Objective This study aimed to characterize the diagnostic and vaccine potential of a novel Mycobacterium tuberculosisantigen Rv0674. Methods To evaluate thediagnostic potential and antigenicity of Rv0674, IgG was eval...Objective This study aimed to characterize the diagnostic and vaccine potential of a novel Mycobacterium tuberculosisantigen Rv0674. Methods To evaluate thediagnostic potential and antigenicity of Rv0674, IgG was evaluated using ELISA and interferon (IFN)-γ was done by using ELISpot assay among TB patients and healthy donors. For immunogenicity evaluation, BALB/c mice were immunized with Rv0674. Cytokine production was determined by cytokine release assay using an ELISA kit, and the antibodies were tested using ELISA. Results The results of serum Elisa tests showed that Rv0674 specific immunoglobulin G (IgG) response was higher in TB patients than negative controls. And Rv0674 had good performance in serological test with sensitivity and specificity of 77.1% and 81.1%, respectively. While it shows poor sensitivity and specificity of 26.23% and 79.69% for IFN-γ tests. In BALB/c mice, Rv0674 adjuvant by DDA/PolyI:C could also induce a high level of IFN-γ, interleukin-2 and interleukin-6 as well as a high IgG titer in both high-and low-dose groups indicating that Rv0674 is essential in humoral and cellular immunity. Moreover, the cytokine profile and IgG isotypecharacterized Rv0674 as a Th1/Th2-mixed-type protective immunity with the predominance of Th1 cytokines. Conclusion Rv0674 may be a good potential candidate for the development of TB serological diagnosis and a new TB vaccine.展开更多
The plasticity and dynamism in the immune responses to both self and environmental stimulation promote the maintenance and adaptation of a system that tends to harmoniously survive and evolve. Fluctuating antigenic fo...The plasticity and dynamism in the immune responses to both self and environmental stimulation promote the maintenance and adaptation of a system that tends to harmoniously survive and evolve. Fluctuating antigenic forces coexist within the immune system and oscillate between order and chaos to the equilibrium. Thus, when mounting a response to internal or environmental antigens, the main host responses can be divided into two immunological categories. The first, a well-adapted mechanism of complex multi-cellular organisms classically known as tolerance, promotes persistent immunological responses. In the second, opposite way, the modulation of inflammatory immune responses occurs, which we call “intolerance”. Tolerance and intolerance can be mediated by humoral molecules, such as inflammatory compounds, complement, and antibodies, and by different cell types, such as sentinel cells, antigen-presenting cells, and cells that orchestrate the immune response. Tolerogenesis is important in vertebrates because it predisposes species to adapt to self and environmental negative-selective forces. This process depends, in large part, on antigenic co-stimulation (AgCS), which operates as a multi-integrated network formed by all immune and non-immune cells of the body that establishes tolerant immunoregulatory interactions from cells to cells and from cells to the environment. Antigenic distribution, quantity, nature, route of administration, and antigenic convergence on co-stimulatory pathways, and concurrent infections, and the presence of microorganisms (commensals and pathogens) in more than one site are important factors for activating AgCS. To conclude, the AgCS route is a natural immune response generated by heterogeneous APC profile with centralized regulation that promote the counterbalance between intolerant e tolerant status, which can have several applications in the medical and biological fields.展开更多
Objective:To investigate the prevalence of microbial antigenic components of circulating immune complexes amongst grades of CD4 T lymphocyte counts in HIV sero positive and seronegative participants.Methods:Polyethele...Objective:To investigate the prevalence of microbial antigenic components of circulating immune complexes amongst grades of CD4 T lymphocyte counts in HIV sero positive and seronegative participants.Methods:Polyethelene glycol(PEG-600) and buffering methods of precipitation and dissociation of immune complexes was used to generate immune solution from sera of 100 HIV sero-positive and 100 HIV sero-negative participants.These were categorized into 3 grades based on CD4 count:】 500 cell/mm,200-499 cell/mm3 and 【200 cell/mm3.The immune solutions were assayed using membrane based immunoassay and antibody titration, along side its unprocessed serum for detection of various microbial antigens and or antibodies. CD4 T cell counts were estimated using Patec Cyflow SL-3 Germany.Results:Antigenic component of immune complexes of various infectious agents was detected in 99 and 70 HIV seropositive and HIV sero-negative participants,respectively.In group A,there were 10 HIV positive participants,including 4(40.0%) had circulating immune complexes(CICs) due to Salmonella species only:1(10.0%) due to Salmonella-Plasmodium falciparum(P.falciparum),SalmonellaP. falciparum-HCV and P.falciparum antigens,respectively.In group B,45(45.4%) HIV seropositive participants with CICs had CD4 T lymphocyte count between 200-499 cells/mm^3.Out of these,20(44.4%) had CICs due to Salmonella species only:9(20%) due to Salmonella-P. falciparum.In group C,there were 44(44.4%) HIV sero-positive participants,including 3(6.8%) due to Salmonella species only:24(54.4%) due to Salmonella-P.falciparum:2(4.5%) due to P. falciparum only.Conclusions:In HIV sero-positive participants,presence of heterogeneity of Salmonella species-P.falciparum antigens was highly incriminated in CD4 count depletion but not homogeneity of malaria parasites antigens.Malaria parasites antigens only were incriminated in CD4^+ count depletion amongst HIV sero-negative participants.Before taking any decision on the management of HIV-1-positive individuals,their malaria and Salmonella paratyphi status should be assessed,but not malaria status alone.展开更多
T cells engineered to express chimeric antigen receptors (CARs) combining an external antibody binding domain with the CD3ζ T cell receptor (TCR) signaling domain for triggering cell activation are being used for imm...T cells engineered to express chimeric antigen receptors (CARs) combining an external antibody binding domain with the CD3ζ T cell receptor (TCR) signaling domain for triggering cell activation are being used for immunotherapeutic targeting of tumor cells in a non-HLA restricted manner. In this study we transduced T cells with a CD19-CAR construct containing a truncated CD34 gene (tCD34) marker and used these to target the B cell antigen CD19 on the surface of a Hodgkin’s lymphoma (HL) cell line (L591) both in vitro and in vivo. Levels of tCD34 expression in transduced peripheral blood mononuclear cells (PBMCs) ranged from 6% - 20% and this was increased to 82% after selection for transduced tCD34+ cells. In vitro cytotoxicity testing on a CD19+ HL cell line (L591) showed specific cell lysis initiated by the CD19-CAR transduced PBMCs. Importantly, CD19-CAR T cells prevented the growth of L591 HL tumor cells when co-injected subcutaneously (sc) in 6/6 severe combined immunodeficient (SCID) mice. There was no evidence of anti-tumor activity when CD19-CAR T cells were infused intravenously (iv) at the same time as L591 HL tumor cells were injected sc. However, 3/6 SCID mice showed tumor rejection within 83 days after iv infusion of CD19-CAR T cells 3 - 9 days after establishment of L591 HL tumors, while all control animals succumbed to tumors within 60 days. Interestingly, immuno-histochemical analysis of L591 HL tumors demonstrated that CD19-CAR T cells were detected not earlier than 11 days after infusion within the tumor mass. These results suggest that CD19 is a potentially attractive target for the immunotherapy of HL.展开更多
AIM： To investigate the role of regulatory T （Treg） cells in CD4^＋ T cell-mediated bladder autoimmune infammation. METHODS： Urothelium-ovalbumin （URO-OVA）/OT-II mice, a double transgenic line that expresses the...AIM： To investigate the role of regulatory T （Treg） cells in CD4^＋ T cell-mediated bladder autoimmune infammation. METHODS： Urothelium-ovalbumin （URO-OVA）/OT-II mice, a double transgenic line that expresses the membrane form of the model antigen （Ag） OVA as a self-Ag on the urothelium and the OVA-specific CD4^＋ T cell receptor specifc for the I-Ab/OVA323-339 epitope in the periphery, were developed to provide an autoimmune environment for investigation of the role of Treg cells in bladder autoimmune infammation. To facilitate Treg cell analysis, we further developed URO-OVA^GFP-Foxp3/OT-II mice, a derived line of URO-OVA/OT-II mice that express the green fuorescent protein （GFP）-forkhead box protein P3 （Foxp3） fusion protein. RESULTS： URO-OVA/OT-II mice failed to develop bladder infammation despite the presence of autoreactive CD4^＋ T cells. By monitoring GFP-positive cells, bladder infltration of CD4^＋ Treg cells was observed in URO-OVA^GFP-Foxp3/OT-II mice. The infiltrating Treg cells were functionally active and expressed Treg cell effector molecule as well as marker mRNAs including transforming growth factor-β, interleukin （IL）-10, fibrinogen-like protein 2, and glucocorticoid-induced tumor necrosis factor receptor （GITR）. Studies further revealed that Treg cells from URO-OVA^GFP-Foxp3/OT-II mice were suppressive and inhibited autoreactive CD4^＋ T cell proliferation and interferon （IFN）-g production in response to OVA Ag stimulation. Depletion of GITR-positive cells led to spontaneous development of bladder infammation and expression of inflammatory factor mRNAs for IFN-γ, IL-6, tumor necrosis factor-α and nerve growth factor in URO-OVA^GFP-Foxp3/OT-II mice. CONCLUSION： Treg cells specifc for bladder epithelial Ag play an important role in immunological homeostasis and the control of CD4^＋ T cell-mediated bladder autoimmune infammation.展开更多
基金National Science and Technology Major Project of China [2018ZX10731301-002]
文摘Objective This study aimed to characterize the diagnostic and vaccine potential of a novel Mycobacterium tuberculosisantigen Rv0674. Methods To evaluate thediagnostic potential and antigenicity of Rv0674, IgG was evaluated using ELISA and interferon (IFN)-γ was done by using ELISpot assay among TB patients and healthy donors. For immunogenicity evaluation, BALB/c mice were immunized with Rv0674. Cytokine production was determined by cytokine release assay using an ELISA kit, and the antibodies were tested using ELISA. Results The results of serum Elisa tests showed that Rv0674 specific immunoglobulin G (IgG) response was higher in TB patients than negative controls. And Rv0674 had good performance in serological test with sensitivity and specificity of 77.1% and 81.1%, respectively. While it shows poor sensitivity and specificity of 26.23% and 79.69% for IFN-γ tests. In BALB/c mice, Rv0674 adjuvant by DDA/PolyI:C could also induce a high level of IFN-γ, interleukin-2 and interleukin-6 as well as a high IgG titer in both high-and low-dose groups indicating that Rv0674 is essential in humoral and cellular immunity. Moreover, the cytokine profile and IgG isotypecharacterized Rv0674 as a Th1/Th2-mixed-type protective immunity with the predominance of Th1 cytokines. Conclusion Rv0674 may be a good potential candidate for the development of TB serological diagnosis and a new TB vaccine.
文摘The plasticity and dynamism in the immune responses to both self and environmental stimulation promote the maintenance and adaptation of a system that tends to harmoniously survive and evolve. Fluctuating antigenic forces coexist within the immune system and oscillate between order and chaos to the equilibrium. Thus, when mounting a response to internal or environmental antigens, the main host responses can be divided into two immunological categories. The first, a well-adapted mechanism of complex multi-cellular organisms classically known as tolerance, promotes persistent immunological responses. In the second, opposite way, the modulation of inflammatory immune responses occurs, which we call “intolerance”. Tolerance and intolerance can be mediated by humoral molecules, such as inflammatory compounds, complement, and antibodies, and by different cell types, such as sentinel cells, antigen-presenting cells, and cells that orchestrate the immune response. Tolerogenesis is important in vertebrates because it predisposes species to adapt to self and environmental negative-selective forces. This process depends, in large part, on antigenic co-stimulation (AgCS), which operates as a multi-integrated network formed by all immune and non-immune cells of the body that establishes tolerant immunoregulatory interactions from cells to cells and from cells to the environment. Antigenic distribution, quantity, nature, route of administration, and antigenic convergence on co-stimulatory pathways, and concurrent infections, and the presence of microorganisms (commensals and pathogens) in more than one site are important factors for activating AgCS. To conclude, the AgCS route is a natural immune response generated by heterogeneous APC profile with centralized regulation that promote the counterbalance between intolerant e tolerant status, which can have several applications in the medical and biological fields.
文摘Objective:To investigate the prevalence of microbial antigenic components of circulating immune complexes amongst grades of CD4 T lymphocyte counts in HIV sero positive and seronegative participants.Methods:Polyethelene glycol(PEG-600) and buffering methods of precipitation and dissociation of immune complexes was used to generate immune solution from sera of 100 HIV sero-positive and 100 HIV sero-negative participants.These were categorized into 3 grades based on CD4 count:】 500 cell/mm,200-499 cell/mm3 and 【200 cell/mm3.The immune solutions were assayed using membrane based immunoassay and antibody titration, along side its unprocessed serum for detection of various microbial antigens and or antibodies. CD4 T cell counts were estimated using Patec Cyflow SL-3 Germany.Results:Antigenic component of immune complexes of various infectious agents was detected in 99 and 70 HIV seropositive and HIV sero-negative participants,respectively.In group A,there were 10 HIV positive participants,including 4(40.0%) had circulating immune complexes(CICs) due to Salmonella species only:1(10.0%) due to Salmonella-Plasmodium falciparum(P.falciparum),SalmonellaP. falciparum-HCV and P.falciparum antigens,respectively.In group B,45(45.4%) HIV seropositive participants with CICs had CD4 T lymphocyte count between 200-499 cells/mm^3.Out of these,20(44.4%) had CICs due to Salmonella species only:9(20%) due to Salmonella-P. falciparum.In group C,there were 44(44.4%) HIV sero-positive participants,including 3(6.8%) due to Salmonella species only:24(54.4%) due to Salmonella-P.falciparum:2(4.5%) due to P. falciparum only.Conclusions:In HIV sero-positive participants,presence of heterogeneity of Salmonella species-P.falciparum antigens was highly incriminated in CD4 count depletion but not homogeneity of malaria parasites antigens.Malaria parasites antigens only were incriminated in CD4^+ count depletion amongst HIV sero-negative participants.Before taking any decision on the management of HIV-1-positive individuals,their malaria and Salmonella paratyphi status should be assessed,but not malaria status alone.
文摘T cells engineered to express chimeric antigen receptors (CARs) combining an external antibody binding domain with the CD3ζ T cell receptor (TCR) signaling domain for triggering cell activation are being used for immunotherapeutic targeting of tumor cells in a non-HLA restricted manner. In this study we transduced T cells with a CD19-CAR construct containing a truncated CD34 gene (tCD34) marker and used these to target the B cell antigen CD19 on the surface of a Hodgkin’s lymphoma (HL) cell line (L591) both in vitro and in vivo. Levels of tCD34 expression in transduced peripheral blood mononuclear cells (PBMCs) ranged from 6% - 20% and this was increased to 82% after selection for transduced tCD34+ cells. In vitro cytotoxicity testing on a CD19+ HL cell line (L591) showed specific cell lysis initiated by the CD19-CAR transduced PBMCs. Importantly, CD19-CAR T cells prevented the growth of L591 HL tumor cells when co-injected subcutaneously (sc) in 6/6 severe combined immunodeficient (SCID) mice. There was no evidence of anti-tumor activity when CD19-CAR T cells were infused intravenously (iv) at the same time as L591 HL tumor cells were injected sc. However, 3/6 SCID mice showed tumor rejection within 83 days after iv infusion of CD19-CAR T cells 3 - 9 days after establishment of L591 HL tumors, while all control animals succumbed to tumors within 60 days. Interestingly, immuno-histochemical analysis of L591 HL tumors demonstrated that CD19-CAR T cells were detected not earlier than 11 days after infusion within the tumor mass. These results suggest that CD19 is a potentially attractive target for the immunotherapy of HL.
基金Supported by The National Institutes of Health to Luo Y,No.RO1DK066079
文摘AIM： To investigate the role of regulatory T （Treg） cells in CD4^＋ T cell-mediated bladder autoimmune infammation. METHODS： Urothelium-ovalbumin （URO-OVA）/OT-II mice, a double transgenic line that expresses the membrane form of the model antigen （Ag） OVA as a self-Ag on the urothelium and the OVA-specific CD4^＋ T cell receptor specifc for the I-Ab/OVA323-339 epitope in the periphery, were developed to provide an autoimmune environment for investigation of the role of Treg cells in bladder autoimmune infammation. To facilitate Treg cell analysis, we further developed URO-OVA^GFP-Foxp3/OT-II mice, a derived line of URO-OVA/OT-II mice that express the green fuorescent protein （GFP）-forkhead box protein P3 （Foxp3） fusion protein. RESULTS： URO-OVA/OT-II mice failed to develop bladder infammation despite the presence of autoreactive CD4^＋ T cells. By monitoring GFP-positive cells, bladder infltration of CD4^＋ Treg cells was observed in URO-OVA^GFP-Foxp3/OT-II mice. The infiltrating Treg cells were functionally active and expressed Treg cell effector molecule as well as marker mRNAs including transforming growth factor-β, interleukin （IL）-10, fibrinogen-like protein 2, and glucocorticoid-induced tumor necrosis factor receptor （GITR）. Studies further revealed that Treg cells from URO-OVA^GFP-Foxp3/OT-II mice were suppressive and inhibited autoreactive CD4^＋ T cell proliferation and interferon （IFN）-g production in response to OVA Ag stimulation. Depletion of GITR-positive cells led to spontaneous development of bladder infammation and expression of inflammatory factor mRNAs for IFN-γ, IL-6, tumor necrosis factor-α and nerve growth factor in URO-OVA^GFP-Foxp3/OT-II mice. CONCLUSION： Treg cells specifc for bladder epithelial Ag play an important role in immunological homeostasis and the control of CD4^＋ T cell-mediated bladder autoimmune infammation.
