TCL1A-expressing B cells are critical for tertiary lymphoid structure formation and the prognosis of oral squamous cell carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a malignant tumor with a poor prognosis. Traditional treatments have limited effectiveness. Regulation of the immune response represents a promising new approach for OSCC treatment. B cells are among the most abundant immune cells in OSCC. However, the role of B cells in OSCC treatment has not been fully elucidated. METHODS: Single-cell RNA sequencing analysis of 13 tissues and 8 adjacent normal tissues from OSCC patients was performed to explore differences in B-cell gene expression between OSCC tissues and normal tissues. We further investigated the relationship between differentially expressed genes and the immune response to OSCC. We utilized tissue microarray data for 146 OSCC clinical samples and RNA sequencing data of 359 OSCC samples from The Cancer Genome Atlas (TCGA) to investigate the role of T-cell leukemia 1 A (TCL1A) in OSCC prognosis. Multiplex immunohistochemistry (mIHC) was employed to investigate the spatial distribution of TCL1A in OSCC tissues. We then investigated the effect of TCL1A on B-cell proliferation and trogocytosis. Finally, lentiviral transduction was performed to induce TCL1A overexpression in B lymphoblastoid cell lines (BLCLs) to verify the function of TCL1A. RESULTS: Our findings revealed that TCL1A was predominantly expressed in B cells and was associated with a better prognosis in OSCC patients. Additionally, we found that TCL1A-expressing B cells are located at the periphery of lymphatic follicles and are associated with tertiary lymphoid structures (TLS) formation in OSCC. Mechanistically, upregulation of TCL1A promoted the trogocytosis of B cells on dendritic cells by mediating the upregulation of CR2, thereby improving antigen-presenting ability. Moreover, the upregulation of TCL1A expression promoted the proliferation of B cells. CONCLUSION: This study revealed the role of B-cell TCL1A expression in TLS formation and its effect on OSCC prognosis. These findings highlight TCL1A as a novel target for OSCC immunotherapy.

Synthesis of a Multi-Stimulus Responsive RGB Fluorescent Organic Molecule Based on Dark Through-Bond Energy Transfer Mechanism.

In this study, a novel multi-stimulus responsive RGB fluorescent organic molecule, RTPE-NH2, was designed and synthesized based on the combination of aggregation-induced emission tetraphenylethylene (TPE) luminophore and acid-responsive fluorescent molecular switch Rhodamine B. RTPE-NH2 exhibits aggregation-induced emission behavior, as well as UV irradiation-stimulus and acid-stimulus responsive fluorescence properties. It could emit orange-red (R), green(G), and blue(B) light in both solution and PMMA film under 365 nm excitation. The dark through-bond energy transfer (DTBET) mechanism was proposed and supported by control experiments and TD-DFT calculations. The synthesis and application of RTPE-NH2 could accelerate the development of organic smart materials with high sensitivity and excellent optical properties.

Sodium aescinate ameliorates chronic neuropathic pain in male mice via suppressing JNK/p38-mediated microglia activation.

OBJECTIVE: A study confirmed that sodium aescinate (SA) can effectively relieve bone cancer pain, but its role in neuropathic pain (NP) remains confused. METHODS: Eighty male mice were randomly divided into four groups: sham+vehicle, sham+SA (40 µg/L, intrathecal injection), chronic contraction injury (CCI)+vehicle, CCI+SA. Behavioral assessments were used to evaluate the locomotor activity and paw withdrawal threshold (PWT) of mice. At the end of the study, spinal cord tissues were collected for histopathological analysis. The JNK/p38 signaling activation, Iba-1 expression, pro-inflammatory cytokines levels, and microglia subtype were assessed by western blotting, immunohistochemical staining, enzyme-linked immunosorbent assay, and flow cytometry with CD86/CD206, respectively. RESULTS: Early treatment with SA delayed the development of mechanical allodynia in CCI mice. Repeated SA treatment could prominently increase the reduction of PWT induced by CCI, and improve the locomotor activity of CCI mice. Mechanically, CCI surgery induced significant up-regulation of p-JNK and p-p38 protein levels, increased number and M1/M2 ratio of microglia, as well as pro-inflammatory factors in the spinal cords of mice, which could be blocked after SA administration. CONCLUSIONS: SA might suppress the activation of microglia and neuroinflammation by selectively inhibiting the JNK/p38 signaling pathway, thereby alleviating CCI-induced NP in male mice.

The interaction of CD300lf and ceramide reduces the development of periodontitis by inhibiting osteoclast differentiation.

AIM: The regulation of osteoclasts (OCs) by inhibitory immunoreceptors maintains bone homeostasis and is considered an important determinant of the extent of periodontal pathology. The aim of this study was to investigate the role of the inhibitory immunoreceptor CD300lf and its ligand ceramide in osteoclastogenesis in periodontitis. MATERIALS AND METHODS: The expression of CD300lf was measured in vitro and in a ligature-induced periodontitis model. The effect of CD300lf ablation on osteoclastogenesis was examined in ligature-retained and ligature removal periodontitis models. The effect of ceramide, the ligand of CD300lf, was examined in osteoclastogenesis in vitro and in vivo by smearing 20 µg of ceramide dissolved in carboxymethylcellulose on teeth and gingiva every other day in an experimental periodontitis model and ligature removal model. RESULTS: CD300lf expression was downregulated during osteoclastogenesis. Ablation of CD300lf in the ligature-induced periodontitis model increased the number of OCs and exacerbated bone damage. Bone resorption caused by CD300lf ablation was reversible following ligature removal. CD300lf-ceramide binding suppressed osteoclastogenesis in vitro and inhibited alveolar bone loss in a mouse periodontitis model. CONCLUSIONS: Our findings reveal that CD300lf-ceramide binding plays a critical negative role in alveolar bone loss in periodontitis by inhibiting OCs differentiation.

Seasonal changes in endoplasmic reticulum stress and steroidogenesis in the ovary of the wild ground squirrels (Citellus dauricus Brandt).

The development of the follicle is accompanied by steroidogenesis and secretion, the endoplasmic reticulum (ER) requires significant synthesis of relevant proteins to support changes in the follicular microenvironment. The aim of this study was to investigate whether seasonal changes in gonadotropins and ovarian steroid hormones in the wild ground squirrels induce endoplasmic reticulum stress (ERS) and changes in ERS-mediated unfolded protein response (UPR) signaling. There were significant seasonal differences in ovarian mass, with values higher in the breeding season and relatively low in the non-breeding season. Histological observations revealed that ovaries in the breeding season had germ cells including primordial follicles, primary follicles, secondary follicles, tertiary follicles, and the corpus luteal, whereas ovaries consisted mainly of primary and secondary follicles in the non-breeding season. Analysis of ovarian transcriptome data showed that 1298 genes were up-regulated in expression and 1432 genes were down-regulated in expression during both periods. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis indicated that these genes were mainly enriched in estrogen signaling pathways, ovarian steroidogenesis and endoplasmic reticulum protein processing pathways. The expression levels of steroidogenic enzymes (P450scc, P450c17, 3ß-HSD, and P450arom) and gonadotropin receptor (FSHR and LHR) were significantly increased during the breeding season compared to the non-breeding season. GRP78 and UPR signaling factors (ATF4, ATF6, XBP1s) associated with ERS were expressed in both seasons. The mRNA expressions of Atf6 and Xbp1s were higher in the breeding season than those of the non-breeding season. Conversely, Atf4 and its downstream homologous protein (Chop) exhibited higher expression during the non-breeding season. In addition, follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol-17ß, and progesterone of serum were significantly higher in the breeding season than those of the non-breeding season. These results suggested that UPR signaling, associated with seasonal changes in ovarian steroidogenesis, was activated during the breeding season and that ERS might be involved in regulating seasonal changes in ovarian steroidogenesis in the wild ground squirrels.

Electric Control of Exchange Bias Effect in FePS3-Fe5GeTe2 van der Waals Heterostructures.

Manipulating the exchange bias (EB) effect using an electronic gate is a significant goal in spintronics. The emergence of van der Waals (vdW) magnetic heterostructures has provided improved means to study interlayer magnetic coupling, but to date, these heterostructures have not exhibited electrical gate-controlled EB effects. Here, we report electrically controllable EB effects in a vdW heterostructure, FePS3-Fe5GeTe2. By applying a solid protonic gate, the EB effects were repeatably electrically tuned. The EB field reaches up to 23% of the coercivity and the blocking temperature ranges from 30 to 60 K under various gate-voltages. The proton intercalations not only tune the average magnetic exchange coupling but also change the antiferromagnetic configurations in the FePS3 layer. These result in a dramatic modulation of the total interface exchange coupling and the resultant EB effects. The study is a significant step toward vdW heterostructure-based magnetic logic for future low-energy electronics.

Crosstalk Defect Detection Method Based on Salient Color Channel Frequency Domain Filtering.

Display crosstalk defect detection is an important link in the display quality inspection process. We propose a crosstalk defect detection method based on salient color channel frequency domain filtering. Firstly, the salient color channel in RGBY is selected by the maximum relative entropy criterion, and the color quaternion matrix of the displayed image is formed with the Lab color space. Secondly, the image color quaternion matrix is converted into the logarithmic spectrum in the frequency domain through the hyper-complex Fourier transform. Finally, Gaussian threshold band-pass filtering and hyper-complex inverse Fourier transform are used to separate the low-contrast defects and background of the display image. The experimental results show that the accuracy of the proposed algorithm reaches 96% for a variety of crosstalk defect detection. Compared with the current advanced defect detection algorithms, the effectiveness of the proposed method for low-contrast crosstalk defect detection is confirmed.

Gate-Controlled Magnetic Phase Transition in a van der Waals Magnet Fe5GeTe2.

Magnetic van der Waals (vdW) materials are poised to enable all-electrical control of magnetism in the two-dimensional limit. However, tuning the magnetic ground state in vdW itinerant ferromagnets by voltage-induced charge doping remains a significant challenge, due to the extremely large carrier densities in these materials. Here, by cleaving the vdW itinerant ferromagnet Fe5GeTe2 (F5GT) into 5.4 nm (around two unit cells), we find that the ferromagnetism (FM) in F5GT can be substantially tuned by the thickness. Moreover, by utilizing a solid protonic gate, an electron doping concentration of above 1021 cm-3 has been exhibited in F5GT nanosheets. Such a high carrier accumulation exceeds that possible in widely used electric double-layer transistors (EDLTs) and surpasses the intrinsic carrier density of F5GT. Importantly, it is accompanied by a magnetic phase transition from FM to antiferromagnetism (AFM). The realization of an antiferromagnetic phase in nanosheet F5GT suggests the promise of applications in high-temperature antiferromagnetic vdW devices and heterostructures.

Gate-Tuned Interlayer Coupling in van der Waals Ferromagnet Fe_{3}GeTe_{2} Nanoflakes.

The weak interlayer coupling in van der Waals (vdW) magnets has confined their application to two dimensional (2D) spintronic devices. Here, we demonstrate that the interlayer coupling in a vdW magnet Fe_{3}GeTe_{2} (FGT) can be largely modulated by a protonic gate. With the increase of the protons intercalated among vdW layers, interlayer magnetic coupling increases. Because of the existence of antiferromagnetic layers in FGT nanoflakes, the increasing interlayer magnetic coupling induces exchange bias in protonated FGT nanoflakes. Most strikingly, a rarely seen zero-field cooled (ZFC) exchange bias with very large values (maximally up to 1.2 kOe) has been observed when higher positive voltages (V_{g}≥4.36 V) are applied to the protonic gate, which clearly demonstrates that a strong interlayer coupling is realized by proton intercalation. Such strong interlayer coupling will enable a wider range of applications for vdW magnets.

The effect of build orientation on the microstructure and properties of selective laser melting Ti-6Al-4V for removable partial denture clasps.

STATEMENT OF PROBLEM: Selective laser melting (SLM) Ti-6Al-4V has been used for removable partial dentures, but the impact of SLM Ti-6Al-4V build orientation is not evident. PURPOSE: The purpose of this in vitro study was to investigate the microstructure and properties of SLM Ti-6Al-4V clasps with different build orientations compared with cast Ti-6Al-4V clasps. MATERIAL AND METHODS: Forty-eight clasps were made from Ti-6Al-4V alloys-by SLM with 3 different build orientations (SLM0, SLM45, and SLM90) and cast (CAST) as a control. The microstructure was investigated by using a metallographic microscope and a confocal laser scanning microscope. The fit and surface roughness of the clasps were measured, and the physical properties were evaluated. In addition, the von Mises stresses in the clasps were calculated by finite element analysis. All specimens were then subjected to insertion and removal tests in artificial saliva to model 5 years of clinical use. After these tests, 3-point bend tests were used to analyze the fracture surface of the clasp arms, which were observed by using a scanning electron microscope. All data were statistically analyzed (α=.05). RESULTS: The microstructure of the Ti-6Al-4V specimens was a comixture of α+ß phases. In addition, growth directions of ß grains were approximately parallel to the build orientation, with acicular α grains present between ß grains. SLM0 and SLM45 had significantly higher roughness than SLM90. Even though the fit was inferior to that of SLM90, SLM0 and SLM45 still performed better than cast specimens (P<.05). The finite element analysis showed that the maximum von Mises stress was located on the middle part of the retainer arms and that the values of the 0.50-mm undercut clasps were much lower than the elastic limit. In addition, the decrease of retentive force in SLM90 clasps was less than that of the CAST group (P<.05). CAST clasps showed brittle fracture, whereas all SLM clasps showed ductile fracture. CONCLUSIONS: The microstructure of SLM Ti-6Al-4V affected the properties of clasps by changing the anisotropy of specimens. Among the tested groups, SLM90 clasps had the best fit, the lowest surface roughness, and the best fatigue resistance. Furthermore, SLM Ti-6Al-4V clasps could be engaged into 0.50-mm undercuts.

High-beam-quality operation of a 2 µm passively Q-switched solid-state laser based on a boron nitride saturable absorber.

Boron nitride (BN) nanosheets were used as the material sources which were coated on a calcium fluoride mirror to achieve a saturable absorber (SA). A passively Q-switched solid-state laser at 2000.5 nm was demonstrated with a Tm,Ho:YAlO3 (Tm,Ho:YAP) crystal and a SA based on BN. An average output power of 650 mW was obtained with a minimum pulse width of 6.3 µs at the pulsed repetition frequency of 41.7 kHz, corresponding to a per pulse energy of 13 µJ. The beam quality factor was measured to be Mx2=1.06 and My2=1.07 at the maximum output power.

A current reference-enhanced strategy endows the GFC in DC microgrids better dynamic response.

The constant voltage strategy (CVS) is more suitable for the small-capacity dc microgrid applications to form the dc bus voltage because it can eliminate the steady voltage deviation. However, its dynamic performance is slowed down by the integrator in the voltage loop and negatively influenced by the load steps. Some advanced methods have been conducted in the literature. However, their principle is to increase the system bandwidth, which is limited because the bandwidth is generally designed about one-tenth of the switching frequency and cannot be increased infinitely. This work pays efforts to increase the gain within the system bandwidth to accelerate the transient response and simultaneously eliminate the influence of the integrator. Therefore, the non-integrator disturbance observer-based (NIDOB) strategy is proposed in this work, and it can feed the load current into the current reference to replace the output of the integrator. Compared to the traditional and non-integrator directly-feedforward (NIDF) strategy, it has a better dynamic performance. Compared to the bandwidth-increased strategy, it does not introduce more noise. The theoretical analysis and experimental results prove its advantages.

Novel Carbon Quantum Dots Precisely Trigger Ferroptosis in Cancer Cells through Antioxidant Inhibition Synergistic Nanocatalytic Activity.

High levels of glutathione (GSH) are an important characteristic of malignant tumors and a significant cause of ineffective treatment and multidrug resistance. Although reactive oxygen species (ROS) therapy has been shown to induce tumor cell death, the strong clearance effect of GSH on ROS significantly reduces its therapeutic efficacy. Therefore, there is a need to develop new strategies for targeting GSH. In this study, novel carbon quantum dots derived from gentamycin (GM-CQDs) were designed and synthesized. On the basis of the results obtained, GM-CQDs contain sp2 and sp3 carbon atoms as well as nitrogen oxygen groups, which decrease the intracellular levels of GSH by downregulating SLC7A11, thereby disrupting redox balance, mediating lipid peroxidation, and inducing ferroptosis. Transcriptome analysis demonstrated that GM-CQDs downregulated the expression of molecules related to GSH metabolism while significantly increasing the expression of molecules related to ferroptosis. The in vivo results showed that the GM-CQDs exhibited excellent antitumor activity and immune activation ability. Furthermore, because of their ideal biological safety, GM-CQDs are highly promising for application as drugs targeting GSH in the treatment of malignant tumors.

Prognostic nutritional index combined with carcinoembryonic antigen and carbohydrate antigen 242 for early prediction of anastomotic leakage after radical gastrectomy for gastric cancer.

OBJECTIVE: To observe the clinical value of prognostic nutritional index (PNI) combined with carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 242 in early prediction of anastomotic leakage after radical gastrectomy for gastric cancer. METHODS: We retrospectively collected clinical data of 350 gastric cancer patients who underwent radical gastrectomy in Gansu Provincial Hospital of Traditional Chinese Medicine between January 2018 and May 2022. According to the occurrence of anastomotic leakage, patients were divided into an occurrence group (n=34) and a non-occurrence group (n=316). The clinical value of PNI combined with CEA and CA242 on the 3rd day after surgery in predicting anastomotic leakage was explored. Lasso regression analysis was used to screen predictive indicators of anastomotic leakage and establish a risk model. RESULTS: In the 350 patients who underwent radical gastrectomy for gastric cancer, anastomotic leakage was observed in 34 cases, with an incidence rate of 9.7%. A higher proportion of patients in the occurrence group exhibited diabetes, hand-sewn anastomosis, advanced tumor node metastasis (TNM) staging, and intraoperative bleeding, when compared to those in the non-occurrence group (P<0.05). Moreover, on the 3rd postoperative day, patients in the occurrence group demonstrated a significantly lower PNI than those in the non-occurrence group, along with elevated levels of CEA and CA242 (P<0.05). The area under the curve (AUC) for PNI, CEA, and CA242 were 0.827, 0.601, and 0.504, respectively, while the AUC for the combination was 0.829. As per the LASSO regression analysis, history of diabetes and PNI were identified as key factors correlating with anastomotic leakage (P<0.05). Employing the risk score formula, we obtained individual risk scores for each sample. Notably, risk scores in the occurrence group significantly surpassed those in the non-occurrence group (P<0.0001). The AUC for the risk score in predicting patient lung infection was 0.854. The internal verification C-index emerged as 0.863 (0.806-0.920), indicating a good model fit. Furthermore, the DeLong test revealed a significantly greater AUC of the risk model, compared to the combination and PNI (P<0.05). CONCLUSION: CEA and CA242 are not promising predictive indicators for anastomotic leakage after surgery in patients with gastric cancer, but the prediction model we established can improve the predictive efficiency of anastomotic leakage in these patients.

Spatiotemporal changes in summer days (SU25) in China from 1961 to 2017 and associated circulation factors.

Understanding the spatiotemporal variations in climate extremes indices, as well as the influencing factors, is critical to the scientific response to climate change. The temporal and spatial variations of SU25 (annual count of days when daily maximum temperature > 25 °C) were discussed in this study, based on daily maximum temperature data from 2398 meteorological stations in China from 1961 to 2017. The contributions of associated large-scale circulation factors to SU25 were quantitatively assessed by using the geographical detector method (GMD). The overall spatial distribution of SU25 was marked by a considerable increase from north to south. The SU25 increased significantly over time, with the national SU25 increasing at a rate of 2.5 days/decade. The Tibet Plateau (TP) had the slowest growth rate, with an average increase rate of 1.4 days/decade. The Hurst values of SU25 in all the subregions were generally high, indicating that most stations of SU25 would continue to increase in the future. Except for TP, the tipping years of other subregions were concentrated in the 1990s, and SU25 increased after the years. Among the large-scale circulation factors affecting SU25 in each subregion, Atlantic Multidecadal Oscillation (AMO) played a major role in SU25 variability. As a whole, the result of the pairwise interaction of each circulation factor was mainly nonlinear enhancement. The joint contributions of multiple factors to SU25 were larger than the contribution of each individual factor.

Graphene-Based Material-Mediated Immunomodulation in Tissue Engineering and Regeneration: Mechanism and Significance.

Tissue engineering and regenerative medicine hold promise for improving or even restoring the function of damaged organs. Graphene-based materials (GBMs) have become a key player in biomaterials applied to tissue engineering and regenerative medicine. A series of cellular and molecular events, which affect the outcome of tissue regeneration, occur after GBMs are implanted into the body. The immunomodulatory function of GBMs is considered to be a key factor influencing tissue regeneration. This review introduces the applications of GBMs in bone, neural, skin, and cardiovascular tissue engineering, emphasizing that the immunomodulatory functions of GBMs significantly improve tissue regeneration. This review focuses on summarizing and discussing the mechanisms by which GBMs mediate the sequential regulation of the innate immune cell inflammatory response. During the process of tissue healing, multiple immune responses, such as the inflammatory response, foreign body reaction, tissue fibrosis, and biodegradation of GBMs, are interrelated and influential. We discuss the regulation of these immune responses by GBMs, as well as the immune cells and related immunomodulatory mechanisms involved. Finally, we summarize the limitations in the immunomodulatory strategies of GBMs and ideas for optimizing GBM applications in tissue engineering. This review demonstrates the significance and related mechanism of the immunomodulatory function of GBM application in tissue engineering; more importantly, it contributes insights into the design of GBMs to enhance wound healing and tissue regeneration in tissue engineering.

Inhibited Crack Development by Compressive Strain in Perovskite Solar Cells with Improved Mechanical Stability.

Metal halide perovskites are promising as next-generation photovoltaic materials, but stability issues are still a huge obstacle to their commercialization. Here, the formation and evolution of cracks in perovskite films during thermal cycling, which affect their mechanical stability, are investigated. Compressive strain is employed to suppress cracks and delamination by in situ formed polymers with low elastic modulus during crystal growth. The resultant devices pass the thermal-cycling qualification (IEC61215:2016), retaining 95% of the initial power conversion efficiency (PCE) and compressive strain after 230 cycles. Meanwhile, the p-i-n devices deliver PCEs of 23.91% (0.0805 cm2 ) and 23.27% (1 cm2 ). The findings shed light on strain engineering with respect to their evolution, which enables mechanically stable perovskite solar cells.

Ag nanoparticles/PbTiO3 with in-situ photocatalytic process and its application in an ultra-sensitive molecularly imprinted hemoglobin detection.

An electrochemical sensor based on loading molecularly imprinted polymers (MIP) on the material surface can improve the specificity towards the object. In this work, a T-shaped PbTiO3 with a high active-exposed (110) facet was prepared by a hydrothermal process. Then, Ag nanoparticles (Ag NPs) modified T-shaped PbTiO3 was obtained by in-situ photocatalytic reduced method under UV irradiation, where a hetero-junction was formed with a well lattice matching between the (111) facet of Ag0 and the (110) facet of PbTiO3. A MIPs modified by Ag nanoparticles (NPs)/PbTiO3 (MAP) electrodes was prepared via electro polymerization process by o-Phenylenediamine (o-PD) in the presence of the template molecule, bovine hemoglobin (BHb), i.e., the detected molecule. The response peak current and concentration of BHb is demonstrated with a good linear relationship in the range of 0.00294-0.41 nM (R2 =0.98), and the detection limit at 0.23 pM (S/N = 3). A heterojunction between Ag NPs and high- active facet of PbTiO3 is beneficial to oxidizing electroactive material ([Fe (CN)6]3-/4-), generating more BHb-imprinting cavities on the modified electrode and improving the sensitivity of sensor. The electrochemical sensor is with a simple, stable structure and high sensitivity to BHb detection. Furthermore, the sensor was successfully applied to detect BHb in the bovine serum samples.

Dynamic changes of exhaustion features in T cells during oral carcinogenesis.

OBJECTIVES: This study aimed to clarify the dynamic changes of exhaustion features in T cells during oral carcinogenesis. MATERIALS AND METHODS: Mice were randomly divided into 4NQO group and control group. The exhaustion features of CD4+ and CD8+ T cells of both groups were detected by flow cytometry. Furthermore, multiplex immunohistochemistry was used to evaluate the expression of inhibitory receptors in human normal, dysplastic, and carcinogenesis tissues. Finally, anti-PD-1 antibody treatment was performed at the early premalignant phase of oral carcinogenesis. RESULTS: The proportion of naive T cells in 4NQO group was lower than those in control group, while the proportion of effector memory T cells was higher in 4NQO group. The expression of inhibitory receptors on CD4+ and CD8+ T cells increased gradually during carcinogenesis. In contrast, the secretion of cytokines by CD4+ and CD8+ T cells decreased gradually with the progression stage. Strikingly, those changes occurred before the onset of oral carcinogenesis. The expression of inhibitory receptors on T cells increased gradually as the human tissues progressed from normal, dysplasia to carcinoma. Interestingly, PD-1 blockade at the early premalignant phase could reverse carcinogenesis progression by restoring T cell function. CONCLUSIONS: T-cell dysfunction was established at the early premalignant phase of oral carcinogenesis; PD-1 blockade at the early premalignant phase can effectively reverse T-cell exhaustion features and then prevent carcinogenesis progression.

TDO2+ myofibroblasts mediate immune suppression in malignant transformation of squamous cell carcinoma.

Characterization of the dynamic change in the immunological landscape during malignant transformation from precancerous lesions to cancerous lesions in squamous cell carcinoma (SCC) is critical for the application of immunotherapy. Here, we performed single-cell RNA-Seq (scRNA-Seq) of 131,702 cells from 13 cancerous tissues of oral squamous cell carcinoma (OSCC), 3 samples of precancerous oral leukoplakia, and 8 adjacent normal samples. We found that tumor-infiltrating CD4+ and CD8+ T cells were functionally inhibited by immunosuppressive ligands expressed on various types of myeloid cells or neutrophils in the process of oral carcinogenesis. Notably, we identified a subset of myofibroblasts that exclusively expressed tryptophan 2,3-dioxygenase (TDO2). These TDO2+ myofibroblasts were located distally from tumor nests, and both CD4+ and CD8+ T cells were enriched around them. Functional experiments revealed that TDO2+ myofibroblasts were more likely to possess the ability for chemotaxis toward T cells but induced the transformation of CD4+ T cells into Tregs and caused CD8+ T cell dysfunction. We further showed that use of the TDO2 inhibitor LM10 attenuated the inhibitory states of T cells, restored the T cell antitumor response, and prevented the progression of OSCC malignant transformation in murine models. Our study reveals a multistep transcriptomic landscape of OSCC and demonstrates that TDO2+ myofibroblasts are potential targets for immunotherapy.