The maturation of dendritic cells(DCs)and infiltration effector T cells in tumor-draining lymph node(tdLN)and tumor tissue are crucial for immunotherapy.Despite constructive progresses have been made with anti-program...The maturation of dendritic cells(DCs)and infiltration effector T cells in tumor-draining lymph node(tdLN)and tumor tissue are crucial for immunotherapy.Despite constructive progresses have been made with anti-programmed death-1(anti-PD1)checkpoint blockade for immunotherapy,the efficacy of PD1/PD-L1 therapy deserves to be improved.Here,we constructed a novel transfersomes based nanovaccine complexed microneedles to enhance anti-PD1 immunotherapy via transdermal immunization for skin tumor therapy.Transfersomes were functionalized with DCs targeting moietyαCD40,co-encapsulated with antigens and adjuvant poly I:C.Moreover,transdermal administration promoted accumulation in tumor-draining lymph nodes(tdLN),which could facilitate cellular uptake,activate DCs maturation and enhance Th1 immune responses.Using a mouse melanoma model,combined therapy of such nanovaccine complexed microneedles with pembrolizumab(αPD1)was able to enhance cytotoxic T lymphocytes activation,promote infiltration and reduce regulatory T cells frequency in tdLN and tumor tissues,which achieved reversion of the immunosuppressive microenvironment into immune activation.This study highlighted the potential of transfersomes based nanovaccines complexed microneedles as an attractive platform for tumor immunotherapy.展开更多
Background:Currently,due to synergy enhancement of anti-tumor effects and potent stimulation of abscopal effects,combination therapy with irradiation and programmed cell death protein 1/programmed death-ligand 1(PD-1/...Background:Currently,due to synergy enhancement of anti-tumor effects and potent stimulation of abscopal effects,combination therapy with irradiation and programmed cell death protein 1/programmed death-ligand 1(PD-1/PD-L1)immune checkpoint inhibition(immuno-radiotherapy,iRT)has revolutionized the therapeutic guidelines.It has been demonstrated that tumor-draining lymph nodes(TDLN)are essential for effective antitumor immunity induced by radiotherapy,immunotherapy,or iRT.Given that the function of TDLN in iRT remains unclear,this study aimed to investigate the function and mechanism of TDLN in iRT-induced abscopal effects.Methods:The function of TDLN was evaluated using unilateral or bilateral MC38 and B16F10 subcutaneous tumor models with or without indicated TDLN.The flow cytometry,multiple immunofluorescence analysis,and NanoString analysis were utilized to detect the composition and function of the immune cells in the primary and abscopal tumor microenvironment.Additionally,we tempted to interrogate the possible mechanisms via RNA-sequencing of tumor-infiltrating lymphocytes and TDLN.Results:TDLN deficiency impaired the control of tumor growth by monotherapy.Bilateral TDLN removal rather than unilateral TDLN removal substantially curtailed iRT-stimulated anti-tumor and abscopal effects.Furthermore,in the absence of TDLN,the infiltration of CD45+and CD8+T cells was substantially reduced in both primary and abscopal tumors,and the anti-tumor function of CD8+T cells was attenuated as well.Additionally,the polarization of tumor-associated macrophages in primary and abscopal tumors were found to be dependent on intact bilateral TDLN.RNA-sequencing data indicated that impaired infiltration and anti-tumor effects of immune cells partially attributed to the altered secretion of components from the tumor microenvironment.Conclusions:TDLN play a critical role in iRT by promoting the infiltration of CD8+T cells and maintaining the M1/M2 macrophage ratio.展开更多
The lymphatic system is important in mounting an immune response to foreign antigens and tumors in humans and animal models. The liver produces a large amount of lymph, and its lymphatic system is divided into three m...The lymphatic system is important in mounting an immune response to foreign antigens and tumors in humans and animal models. The liver produces a large amount of lymph, and its lymphatic system is divided into three major components： the portal, sublobular and superficial lymphatic vessels. Despite the fact that mice are the most commonly used laboratory animals, detailed descriptions of the anatomical location and function of the lymph nodes （LNs） that drain the liver are surprisingly absent. In this study, we found that the portal and celiac LNs adjacent to mouse liver were stained with Evans blue within 5-8 min. Enhanced green fluorescence protein （EGFP）-positive cells from the liver also drained into the two aforementioned LNs. These data indicate that the portal and celiac LNs drain the mouse liver. Lymphadenectomy of the identified liver-draining LNs resulted in hepatitis B virus （HBV） persistence in immunocompetent mice compared with the sham group. In addition, the frequencies of CD8＋ T cells and dendritic cells （DCs） increased significantly in the liver-draining LNs after hydrodynamic injection of HBV plasmid. Liver-draining LN cells in HBV plasmid-injected mice also showed significant antigen-specific proliferation in response to stimulation with recombinant hepatitis B core antigen in vitro. Adoptive transfer of these cells into Rag1-/- mice induced a reduction in the serum concentration of hepatitis B surface antigen （HBsAg） compared to liver-draining LN cells in uninjected mice. Altogether our data characterize the liver-draining LNs and provide evidence that the liver-draining LNs induce an anti-HBV-specific immune response responsible for HBV clearance.展开更多
To find a feasible method for the stimulation of tumor-draining lymph node （TDLN） cells in preparation for use in the clinic, the CTL activity of TDLN cells induced by different stimuli （IL-2 alone, IL-2 ＋ autolog...To find a feasible method for the stimulation of tumor-draining lymph node （TDLN） cells in preparation for use in the clinic, the CTL activity of TDLN cells induced by different stimuli （IL-2 alone, IL-2 ＋ autologous tumor antigen （atAg）, IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg） was measured by maximal LDH enzyme release. The mechanisms were explored by the observation of morphology and the detection of CD83^＋ TDLN cells. The expansion of TDLN cells by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was significantly higher than that by IL-2 alone or IL-2 ＋ atAg （p 〈 0.01）. Antitumor CTL activity of TDLN cells induced by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was significantly higher than those of other groups. The number of CD83^＋ cells within the TDLN population treated with IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was significantly elevated. The method of stimulating TDLN cells by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was better than the stimulation with IL-2 or IL-2 ＋ atAg. TDLN cells induced by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg produced more dendritic cells （DCs）. In our study, we established a system that T cells and DCs were stimulated together ex vivo, which was easy to conduct and produce promising results. It provided a new method for improving TDLN cell antitumor activity which might be used in the clinical cancer therapy. Cellular ＆ Molecular Immunology. 2008;5（4）：307-313.展开更多
基金work was supported by the National Natural Science Foundation of China(No.31670972)the Taishan Scholar Program,China.
文摘The maturation of dendritic cells(DCs)and infiltration effector T cells in tumor-draining lymph node(tdLN)and tumor tissue are crucial for immunotherapy.Despite constructive progresses have been made with anti-programmed death-1(anti-PD1)checkpoint blockade for immunotherapy,the efficacy of PD1/PD-L1 therapy deserves to be improved.Here,we constructed a novel transfersomes based nanovaccine complexed microneedles to enhance anti-PD1 immunotherapy via transdermal immunization for skin tumor therapy.Transfersomes were functionalized with DCs targeting moietyαCD40,co-encapsulated with antigens and adjuvant poly I:C.Moreover,transdermal administration promoted accumulation in tumor-draining lymph nodes(tdLN),which could facilitate cellular uptake,activate DCs maturation and enhance Th1 immune responses.Using a mouse melanoma model,combined therapy of such nanovaccine complexed microneedles with pembrolizumab(αPD1)was able to enhance cytotoxic T lymphocytes activation,promote infiltration and reduce regulatory T cells frequency in tdLN and tumor tissues,which achieved reversion of the immunosuppressive microenvironment into immune activation.This study highlighted the potential of transfersomes based nanovaccines complexed microneedles as an attractive platform for tumor immunotherapy.
基金the Academic Promotion Program of Shandong First Medical Univer-sity(2019ZL002)Research Unit of Radiation Oncology,Chinese Academy of Medical Sciences(2019RU071)+2 种基金the foundation of National Natural Science Foundation of China(81627901,81972863,82030082 and 31900649)the foundation of Natural Science Foundation of Shandong(ZR201911040452 and ZR2019LZL018)the Cancer Pre-vention and Treatment Joint Fund of Natural Science Foundation of Shandong Province(ZR2020LZL014).
文摘Background:Currently,due to synergy enhancement of anti-tumor effects and potent stimulation of abscopal effects,combination therapy with irradiation and programmed cell death protein 1/programmed death-ligand 1(PD-1/PD-L1)immune checkpoint inhibition(immuno-radiotherapy,iRT)has revolutionized the therapeutic guidelines.It has been demonstrated that tumor-draining lymph nodes(TDLN)are essential for effective antitumor immunity induced by radiotherapy,immunotherapy,or iRT.Given that the function of TDLN in iRT remains unclear,this study aimed to investigate the function and mechanism of TDLN in iRT-induced abscopal effects.Methods:The function of TDLN was evaluated using unilateral or bilateral MC38 and B16F10 subcutaneous tumor models with or without indicated TDLN.The flow cytometry,multiple immunofluorescence analysis,and NanoString analysis were utilized to detect the composition and function of the immune cells in the primary and abscopal tumor microenvironment.Additionally,we tempted to interrogate the possible mechanisms via RNA-sequencing of tumor-infiltrating lymphocytes and TDLN.Results:TDLN deficiency impaired the control of tumor growth by monotherapy.Bilateral TDLN removal rather than unilateral TDLN removal substantially curtailed iRT-stimulated anti-tumor and abscopal effects.Furthermore,in the absence of TDLN,the infiltration of CD45+and CD8+T cells was substantially reduced in both primary and abscopal tumors,and the anti-tumor function of CD8+T cells was attenuated as well.Additionally,the polarization of tumor-associated macrophages in primary and abscopal tumors were found to be dependent on intact bilateral TDLN.RNA-sequencing data indicated that impaired infiltration and anti-tumor effects of immune cells partially attributed to the altered secretion of components from the tumor microenvironment.Conclusions:TDLN play a critical role in iRT by promoting the infiltration of CD8+T cells and maintaining the M1/M2 macrophage ratio.
基金This work was supported by the Ministry of Science and Technology of China （973 Basic Science Project 2013CB944902）, the Natural Science Foundation of China （Nos. 91029303, 31021061） and the National Science and Technology Major Projects （2013ZX10002002-002）. We thank Professor Pei-Jer Chen for providing the pAAV-HBV plasmid.
文摘The lymphatic system is important in mounting an immune response to foreign antigens and tumors in humans and animal models. The liver produces a large amount of lymph, and its lymphatic system is divided into three major components： the portal, sublobular and superficial lymphatic vessels. Despite the fact that mice are the most commonly used laboratory animals, detailed descriptions of the anatomical location and function of the lymph nodes （LNs） that drain the liver are surprisingly absent. In this study, we found that the portal and celiac LNs adjacent to mouse liver were stained with Evans blue within 5-8 min. Enhanced green fluorescence protein （EGFP）-positive cells from the liver also drained into the two aforementioned LNs. These data indicate that the portal and celiac LNs drain the mouse liver. Lymphadenectomy of the identified liver-draining LNs resulted in hepatitis B virus （HBV） persistence in immunocompetent mice compared with the sham group. In addition, the frequencies of CD8＋ T cells and dendritic cells （DCs） increased significantly in the liver-draining LNs after hydrodynamic injection of HBV plasmid. Liver-draining LN cells in HBV plasmid-injected mice also showed significant antigen-specific proliferation in response to stimulation with recombinant hepatitis B core antigen in vitro. Adoptive transfer of these cells into Rag1-/- mice induced a reduction in the serum concentration of hepatitis B surface antigen （HBsAg） compared to liver-draining LN cells in uninjected mice. Altogether our data characterize the liver-draining LNs and provide evidence that the liver-draining LNs induce an anti-HBV-specific immune response responsible for HBV clearance.
文摘To find a feasible method for the stimulation of tumor-draining lymph node （TDLN） cells in preparation for use in the clinic, the CTL activity of TDLN cells induced by different stimuli （IL-2 alone, IL-2 ＋ autologous tumor antigen （atAg）, IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg） was measured by maximal LDH enzyme release. The mechanisms were explored by the observation of morphology and the detection of CD83^＋ TDLN cells. The expansion of TDLN cells by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was significantly higher than that by IL-2 alone or IL-2 ＋ atAg （p 〈 0.01）. Antitumor CTL activity of TDLN cells induced by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was significantly higher than those of other groups. The number of CD83^＋ cells within the TDLN population treated with IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was significantly elevated. The method of stimulating TDLN cells by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg was better than the stimulation with IL-2 or IL-2 ＋ atAg. TDLN cells induced by IL-2 ＋ GM-CSF ＋ IL-4 ＋ atAg produced more dendritic cells （DCs）. In our study, we established a system that T cells and DCs were stimulated together ex vivo, which was easy to conduct and produce promising results. It provided a new method for improving TDLN cell antitumor activity which might be used in the clinical cancer therapy. Cellular ＆ Molecular Immunology. 2008;5（4）：307-313.
