Objective To design and construct the eukaryotic expression vector which expresses Anti-CD3 ScFv-B7.1 fusion molecules and predict the biological characteristics, the rationality and feasibility of the spacer. Methods...Objective To design and construct the eukaryotic expression vector which expresses Anti-CD3 ScFv-B7.1 fusion molecules and predict the biological characteristics, the rationality and feasibility of the spacer. Methods To analyze the flexibility, Hoop & Woods hydrophilicity and the epitope of Anti-CD3 ScFv-B7.1 fusion molecule at secondary structure level by computer simulation utilizing the GoldKey software. Results By comparing with Anti-CD3 ScFv and B7.1 respectively, it shows that Anti-CD3 ScFv-B7.1 fusion molecules can form correct secondary structure with the linking of the spacer, the fusion does not change the original hydrophilicity and epitopes of both Anti-CD3 ScFv and B7.1, no new epitopes emerge; The spacer is flexible and shows low antigenicity. Conclusion The design of Anti-CD3 ScFv-B7.1 fusion molecule are rational and feasible, the expressed fusion protein could retain the maximum biological activity and the function of both Anti-CD3 ScFv and B7.1.展开更多
The precise mechanism underlying the effects of anti-CD4 antibody and calcium ions(Ca^(2+)) in peanut allergy remains unknown.C3 H/HeJ mice sensitized with peanut protein extract(PPE)were injected with anti-CD4 antibo...The precise mechanism underlying the effects of anti-CD4 antibody and calcium ions(Ca^(2+)) in peanut allergy remains unknown.C3 H/HeJ mice sensitized with peanut protein extract(PPE)were injected with anti-CD4 antibodies for 4 weeks.Stimulation with PPE increased the specific immunoglobulin E(IgE),cytokine,histamine,and mMcp-1 levels,upregulated decorin(Dcn)expression,induced Ca^(2+) inflow in the spleen,and augmented the expression of the transcription factors GATA-3 and Foxp3,which resulted in Th2 and Treg cell activation.Notably,the Ca^(2+) levels were positively correlated with the histamine,interleukin(IL)-4,IL-5,and IL-13 levels,and negatively correlated with IL-10 levels.However,administration of anti-CD4 antibodies markedly alleviated allergic symptoms,activated T cells,and reduced Ca^(2+) inflow,cytokine,histamine,mMcp-1,and the IgHG3,CXCLI2,MMP2 and FABP4 gene.Our results indicated that anti-CD4 antibodies can ameliorate PPE-induced allergy,which is probably related to the suppression of Ca^(2+) inflow,and inhibiting histamine,cytokine and IgHG3,CXCL12,MMP2,and FABP4,thus exerting a protective effect against PPEsensitized food allergy.展开更多
To evaluate the effect of anti-HER-2 × anti-CD3 bi-specific antibody（BsAb） on the growth of HER-2/neu-expressing human gastric carcinoma in vitro and in vivo, an MTT assay was carried out to test the inhibitive...To evaluate the effect of anti-HER-2 × anti-CD3 bi-specific antibody（BsAb） on the growth of HER-2/neu-expressing human gastric carcinoma in vitro and in vivo, an MTT assay was carried out to test the inhibitive rates of herceptin, anti-CD3 and BsAb antibodies on SGC-7901 gastric carcinoma cells. Immunocytochemistry methods were used to test the HER-2 level of SGC-7901. Nude mice models were employed to test the effect of HER-2 CD3 BsAh combined with effector ceils（ peripheral blood lymphatic cells of healthy human beings） on the growth of tumors in animals. Compared with that of the untreated control group, the tumor cell growth rates in vitro and in vivo will both be significantly inhibited when treated with effector cells combined with anti-CD3 McAb, herceptin or HER2 CD3 BsAb （p 〈0. 05）, and the growth inhibition is the most remarkable in the group treated with HER2 CD3 BsAb combined with effector cells. The growth of tumor xenografts will also be significantly inhibited in the group treated with HER2 CD3 BsAb combined with effector cells when compared with that in the group treated with anti-CD3 McAb or the group treated with herceptin combined with effector cells（p 〈0. 05）. We can conclude that HER-2/neu is possibly a useful target for immunotherapy of gastric carcinoma, and anti-HER2 × anti-CD3 BsAb has evident anti-tumor efficacy both, in vitro and in vivo.展开更多
The proliferation of splenocytes from healthy adults was induced by anti-CD3 McAb,PHA and IL-2.The proliferative capability and anti-tumor activity as well as phenotypes of the splenocytes cultured in different medium...The proliferation of splenocytes from healthy adults was induced by anti-CD3 McAb,PHA and IL-2.The proliferative capability and anti-tumor activity as well as phenotypes of the splenocytes cultured in different medium systems were studied.The results showed that anti-CD3 McAb and PHA not only enhanced the proliferation of splenocytes induced by IL-2,but also produced synergism if used simultaneously.The expressions of CD4 and Tac of cellular surface markers were increased after splenocytes were induced by anti-CD3 McAb and PHA.The results of anti-tumor activity of LAK cells suggested that PHA had the capability to promote anti-tumor activity of LAK cells by both direct and in direct pathways,but anti-CD3 McAb indirectly promoted anti-tumor activity of LAK cellsby enhanctng splenocyte proliferation.展开更多
BACKGROUND Islets of Langerhans beta cells diminish in autoimmune type 1 diabetes mellitus(T1DM).Teplizumab,a humanized anti-CD3 monoclonal antibody,may help T1DM.Its long-term implications on clinical T1DM developmen...BACKGROUND Islets of Langerhans beta cells diminish in autoimmune type 1 diabetes mellitus(T1DM).Teplizumab,a humanized anti-CD3 monoclonal antibody,may help T1DM.Its long-term implications on clinical T1DM development,safety,and efficacy are unknown.AIM To assess the effectiveness and safety of teplizumab as a therapeutic intervention for individuals with T1DM.METHODS A systematic search was conducted using four electronic databases(PubMed,Embase,Scopus,and Cochrane Library)to select publications published in peerreviewed journals written in English.The odds ratio(OR)and risk ratio(RR)were calculated,along with their 95%CI.We assessed heterogeneity using Cochrane Q and I2 statistics and the appropriate P value.RESULTS There were 8 randomized controlled trials(RCTs)in the current meta-analysis with a total of 1908 T1DM patients from diverse age cohorts,with 1361 patients receiving Teplizumab and 547 patients receiving a placebo.Teplizumab was found to have a substantial link with a decrease in insulin consumption,with an OR of 4.13(95%CI:1.72 to 9.90).Teplizumab is associated with an improved Cpeptide response(OR 2.49;95%CI:1.62 to 3.81)and a significant change in Glycated haemoglobin A1c(HbA1c)levels in people with type 1 diabetes[OR 1.75(95%CI:1.03 to 2.98)],and it has a RR of 0.71(95%CI:0.53 to 0.95).CONCLUSION In type 1 diabetics,teplizumab decreased insulin consumption,improved C-peptide response,and significantly changed HbA1c levels with negligible side effects.Teplizumab appears to improve glycaemic control and diabetes management with good safety and efficacy.展开更多
Regulatory T cells （Treg） play important roles in immune system homeostasis, and may also be involved in tumor immunotolerance by suppressing Th1 immune response which is involved in anti-tumor immunity. We have pre...Regulatory T cells （Treg） play important roles in immune system homeostasis, and may also be involved in tumor immunotolerance by suppressing Th1 immune response which is involved in anti-tumor immunity. We have previously reported that immunization with attenuated activated autologous T cells leads to enhanced anti-tumor immunity and upregulated Thl responses in vivo. However, the underlying molecular mechanisms are not well understood. Here we show that Treg function was significantly downregulated in mice that received immunization of attenuated activated autologous T cells. We found that Foxp3 expression decreased in CD4＋CD25＋ T cells from the immunized mice. Moreover, CD4＋CD25＋Foxp3＋ Treg obtained from immunized mice exhibited diminished immunosuppression ability compared to those from naive mice. Further analysis showed that the serum of immunized mice contains a high level ofanti-CD25 antibody （about 30 ng/ml, p〈0.01 vs controls）. Consistent with a role ofanti-CD25 response in the downregulation of Treg, adoptive transfer of serum from immunized mice to naive mice led to a significant decrease in Treg population and function in recipient mice. The triggering of anti-CD25 response in immunized mice can be explained by the fact that CD25 was induced to a high level in the ConA activated autologous T cells used for immunization. Our results demonstrate for the first time that immunization with attenuated activated autologous T cells evokes anti-CD25 antibody production, which leads to impeded CD4＋CD25＋Foxp3＋ Treg expansion and function in vivo. We suggest that dampened Treg function likely contributes to enhanced Thl response in immunized mice and is at least part of the mechanism underlying the boosted anti-tumor immunity.展开更多
Type 1 diabetes(T1D) is an autoimmune disease that results from the destruction of insulin-producing cells by autoreactive T cells,leading to lifelong dependency on insulin therapy and increased risk of long-term card...Type 1 diabetes(T1D) is an autoimmune disease that results from the destruction of insulin-producing cells by autoreactive T cells,leading to lifelong dependency on insulin therapy and increased risk of long-term cardiovascular complications.Here we take the opportunity of the 20thanniversary of the generation of the BDC2.5 TCR transgenic non-obese diabetic(NOD) mouse model,to provide a brief overview of the significant progress that has been made in understanding the role of T cells in the disease pathogenesis period.This included development of hundreds of reagents that block or even reverse new-onset disease by directly or indirectly controlling T cells.We also reflect on the sobering fact that none of these strategies has shown significant efficacy in clinical trials and discuss potential reasons hindering translation of the preclinical findings into successful therapeutic strategies and potential ways forward.展开更多
Atherosclerosis is a significant contributor to global cardiovascular disease.Reducing the formation of atherosclerotic plaque effectively can lead to a decrease in cardiovascular diseases.Therefore,controlling macrop...Atherosclerosis is a significant contributor to global cardiovascular disease.Reducing the formation of atherosclerotic plaque effectively can lead to a decrease in cardiovascular diseases.Therefore,controlling macrophage function is crucial.This study presents the creation of a bifunctional nanoparticle that is specific to macrophages to achieve intracellular and extracellular synergistic therapy for restoring macrophage functions.The nanoparticle is conjugated with anti-CD47 antibody to modulate extracellular CD47-SIRPαphagocytic signaling axis on the outer surface of macrophages and encapsulates the NLRP3 inhibitor(CY-09)to regulate intracellular inflammation response of macrophages.The results showed that the nanoparticles accumulate in the atherosclerotic plaque,alter macrophage phagocytosis,inhibit NLRP3 inflammasome activation,and decrease the plaque burden in Apoe^(-/-)mice whilst ensuring safety.Examination of single-cell RNA sequencing indicates that this multifunctional nanoparticle decreases the expression of genes linked to inflammation and manages inflammatory pathways in the plaque lesion.This study proposes a synergistic therapeutic approach that utilizes a bifunctional nanoparticle,conjugated with anti-CD47,to regulate the microenvironment of plaques.展开更多
文摘Objective To design and construct the eukaryotic expression vector which expresses Anti-CD3 ScFv-B7.1 fusion molecules and predict the biological characteristics, the rationality and feasibility of the spacer. Methods To analyze the flexibility, Hoop & Woods hydrophilicity and the epitope of Anti-CD3 ScFv-B7.1 fusion molecule at secondary structure level by computer simulation utilizing the GoldKey software. Results By comparing with Anti-CD3 ScFv and B7.1 respectively, it shows that Anti-CD3 ScFv-B7.1 fusion molecules can form correct secondary structure with the linking of the spacer, the fusion does not change the original hydrophilicity and epitopes of both Anti-CD3 ScFv and B7.1, no new epitopes emerge; The spacer is flexible and shows low antigenicity. Conclusion The design of Anti-CD3 ScFv-B7.1 fusion molecule are rational and feasible, the expressed fusion protein could retain the maximum biological activity and the function of both Anti-CD3 ScFv and B7.1.
基金supported by the National Natural Science Foundation of China(31972185)。
文摘The precise mechanism underlying the effects of anti-CD4 antibody and calcium ions(Ca^(2+)) in peanut allergy remains unknown.C3 H/HeJ mice sensitized with peanut protein extract(PPE)were injected with anti-CD4 antibodies for 4 weeks.Stimulation with PPE increased the specific immunoglobulin E(IgE),cytokine,histamine,and mMcp-1 levels,upregulated decorin(Dcn)expression,induced Ca^(2+) inflow in the spleen,and augmented the expression of the transcription factors GATA-3 and Foxp3,which resulted in Th2 and Treg cell activation.Notably,the Ca^(2+) levels were positively correlated with the histamine,interleukin(IL)-4,IL-5,and IL-13 levels,and negatively correlated with IL-10 levels.However,administration of anti-CD4 antibodies markedly alleviated allergic symptoms,activated T cells,and reduced Ca^(2+) inflow,cytokine,histamine,mMcp-1,and the IgHG3,CXCLI2,MMP2 and FABP4 gene.Our results indicated that anti-CD4 antibodies can ameliorate PPE-induced allergy,which is probably related to the suppression of Ca^(2+) inflow,and inhibiting histamine,cytokine and IgHG3,CXCL12,MMP2,and FABP4,thus exerting a protective effect against PPEsensitized food allergy.
文摘To evaluate the effect of anti-HER-2 × anti-CD3 bi-specific antibody（BsAb） on the growth of HER-2/neu-expressing human gastric carcinoma in vitro and in vivo, an MTT assay was carried out to test the inhibitive rates of herceptin, anti-CD3 and BsAb antibodies on SGC-7901 gastric carcinoma cells. Immunocytochemistry methods were used to test the HER-2 level of SGC-7901. Nude mice models were employed to test the effect of HER-2 CD3 BsAh combined with effector ceils（ peripheral blood lymphatic cells of healthy human beings） on the growth of tumors in animals. Compared with that of the untreated control group, the tumor cell growth rates in vitro and in vivo will both be significantly inhibited when treated with effector cells combined with anti-CD3 McAb, herceptin or HER2 CD3 BsAb （p 〈0. 05）, and the growth inhibition is the most remarkable in the group treated with HER2 CD3 BsAb combined with effector cells. The growth of tumor xenografts will also be significantly inhibited in the group treated with HER2 CD3 BsAb combined with effector cells when compared with that in the group treated with anti-CD3 McAb or the group treated with herceptin combined with effector cells（p 〈0. 05）. We can conclude that HER-2/neu is possibly a useful target for immunotherapy of gastric carcinoma, and anti-HER2 × anti-CD3 BsAb has evident anti-tumor efficacy both, in vitro and in vivo.
文摘The proliferation of splenocytes from healthy adults was induced by anti-CD3 McAb,PHA and IL-2.The proliferative capability and anti-tumor activity as well as phenotypes of the splenocytes cultured in different medium systems were studied.The results showed that anti-CD3 McAb and PHA not only enhanced the proliferation of splenocytes induced by IL-2,but also produced synergism if used simultaneously.The expressions of CD4 and Tac of cellular surface markers were increased after splenocytes were induced by anti-CD3 McAb and PHA.The results of anti-tumor activity of LAK cells suggested that PHA had the capability to promote anti-tumor activity of LAK cells by both direct and in direct pathways,but anti-CD3 McAb indirectly promoted anti-tumor activity of LAK cellsby enhanctng splenocyte proliferation.
文摘BACKGROUND Islets of Langerhans beta cells diminish in autoimmune type 1 diabetes mellitus(T1DM).Teplizumab,a humanized anti-CD3 monoclonal antibody,may help T1DM.Its long-term implications on clinical T1DM development,safety,and efficacy are unknown.AIM To assess the effectiveness and safety of teplizumab as a therapeutic intervention for individuals with T1DM.METHODS A systematic search was conducted using four electronic databases(PubMed,Embase,Scopus,and Cochrane Library)to select publications published in peerreviewed journals written in English.The odds ratio(OR)and risk ratio(RR)were calculated,along with their 95%CI.We assessed heterogeneity using Cochrane Q and I2 statistics and the appropriate P value.RESULTS There were 8 randomized controlled trials(RCTs)in the current meta-analysis with a total of 1908 T1DM patients from diverse age cohorts,with 1361 patients receiving Teplizumab and 547 patients receiving a placebo.Teplizumab was found to have a substantial link with a decrease in insulin consumption,with an OR of 4.13(95%CI:1.72 to 9.90).Teplizumab is associated with an improved Cpeptide response(OR 2.49;95%CI:1.62 to 3.81)and a significant change in Glycated haemoglobin A1c(HbA1c)levels in people with type 1 diabetes[OR 1.75(95%CI:1.03 to 2.98)],and it has a RR of 0.71(95%CI:0.53 to 0.95).CONCLUSION In type 1 diabetics,teplizumab decreased insulin consumption,improved C-peptide response,and significantly changed HbA1c levels with negligible side effects.Teplizumab appears to improve glycaemic control and diabetes management with good safety and efficacy.
基金This work was supported by National Natural Science Foundation of China(No.30671945)Science and Technology Commission of Shanghai Municipality(Nos.06JC14044,05ZR14055,054319928,04DZ14902)+2 种基金Shanghai Municipal Education(No.05BZ26)Shanghai Leading Academic Discipline Project(T0206)Science Foundation of Shanghai Institute of Immunology(No.07-A04,to Ningli Li).
文摘Regulatory T cells （Treg） play important roles in immune system homeostasis, and may also be involved in tumor immunotolerance by suppressing Th1 immune response which is involved in anti-tumor immunity. We have previously reported that immunization with attenuated activated autologous T cells leads to enhanced anti-tumor immunity and upregulated Thl responses in vivo. However, the underlying molecular mechanisms are not well understood. Here we show that Treg function was significantly downregulated in mice that received immunization of attenuated activated autologous T cells. We found that Foxp3 expression decreased in CD4＋CD25＋ T cells from the immunized mice. Moreover, CD4＋CD25＋Foxp3＋ Treg obtained from immunized mice exhibited diminished immunosuppression ability compared to those from naive mice. Further analysis showed that the serum of immunized mice contains a high level ofanti-CD25 antibody （about 30 ng/ml, p〈0.01 vs controls）. Consistent with a role ofanti-CD25 response in the downregulation of Treg, adoptive transfer of serum from immunized mice to naive mice led to a significant decrease in Treg population and function in recipient mice. The triggering of anti-CD25 response in immunized mice can be explained by the fact that CD25 was induced to a high level in the ConA activated autologous T cells used for immunization. Our results demonstrate for the first time that immunization with attenuated activated autologous T cells evokes anti-CD25 antibody production, which leads to impeded CD4＋CD25＋Foxp3＋ Treg expansion and function in vivo. We suggest that dampened Treg function likely contributes to enhanced Thl response in immunized mice and is at least part of the mechanism underlying the boosted anti-tumor immunity.
基金Supported by The NIH(1R56AI099027 and 1R01AI099027-01)American Heart Association(10GRNT4200003)
文摘Type 1 diabetes(T1D) is an autoimmune disease that results from the destruction of insulin-producing cells by autoreactive T cells,leading to lifelong dependency on insulin therapy and increased risk of long-term cardiovascular complications.Here we take the opportunity of the 20thanniversary of the generation of the BDC2.5 TCR transgenic non-obese diabetic(NOD) mouse model,to provide a brief overview of the significant progress that has been made in understanding the role of T cells in the disease pathogenesis period.This included development of hundreds of reagents that block or even reverse new-onset disease by directly or indirectly controlling T cells.We also reflect on the sobering fact that none of these strategies has shown significant efficacy in clinical trials and discuss potential reasons hindering translation of the preclinical findings into successful therapeutic strategies and potential ways forward.
基金supported by the National Natural Science Foundation of China(82202310,82170375 and 11902211)Key Research and Development Project of Science&Technology Department of Sichuan Province(2022ZDZX0020)+4 种基金Nature Science Foundation of Sichuan Province(2023NSFSC1630 and 2023NSFSC1339)1⋅3⋅5 project for disciplines of excellence-Clinical Research Fund,West China Hospital,Sichuan UniversityChina Postdoctoral Science Foundation(2022M712254)The Fundamental Research Funds for the Central Universities(2022SCU12049)Post-Doctor Research Project,West China Hospital,Sichuan University(2021HXBH068)。
文摘Atherosclerosis is a significant contributor to global cardiovascular disease.Reducing the formation of atherosclerotic plaque effectively can lead to a decrease in cardiovascular diseases.Therefore,controlling macrophage function is crucial.This study presents the creation of a bifunctional nanoparticle that is specific to macrophages to achieve intracellular and extracellular synergistic therapy for restoring macrophage functions.The nanoparticle is conjugated with anti-CD47 antibody to modulate extracellular CD47-SIRPαphagocytic signaling axis on the outer surface of macrophages and encapsulates the NLRP3 inhibitor(CY-09)to regulate intracellular inflammation response of macrophages.The results showed that the nanoparticles accumulate in the atherosclerotic plaque,alter macrophage phagocytosis,inhibit NLRP3 inflammasome activation,and decrease the plaque burden in Apoe^(-/-)mice whilst ensuring safety.Examination of single-cell RNA sequencing indicates that this multifunctional nanoparticle decreases the expression of genes linked to inflammation and manages inflammatory pathways in the plaque lesion.This study proposes a synergistic therapeutic approach that utilizes a bifunctional nanoparticle,conjugated with anti-CD47,to regulate the microenvironment of plaques.
