0.026). We did not find any significant correlation of iN KT cells with OS(P=0.19),albeit we observed a trend to a longer survival after 10 mo in patients with iN KT above median basal level(0.382 cells/microliter). Correlation of OS and progression-free survival(PFS) with interesting SNPs involved in ADCC ability revealed not to be significant. Patients carrying alleles both with A in FCGR2 A and TT in FCGR3A presented a trend of longer PFS(median 9 vs 5 mo;P=0.064). Chemotherapy impacted both iN KT cells and ADCC activity. Their prognostic values get lost when we analysed them after 2 and 4 mo of treatment.CONCLUSION: Our results suggest a link between iN KT cells,basal ADCC activity,genotypes in FCGR2A and FCGR3A,and efficacy of cetuximab in KRAS wt mC RC patients.展开更多
Objective:To delineate immunomodulatory role of histamine on antibody generation pr of ile in rabbit in the present dose-dependent histamine study.Methods:The cohort comprised of three groups(Ⅲ,ⅣandⅤ),containing si...Objective:To delineate immunomodulatory role of histamine on antibody generation pr of ile in rabbit in the present dose-dependent histamine study.Methods:The cohort comprised of three groups(Ⅲ,ⅣandⅤ),containing six rabbits each,and received subcutaneous histamine 50μg/kg×bis in die(b.i.d.),100μg/kg×b.i.d.and 200μg/kg X b.i.d.,respectively for 10 days (starting from the 1st day).They were subsequently immunized on the 3rd day with intravenous injection of sheep blood cell(SRBC)(1×10 cells/mL).GroupⅡ(positive control)(n=6) received vehicle(sterile distilled water) and immunized at day 3 similarly while groupⅠ(negative control) (n=6) remained non-immunized and received only vehicle.All experimentations were performed in triplicate.Blood samples were collected on pre-immunization(pre-I)(day 0),as well as on days 7-,14-,21-,28- and 58- post-immunization(post-I).Immunological parameters[total immunoglobulins(Igs),IgM and IgG]were analyzed by enzyme linked immunosorbent assay (ELISA) technique.Results:Histamine could influence a detectable antibody response to SRBC as early as day 7-post-I,which lasted until day 58- post-I.The results were found statistically significant(P【 0.05).Conclusions:Our results provide evidence that histamine has a short-term effect on antibody generation(until its presence in the body),and the antibody generation titer in vivo were affected by the concentration of histamine.展开更多
Objective: Our group has previously observed that in patients with small-cell lung cancers(SCLCs), the expression of a tumor antigen, glioma big potassium(g BK) ion channel, is higher at the time of death than when th...Objective: Our group has previously observed that in patients with small-cell lung cancers(SCLCs), the expression of a tumor antigen, glioma big potassium(g BK) ion channel, is higher at the time of death than when the cancer is first treated by surgical resection. This study aimed to determine whether this dichotomy was common in other potential lung tumor antigens by examining the same patient samples using our more extensive profile analysis of tumor-antigen precursor protein(TAPP). We then tested the hypothesis that therapeutic intervention may inadvertently cause this increased g BK production.Methods: SCLC samples(eight surgical resections and three autopsy samples) and three control lungs were examined by quantitative real-time polymerase chain reaction for 42 potential TAPPs that represent potential T-cell-mediated immunological targets. Results: Twenty-two TAPP m RNAs displayed the same profile as g BK, i.e., more m RNAs were expressed at autopsy than in their surgical counterparts. B-cyclin and mouse double minute 2, human homolog of P53-binding protein were elevated in both autopsy and surgical specimens above the normal-lung controls. When HTB119 cells were incubated with doxorubicin, g BK was strongly induced, as confirmed by intracellular flow cytometry with a g BK-specific antibody.Conclusion: Our findings suggested that more immunological targets became available as the tumor responded to chemotherapy and proceeded toward its terminal stages.展开更多
The gene encoding the heavy-and light-chain Fv regions of monoclonal antibody PS-9,which recognizes a cancer-associated antigen S-Tn on the most adenocarcinoma,was clonedby PCR techniques.The light and heavy chains we...The gene encoding the heavy-and light-chain Fv regions of monoclonal antibody PS-9,which recognizes a cancer-associated antigen S-Tn on the most adenocarcinoma,was clonedby PCR techniques.The light and heavy chains were connected by a flexible linker to form asingle chain variable fragment(ScFv)gene with 720bp,which was in turn fused topCANTAB 5 phage.The single chain Fv was expressed as fusion protein displayed on thephage surface.The phagemid is used to transform compepent E.Coli TG1 cells,then infect-ed with M13K07 helper phage to rescue the phagemid and antibody ScFv gene.All rand-mized 12 clones were shown reacting with colon cancer cell line Ls174t,which expresses S-Tnantigen.The recombinant phage has been infected E.Coli HB2151 cells to produe soluble an-tibody,which can be used for immunodetection and immunotherapy for cancer.展开更多
In this paper,we present the effective distance between T-cell and B-cell in an immune system using Stop and Wait(S/W)Automatic Repeat Request(ARQ).The concentration of the molecules can be increased by increasing the...In this paper,we present the effective distance between T-cell and B-cell in an immune system using Stop and Wait(S/W)Automatic Repeat Request(ARQ).The concentration of the molecules can be increased by increasing the transmitting number of molecules but it may reduce the performance of communication due to higher collision or interference with other molecules.It is also reported in the literature that the concentration of the emitted molecules reduces if the distance from Transmitter(Tx)to Receiver(Rx)increases.Thus,this paper mainly focuses on enhancing the receiver’s capture probability and higher successful complete transmission of the desired molecules by obtaining the effective distance from T-cell to B-cell.In order to find the effective distance,T-cell transmits the molecules 1(Interleukins-2)to B-cell,upon successful reception of molecules 1,antibodies(molecules 2)transmit back to T-cell.Then,the effective distance of an immune system can be obtained after T-cell detects the concentration of the molecules 2 with respect to time.Different schemes of S/W ARQ protocols have implemented in Molecular Communication(MC)but it requires retransmission of duplicate copies due to the lack of addressing an effective distance.Thus,the simulations are performed in MATLAB and the results obtain higher capture probability and also successful complete transmission of the desired molecules.展开更多
基金the Fondazione Veronesi that granted Daniela Vivenza and Martino Monteverde with PostDoctoral Fellowship Veronesithe Fondazione Cassa Risparmio of Cuneo for partially supporting the study
文摘AIM:To investigate the prognostic role of invariant natural killer T(iNKT) cells and antibody-dependent cell-mediated cytotoxicity(ADCC) in wild type KRAS metastatic colorectal cancer(mC RC) patients treated with cetuximab.METHODS: Forty-one KRAS wt mC RC patients,treated with cetuximab and irinotecan-based chemotherapy in Ⅱ and Ⅲ lines were analyzed. Genotyping of single nucleotide polymorphism(SNP)s in the FCGR2A,FCGR3A and in the 3' untranslated regions of KRAS and mutational analysis for KRAS,BRAF and NRAS genes was determined either by sequencing or allelic discrimination assays. Enriched NK cells were obtained from lymphoprepperipheral blood mononuclear cell and iN KT cells were defined by co-expression of CD3,TCRVα24,TCRVβ11. ADCC was evaluated as ex vivo NK-dependent activity,measuring lactate dehydrogenase release.RESULTS: At basal,mCRC patients performing ADCC activity above the median level(71%) showed an improved overall survival(OS) compared to patients with ADCC below(median 16 vs 8 mo;P=0.026). We did not find any significant correlation of iN KT cells with OS(P=0.19),albeit we observed a trend to a longer survival after 10 mo in patients with iN KT above median basal level(0.382 cells/microliter). Correlation of OS and progression-free survival(PFS) with interesting SNPs involved in ADCC ability revealed not to be significant. Patients carrying alleles both with A in FCGR2 A and TT in FCGR3A presented a trend of longer PFS(median 9 vs 5 mo;P=0.064). Chemotherapy impacted both iN KT cells and ADCC activity. Their prognostic values get lost when we analysed them after 2 and 4 mo of treatment.CONCLUSION: Our results suggest a link between iN KT cells,basal ADCC activity,genotypes in FCGR2A and FCGR3A,and efficacy of cetuximab in KRAS wt mC RC patients.
基金University Grants Commission, New Delhi,India for providing UGC Fellowship[UGC letter DON F.19-33/2006(CU)]M.Shahid is grateful to Department of Science & Technology,Ministry of Science & Technology,Government of India for awarding"Young Scientist Project Award"(FT/SR-L-111/2006)
文摘Objective:To delineate immunomodulatory role of histamine on antibody generation pr of ile in rabbit in the present dose-dependent histamine study.Methods:The cohort comprised of three groups(Ⅲ,ⅣandⅤ),containing six rabbits each,and received subcutaneous histamine 50μg/kg×bis in die(b.i.d.),100μg/kg×b.i.d.and 200μg/kg X b.i.d.,respectively for 10 days (starting from the 1st day).They were subsequently immunized on the 3rd day with intravenous injection of sheep blood cell(SRBC)(1×10 cells/mL).GroupⅡ(positive control)(n=6) received vehicle(sterile distilled water) and immunized at day 3 similarly while groupⅠ(negative control) (n=6) remained non-immunized and received only vehicle.All experimentations were performed in triplicate.Blood samples were collected on pre-immunization(pre-I)(day 0),as well as on days 7-,14-,21-,28- and 58- post-immunization(post-I).Immunological parameters[total immunoglobulins(Igs),IgM and IgG]were analyzed by enzyme linked immunosorbent assay (ELISA) technique.Results:Histamine could influence a detectable antibody response to SRBC as early as day 7-post-I,which lasted until day 58- post-I.The results were found statistically significant(P【 0.05).Conclusions:Our results provide evidence that histamine has a short-term effect on antibody generation(until its presence in the body),and the antibody generation titer in vivo were affected by the concentration of histamine.
文摘Objective: Our group has previously observed that in patients with small-cell lung cancers(SCLCs), the expression of a tumor antigen, glioma big potassium(g BK) ion channel, is higher at the time of death than when the cancer is first treated by surgical resection. This study aimed to determine whether this dichotomy was common in other potential lung tumor antigens by examining the same patient samples using our more extensive profile analysis of tumor-antigen precursor protein(TAPP). We then tested the hypothesis that therapeutic intervention may inadvertently cause this increased g BK production.Methods: SCLC samples(eight surgical resections and three autopsy samples) and three control lungs were examined by quantitative real-time polymerase chain reaction for 42 potential TAPPs that represent potential T-cell-mediated immunological targets. Results: Twenty-two TAPP m RNAs displayed the same profile as g BK, i.e., more m RNAs were expressed at autopsy than in their surgical counterparts. B-cyclin and mouse double minute 2, human homolog of P53-binding protein were elevated in both autopsy and surgical specimens above the normal-lung controls. When HTB119 cells were incubated with doxorubicin, g BK was strongly induced, as confirmed by intracellular flow cytometry with a g BK-specific antibody.Conclusion: Our findings suggested that more immunological targets became available as the tumor responded to chemotherapy and proceeded toward its terminal stages.
基金Supported by Grant of Medical Science from the Ministry of Health of PLA.
文摘The gene encoding the heavy-and light-chain Fv regions of monoclonal antibody PS-9,which recognizes a cancer-associated antigen S-Tn on the most adenocarcinoma,was clonedby PCR techniques.The light and heavy chains were connected by a flexible linker to form asingle chain variable fragment(ScFv)gene with 720bp,which was in turn fused topCANTAB 5 phage.The single chain Fv was expressed as fusion protein displayed on thephage surface.The phagemid is used to transform compepent E.Coli TG1 cells,then infect-ed with M13K07 helper phage to rescue the phagemid and antibody ScFv gene.All rand-mized 12 clones were shown reacting with colon cancer cell line Ls174t,which expresses S-Tnantigen.The recombinant phage has been infected E.Coli HB2151 cells to produe soluble an-tibody,which can be used for immunodetection and immunotherapy for cancer.
基金“Visvesvaraya Ph. D Scheme”, Govt. of India to give us an opportunity to complete this paper with their financial support
文摘In this paper,we present the effective distance between T-cell and B-cell in an immune system using Stop and Wait(S/W)Automatic Repeat Request(ARQ).The concentration of the molecules can be increased by increasing the transmitting number of molecules but it may reduce the performance of communication due to higher collision or interference with other molecules.It is also reported in the literature that the concentration of the emitted molecules reduces if the distance from Transmitter(Tx)to Receiver(Rx)increases.Thus,this paper mainly focuses on enhancing the receiver’s capture probability and higher successful complete transmission of the desired molecules by obtaining the effective distance from T-cell to B-cell.In order to find the effective distance,T-cell transmits the molecules 1(Interleukins-2)to B-cell,upon successful reception of molecules 1,antibodies(molecules 2)transmit back to T-cell.Then,the effective distance of an immune system can be obtained after T-cell detects the concentration of the molecules 2 with respect to time.Different schemes of S/W ARQ protocols have implemented in Molecular Communication(MC)but it requires retransmission of duplicate copies due to the lack of addressing an effective distance.Thus,the simulations are performed in MATLAB and the results obtain higher capture probability and also successful complete transmission of the desired molecules.
