A key question in the coronavirus disease 2019(COVID-19)pandemic is the duration of specific T cell responses against the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)post primary infection,which is diff...A key question in the coronavirus disease 2019(COVID-19)pandemic is the duration of specific T cell responses against the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)post primary infection,which is difficult to address due to the large-scale COVID-19 vaccination and re-exposure to the virus.Here,we conducted an analysis of the long-term SARS-CoV-2-specific T cell responses in a unique cohort of convalescent individuals(CIs)that were among the first to be infected worldwide and without any possible antigen re-exposure since then.The magnitude and breadth of SARS-CoV-2-specific T cell responses correlated inversely with the time that had elapsed from disease onset and the age of those CIs.The mean magnitude of SARS-CoV-2-specific CD4 and CD8 T cell responses decreased about 82%and 76%,respectively,over the time period of ten months after infection.Accordingly,the longitudinal analysis also demonstrated that SARS-CoV-2-specific T cell responses waned significantly in 75%of CIs during the follow-up.Collectively,we provide a comprehensive characterization of the long-term memory T cell response in CIs,suggesting that robust SARS-CoV-2-specific T cell immunity post primary infection may be less durable than previously expected.展开更多
BACKGROUND The global outbreak of human severe acute respiratory syndrome coronavirus(SARS-CoV)-2 infection represents an urgent need for readily available,accurate and rapid diagnostic tests.Nucleic acid testing of r...BACKGROUND The global outbreak of human severe acute respiratory syndrome coronavirus(SARS-CoV)-2 infection represents an urgent need for readily available,accurate and rapid diagnostic tests.Nucleic acid testing of respiratory tract specimens for SARS-CoV-2 is the current gold standard for diagnosis of coronavirus disease 2019(COVID-19).However,the diagnostic accuracy of reverse transcription polymerase chain reaction(RT-PCR)tests for detecting SARS-CoV-2 nucleic acid may be lower than optimal.The detection of SARS-CoV-2-specific antibodies should be used as a serological non-invasive tool for the diagnosis and management of SARS-CoV-2 infection.AIM To investigate the diagnostic value of SARS-CoV-2 IgM/IgG and nucleic acid detection in COVID-19.METHODS We retrospectively analyzed 652 suspected COVID-19 patients,and 206 non-COVID-19 patients in Wuhan Integrated TCM and Western Medicine Hospital.Data on SARS-CoV-2 nucleic acid tests and serum antibody tests were collected to investigate the diagnostic value of nucleic acid RT-PCR test kits and immunoglobulin(Ig)M/IgG antibody test kits.The j2 test was used to compare differences between categorical variables.A 95%confidence interval(CI)was provided by the Wilson score method.All analyses were performed with IBM SPSS Statistics version 22.0(IBM Corp.,Armonk,NY,United States).RESULTS Of the 652 suspected COVID-19 patients,237(36.3%)had positive nucleic acid tests,311(47.7%)were positive for IgM,and 592(90.8%)were positive for IgG.There was a significant difference in the positive detection rate between the IgM and IgG test groups(P<0.001).Using the RT-PCR results as a reference,the specificity,sensitivity,and accuracy of IgM/IgG combined tests for SARS-CoV-2 infection were 98.5%,95.8%,and 97.1%,respectively.Of the 415 suspected COVID-19 patients with negative nucleic acid test results,366 had positive IgM/IgG tests with a positive detection rate of 88.2%.CONCLUSION Our data indicate that serological IgM/IgG antibody combined test had high sensitivity and specificity for the diagnosis of SARS-CoV-2 infection,and can be used in combination with RT-PCR for the diagnosis of SARS-CoV-2 infection.展开更多
To editor:In March 2020,the World Health Organization officially announced the coronavirus disease 2019(COVID-19)pandemic,also known as the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)outbreak,as a seri...To editor:In March 2020,the World Health Organization officially announced the coronavirus disease 2019(COVID-19)pandemic,also known as the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)outbreak,as a serious global health crisis due to the alarming levels of morbidity and mortality associated with the virus worldwide.1 SARS-CoV-2 infected individuals may remain asymptomatic in the early stage,but as the disease progresses,it can lead to severe pneumonia,dyspnea,organ dysfunction,and even death.As of November 2023,COVID-19 has affected more than 772 million people and led to the deaths of more than six million individuals globally.2 During pregnancy,women with symptomatic SARS-CoV-2 infection face an increased risk of developing severe complications,with elevated rates of hospital admission,intensive care unit admission,and death.展开更多
During viral infections,antibodies and T cells act together to prevent pathogen spread and remove virus-infected cells.Virusspecific adaptive immunity can,however,also trigger pathological processes characterized by l...During viral infections,antibodies and T cells act together to prevent pathogen spread and remove virus-infected cells.Virusspecific adaptive immunity can,however,also trigger pathological processes characterized by localized or systemic inflammatory events.The protective and/or pathological role of virus-specific T cells in SARS-CoV-2 infection has been the focus of many studies in COVID-19 patients and in vaccinated individuals.Here,we review the works that have elucidated the function of SARS-CoV-2-specific T cells in patients and in vaccinated individuals.Understanding whether SARS-CoV-2-specific T cells are more linked to protection or pathogenesis is pivotal to define future therapeutic and prophylactic strategies to manage the current pandemic.展开更多
Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is an ongoing pandemic that poses a great threat to human health worldwide.As the humoral immune response plays ...Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is an ongoing pandemic that poses a great threat to human health worldwide.As the humoral immune response plays essential roles in disease occurrence and development,understanding the dynamics and characteristics of virus-specific humoral immunity in SARS-CoV-2-infected patients is of great importance for controlling this disease.In this review,we summarize the characteristics of the humoral immune response after SARS-CoV-2 infection and further emphasize the potential applications and therapeutic prospects of SARSCoV-2-specific humoral immunity and the critical role of this immunity in vaccine development.Notably,serological antibody testing based on the humoral immune response can guide public health measures and control strategies;however,it is not recommended for population surveys in areas with very low prevalence.Existing evidence suggests that asymptomatic individuals have a weaker immune response to SARS-CoV-2 infection,whereas SARS-CoV-2-infected children have a more effective humoral immune response than adults.The correlations between antibody(especially neutralizing antibody)titers and protection against SARS-CoV-2 reinfection should be further examined.In addition,the emergence of cross-reactions among different coronavirus antigens in the development of screening technology and the risk of antibody-dependent enhancement related to SARS-CoV-2 vaccination should be given further attention.展开更多
Background:T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibition motif domains(TIGIT),an inhibitory receptor expressed on T cells,plays a dysfunctional role in antiviral infection and ...Background:T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibition motif domains(TIGIT),an inhibitory receptor expressed on T cells,plays a dysfunctional role in antiviral infection and antitumor activity.However,it is unknown whether TIGIT expression on T cells influences the immunological effects of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)inactivated vaccines.Methods:Forty-five people living with HIV(PLWH)on antiretroviral therapy(ART)for more than two years and 31 healthy controls(HCs),all received a third dose of a SARS-CoV-2 inactivated vaccine,were enrolled in this study.The amounts,activation,proportion of cell subsets,and magnitude of the SARS-CoV-2-specific immune response of TIGIT^(+)CD4^(+)and TIGIT^(+)CD8^(+)T cells were investigated before the third dose but 6 months after the second vaccine dose(0W),4 weeks(4W)and 12 weeks(12W)after the third dose.Results:Compared to that in HCs,the frequency of TIGIT^(+)CD8^(+)T cells in the peripheral blood of PLWH increased at 12W after the third dose of the inactivated vaccine,and the immune activation of TIGIT^(+)CD8^(+)T cells also increased.A decrease in the ratio of both T naïve(T_(N))and central memory(T_(CM))cells among TIGIT^(+)CD8^(+)T cells and an increase in the ratio of the effector memory(T_(EM))subpopulation were observed at 12W in PLWH.Interestingly,particularly at 12W,a higher proportion of TIGIT^(+)CD8^(+)T cells expressing CD137 and CD69 simultaneously was observed in HCs than in PLWH based on the activation-induced marker assay.Compared with 0W,SARS-CoV-2-specific TIGIT^(+)CD8^(+)T-cell responses in PLWH were not enhanced at 12W but were enhanced in HCs.Additionally,at all time points,the SARS-CoV-2-specific responses of TIGIT^(+)CD8^(+)T cells in PLWH were significantly weaker than those of TIGIT-CD8^(+)T cells.However,in HCs,the difference in the SARS-CoV-2-specific responses induced between TIGIT^(+)CD8^(+)T cells and TIGIT-CD8^(+)T cells was insignificant at 4W and 12W,except at 0W.Conclusions:TIGIT expression on CD8^(+)T cells may hinder the T-cell immune response to a booster dose of an inactivated SARS-CoV-2 vaccine,suggesting weakened resistance to SARS-CoV-2 infection,especially in PLWH.Furthermore,TIGIT may be used as a potential target to increase the production of SARS-CoV-2-specific CD8^(+)T cells,thereby enhancing the effectiveness of vaccination.展开更多
Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)-induced cytokine storm is closely associated with coronavirus disease 2019(COVID-19)severity and lethality.However,drugs that are effective against inflammat...Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)-induced cytokine storm is closely associated with coronavirus disease 2019(COVID-19)severity and lethality.However,drugs that are effective against inflammation to treat lethal COVID-19 are still urgently needed.Here,we constructed a SARS-CoV-2 spike protein-specific CAR,and human T cells infected with this CAR(SARS-CoV-2-S CAR-T)and stimulated with spike protein mimicked the T-cell responses seen in COVID-19 patients,causing cytokine storm and displaying a distinct memory,exhausted,and regulatory T-cell phenotype.THP1 remarkably augmented cytokine release in SARS-CoV-2-S CAR-T cells when they were in coculture.Based on this"two-cell"(CAR-T and THP1 cells)model,we screened an FDA-approved drug library and found that felodipine,fasudil,imatinib,and caspofungin were effective in suppressing the release of cytokines,which was likely due to their ability to suppress the NF-kB pathway in vitro.Felodipine,fasudi,imatinib,and caspofungin were further demonstrated,although to different extents,to attenuate lethal inflammation,ameliorate severe pneumonia,and prevent mortality in a SARS-CoV-2-infected Syrian hamster model,which were also linked to their suppressive role in inflammation.In summary,we established a SARS-CoV-2-specific CAR-T-cell model that can be utilized as a tool for anti-inflammatory drug screening in a fast and high-throughput manner.The drugs identified herein have great potential for early treatment to prevent COVID-19 patients from cytokine storm-induced lethality in the clinic because they are safe,inexpensive,and easily accessible for immediateuseinmostcountries.展开更多
目的评价原型株SARS-CoV-2灭活疫苗免疫BALB/c小鼠后对Delta株病毒的体液及细胞免疫效果,为现有疫苗对变异株的保护效果评价以及研发更加安全有效的疫苗提供参考。方法将SARS-CoV-2灭活疫苗经腹腔免疫雌性BALB/c小鼠2次,间隔14 d,以免疫...目的评价原型株SARS-CoV-2灭活疫苗免疫BALB/c小鼠后对Delta株病毒的体液及细胞免疫效果,为现有疫苗对变异株的保护效果评价以及研发更加安全有效的疫苗提供参考。方法将SARS-CoV-2灭活疫苗经腹腔免疫雌性BALB/c小鼠2次,间隔14 d,以免疫PBS作为对照,每组10只。初次免疫后第7、14、21、28、35和42天采集血清,间接ELISA法检测血清中针对Delta株病毒S和N蛋白的结合抗体效价,微量中和试验检测针对Delta株病毒的中和抗体效价。初次免疫后第42天,取小鼠脾脏,进行Elispot检测,评价细胞免疫水平。结果初次免疫后第7天即可检测到S蛋白结合抗体,加强免疫后抗体效价进一步升高,至第21天抗体几何平均滴度(geometric mean titer,GMT)为89144;而初次免疫后N蛋白结合抗体水平较低,加强免疫后迅速升高,与S蛋白抗体水平相当。初次免疫后第7、14天小鼠中和抗体阳转数为4/10和8/10,加强免疫后全部小鼠抗体阳转,中和抗体GMT达391。初次免疫后第42天,疫苗组IFNγ和IL-2平均斑点数均显著高于对照组(t分别为8.094和13.08,P均<0.0001)。结论SARS-CoV-2灭活疫苗2次免疫能够有效刺激小鼠产生针对Delta株病毒的体液免疫和细胞免疫。展开更多
基金supported by the National Natural Science Foundation of China (92169105,82172256,81861138044,91742114 and M-0060)the Fundamental Research Funds for the Central Universities (2020kfyXGYJ028,2020kfyXGYJ046 and 2020kfyXGYJ016)+2 种基金the National Science and Technology Major Project (2017ZX10202203-007-006,2017ZX10202202-001-009,2017ZX10202202-002-008,2017ZX10202201-002-003)the Deutsche Forschungsgemeinschaft (DI 714/22-1,ZE 893/2-1,and RTG1949/2)the Medical Faculty of the University of Duisburg-Essen and Stiftung Universiatsmedizin,University Hospital Essen,Germany,and the Tongji-Rongcheng Center for Biomedicine,Huazhong University of Science and Technology。
文摘A key question in the coronavirus disease 2019(COVID-19)pandemic is the duration of specific T cell responses against the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)post primary infection,which is difficult to address due to the large-scale COVID-19 vaccination and re-exposure to the virus.Here,we conducted an analysis of the long-term SARS-CoV-2-specific T cell responses in a unique cohort of convalescent individuals(CIs)that were among the first to be infected worldwide and without any possible antigen re-exposure since then.The magnitude and breadth of SARS-CoV-2-specific T cell responses correlated inversely with the time that had elapsed from disease onset and the age of those CIs.The mean magnitude of SARS-CoV-2-specific CD4 and CD8 T cell responses decreased about 82%and 76%,respectively,over the time period of ten months after infection.Accordingly,the longitudinal analysis also demonstrated that SARS-CoV-2-specific T cell responses waned significantly in 75%of CIs during the follow-up.Collectively,we provide a comprehensive characterization of the long-term memory T cell response in CIs,suggesting that robust SARS-CoV-2-specific T cell immunity post primary infection may be less durable than previously expected.
基金Natural Science Foundation of Hubei Province,China,No.2016CFB596and Wuhan City Medical Research Project,China,No.WX17Q39 and No.WX15B14.
文摘BACKGROUND The global outbreak of human severe acute respiratory syndrome coronavirus(SARS-CoV)-2 infection represents an urgent need for readily available,accurate and rapid diagnostic tests.Nucleic acid testing of respiratory tract specimens for SARS-CoV-2 is the current gold standard for diagnosis of coronavirus disease 2019(COVID-19).However,the diagnostic accuracy of reverse transcription polymerase chain reaction(RT-PCR)tests for detecting SARS-CoV-2 nucleic acid may be lower than optimal.The detection of SARS-CoV-2-specific antibodies should be used as a serological non-invasive tool for the diagnosis and management of SARS-CoV-2 infection.AIM To investigate the diagnostic value of SARS-CoV-2 IgM/IgG and nucleic acid detection in COVID-19.METHODS We retrospectively analyzed 652 suspected COVID-19 patients,and 206 non-COVID-19 patients in Wuhan Integrated TCM and Western Medicine Hospital.Data on SARS-CoV-2 nucleic acid tests and serum antibody tests were collected to investigate the diagnostic value of nucleic acid RT-PCR test kits and immunoglobulin(Ig)M/IgG antibody test kits.The j2 test was used to compare differences between categorical variables.A 95%confidence interval(CI)was provided by the Wilson score method.All analyses were performed with IBM SPSS Statistics version 22.0(IBM Corp.,Armonk,NY,United States).RESULTS Of the 652 suspected COVID-19 patients,237(36.3%)had positive nucleic acid tests,311(47.7%)were positive for IgM,and 592(90.8%)were positive for IgG.There was a significant difference in the positive detection rate between the IgM and IgG test groups(P<0.001).Using the RT-PCR results as a reference,the specificity,sensitivity,and accuracy of IgM/IgG combined tests for SARS-CoV-2 infection were 98.5%,95.8%,and 97.1%,respectively.Of the 415 suspected COVID-19 patients with negative nucleic acid test results,366 had positive IgM/IgG tests with a positive detection rate of 88.2%.CONCLUSION Our data indicate that serological IgM/IgG antibody combined test had high sensitivity and specificity for the diagnosis of SARS-CoV-2 infection,and can be used in combination with RT-PCR for the diagnosis of SARS-CoV-2 infection.
文摘To editor:In March 2020,the World Health Organization officially announced the coronavirus disease 2019(COVID-19)pandemic,also known as the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)outbreak,as a serious global health crisis due to the alarming levels of morbidity and mortality associated with the virus worldwide.1 SARS-CoV-2 infected individuals may remain asymptomatic in the early stage,but as the disease progresses,it can lead to severe pneumonia,dyspnea,organ dysfunction,and even death.As of November 2023,COVID-19 has affected more than 772 million people and led to the deaths of more than six million individuals globally.2 During pregnancy,women with symptomatic SARS-CoV-2 infection face an increased risk of developing severe complications,with elevated rates of hospital admission,intensive care unit admission,and death.
基金This study is supported by the Singapore Ministry of Health’s National Medical Research Council under its COVID-19 Research Fund(COVID19RF3-0060)the Singapore Ministry of Health’s National Medical Research Council MOH-000019(MOH-StaR17Nov-0001)the National Research Foundation,Singapore(NRFCRP17-2017-06).
文摘During viral infections,antibodies and T cells act together to prevent pathogen spread and remove virus-infected cells.Virusspecific adaptive immunity can,however,also trigger pathological processes characterized by localized or systemic inflammatory events.The protective and/or pathological role of virus-specific T cells in SARS-CoV-2 infection has been the focus of many studies in COVID-19 patients and in vaccinated individuals.Here,we review the works that have elucidated the function of SARS-CoV-2-specific T cells in patients and in vaccinated individuals.Understanding whether SARS-CoV-2-specific T cells are more linked to protection or pathogenesis is pivotal to define future therapeutic and prophylactic strategies to manage the current pandemic.
基金supported by the Major National S&T Program Grant(2017ZX10202203 and 2017ZX10302201 to AH)the Science&Technology Commission of China,the Emergency Project(cstc2020jscx-fyzx0053 to AH)+3 种基金the Science&Technology Commission of Chongqing,the National Natural Science Foundation of China(82002131 to YL)the Natural Science Foundation Project of CQ CSTC(cstc2020jcyjmsxmX0081 to YL)the foundation(KJCXZD2020018 to YL,CYS21244 to JZ)the Chongqing Municipal Education Commission,and the COVID-19 Emergency Project(CQMUNCP0207 to YL)from Chongqing Medical University。
文摘Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is an ongoing pandemic that poses a great threat to human health worldwide.As the humoral immune response plays essential roles in disease occurrence and development,understanding the dynamics and characteristics of virus-specific humoral immunity in SARS-CoV-2-infected patients is of great importance for controlling this disease.In this review,we summarize the characteristics of the humoral immune response after SARS-CoV-2 infection and further emphasize the potential applications and therapeutic prospects of SARSCoV-2-specific humoral immunity and the critical role of this immunity in vaccine development.Notably,serological antibody testing based on the humoral immune response can guide public health measures and control strategies;however,it is not recommended for population surveys in areas with very low prevalence.Existing evidence suggests that asymptomatic individuals have a weaker immune response to SARS-CoV-2 infection,whereas SARS-CoV-2-infected children have a more effective humoral immune response than adults.The correlations between antibody(especially neutralizing antibody)titers and protection against SARS-CoV-2 reinfection should be further examined.In addition,the emergence of cross-reactions among different coronavirus antigens in the development of screening technology and the risk of antibody-dependent enhancement related to SARS-CoV-2 vaccination should be given further attention.
基金supported by Beijing Natural Science Foundation(No.L222068 to Bin Su)the National Natural Science Foundation of China(NSFC,No.82272319 to Hu Wu,and No.81974303 to Bin Su)+3 种基金the High-Level Public Health Specialized Talents Project of Beijing Municipal Health Commission(No.2022-2-018 to Bin Su,and No.2022-1-007 to Tong Zhang)the Climbing the peak(Dengfeng)Talent Training Program of Beijing Hospitals Authority(No.DFL20191701 to Tong Zhang)the Beijing Health Technologies Promotion Program(No.BHTPP202002 to Tong Zhang)Beijing Key Laboratory for HIV/AIDS Research(No.BZ0089).
文摘Background:T-cell immunoreceptor with immunoglobulin and immunoreceptor tyrosine-based inhibition motif domains(TIGIT),an inhibitory receptor expressed on T cells,plays a dysfunctional role in antiviral infection and antitumor activity.However,it is unknown whether TIGIT expression on T cells influences the immunological effects of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)inactivated vaccines.Methods:Forty-five people living with HIV(PLWH)on antiretroviral therapy(ART)for more than two years and 31 healthy controls(HCs),all received a third dose of a SARS-CoV-2 inactivated vaccine,were enrolled in this study.The amounts,activation,proportion of cell subsets,and magnitude of the SARS-CoV-2-specific immune response of TIGIT^(+)CD4^(+)and TIGIT^(+)CD8^(+)T cells were investigated before the third dose but 6 months after the second vaccine dose(0W),4 weeks(4W)and 12 weeks(12W)after the third dose.Results:Compared to that in HCs,the frequency of TIGIT^(+)CD8^(+)T cells in the peripheral blood of PLWH increased at 12W after the third dose of the inactivated vaccine,and the immune activation of TIGIT^(+)CD8^(+)T cells also increased.A decrease in the ratio of both T naïve(T_(N))and central memory(T_(CM))cells among TIGIT^(+)CD8^(+)T cells and an increase in the ratio of the effector memory(T_(EM))subpopulation were observed at 12W in PLWH.Interestingly,particularly at 12W,a higher proportion of TIGIT^(+)CD8^(+)T cells expressing CD137 and CD69 simultaneously was observed in HCs than in PLWH based on the activation-induced marker assay.Compared with 0W,SARS-CoV-2-specific TIGIT^(+)CD8^(+)T-cell responses in PLWH were not enhanced at 12W but were enhanced in HCs.Additionally,at all time points,the SARS-CoV-2-specific responses of TIGIT^(+)CD8^(+)T cells in PLWH were significantly weaker than those of TIGIT-CD8^(+)T cells.However,in HCs,the difference in the SARS-CoV-2-specific responses induced between TIGIT^(+)CD8^(+)T cells and TIGIT-CD8^(+)T cells was insignificant at 4W and 12W,except at 0W.Conclusions:TIGIT expression on CD8^(+)T cells may hinder the T-cell immune response to a booster dose of an inactivated SARS-CoV-2 vaccine,suggesting weakened resistance to SARS-CoV-2 infection,especially in PLWH.Furthermore,TIGIT may be used as a potential target to increase the production of SARS-CoV-2-specific CD8^(+)T cells,thereby enhancing the effectiveness of vaccination.
基金Fundamental Research Funds for the Central Universities(20720200104)to LXthe Ministry of Science and Technology of China(2020YFA0112300 and 2020YFA0803600)+3 种基金the National Natural Science Foundation of China(82125028,U22A20320,91953114,31871319,81761128015,and 81861130370)the Natural Science Foundation of Fujian Province of China(2020J02004)the Fundamental Research Funds for the Central University(20720190145 and 20720220003)to WLthe China Postdoctoral Science Foundation(2022M720119)to ZZZ.Part of Fig.6 was drawnbyusingFigdraw.
文摘Severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)-induced cytokine storm is closely associated with coronavirus disease 2019(COVID-19)severity and lethality.However,drugs that are effective against inflammation to treat lethal COVID-19 are still urgently needed.Here,we constructed a SARS-CoV-2 spike protein-specific CAR,and human T cells infected with this CAR(SARS-CoV-2-S CAR-T)and stimulated with spike protein mimicked the T-cell responses seen in COVID-19 patients,causing cytokine storm and displaying a distinct memory,exhausted,and regulatory T-cell phenotype.THP1 remarkably augmented cytokine release in SARS-CoV-2-S CAR-T cells when they were in coculture.Based on this"two-cell"(CAR-T and THP1 cells)model,we screened an FDA-approved drug library and found that felodipine,fasudil,imatinib,and caspofungin were effective in suppressing the release of cytokines,which was likely due to their ability to suppress the NF-kB pathway in vitro.Felodipine,fasudi,imatinib,and caspofungin were further demonstrated,although to different extents,to attenuate lethal inflammation,ameliorate severe pneumonia,and prevent mortality in a SARS-CoV-2-infected Syrian hamster model,which were also linked to their suppressive role in inflammation.In summary,we established a SARS-CoV-2-specific CAR-T-cell model that can be utilized as a tool for anti-inflammatory drug screening in a fast and high-throughput manner.The drugs identified herein have great potential for early treatment to prevent COVID-19 patients from cytokine storm-induced lethality in the clinic because they are safe,inexpensive,and easily accessible for immediateuseinmostcountries.
文摘目的评价原型株SARS-CoV-2灭活疫苗免疫BALB/c小鼠后对Delta株病毒的体液及细胞免疫效果,为现有疫苗对变异株的保护效果评价以及研发更加安全有效的疫苗提供参考。方法将SARS-CoV-2灭活疫苗经腹腔免疫雌性BALB/c小鼠2次,间隔14 d,以免疫PBS作为对照,每组10只。初次免疫后第7、14、21、28、35和42天采集血清,间接ELISA法检测血清中针对Delta株病毒S和N蛋白的结合抗体效价,微量中和试验检测针对Delta株病毒的中和抗体效价。初次免疫后第42天,取小鼠脾脏,进行Elispot检测,评价细胞免疫水平。结果初次免疫后第7天即可检测到S蛋白结合抗体,加强免疫后抗体效价进一步升高,至第21天抗体几何平均滴度(geometric mean titer,GMT)为89144;而初次免疫后N蛋白结合抗体水平较低,加强免疫后迅速升高,与S蛋白抗体水平相当。初次免疫后第7、14天小鼠中和抗体阳转数为4/10和8/10,加强免疫后全部小鼠抗体阳转,中和抗体GMT达391。初次免疫后第42天,疫苗组IFNγ和IL-2平均斑点数均显著高于对照组(t分别为8.094和13.08,P均<0.0001)。结论SARS-CoV-2灭活疫苗2次免疫能够有效刺激小鼠产生针对Delta株病毒的体液免疫和细胞免疫。
