Oxidative potential associated with reactive oxygen species of size-resolved particles: The important role of the specific sources.

Oxidative potential (OP) is a predictor of particulate matter (PM) toxicity. Size-resolved PM and its components that influence OP values can be generated from several sources. However, There is little research have attempted to determine the PM toxicity generated from specific sources. This paper studied the OP characterization and reactive oxygen species (ROS) formation of particles from specific sources and their effects on human health. OP associated with ROS of size-resolved particles was analyzed by using dithiothreitol (DTT) method and electron paramagnetic resonance (EPR) spectroscopy technology. And OP and ROS deposition of specific source PM were calculated for health through the Multi-path particle deposition (MPPD) model. The results evidenced that the highest water-soluble OP (OPws) from traffic sources (OPm: 104.50 nmol min-1·ug-1; OPv: 160.15 nmol min-1·m-3) and the lowest from ocean sources (OPm: 22.25 nmol⋅min-1⋅ug-1; OPv: 54.16 nmol min-1·m-3). The OPws allocation in PM from different sources all have a unimodal pattern range from 0.4 to 3.2 µm. ROS (·OH) displayed the uniform trend as PM OPws, indicating that PM< 3.2 is the major contributor to adverse health impacts for size-resolved PM because of its enhanced oxidative activity compared with PM> 3.2. Furthermore, this study predicted the DTT consumption of PM were assigned to different components. Most DTT losses are attributed to the transition metals. For specific sources, transition metals dominates DTT losses, accounting for 38%-80% of DTT losses from different sources, followed by Hulis-C, accounting for 1%-10%. MPPD model calculates that over 66% of pulmonary DTT loss comes by PM< 3.2, and over 71% of pulmonary ROS generation from PM< 3.2. Among these sources of pollution, traffic emissions are the primary contributors to reactive oxygen species (ROS) in environmental particulate matter (PM). Therefore, emphasis should be placed on controlling traffic emissions, especially in coastal areas.

Oxidative potential, environmentally persistent free radicals and reactive oxygen species of size-resolved ambient particles near highways.

Particulate matter (PM) is a group of atmospheric pollutants with an uncertain toxicity, particularly when collected near highways. This study examined the oxidative potential (OP) of, as well as the environmentally persistent free radicals (EPFRs) and reactive oxygen species (ROS) present in PM samples collected near highways in Xiamen, China. Our findings revealed that PM had a relatively high OP, ranging from 3.8 to 18.5 nmol/min/µg, surpassing values reported in previous research. The oxidative potential of the water-insoluble fraction (OPWIS), which accounted for 68% of the total oxidative potential (OPTotal), demonstrated rapid toxicity, whereas the oxidative potential of the water-soluble fraction (OPWS) displayed a steadier toxicity release pattern. The primary free radicals detected in PM were oxygen-centered. The measured concentration of EPFRs was 6.073 × 1014 spins/m3, which is lower than that reported in previous studies, possibly because of the high relative humidity of the road environment in Xiamen. We also investigated the interaction between PM and water near highways and observed the generation of R and OH radicals. Additionally, we analysed the sample composition and evaluated the contributions of the different components to OPTotal. Transition metals (Fe, Cu, and Zn) were identified as the major contributors, accounting for 33.2% of the OPTotal. The positive correlation observed between EPFRs and ROS suggests that EPFRs may be involved in ROS generation. The correlation analysis indicated that the oxidative potential measured using the DTT method (OPDTT) could serve as an indicator of ROS generation. Finally, based on the relationship between OPDTT, EPFRs, and ROS, we propose that reducing the emission of transition metals, particularly Fe, represents an effective control measure for mitigating PM toxicity near highways.

The overlooked formation of environmentally persistent free radicals on particulate matter collected from biomass burning under light irradiation.

BACKGROUND: The illumination process may be an important contributor to environmentally persistent free radicals (EPFRs) in atmospheric particles, but the ability of light to generate EPFRs in combustion products remains unclear. OBJECTIVE: This paper studies the characteristics and formation mechanism of EPFRs in combustion particles after photoexcitation. METHOD: The secondary photochemical processes and the generation and decay capability of EPFRs in size-resolved (<10 µm) biomass combustion particles were analysed by electron paramagnetic resonance (EPR) spectroscopy. RESULT: Our results indicated that secondary EPFRs can be generated after illumination and the produced EPFRs have a lifetime of approximately 1 day. The content of secondary EPFRs after light exposure increased by 20 %-30 % compared to that of the original EPFRs. Through the analysis of components of different polarities, it was found that non-extractable substances were the main contributors to secondary EPFRs (75 %), followed by extractable organics. This study showed that metal species and quinones are important precursors for the formation of secondary EPFRs from non-extractable and extractable PM components, respectively. We found that O2 molecules are an important factor for the formation of secondary EPFRs from organic substances without oxygen functional groups. CONCLUSIONS: This study presents information about the effects of light and O2 on the generation of EPFRs, and the unstable nature of secondary EPFRs has important implications for assessing the health risks of atmospheric particles.

Sequential administration of anti-PD-1 and anti-Tim-3 combined with an SA-GM-CSF-anchored vaccine overcomes adaptive immune resistance to reject established bladder cancer.

Program death receptor-1 (PD-1) and T-cell immunoglobulin and mucin domain-containing protein-3 (Tim-3) play an important role in tumor immune evasion. PD-1 blockade could produce an effective anti-tumor effect but the response rate was low due to lacking of tumor infiltrating lymphocytes (TILs) and existing of other negative regulatory pathways. Streptavidin(SA)-GM-CSF surface-anchored tumor cells vaccine could induce specific anti-tumor immune response. However, this vaccine failed to induce regression of established tumor because it also up-regulated PD-1 expression on tumor cells dependent on IFNγ and up-regulated PD-1/Tim-3 expression on CD8+ TILs. Subsets of CD8+ TILs assay showed that PD-1 expression was closely associated with CD8+ TILs exhaustion, and Tim-3 expression was closely correlated with secretion function but not proliferation of CD8+ TILs. Sequential administration of anti-PD-1 and anti-Tim-3 could further improve the efficacy of SA-GM-CSF-anchored vaccine therapy, and tumor regression was noted in over 50%. This triple therapy improves the specific cytotoxic activity and decreased the apoptosis of CD8+ TILs. These findings indicated that this triple therapy could induce a more robust anti-tumor immune response.

The associations of lens power with age, axial length and type 2 diabetes mellitus in Chinese adults aged 50 and above.

BACKGROUND: To investigate the associations of lens power with age, axial length (AL), and Type 2 diabetes mellitus (DM) in Chinese adults aged 50 and above. METHODS: Random clustering sampling was used to identify adults aged 50 years and above in urban regions of Shanghai. The participants underwent a comprehensive ophthalmic examination including subjective refraction, autorefraction, and IOL-Master. The crystalline lens power was calculated using Bennett's formula. RESULTS: A total of 4177 adults were included. A linear decrease in lens power was observed both with age and with AL, followed by a stop of lens power loss after the age of 70 or when AL ≥ 25 mm, respectively. Participants with Type 2 DM presented higher lens power (0.43 diopter (D), p < 0.001) and thicker lens thickness (0.06 mm, p < 0.001). In multivariate regression models, there was a positive correlation between lens power and Type 2 DM when age < 75 years (p < 0.001) or AL < 25 mm (p < 0.001) after adjusting for other factors, while no significant association was found in participants aged ≥ 75 years (p = 0.122) or with AL ≥ 25 mm (p = 0.172). CONCLUSIONS: The lens power in adults aged 50 and above exhibited two stages with age and with AL. Type 2 DM caused an increase in lens power, which was not seen in participants aged ≥ 75 years or with AL ≥ 25 mm.

Review of Huang-huai sheep, a new multiparous mutton sheep breed first identified in China.

Huang-huai sheep are a new multiparous mutton sheep breed that has been cultivated by domestic scientific research institutes, governments, and sheep farms in China. Huang-huai sheep were bred using Dorper sheep as a sire and Small-tailed Han sheep as a dam. The breeding of Huang-huai sheep started in 2003, and three stages have been carried out: crossbreeding innovation, fixation in a two-way-crossbred closed flock, and herd propagation. A pilot test of Huang-huai sheep was conducted on 6 sheep farms from 2017 to 2018, and hereditary properties and production performance were evaluated in 2019. Huang-huai sheep were identified on site by the National Livestock and Poultry Resources Committee of China in December 2019 and approved as a new multiparous mutton sheep breed in China. The genetic distance showed that Huang-huai sheep are most closely related to Dorper sheep, Luxi black-headed sheep, and Small-tailed Han sheep, but the genetic distances are subspecies (0.02-0.20) each other. The body weights of adult Huang-huai sheep are 98.1 ± 5.2 kg (♂) and 71.7 ± 3.5 kg (♀), and those of 6-month-old Huang-huai sheep are 58.50 ± 6.55 kg (♂) and 52.45 ± 5.67 kg (♀). The slaughter rates of 6-month-old sheep are 56.02 ± 1.25% (♂) and 53.19 ± 1.19% (♀). The estrus cycle of Huang-huai sheep is 19.32 ± 2.8 days, the first estrus cycle occurs at 168 ± 12 days, the annual lambing rate of ewes is 252.82% ± 10.69%, the survival rate of lambs is 95.79 ± 0.95%, and the number of weaned lambs per ewe per year is 2.38 ± 0.14. The growth performance, carcass quality, and reproductive performance of Huang-huai sheep have been improved, resulting in considerable economic and social benefits and broader market prospects. This breed represents a new multiparous mutton sheep breed adapted for industrial sheep farms in China.

Regional consistency and sample size considerations in a multiregional equivalence trial.

The main objective of a confirmatory multiregional clinical trial (MRCT) is to demonstrate the overall efficacy of test drugs in all participating regions as well as to evaluate the possibility of extrapolating the overall results to each region. With the emergence of the demands of biosimilar drugs development, some guidelines recommended using equivalence design to demonstrate the comparability of efficacy between biosimilar and reference drugs. Previous discussions about assessing regional consistency in MRCT are mainly focused on superiority or non-inferiority designs, while the extensions to equivalence designs were limited. In this work, we proposed a flexible regional consistency criterion for the MRCT with equivalence design. Based on this criterion, sample size determination and sample allocation were also discussed.

Insertional mutagenesis of Brachypodium distachyon using the Tnt1 retrotransposable element.

Brachypodium distachyon is an annual C3 grass used as a monocot model system in functional genomics research. Insertional mutagenesis is a powerful tool for both forward and reverse genetics studies. In this study, we explored the possibility of using the tobacco retrotransposon Tnt1 to create a transposon-based insertion mutant population in B. distachyon. We developed transgenic B. distachyon plants expressing Tnt1 (R0) and in the subsequent regenerants (R1) we observed that Tnt1 actively transposed during somatic embryogenesis, generating an average of 6.37 insertions per line in a population of 19 independent R1 regenerant plants analyzed. In seed-derived progeny of R1 plants, Tnt1 segregated in a Mendelian ratio of 3:1 and no new Tnt1 transposition was observed. A total of 126 flanking sequence tags (FSTs) were recovered from the analyzed R0 and R1 lines. Analysis of the FSTs showed a uniform pattern of insertion in all the chromosomes (1-5) without any preference for a particular chromosome region. Considering the average length of a gene transcript to be 3.37 kb, we estimated that 29 613 lines are required to achieve a 90% possibility of tagging a given gene in the B. distachyon genome using the Tnt1-based mutagenesis approach. Our results show the possibility of using Tnt1 to achieve near-saturation mutagenesis in B. distachyon, which will aid in functional genomics studies of other C3 grasses.

Increased Tim-3 expression on TILs during treatment with the Anchored GM-CSF vaccine and anti-PD-1 antibodies is inversely correlated with response in prostate cancer.

Programmed death receptor-1 (PD-1) and T cell immunoglobulin and mucin domain-containing protein-3 (Tim-3) play important roles in tumor immune evasion. PD-1 blockade could produce an effective antitumor effect in many solid tumors except prostate cancer (PCa) because of rare programmed death ligand-1 (PD-L1) expression on PCa cells. Streptavidin (SA)-GM-CSF surface-anchored tumor cell (Anchored GM-CSF) vaccines could increase the number of tumor-infiltrating lymphocytes (TILs) and induce specific antitumor immune responses. The Anchored-GM-CSF vaccine and anti-PD-1 antibodies exerted synergistic effects in mouse models of PCa metastasis. However, the response rate was low due to the presence of other negative regulatory pathways. Tim-3 expression could be upregulated at resistance to combination therapy with anti-PD-1 antibodies and the Anchored GM-CSF vaccine. Sequential administration of anti-PD-1 and anti-Tim-3 antibodies could further improve the efficacy of the Anchored GM-CSF vaccine therapy, and tumor regression was noted in over 60% of animals. This triple therapy improved the specific cytotoxic activity, proliferation and secretion of CD8+ TILs and reduced the production of tumor-promoting cytokines. These findings indicated that this triple therapy could induce a robust antitumor immune response in mouse models of PCa.

LncARSR sponges miR-129-5p to promote proliferation and metastasis of bladder cancer cells through increasing SOX4 expression.

Emerging evidences have indicated that long non-coding RNAs (lncRNAs) are potential biomarkers, playing important roles in the development of cancer. LncRNA Activated in RCC with Sunitinib Resistance (lncARSR) is a novel lncRNA that functions as a potential biomarker and is involved in the progression of cancers. However, the clinical significance and molecular mechanism of lncARSR in bladder cancer (Bca) remains unknow. In this study, we discovered that lncARSR was significantly up-regulated in bladder cancer. In addition, increased expression of lncARSR was positively correlated with higher histological grade and larger tumor size. Further experiments demonstrated that suppression of lncARSR attenuated the proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) process of Bca cells. Mechanistically, lncARSR was mainly located in the cytoplasm and acted as a miRNA sponge to positively modulate the expression of Sex-determining region Y-related high-mobility-group box transcription factor 4 (SOX4) via sponging miR-129-5p and subsequently promoted the proliferation and metastasis of Bca cells, thus playing an oncogenic role in Bca pathogenesis. In conclusion, our study indicated that lncARSR plays a critical regulatory role in Bca cells and lncARSR may serve as a potential diagnostic biomarker and therapeutic target for bladder cancer.

Concordance of chronic conditions among the household members in Shanghai: a cross-sectional study.

OBJECTIVES: Members living in the same household tend to share some similar behaviours and environment. We want to quantitatively assess the associations of chronic conditions to investigate the concordance of disease status among the household members. SETTING: Shanghai, China. PARTICIPANTS: Our data were from the fifth Health Service Survey in Shanghai in 2013. 12 002 households with 31 531 residents were selected in this survey by using a three-stage, stratified, random sampling method. OUTCOME MEASURES: Five highly prevalent chronic conditions, namely hypertension, diabetes, ischaemic heart disease (IHD), cerebrovascular disease (CVD) and obesity were chosen. The generalised estimating equations (GEE) model was used to estimate the associations adjusted for age, gender, education status, health insurance status, smoking and drinking. Using a subsample of adult children with parents' chronic conditions as the key risk factor and a subsample of wives with the chronic conditions of the husband as key risk factor, we reran our GEE models to explore chronic condition concordance within these relationships. RESULTS: A total of 10 198 households with 27 010 adult participants were included. Using all adult household members, we found positive statistically significant associations between one's chronic conditions and the same disease status of their household members (hypertension (OR=3.26, 95% CI 3.02 to 3.52); diabetes (OR=1.68, 95% CI 1.40 to 2.01); IHD (OR=5.31, 95% CI 3.56 to 7.92); CVD (OR=3.40, 95% CI 1.99 to 5.80); obesity (OR=3.41, 95% CI 2.34 to 4.96)). The results of analysing ad-child subsample and spouse subsample also showed similar associations. Moreover, the potential concordance of different chronic conditions was found between hypertension and diabetes. CONCLUSIONS: We found chronic condition concordance within households. This study provides evidence that the chronic conditions of other members of a household may be a significant risk factor for a household member's own health.

Combination immunotherapy with interleukin-2 surface-modified tumor cell vaccine and programmed death receptor-1 blockade against renal cell carcinoma.

Immunotherapy may be an effective way to prevent postoperative recurrence of renal cell carcinoma. Streptavidin-interleukin-2 (SA-IL-2) surface-modified tumor cell vaccine developed through our protein-anchor technology could induce specific antitumor T-cell responses, but this immunotherapy cannot completely eradicate the tumor. These effector T cells highly expressed programmed death receptor-1 (PD-1), and the expression of programmed death ligand-1 (PD-L1) in the tumor environment also was upregulated after SA-IL-2-modified vaccine therapy. PD-1/PD-L1 interaction promotes tumor immune evasion. Adding PD-1 blockade to SA-IL-2-modified vaccine therapy increased the number of CD4+ , CD8+ and CD8+ interferon-γ+ but not CD4+ Foxp3+ T cells. PD-1 blockade could rescue the activity of tumor-specific T lymphocytes induced by the SA-IL-2-modified vaccine. Combination therapy delayed tumor growth and protected mice against a second Renca cells but not melanoma cells challenge. Taken together, PD-1 blockade could reverse immune evasion in the treatment with SA-IL-2-modified vaccine, and eventually induce a stronger specific antitumor immune response against renal cell carcinoma.

Impact of hypertension on health-related quality of life among different age subgroups in Shanghai: the subpopulation treatment effect pattern plot analysis.

The aim of our study was to investigate the effect of hypertension on health-related quality of life (HRQoL) among different age subgroups of people in Shanghai using subpopulation treatment effect pattern plot (STEPP) methodology. We utilized data from the fifth Health Service Survey of Shanghai, 2013, which utilizes a cross-sectional study design. The participants were selected into the survey by using a three-stage, stratified, random sampling method. HRQoL was evaluated by the EuroQol five-dimensional 3 level (EQ-5D-3L) questionnaire, and the EuroQol-visual analog scales (EQ-VAS) score was the main outcome. A generalized estimating equations (GEE) model adjusted for socio-demographic covariates was used to determine the effect of hypertension on HRQoL. STEPP analysis was performed to explore the effect of hypertension within overlapping age subpopulations. Subgroup analyses for gender were conducted for the main outcome. A total of 28,730 residents who were 18 years or older were included in our study. The results of the multivariate GEE model showed that hypertension negatively affected HRQoL in the study population (estimate = -1.85, p < 0.0001). According to the STEPP analysis, we found that the EQ-VAS score in the hypertension group was lower than that in non-hypertension group for every age group. Additionally, the results of subgroup analyses indicated that the difference of score between two groups was larger among young women. When compared to respondents without hypertension, respondents with hypertension experienced lower HRQoL regardless of gender or any range of age. Furthermore, the impact of hypertension on HRQoL of young women might be more obvious.

SIRT3 Inactivation Promotes Acute Kidney Injury Through Elevated Acetylation of SOD2 and p53.

BACKGROUND: The deactivation of SIRT3, a novel deacetylase located in mitochondria, can aggravate multiple organ dysfunction. However, the role of SIRT3 and its downstream targets in ischemia/reperfusion (I/R)-induced acute kidney injury (AKI) remain unknown. MATERIALS AND METHODS: I/R was reproduced in a rat model using a clamp placed on the left and right renal pedicles for 40 min. The rats were intraperitoneally injected with either the vehicle or a selective SIRT3 inhibitor (3-TYP) and scarified at different time points (4, 8, and 24 h after I/R). A portion of the renal tissue was extracted for histological analysis, and another portion was collected for the isolation of renal tubular epithelial cells for Western blotting, SOD2 and SIRT3 activity, cell apoptosis, and the determination of oxidative stress. RESULTS: The I/R-induced AKI model was successfully reproduced and SIRT3 activity was considerably reduced than control (sham operated) group, accompanied by increased acetylation of SOD2 and p53, as well as their elevated physical interaction in extracted mitochondrial protein (all P values < 0.05). Moreover, SIRT3 suppression by 3-TYP treatment (comparing with the vehicle treatment group) aggravated AKI, as evidenced by increased indicators of oxidative stress (increased mitochondrial red fluorescence MitoSOX and decreased reduced glutathione/oxidized glutathione ratio, all P values < 0.01). CONCLUSIONS: The elevation of SOD2 and p53 protein acetylation in the mitochondria of renal tubular epithelial cells is an important signaling event in the pathogenesis of I/R-induced AKI. Thus, deacetylase SIRT3 may be an upstream regulator of both SOD2 and p53, and the SIRT3 deactivation may aggravate AKI.

PD-1 Blockade Overcomes Adaptive Immune Resistance in Treatment with Anchored-GM-CSF Bladder Cancer Cells Vaccine.

Purpose: To investigate whether PD-L1 mediated adaptive resistance could occur in treatment with Anchored-GM-CSF vaccine and whether PD-1 blockade combined with Anchored-GM-CSF vaccine could induce a greater anti-tumor immune response than either immunotherapy alone. Materials and Methods: After establishing long-established subcutaneous metastasis bladder cancer models, mice were treated with Anchored-GM-CSF vaccine and/or anti-PD-1 antibody. T-lymphocyte-cytotoxicity, flow cytometric analysis, immunohistochemical, immunofluorescence staining, CD8+ -T cell apoptosis and enzyme-linked immunosorbent assays were performed to evaluate the efficacy of combination therapy with anchored-GM-CSF vaccine and PD-1 blockade and to explore the related mechanism. Results: Anchored-GM-CSF vaccine could significantly increase the number of mature DCs and up-regulate PD-L1 expression dependent on IFN-γ released from CD8+ T cells. Anchored-GM-CSF vaccine combined with anti-PD-1 antibody could effectively inhibit tumor growth and even cause regression of the established tumor. More CD4+ and CD8+ T cells appeared at tumor sites and in peripheral blood in the combination therapy group than in the control groups. Splenocytes from mice of the combination therapy group exhibited the most potent cytotoxicity to MB49 cells. Apoptotic assays showed that PD-1 blockade could significantly reduce CD8+ T cells apoptosis. Conclusions: Anchored-GM-CSF vaccines and anti-PD-1 antibodies have synergistic effects in metastatic bladder cancer treatment. PD-1 blockade can overcome immune resistance in treatment with the Anchored-GM-CSF vaccine, while Anchored-GM-CSF vaccine can enhance the efficacy of PD-1 blockade therapy.

Protective effect of electro-acupuncture on liver ischemia-reperfusion injury in rats.

Liver ischemia-reperfusion injury is an important clinical complication in which excessive inflammation is a key factor; however, few studies have provided effective means of its regulation. As previous studies suggested that electro-acupuncture (EA) is able control excessive inflammation, the present study aimed to explore its effects on liver ischemia-reperfusion injury in experimental rats. The animals were randomly divided into surgery and sham groups, which were further divided into four sub-groups, including a non-treatment (NT), a non-point acupuncture (NPA), the non-selective nicotinic acetylcholine receptor (AChR) agonist 1,1-dimethyl-4-phenyl L-pioperazinium iodide (DMPPI) and an EA group. The alanine aminotransferase (ALT), serum cytokine and myeloperoxidase (MP) levels and the tissue pathology were evaluated after 90 min of ischemia followed by a 4, 8 or 24 h reperfusion. The results demonstrated that EA and DMPPI suppressed serum ALT elevation at 4 and 8 h reperfusion, whereas NPA did not. I/R induced hepatocellular necrosis, and cytoplasmic vacuolization and sinusoidal congestion was ameliorated by EA treatment after an 8 and 24 h reperfusion. In addition, EA also inhibited liver neutrophil accumulation, evidenced by a decreased MPO level at 8 h reperfusion. EA also suppressed the release of serum inflammatory factors TNF-α and IL-6 for the duration of reperfusion. However, little influence on IL-10 was observed. Mechanistically, vagus block by subphrenic vagotomy or mecamylamine hydrochloride abolished EA effect on liver damage, neutrophil accumulation and inflammatory factor release. In conclusion, it was demonstrated that EA protects the liver against I/R induced injury by inhibiting the inflammatory response, which is associated with the vagus.

Norcantharidin enhances antitumor immunity of GM-CSF prostate cancer cells vaccine by inducing apoptosis of regulatory T cells.

Norcantharidin (NCTD) is a promising antitumor drug with low toxicity. It was reported to be able to regulate immunity, but the mechanism is not yet clear. Here we explored whether NCTD could enhance the antitumor immunity induced by prostate cancer cell vaccine. The results of the in vitro study showed that NCTD induced apoptosis and inhibited proliferation of regulatory T cells (Tregs). Mechanistic research showed that NCTD inhibited Akt activation and activated FOXO1 transcription, resulting in a pro-apoptotic effect. The results of the in vivo study showed that more tumor-infiltrating Tregs existed within peripheral blood and tumor tissue after treatment with the vaccine. Adding NCTD to vaccine treatment could decrease the number of tumor-infiltrating Tregs and increase the number of CD4+ and CD8+ T cells. Combination therapy with NCTD and vaccine was more effective in inhibiting tumor growth than the vaccine alone. In general, this is the first report that NCTD could induce apoptosis of Tregs and enhance the vaccine-induced immunity.

PD-1 blockade enhances the antitumor efficacy of GM-CSF surface-modified bladder cancer stem cells vaccine.

Eliminating cancer stem cells (CSCs) is a key issue in eradicating tumor. The streptavidin-granulocyte-macrophage-colony stimulating factor (SA-GM-CSF) surface-modified bladder CSCs vaccine previously developed using our protein-anchor technology could effectively induce specific immune response for eliminating CSCs. However, program death receptor-1 (PD-1)/program death ligand 1 (PD-L1) signaling in tumor microenvironment results in tumor-adaptive immune resistance. Although the CSCs vaccine could increase the number of CD8+ T cells, a part of these CD8+ T cells expressed PD-1. Moreover, the CSCs vaccine upregulated the PD-L1 expression of tumor cells, resulting in immune resistance. Adding PD-1 blockade to the CSCs vaccine therapy increased the population of CD4+ , CD8+ and CD8+ IFN-γ+ but not CD4+ Foxp3+ T cells and induced the highest production of IFN-γ. PD-1 blockade could effectively enhance the functions of tumor-specific T lymphocytes generated by the CSCs vaccine. This combination therapy improved the cure rate among mice and effectively protected the mice against a second CSCs cell challenge, but not a RM-1 cell challenge. These results indicate that PD-1 blockade combined with the GM-CSF-modified CSCs vaccine effectively induced a strong and specific antitumor immune response against bladder cancer.

Depletion of regulatory T cells by anti-ICOS antibody enhances anti-tumor immunity of tumor cell vaccine in prostate cancer.

ICOS+Treg cells exert important immunosuppressive effects in tumor immunity. We adopt a combination approach of ICOS+Treg cells depletion with tumor cell vaccine to evaluate anti-tumor immunity in mouse prostate cancer model. Streptavidin (SA)-mGM-CSF surface-modified RM-1 cells were prepared as the vaccine and the mouse subcutaneous prostate tumor model was used to evaluate the immunity. Tumor growth, flow cytometry, immunohistochemistry, immunofluorescence and enzyme linked immunosorbent assay (ELISA) were performed to evaluate the therapeutic effects. Our results demonstrated that SA-mGM-CSF vaccine was prepared successfully and tumor growth was inhibited. The tumor size in the combination group was much smaller than that in the vaccine with IgG mAb group. The portions of dendritic cells, CD8+ and CD4+T cells in the mice blood and tumor tissues were increased after treatment with vaccine. There were more immune-suppressing Tregs infiltrated into tumor after treatment with tumor cell vaccine, and ICOS blocking could deplete the infiltrated Tregs, and T lymphocytes increased more dramatically in the combination therapy group. The concentrations of interferon-γ were increased in all vaccine group, the concentrations of Interleukin-10 and Interleukin-4 were much lower in the combination group. Our study demonstrated that ICOS blocking could deplete the tumor-infiltrated ICOS+Treg cells. Combining GM-CSF surface-modified RM-1 cell vaccine with Anti-ICOS antibody could induce better antitumor immunity than a vaccine alone.

PD-1/PD-L1 blockade enhances the efficacy of SA-GM-CSF surface-modified tumor vaccine in prostate cancer.

Program death receptor-1 (PD-1)/program death ligand 1 (PD-L1) signaling plays an important role in tumor adaptive immune resistance. The streptavidin-granulocyte-macrophage colony stimulating factor (SA-GM-CSF) surface-modified tumor cells vaccine developed through our novel protein-anchor technology could significantly promote the activation of dendritic cells. Although GM-CSF vaccine could significantly increase the number of tumor-specific CD8+T-cells, the majority of these CD8+T-cells expressed PD-1. Moreover, GM-CSF vaccine up-regulated the PD-L1 expression of tumor cells, resulting in immune resistance. Adding PD-1/PD-L1 blockade to GM-CSF vaccine therapy could significantly increase the population of CD4+ T, CD8+ T and CD8+ IFN-γ+ T but not CD4+ Foxp3+ T-cells and induced the highest production of IFN-γ. PD-1/PD-L1 blockade could effectively rescue the tumor-specific T lymphocytes generated by the GM-CSF vaccine, resulting in consistent tumor rejection. Taken together, PD-1/PD-L1 blockade combined with SA-GM-CSF-modified vaccine could effectively induce a strong specific antitumor immune response against prostate cancer.