Effect of Chemotherapy on Peripheral Blood DC Cells and Related Immune Cytokines in Patients with Non-Small Cell Lung Cancer

Objective: To analyze the effects of chemotherapy on peripheral blood DC cells and related immune cytokines (NKG2D, DC cells, TNF-a, IFN-r, HMGB-1) in patients with non-small cell lung cancer (NSCLC). Methods: Ninety-five NSCLC patients who attended the Oncology Department of the Affiliated Hospital of Chengde Medical College from September 2018 to February 2021 were selected as the research objects, and the changes in the expression levels of DC cells, NKG2D, TNF-a, IFN-r, HMGB-1 in the peripheral blood of patients at different time points (before chemotherapy, after the first chemotherapy, and after the second chemotherapy) were analyzed, and the correlation between DC cells in blood and NKG2D, TNF-a, IFN-r, HMGB-1 at each time point was explored. Results: The expression levels of NKG2D, TNF-a, IFN-r, and HMGB-1 in the peripheral blood of the patient before chemotherapy, after the first chemotherapy, and after the second chemotherapy gradually decreased, and there was no significant change in DC cells, except for DC cells at different times. The difference between each factor of each point was statistically significant (all P < 0.05). Pearson correlation analysis showed that there was no correlation between peripheral blood DC cells of patients at different time points and other factors. Conclusion: The decrease of other immune cytokines except DC cells in peripheral blood of patients with NSCLC after chemotherapy may be one of the mechanisms by which the patient’s immune function is suppressed. There is no correlation between DC cells and other factors.

Serum IL-10 from systemic lupus erythematosus patients suppresses the differentiation and function of monocyte-derived dendritic cells

The role played by cytokines, other than interferon （IFN）-a, in the differentiation and function of dendritic cells （DCs） in systemic lupus erythematosus （SLE）, remains unclear. Serum interleukin-10 （IL-10） levels are generally elevated in SLE patients, which might modulate the differentiation of DCs. In this study, DCs were induced from monocytes either by transendothelial trafficking or by culture with granulocyte-macrophage colony-stimulating factor （GM-CSF） ＋ IL-4 ＋ tumor necrosis factor （TNF）-a. Both systems were used to investigate the effects of elevated serum IL-10 level on DC differentiation in SLE patients. The results showed that monocyte-derived DCs induced by either SLE serum or exogenous IL-10 reduced the expression of human leukocyte antigen （HLA）-DR and CD80, decreased IL-12p40 level, and increased IL-10 level, and exhibited an impaired capacity to stimulate allogenic T-cell proliferation. These results indicate that serum IL-10 may be involved in the pathogenesis of SLE by modulating the differentiation and function of DCs.

Immune responses of dendritic cells combined with tumor-derived autophagosome vaccine on hepatocellular carcinoma

Background： To induce and collect tumor-derived autophagosomes （DRibbles） from tumor cells as an antitumor vaccine by inhibiting the functions of proteasomes and lysosomes. Methods： Dendritic cells （DCs） generated from peripheral blood mononuclear cell （PBMC） of hepatocellular carcinoma （HCC） patients were cocultured with DRibbles, and then surface molecules of DCs, as well as surface molecules on DCs, were determined by flow cytometry. Meanwhile, immune responses of the DCs-DRibbles were examined by mixed lymphocyte reactions. Results： DRibbles significantly induced the expression of CD80, CD83, CD86 and HLA-DR on DCs. The enzyme-linked immunosorbnent assay （ELISA） showed that IFN-γ, levels after vaccination increased than before in most patients, but CDS＋ proportion of PBMC increased only in nine patients. Higher levels of IFN-γ, were detected in the CD8＋ cells than CD4＋ T cells. These results suggested that DCs-DRibbles vaccine could induce antigen-specific cellular immune response on HCC and could prime strong CD8＋ T cell responses, supporting it as a tumor vaccine candidate. Conclusions： Our results demonstrate that HCC/DRibbles-pulsed DCs immunotherapy might be deployed as an effective antitumor vaccine for HCC immunotherapy in clinical trials.

A METHOD TO GENERATE MATURE DENDRITIC CELLS FROM CRYOPRESERVED PBMC

Objective: To established methods for cryopreserving peripheral blood mononuclear cells (PBMCs) andproducing DCs from cryopreserved PBMCs. Methods:Mature DCs were generated from cryopreserved PBMCs by using IL-4, GM-CSF, TNF-a, IL-1b, IL-6, pgE2 and LPS. The phenotype of the resultant DCs was investigated by flow cytometry. The functions of the resultant DCs were verified by Elispot assay. Results: The resultant DCs expressed high levels of HLA ABC, HLA DR, costimulatory molecules and the DC maturation marker CD83. The mature DCs wegenerated from frozen PBMCs were able to prime CD8 T cells into long term IFN-g producing peptide specific CTL. Conclusion: The DCs we developed from cryopreservedPBMC were fully mature and had the capability tostimulate immune reaction. Thus, we developed a method to generate functional mature DC from cryopreservedPBMC.

Effects of treatment time and temperature on the DC corona pretreatment performance of waste activated sludge

In order to improve the anaerobic digestion efficiency of waste activated sludge（WAS）,a pretreatment procedure should be carried out so as to disrupt the microbial cell structure,thus releasing intracellular organic matters.In this paper,a corona discharge triggered by a DC voltage was employed to pre-treat WAS for various time periods under different temperatures.The magnitude of the DC voltage was 4 k V at both negative and positive polarities.The changes in the soluble chemical oxygen demand,phosphorus and nitrogen content,and p H value within the WAS were utilized to estimate the pretreatment performance of the DC corona.It was found that with increasing treatment time,the pretreatment efficiency tends to be reduced.With increased temperature,the pretreatment efficiency appears to be better.It is suggested that the oxidative species and the active particles generated in the corona discharge play an important role in disrupting the microbial cell structure,which is dependent upon the treatment time and the temperature.

树突状细胞在溃疡性结肠炎发病中的价值与途径分析

树突状细胞(dendritic cells,DC)作为目前发现的功能最强的专职抗原提呈细胞,其功能紊乱是溃疡性结肠炎(Ulcerative Colitis,UC)发病的主要原因,其多可从通过共刺激分子的表达参与Th细胞分化参与Th17/Treg细胞间平衡调节肠黏膜免疫状态,通过分泌细胞因子参与Th1/Th2细胞间平衡调节黏膜免疫状态等多个角度参与溃疡性结肠炎的发病,且通过干预树突状细胞功能可以有效治疗溃疡性结肠炎,提示调控树突状细胞活化和功能可能是有效治疗溃疡性结肠炎的有效策略之一。

Study on the Changes of Immune Factors in Different Stages of Non-Small Cell Lung Cancer Chemotherapy

Objective: To analyze various immune cytokines (NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-a, IFN-r) and peripheral blood of patients with non-small cell lung cancer (NSCLC) at different times after chemotherapy. Changes in CD4+, CD8+, Th17 and IgG, IgM, and IgA levels. Methods: A total of 118 NSCLC patients who attended the Oncology Department of the Affiliated Hospital of Chengde Medical College from September 2018 to September 2021 were selected as the research objects, and the patients were analyzed at different time points (before chemotherapy, after the first chemotherapy, and after the second chemotherapy). The effects of NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-A, IFN-r, CD4+, CD8+ Th17, IgG, IgM and IgA levels in peripheral blood at different time points (before chemotherapy, after the first chemotherapy and after the second chemotherapy) were analyzed. The changes of NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-A, IFN-r and the levels of CD4+, CD8+ Th17, IgG, IgM and IgA in peripheral blood were compared at each time point. Results: NKG2D, IL-12, IL-15, IL-18, TNF-a, IFN-r gradually decreased before chemotherapy, one week after chemotherapy, and two weeks after chemotherapy, the difference was statistically significant, but DC cells were not significant Variety. CD4+ and CD8+ both increased significantly, and the levels of Th17, IgG, IgM, and IgA gradually decreased. Conclusion: In the course of chemotherapy, all immune factors except DC cells were significantly decreased compared with those before chemotherapy, and the decrease of immune factors except DC cells was positively correlated with the length of chemotherapy cycle. If additional immunotherapy is needed, it should be carried out in the early stage of chemotherapy.

Adoptive transfer of FTY720-treated immature bone marrowderived dendritic cells(BMDCs) significantly reduced the spontaneous resorption rate in the CBA/J ×DBA/2 mouse model

Objective To investigate the effect of FTY720-treated immature bone marrow-derived dendritic cells（BMDCs） on the embryo resorption rate in the CBA/J× DBA/2 abortion mouse model.Methods The dendritic cells（DCs） were derived from bone marrow of DBA/2 mice, and then co-cultured with FTY720. The abortion mouse models were established by mating female CBA/J mice with DBA/2 mice. Via the CBA/J×DBA/2 abortion mouse model, six groups were established, group A： normal pregnancy model; group B： abortion mouse model with no treatment; group C： abortion mouse model injected with DC culture medium（DCCM）; group D： abortion mouse model injected with DC; group E： abortion mouse model injected with FTY720; group F： abortion model mouse injected with FTY720-DC. The differences were compared in the embryo resorption rates of the CBA/J ×DBA/2 abortion mouse model treated with FTY720-DC or different controls observed on gestation day 12 to 14, and then the microenvironment in murine pregnancy was investigated.Results The embryo resorption rate was statistically significantly decreased in group D and group E when they compared with group B and group C（P〈0.05, respectively）.Furthermore, the embryo resorption rate in group F showed a statistically significant decrease when compared with the other groups except group A（P〈0.01）. These resultssuggest that FTY720-DCs possess a notable advantage over DCs or FTY720 in reducing the embryo resorption rate of the abortion mouse model. The percentage of Th17（IL-17＋CD4＋T cells） in peripheral blood mononuclear cell（PBMC） in the abortion mouse model was 4.35%±0.34% before treated with FTY720-DC, and was1.34%±0.28% after treated with FTY720-DC（P〈0.01）. The percentage of Tregs（CD4~＋CD25~＋Foxp3~＋T cells） in PBMC was significantly increased in group F（8.35%±1.80%） as compared with group B（2.68%±0.65%）（P〈0.01）.Conclusion Adoptive transfer of FTY720-DC can statistically significantly reduce the embryo resorption rate in the CBA/J×DBA/2 abortion mouse model. The lower embryo resorption rate in the FTY720-DC treated abortion mouse model may be caused by the imbalance of Treg/Th17.

Differential centrifugation enhances the anti-tumor immune effect of tumor lysate-pulsed dendritic cell vaccine against glioblastoma

Objective This study aimed to improve the antitumor immunocompetence of a tumor lysate-pulsed dendritic cell(DC)vaccine through differential centrifugation and provide a theoretical basis for its clinical application in glioblastoma.Methods Peripheral blood mononuclear cells were extracted using Ficoll-Paque PLUS and induced into mature DCs in vitro with a cytokine cocktail.The modified tumor lysate was generated by differential centrifugation.The maturity markers of DCs in each group,namely the modified tumor lysate,tumor lysate,and negative and positive control groups,were assessed using flow cytometry.Furthermore,their ability to stimulate lymphocyte proliferation and in vitro antitumor effects were assessed using Cell Trace TM CFSE.IFN-γsecretion levels were measured with ELISA.Intracellular reactive oxygen species were measured using 2’,7’-dichlorofluorescein diacetate(DCFDA)staining.The results were statistically analyzed using an unpaired Student’s t-test and were considered significant at P<0.05.Results Compared with tumor lysate-pulsed DCs,modified tumor lysate-pulsed DCs had a higher expression of maturity markers:CD1a(7.38±0.53%vs.4.47±0.75%)and CD83(19.81±4.09%vs.9.64±1.50%),were better capable of stimulating lymphocyte proliferation[proliferation index(PI):8.54±0.16 vs.7.35±0.05],secreting IFN-γ,and inducing stronger in-vitro cytotoxic T lymphocyte(CTL)cytotoxicity against glioblastoma cells.In addition,we found that the level of ROS in modified tumor lysate-pulsed DCs was lower than that in tumor lysate-pulsed DCs.Conclusion Differential centrifugation of tumor lysates can improve the antitumor immunocompetence of DC vaccines,and reactive oxygen species may be the key to affecting DC function in the whole tumor lysate.

Enhancing the treatment effects of tumor cell purified autogenous heat shock protein 70-peptide complexes on HER-3-overexpressing breast cancer

Objective The aim of this study was to enhance the treatment effect of tumor purified autogenous heat shock protein 70-peptide complexes(HSP70-PCs)on HER-3-overexpressing breast cancer.Methods In this study,we first studied the expression of HER-3 in breast cancer tissues and its relationship with patient characteristics.We then purified HSP70-PCs from primary breast cancer cells with different HER-2 and HER-3 expression profiles and determined the cytotoxicity of autogenous dendritic cells(DCs)and CD8+T cells induced by these complexes.Third,recombinant human HSP70-HER-3 protein complexes were used to inhibit the autogenous HSP70-PCs purified from HER-3-overexpressing breast cancer cells,and the resulting immunological response was examined.Results The results show that HSP70-PCs can be combined with recombinant HSP70-HER-3 protein complexes to induce stronger immunological responses than autogenous HSP70-PCs alone and that these treatments induce autogenous CD8+T cell killing of HER-3-positive breast cancer cells.Conclusion These findings provide a new direction for HSP70-DC-based immunotherapy for patients with HER-3-overexpressing breast cancer.

Dendritic cells maturation facilitated by group-adjustable lipopolysaccharide analogues synthesized via RAFT polymerization

Transforming immature DCs into mature state to activate cellular immunity is a critical step in initiating immunoprophylaxis and immunotherapy.Lipopolysaccharides(LPS)can promote DCs maturation by binding receptor on DCs surface,but their clinical application is limited due to biological toxicity.Although many LPS analogues have been developed,complex synthesis and purification hinder their practical application.Here,we propose a novel and simple strategy to synthesize LPS analogues with adjustable structural units.Using monomer units similar to the key functional groups of LPS,we synthesize LPS analogues with different group ratios by RAFT polymerization.The obtained analogues have little negative effect on cell viability.Compared with LPS,the analogues show greater promoting effect on DCs maturation.And the analogues can be applied to different scenarios since the degrees of promoting DCs maturation by LPS analogues with different group ratios are different.This strategy provides a new direction for synthesizing LPS analogues,and it has the potential to produce LPS analogues on a large scale with tunable promoting DCs maturation effect.

Nanomedicine-mediated ubiquitination inhibition boosts antitumor immune response via activation of dendritic cells

Tumor immunotherapy as a promising method for tumor treatment received tremendous attention. However, the problem of low clinical response rate still needs to be solved, especially in the poorly immunogenic tumors. The enhancement of tumor antigens presentation can effectively activate dendritic cells (DCs) and improve the tumor immunotherapy. In this work, TAK-243 as an inhibitor of the ubiquitin activating enzyme (UAE), was fabricated into cationic lipid-assisted nanoparticle (CLANTAK-243). The obtained CLANTAK-243 could act as an effective tumor immunotherapy enhancer to promote the maturation of DCs as well as antigen presentation, which obviously stimulated the T cells activation and proliferation. Such CLANTAK-243 injected intravenously could well trigger immune response to tumor cells in vivo. Importantly, mice treated with CLANTAK-243 could obtain a long immune memory effect to protect themselves from re-challenged tumor cells. Therefore, this work presented an effective immunotherapy strategy for poorly immunogenic tumor.

Cytokines mRNA in bone marrow-derived dendritic cells in asthmatic mouse

By inducing and amplifying dendritic cells(DCs)derived from the bone marrow of asthma murine in vitro,cytokines mRNA were expressed,and the functions of DCs were investigated.Cells isolated from murine bone marrow have been cultured with rmGM-CSF and rmIL-4,and the expression of cytokines mRNA was determined by ribonuclease protection assay combined with multi-probe templates.Large numbers of DCs have been obtained from bone marrow,and they expressed interleukin-13(IL-13),interleukin-9(IL-9),and interleukin-3(IL-3)mRNA.Moreover,the level of IL-13 mRNA and IL-9 mRNA ex-pressed by DCs in asthmatic mice was significantly higher than those in the control groups(P<0.05).But,the level of IL-3 mRNA showed no discrepancy between the two groups(P>0.05).DCs are very important in the forming and developing of asthma,which implies that the therapy targeted at DCs will possibly become a new goal.

Mucosal delivery of nanovaccine strategy against COVID-19 and its variants

Despite the global administration of approved COVID-19 vaccines(e.g.,ChAdOx1 nCoV-19?,mRNA-1273?,BNT162b2?),the number of infections and fatalities continue to rise at an alarming rate because of the new variants such as Omicron and its subvariants.Including COVID-19 vaccines that are licensed for human use,most of the vaccines that are currently in clinical trials are administered via parenteral route.However,it has been proven that the parenteral vaccines do not induce localized immunity in the upper respiratory mucosal surface,and administration of the currently approved vaccines does not necessarily lead to sterilizing immunity.This further supports the necessity of a mucosal vaccine that blocks the main entrance route of COVID-19:nasal and oral mucosal surfaces.Understanding the mechanism of immune regulation of M cells and dendritic cells and targeting them can be another promising approach for the successful stimulation of the mucosal immune system.This paper reviews the basic mechanisms of the mucosal immunity elicited by mucosal vaccines and summarizes the practical aspects and challenges of nanotechnology-based vaccine platform development,as well as ligand hybrid nanoparticles as potentially effective target delivery agents for mucosal vaccines.

Mathematical analysis of Hepatitis C Virus infection model in the framework of non-local and non-singular kernel fractionalderivative

In this paper,we study a mathematical model of Hepatitis C Virus(HCV)infection.We present a compartmental mathematical model involving healthy hepatocytes,infected hepatocytes,non-activated dendritic cells,activated dendritic cells and cytotoxic T lymphocytes.The derivative used is of non-local fractional order and with non-singular kernel.The existence and uniqueness of the system is proven and its stability is analyzed.Then,by applying the Laplace Adomian decomposition method for the fractional derivative,we present the semi-analytical solution of the model.Finally,some numerical simulations are performed for concrete values of the parameters and several graphs are plotted to reveal the qualitative properties of the solutions.

A Recombinant Rabies Virus Expressing Fms-like Tyrosine Kinase 3 Ligand(Flt3L) Induces Enhanced Immunogenicity in Mice

Rabies is a zoonotic disease that still causes 59,000 human deaths each year,and rabies vaccine is the most effective way to control the disease.Our previous studies suggested that the maturation of DC plays an important role in enhancing the immunogenicity of rabies vaccine.Flt3L has been reported to own the ability to accelerate the DC maturation,therefore,in this study,a recombinant rabies virus expressing mouse Flt3L,designated as LBNSE-Flt3L,was constructed,and its immunogenicity was characterized.It was found that LBNSE-FU3L could enhance the maturation of DC both in vitro and in vivo,and significantly more TFH cells and Germinal Center B(GC B)cells were generated in mice immunized with LBNSE-FU3L than those immunized with the parent virus LBNSE.Consequently,expressing of Flt3L could elevate the level of virus-neutralizing antibodies(VNA)in immunized mice which provides a better protection from a lethal rabies virus challenge.Taken together,our study extends the potential of Flt3L as a good adjuvant to develop novel rabies vaccine by enhancing the VNA production through activating the DC—Tfh^GC B axis in immunized mice.