Prediction of Th17/Treg cell balance on length of stay in intensive care units of patients with sepsis.

Background: Prolonged length of stay (LOS) of sepsis can drain a hospital's material and human resources. This study investigated the correlations between T helper type 17 (Th17) and regulatory T (Treg) balance with LOS in sepsis. Methods: A prospective clinical observational study was designed in Changhai Hospital affiliated to Naval Medical University in Shanghai, China, from January to October 2020. The patients diagnosed with sepsis and who met the inclusion and exclusion criteria were recruited and whether the levels of cytokines, procalcitonin, subtypes, and biomarkers of T cells in the peripheral blood were detected. We analyzed the correlation between these and LOS. Results: Sixty septic patients were classified into two groups according to whether their intensive care unit (ICU) stay exceeded 14 days. The patients with LOS ≥14 days were older ([72.6±7.5] years vs. [63.3±10.4] years, P=0.015) and had higher Sequential Organ Failure Assessment (SOFA) (median [interquartile range]: 6.5 [5.0-11.0] vs. 4.0 [3.0-6.0], P=0.001) and higher Acute Physiology and Chronic Health Evaluation (APACHE) II scores (16.0 [13.0-21.0] vs. 8.5 [7.0-14.0], P=0.001). There was no difference in other demographic characteristics and cytokines, interleukin-6, tumor necrosis factor-α, and interleukin-10 between the two groups. The Th17/Treg ratio of sepsis with LOS <14 days was considerably lower (0.48 [0.38-0.56] vs. 0.69 [0.51-0.98], P=0.001). For patients with LOS ≥14 days, the area under the receiver operating characteristic curve for the Th17/Treg ratio was 0.766. It improved to 0.840 and 0.850 when combined with the SOFA and APACHE II scores, respectively. Conclusions: The Th17/Treg ratio was proportional to septic severity and can be used as a potential predictor of ICU stay in sepsis, presenting a new option for ICU practitioners to better care for patients with sepsis.

Hydrogel Crosslinked with Nanoparticles for Prevention of Surgical Hemorrhage and Recurrence of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is acknowledged as an immunosuppressive neoplasm, whereby the inactive microenvironment facilitates immune tolerance and evasion of HCC. Post-surgical resected liver cancer exhibits a proclivity for relapse, rendering prevention of recurrence challenging as it may transpire at any point subsequent to surgery. Among the various anti-recurrence interventions, the primary clinical approach involving the administration of regimens atezolizumab and bevacizumab (A+T) is deemed the most efficacious in reversing the tumor microenvironment, albeit still lacking in complete satisfaction. Therefore, the objective is to utilize a recently developed block copolymer as a protective carrier for two specific monoclonal antibody drugs. Subsequently, a modified hemostatic hydrogel will be synthesized for application during hepatic surgery. The immunotherapy impact of this approach is significantly prolonged and intensified due to the combined hemostasis properties and controlled release of the constituents within the synthesized nanocomposite hydrogel. Furthermore, these nanocomposite hydrogels exhibit remarkable efficacy in preventing postoperative wound bleeding and substantially enhancing the safety of liver cancer resection. This research on the anti-recurrence hydrogel system presents a novel therapeutic approach for addressing local recurrence of liver cancer, potentially offering a substantial contribution to the field of surgical treatment for liver cancer in the future.

Application of a Chitosan-based Active Packaging Film Prepared with Cell-free Supernatant of Lacticaseibacillus paracasei ALAC-4 in Mongolian Cheese Preservation.

Fungal spoilage of food is a worldwide concern prompting the development of many antimicrobial agents and applications. In this study, the cell-free supernatant (CFS) of Lacticaseibacillus paracasei ALAC-4 had a significant inhibition effect on fungi. The CFS with antifungal activities were combined with chitosan (CS) matrix to prepare an active packaging CS-CFS films by using a solvent casting method and used for the packaging of Mongolian cheese for 15 days during storage at 4 ± 1℃. The optimized formulation of the film were 1.25% (w/v) chitosan, 1.75% (w/v) gelatin, 0.3% (v/v) glycerol, and 9.6% (w/v) CFS. It was found that CS-CFS films exhibited strong antifungal activities against molds and yeasts, especially Candida albicans, and also had excellent mechanical properties. Additionally, FTIR spectroscopy indicated that hydrogen bonds between the CFS and CS formed, and there was a smooth surface, compact cross-section observed in SEM morphologies of CS-CFS films. Furthermore, CS-CFS film also displayed a strong antifungal effect against molds and yeasts on cheese surface. These results suggest that the chitosan-based CS-CFS film has a promising application for Mongolian cheese and food preservation.

Generation and characterization of QLS22001, a humanized monoclonal antibody that neutralizes IL-17A and IL-17F with an extended half-life.

Therapeutic intervention to block IL-17A signaling has proven to be an effective treatment for numerous autoimmune diseases, including psoriasis, psoriatic arthritis, and axial spondylarthritis. Among the IL-17 family members, IL-17F, which shares 55% sequence homology with IL-17A, has been reported to functionally overlap with IL-17A in many inflammatory diseases. In this study, we describe the generation and characterization of QLS22001, a humanized monoclonal IgG1 antibody with an extended half-life and high affinity for both IL-17A and IL-17F. QLS22001 effectively blocks IL-17A and IL-17F mediated signaling pathways both in vitro and in vivo. Briefly, the YTE (M225Y/S254T/T256E) modification was introduced into the Fc fragment of QLS22001 WT Fc to prolong its half-life, and the resulting construct was named QLS22001. Functionally, it significantly inhibits IL-17A- and IL-17F-stimulated signaling in cell-based IL-6 release and reporter assays. The dual neutralization of the endogenous IL-17A and IL-17F produced by Th17 cells, as opposed to the selective blockade of IL-17A alone, results in a greater suppression of inflammatory cytokine secretion, according to in vitro blockade assays. Furthermore, in an in vivo mouse pharmacodynamic study, QLS22001 blocked human IL-17A-induced mouse keratinocyte chemoattractant (KC) release. In cynomolgus monkey pharmacokinetics evaluation, QLS22001 showed linear pharmacokinetic characteristics with a mean half-life of 31.2 days, while its parent antibody, QLS22001 WT Fc, had a mean half-life of 17.2 days. In addition, QLS22001 does not induce cytokine release in a human whole-blood assay. Collectively, these data provide a comprehensive preclinical characterization of QLS22001 and support its clinical development.

Joint effect of maternal pre-pregnancy body mass index and folic acid supplements on gestational diabetes mellitus risk: a prospective cohort study.

BACKGROUND: The joint effect of folic acid (FA) supplements and maternal pre-pregnancy body mass index (BMI) on gestational diabetes mellitus (GDM) has not been fully addressed. This study aimed to examine the joint effect of FA supplements and pre-pregnancy BMI on GDM. METHODS: Pregnant women at 4 to 14 weeks of gestation (n = 3186) were recruited during their first prenatal visit in Qingdao from May 1, 2019, to June 27, 2021. The main outcome was GDM at 24-28 weeks' gestation. Screening was based on 75 g 2-hour oral glucose tolerance (OGTT), a fasting glucose ≥ 5.1 mmol/L, or a 1-hour result ≥ 10.0 mmol/L, or a 2-hour result ≥ 8.5 mmol/L. The interactive effect of FA supplements and pre-pregnancy BMI on GDM was examined using logistic regression analysis and ratio of odds ratios (ROR) was used to compare subgroup differences. RESULTS: Overall, 2,095 pregnant women were included in the analysis, and GDM incidence was 17.76%. Compared with women with pre-pregnancy BMI lower than 25.0 kg/m2 and FA-Sufficient supplements ≥ 400 µg/day (FA-S) population, the adjusted odds ratios (aORs) of FA-S and FA-Deficiency supplements < 400 µg/d (FA-D) were 3.57 (95% confidence interval [CI]: 2.02-6.34) and 10.82 (95% CI: 1.69-69.45) for the obese women (BMI ≥ 30.0 kg/m2), and the aORs of FA-S and FA-D were 2.17 (95% CI: 1.60-2.95) and 3.27 (95% CI: 1.55-6.92) for overweight women (25.0 kg/m2 ≤ BMI < 30.0 kg/m2). However, the risk of GDM did not differ significantly between the FA-D and the FA-S group in pre-pregnancy obese women (ROR = 2.70, 95%CI: 0.47-2.30), or overweight women (ROR = 0.66, 95%CI: 0.30-1.49). After further stratification of FA supplementation time, F-D and FA-S in obese women showed an interaction when FA supplement intake time < 3 months. However, there was no significant difference between subgroups (ROR = 1.63, 95% CI: 0.37-7.04). CONCLUSION: Maternal pre-pregnancy BMI was associated with the incidence of GDM, the dose of FA supplementation from pre-pregnancy to early pregnancy was not found to be related to the incidence of GDM. The dosage of FA supplement was not associated with GDM irrespective of maternal pre-pregnancy BMI.

Does recurrent gestational diabetes mellitus increase the risk of preterm birth? A population-based cohort study.

AIMS: To investigate whether recurrent gestational diabetes mellitus (GDM) is associated with an increased risk of preterm birth. METHODS: We conducted a prospective population-based cohort study covering all live singleton births born to nulliparous and multiparous mothers aged 20 years and older in Qingdao, from 2018 to 2020 (n = 105,528). Preterm birth (<37 gestational weeks) was classified into moderate preterm birth (32-36 weeks of gestation) and very preterm birth (<32 weeks). Logistic regression analysis was performed to estimate the risk and severity of prematurity in relation to parity among mothers with previous GDM, current GDM, and recurrent GDM (previous and current GDM), using mothers without GDM as the reference group. Z-test and ratio of odds ratios (ROR) were used to determine subgroup differences. RESULTS: Maternal GDM increased the risk of preterm birth in both nullipara (ORadj = 1.28, 95 %CI: 1.14-1.45) and multipara (ORadj = 1.26, 95 %CI: 1.14-1.40). However, the risk of premature delivery in multiparous mothers with recurrent GDM and those with current GDM did not differ significantly, with a ROR of 0.89 (95 %CI: 0.71-1.12). The risk of recurrent GDM on preterm birth was most pronounced among multiparous mothers with pre-pregnancy BMI above 30 kg/m2 (ORadj = 2.18, 95 %CI: 1.25-3.82) as compared with those with current GDM alone (ROR = 2.20, 95 %CI: 1.07-4.52). The risk of GDM for moderate preterm birth was similar to that of overall preterm birth. In contrast, GDM was not associated with very preterm birth irrespective of parity (all P values > 0.05). CONCLUSIONS: Maternal GDM increased the risk of preterm birth in nullipara and multipara, whereas recurrent GDM was not associated with a further increase in the risk of prematurity in multiparous mothers. Maternal GDM did not contribute to very preterm birth irrespective of parity. Our findings can be useful for facilitating more targeted preventive strategies for adverse pregnancy outcomes.

Establishment of early diagnosis models for cervical precancerous lesions using large-scale cervical cancer screening datasets.

BACKGROUND: Human papilloma virus (HPV) DNA test was applied in cervical cancer screening as an effective cancer prevention strategy. The viral load of HPV generated by different assays attracted increasing attention on its potential value in disease diagnosis and progression discovery. METHODS: In this study, three HPV testing datasets were assessed and compared, including Hybrid Capture 2 (n = 31,954), Aptima HPV E6E7 (n = 3269) and HPV Cobas 4800 (n = 13,342). Logistic regression models for diagnosing early cervical lesions of the three datasets were established and compared. The best variable factor combination (VL + BV) and dataset (HC2) were used for the establishment of six machine learning models. Models were evaluated and compared, and the best-performed model was validated. RESULTS: Our results show that viral load value was significantly correlated with cervical lesion stages in all three data sets. Viral Load and Bacterial Vaginosis were the best variable factor combination for logistic regression model establishment, and models based on the HC2 dataset performed best compared with the other two datasets. Machine learning method Xgboost generated the highest AUC value of models, which were 0.915, 0.9529, 0.9557, 0.9614 for diagnosing ASCUS higher, ASC-H higher, LSIL higher, and HSIL higher staged cervical lesions, indicating the acceptable accuracy of the selected diagnostic model. CONCLUSIONS: Our study demonstrates that HPV viral load and BV status were significantly associated with the early stages of cervical lesions. The best-performed models can serve as a useful tool to help diagnose cervical lesions early.

Markov State Models and Molecular Dynamics Simulations Reveal the Conformational Transition of the Intrinsically Disordered Hypervariable Region of K-Ras4B to the Ordered Conformation.

K-Ras4B, the most frequently mutated Ras isoform in human tumors, plays a vital part in cell growth, differentiation, and survival. Its tail, the C-terminal hypervariable region (HVR), is involved in anchoring K-Ras4B at the cellular plasma membrane and in isoform-specific protein-protein interactions and signaling. In the inactive guanosine diphosphate-bound state, the intrinsically disordered HVR interacts with the catalytic domain at the effector-binding region, rendering K-Ras4B in its autoinhibited state. Activation releases the HVR from the catalytic domain, with its ensemble favoring an ordered α-helical structure. The large-scale conformational transition of the HVR from the intrinsically disordered to the ordered conformation remains poorly understood. Here, we deploy a computational scheme that integrates a transition path-generation algorithm, extensive molecular dynamics simulation, and Markov state model analysis to investigate the conformational landscape of the HVR transition pathway. Our findings reveal a stepwise pathway for the HVR transition and uncover several key conformational substates along the transition pathway. Importantly, key interactions between the HVR and the catalytic domain are unraveled, highlighting the pathogenesis of K-Ras4B mild mutations in several congenital developmental anomaly syndromes. Together, these findings provide a deeper understanding of the HVR transition mechanism and the regulation of K-Ras4B activity at an atomic level.

Mechanistic Insights into the Long-range Allosteric Regulation of KRAS Via Neurofibromatosis Type 1 (NF1) Scaffold Upon SPRED1 Loading.

Allosteric regulation is the most direct and efficient way of regulating protein function, wherein proteins transmit the perturbations at one site to another distinct functional site. Deciphering the mechanism of allosteric regulation is of vital importance for the comprehension of both physiological and pathological events in vivo as well as the rational allosteric drug design. However, it remains challenging to elucidate dominant allosteric signal transduction pathways, especially for large and multi-component protein machineries where long-range allosteric regulation exits. One of the quintessential examples having long-range allosteric regulation is the ternary complex, SPRED1-RAS-neurofibromin type 1 (NF1, a RAS GTPase-activating protein), in which SPRED1 facilitates RAS-GTP hydrolysis by interacting with NF1 at a distal, allosteric site from the RAS binding site. To address the underlying mechanism, we performed extensive Gaussian accelerated molecular dynamics simulations and Markov state model analysis of KRAS-NF1 complex in the presence and absence of SPRED1. Our findings suggested that SPRED1 loading allosterically enhanced KRAS-NF1 binding, but hindered conformational transformation of the NF1 catalytic center for RAS hydrolysis. Moreover, we unveiled the possible allosteric pathways upon SPRED1 binding through difference contact network analysis. This study not only provided an in-depth mechanistic insight into the allosteric regulation of KRAS by SPRED1, but also shed light on the investigation of long-range allosteric regulation among complex macromolecular systems.

Maternal gestational diabetes in singleton pregnancies conceived by ART may be modified by periconceptional B vitamins.

Background: The risk of maternal gestational diabetes mellitus (GDM) may be influenced by pregnancies conceived through assisted reproductive technology (ART). However, the influence of the dosage of B vitamins (folate, vitamin B6 and vitamin B12) on GDM weren't considered. Thus, we hypothesized that periconceptional B vitamins could modify maternal GDM in singleton pregnancies conceived by ART. Methods: This study is a prospective cohort study using data from 3,252 women with singleton pregnancies and received a 75 g oral glucose tolerance test (OGTT) at 24-28 weeks of gestation. We included an interaction term in the multivariable logistic and linear regression models, respectively, to test our hypothesis. Results: Women who underwent ART were significantly associated with the incidence of GDM compared with spontaneous pregnancy women. The adjusted odds ratio (aOR) was 1.59, and the 95% confidence interval (CI) was 1.08-2.34. ART pregnancies also elevated OGTT (oral glucose tolerance test) 1-h blood glucose levels and OGTT 2-h blood glucose levels (P < 0.05). A positive association between dietary vitamin B6 (aOR = 1.60, 95% CI: 1.13-2.27), dietary vitamin B12 (aOR = 1.88, 95% CI: 1.34-2.64) and dietary folate (aOR = 1.66, 95% CI: 1.19-2.32) with GDM risk comparing the highest to the lowest quartile (all P trend < 0.001). The aORs of GDM for inadequate (< 400 µg/day), adequate (400-800 µg/day), and excessive (> 800 µg/day) supplemental folate intake were 1.00, 0.93, and 1.30, respectively (P trend = 0.033). Since only the supplemental folate illustrates a statistically significant interaction with ART (P for interaction < 0.05), the association between ART and GDM and OGTT blood glucose levels stratifying by supplemental folate were further evaluated. These increased risks of GDM (aOR = 1.62, 95% CI: 1.39-3.39) and the regression coefficients (ß) of 1-h blood glucose (ß = 0.76, 95% CI: 0.39-1.13) and 2-h blood glucose (ß = 0.60, 95% CI: 0.29-0.92) in the multiple linear regression model were significant only in the ART group with excessive supplemental folate (> 800 µg/day). Conclusion: The risk of GDM is significantly elevated, particularly among those women who conceived ART with the intake of excessive supplemental folate (> 800 µg/day).

Flame Retardancy Behaviors of Flexible Polyurethane Foam Based on Reactive Dihydroxy P-N-containing Flame Retardants.

Green and environment-friendly high-efficiency flame retardants (FRs) are crucial to polymer FR modification. Here, a green FR 2-((bis(2-hydroxyethyl)amino)methyl)-5,5-dimethyl-1,3,2-dioxaphosphinane 2-oxide (HAMPP) was synthesized. The HAMPP was incorporated with a cyclic phosphorus structure, which will readily carbonize to inhibit or prevent further combustion. Moreover, the HAMPP contains dihydroxy reactive groups that can be used as a monomer in the polymerization reaction to obtain the main chain containing phosphorus polymer. Research studies on FRs were based on flexible polyurethane foam (PU-HAMPPs). The limiting oxygen index value of PU foam with 10% HAMPP could reach 23.7%, passing a UL-94 V-0 rating together. With the addition of HAMPP, the peak heat release rate of PU foam decreased significantly, the decomposition temperature increased, the heat release capacity reduced by 31%, and the char yield increased by 42%. The chemical composition and morphology of the char residual have been studied and analyzed thoroughly. We find that HAMPP forms a molten viscous protective layer uniformly on the material surface and releases some incombustible gases. These indicated that the FR exploited both condensed-phase and gas-phase flame retardancy mechanisms. Besides, the addition of FRs improved the mechanical properties.

The possible role of visceral fat in early pregnancy as a predictor of gestational diabetes mellitus by regulating adipose-derived exosomes miRNA-148 family: protocol for a nested case-control study in a cohort study.

BACKGROUND: Gestational diabetes mellitus (GDM) has become alarming public health concern. It is associated with adverse pregnancy outcomes and increased risk of postpartum type 2 diabetes. Pre-pregnant body mass index (BMI), waist circumference and other anthropometric parameters have been proposed to predict GDM. However, visceral fat thickness can better reflect the distribution of body fat, and may more accurately predict the risk of GDM. Visceral fat thickness may lead to insulin resistance by regulating the adipose-derived exosomes miRNA-148 family, which affect the development of GDM. Evidence from prospective cohort studies on visceral fat thickness as a predictor of GDM and the possible mechanisms is still insufficient. METHODS: In this prospective cohort study, we will recruit 3000 women at first antenatal visit between 4 and 12 weeks of gestation. Baseline socio-demographic factors and visceral fat thickness will be assessed by questionnaire form and the ultrasonic measurement, respectively. At 20 weeks of gestation, 10 ml blood samples will be drawn and we will extract adipose-derived exosomes miRNA on the basis of nested case-control study. GDM will be screened at 24-28 weeks' gestation and the expression of miRNA-148 family between pregnant women with GDM and without GDM will be analyzed. Intermediary analysis will be used to investigate whether visceral fat thickness can predict GDM by regulating adipose-derived exosomes miRNA-148 family. DISCUSSION: We hypothesized that visceral fat thickness may predict GDM by regulating the miRNA-148 family of adipose-derived exosomes. The findings of the study will assist in further clarifying the pathophysiological mechanism of GDM, it will also provide technical support for effective screening of high-risk pregnant women with GDM.

Liquid Transport in Fibrillar Channels of Ion-Associated Cellular Nanowood Foams.

A mechanical disintegration of waste wood biomass and freeze-induced assembly of colloidal nanowood were effectively deployed to explore ion-associated cellular foams (NWFs) with unidirectional channels. Under the assistance of inorganic ions, the as-fabricated foams were significantly enhanced in physical stability, compressive strength, flame retardancy, and thermal barrier, accounting for the tuning effects of pores and channels, surface charges, and microphase interaction by ion effects and freeze orientation. As a result, the vascular-like ion-doped channels benefited from quick capillary liquid transport. Under 1 sun illumination, NWF-V as a 3-D evaporator exhibited a high evaporation rate of 1.50 kg m-2 h-1 and a conversion efficiency of up to 88.9% for seawater desalination. Dramatically, an average of 12.5 kg m-2 of fresh water could be generated on each sunny day by outdoor NWFs for durability beyond 15 days. Under the drive of fuel combustion, an efficient conveying of ethanol or pump oil could be at rates of 0.44 and 0.26 mL min-1, respectively. Moreover, combustion flame with variable color was generated according to the doping cations in NWFs. Therefore, sustainable, green, facile, and multifunctional wood-based cellular foams could be tailored, scaled-up, and applied as color flame burners or desalination evaporators under combustion or solar drive in the energy and environment fields.

Mesophase transition of cellulose nanocrystals aroused by the incorporation of two cellulose derivatives.

Cellulose nanocrystals (CNCs) per their twisting structure and high aspect ratio and charged surface property are increasingly receiving great attention in chiral photonic crystal and pigment fabrication. However, the cholesteric mesophases of CNCs is unstable and easily destroyed by the additives with high Mw. In this work, hydroxypropyl cellulose (HPC) and carboxymethyl cellulose (CMC) are incorporated into CNCs for a continuous mesophase transition monitoring. We investigated the effects of HPC and CMC on the properties of CNCs with respect to the morphology, mesophase, rheology, and structure-color properties. Our results showed that the addition of CMC (≥ 1 wt%) prevented the formation of a continuous cholesteric phase but resulting in a fast gelation due to the strong repulsion between CMC and CNCs. Alternatively, the cholesteric phase was well-preserved in the CNC/HPC in which HPC (< 10 wt%) served as an efficient tuner of phase transition, color hue and rheology properties.

Boosting electrocatalytic hydrogen evolution by plasmon-driven hot-electron excitation.

High-performance catalysts for electrocatalytic and photoelectrochemical water splitting hold great promise for renewable energy conversion and storage. Herein, using porous N-doped carbon supported Au nanoparticles as catalysts, we demonstrate that the photon-induced localized surface plasmon resonance (LSPR) excitation on Au nanoparticles dramatically improves the hydrogen evolution reaction (HER), leading to a more than 4-fold increase of current and meanwhile affording a markedly decreased overpotential of 99 mV at a current density of 10 mA cm-2. The HER enhancement can be largely attributed to the efficient charge transfer of N-doped carbon that fastens the injection of hot electrons from plasmonic Au nanoparticles. This study highlights the increase of HER catalysis efficiency by plasmonic excitation and could provide new avenues towards the design of higher energy conversion catalytic water splitting systems with the assistance of light energy.

Tim-3 expression represents dysfunctional tumor infiltrating T cells in renal cell carcinoma.

PURPOSE: Renal cell carcinoma (RCC) is the most common cancer of kidney. Evidences have shown that RCC is sensitive to various immunotherapies. Tim-3 plays a role in suppressing Th1-mediated immune responses. However, no study has yet examined the effect of Tim-3 on tumor infiltrating lymphocytes (TILs) in RCC. METHODS: We investigated the expression and function of Tim-3 on TIL CD4+ T cells and TIL CD8+ T cells from 30 RCC patients. RESULTS: Levels of Tim-3 were significantly increased on both TIL CD4+ T cells and TIL CD8+ T cells and were associated with higher stages of the cancer. Also, GATA-3 and interferon gamma (IFN-γ) were down-regulated, whereas T-bet was up-regulated in TIL Tim-3+ T cells, indicating that Tim-3 expression defined a population of dysfunctional TIL Th1/Tc1 cells. Mechanism analyses showed that TIL Tim-3-expressing CD8+ T cells exhibited impaired Stat5 and p38 signaling pathway. Blocking the Tim-3 pathway restored cell proliferation and increased IFN-γ production in TIL CD4+ and CD8+ T cells of RCC. CONCLUSIONS: These results suggest that Tim-3 may be used as a novel target for increasing immune responses in RCC tumor microenvironment.

Interleukin 10-expressing B cells inhibit tumor-infiltrating T cell function and correlate with T cell Tim-3 expression in renal cell carcinoma.

Renal cell carcinoma is among the leading causes of cancer-related death and was found to induce IL-10. We started by focusing on IL-10-secreting cells in tumor-infiltrating lymphocytes in renal cell carcinoma patients and observed that both CD3(+) T cells and CD19(+) B cells contributed to an elevated IL-10 expression. We then focused on IL-10-expressing B cells, and found that compared to non-IL-10-producing B cells, the IL-10-expressing B cells had significantly lower levels of CD19 and CD20 expression, a lack of IgM and IgD expression, while the level of CD27 was elevated. Moreover, culturing under unstimulated conditions resulted in higher antibody production by these IL-10-producing B cells than their peripheral blood counterparts, which strongly suggested that they are plasmablast-differentiating cells. Both IgA and IgG subtypes were found but IgA had a higher relative abundance in the tumor-infiltrating fraction. We then observed inverse correlations between the frequency of IL-10-producing B cells and pro-inflammatory cytokine-producing T cells and T cell proliferation. The expression of T cell exhaustion marker Tim-3, however, was upregulated in patients with high frequencies of IL-10-producing B cells. Moreover, supernatant from tumor B cells suppressed T cell inflammation. In addition, frequencies of IL-10-producing tumor-infiltrating B cells were inversely correlated with resected tumor size, and were higher in later stage tumors. Together, our data demonstrated that IL-10-producing B cells had plasmablast-differentiating phenotype, and could contribute to T cell immunosuppression in renal cell carcinoma.