Immune Regulation and Immune Therapy in Melanoma: Review with Emphasis on CD155 Signalling.

Melanoma is commonly diagnosed in a younger population than most other solid malignancies and, in Australia and most of the world, is the leading cause of skin-cancer-related death. Melanoma is a cancer type with high immunogenicity; thus, immunotherapies are used as first-line treatment for advanced melanoma patients. Although immunotherapies are working well, not all the patients are benefitting from them. A lack of a comprehensive understanding of immune regulation in the melanoma tumour microenvironment is a major challenge of patient stratification. Overexpression of CD155 has been reported as a key factor in melanoma immune regulation for the development of therapy resistance. A more thorough understanding of the actions of current immunotherapy strategies, their effects on immune cell subsets, and the roles that CD155 plays are essential for a rational design of novel targets of anti-cancer immunotherapies. In this review, we comprehensively discuss current anti-melanoma immunotherapy strategies and the immune response contribution of different cell lineages, including tumour endothelial cells, myeloid-derived suppressor cells, cytotoxic T cells, cancer-associated fibroblast, and nature killer cells. Finally, we explore the impact of CD155 and its receptors DNAM-1, TIGIT, and CD96 on immune cells, especially in the context of the melanoma tumour microenvironment and anti-cancer immunotherapies.

Prognostic Effect of Tertiary Lymphoid Structures in Epstein-Barr Virus-Associated Gastric Carcinomas Measured by Digital Image Analysis.

Epstein-Barr virus-associated gastric carcinoma (EBVaGC) is characterized by prominent tumor-infiltrating lymphocytes (TILs) and has a favorable prognosis. Tertiary lymphoid structures (TLS), characterized by ectopic aggregated lymphocytes with high-endothelial venules (HEV), are associated with favorable outcomes in various solid tumors. We hypothesized that EBVaGC, characterized by intense TILs, may be closely associated with TLS or HEV. To test this hypothesis, we digitally analyzed the TLS, HEV, and TILs in 73 surgically resected advanced EBVaGCs. For HEV, dual MECA-79 and CD31 dual immunohistochemistry were performed, and the ectopic expression of MECA-79 in tumor cells was measured. In 73 patients with EBVaGC, a high-TLS ratio was found in 29 (39.7%) cases, high-tumor-associated HEV density in 44 (60.3%) cases, and high-CD8+ TIL density in 38 (52.1%) cases. Ectopic tumor expression of MECA-79 was observed in 36 patients (49.3%) cases. A low-TLS ratio and tumor-associated HEV density were significantly associated with lymph node metastasis (P =.005 and.042, respectively). Ectopic MECA-79 expression was significantly associated with lymph node metastasis (P =.003). Patients with a low-TLS ratio (P =.038), low-HEV density (P =.042), and ectopic tumor MECA-79 expression (P =.032) had significantly worse prognoses. In conclusion, TLS ratio and HEV density affect the survival of patients with EBVaGC and may be related to the immune response that interrupts lymph node metastasis.

Effect of multiple quantum well periods on structural properties and performance of extended short-wavelength infrared LEDs.

We present research on the role of multiple quantum well periods in extended short-wavelength infrared InGaAs/InAsPSb type-I LEDs. The fabricated LEDs consisted of 6, 15, and 30 quantum well periods, and we evaluated the structural properties and device performance through a combination of theoretical simulations and experimental characterization. The strain and energy band offset was precisely controlled by carefully adjusting the composition of the InAsPSb quaternary material, achieving high valence and conduction band offsets of 350 meV and 94 meV, respectively. Our LEDs demonstrated a high degree of relaxation of 94-96 %. Additionally, we discovered that the temperature-dependent dark current characterization attributed to generation-recombination and trap-assign tunneling, with trap-assign tunneling being more dominant at lower current injections. Electroluminescence analysis revealed that the predominant emission mechanism of the LEDs originated from localized exciton and free exciton radiative recombination, which the 30 quantum wells LED exhibited the highest contribution of the localized exciton/free exciton radiative recombination. We observed that increasing the quantum well periods from 6 to 15 led to an increase in the 300 K electroluminescence intensity of the LED. However, extending the quantum well period to 30 resulted in a decline in emission intensity due to the degradation of the epitaxial film quality.

Mesonephric-like Adenocarcinoma of the Uterine Corpus: Clinicopathological and Prognostic Significance of L1 Cell Adhesion Molecule (L1CAM) Over-expression.

BACKGROUND/AIM: The expression of L1 cell adhesion molecule (L1CAM) in uterine mesonephric-like adenocarcinoma (MLA) remains understudied. Our aim was to explore the L1CAM expression in uterine MLA, delving into its clinicopathological implications and prognostic significance. PATIENTS AND METHODS: We conducted L1CAM immunostaining in MLA, endometrioid carcinoma (EC), and serous carcinoma (SC), compared L1CAM expression across these histological types, and probed the relationship between L1CAM expression and the clinicopathological features and outcomes of patients with MLA. RESULTS: High L1CAM expression was evident in 15 of 28 MLA cases (53.6%). This rate surpassed that of EC (7.5%) but was less than that of SC (78.9%). A high L1CAM expression correlated with initial distant metastasis, advanced initial stage, lung metastasis, and the recurrence of MLA. L1CAM-high MLA exhibited worse disease-free and overall survival than L1CAM-low MLA. CONCLUSION: L1CAM over-expression was observed in more than half of the MLA cases, and was associated with aggressive clinicopathological traits and adverse outcomes in patients with uterine MLA.

Formation of Antifouling Brushes on Various Substrates Using a Melanin-Inspired Initiator Film.

In this study, we developed a substrate-independent initiator film that can undergo surface-initiated polymerization to form an antifouling brush. Inspired by the melanogenesis found in nature, we synthesized a tyrosine-conjugated bromide initiator (Tyr-Br) that contains phenolic amine groups as the dormant coating precursor and α-bromoisobutyryl groups as the initiator. The resultant Tyr-Br was stable under ambient air conditions and underwent melanin-like oxidation only in the presence of tyrosinase to form an initiator film on various substrates. Subsequently, an antifouling polymer brush was formed using air-tolerant activators regenerated by electron transfer for atom transfer radical polymerization (ARGET ATRP) of zwitterionic carboxybetaine. The entire surface coating procedure, including the initiator layer formation and ARGET ATRP, occurred under aqueous conditions and did not require organic solvents or chemical oxidants. Therefore, antifouling polymer brushes can be feasibly formed not only on experimentally preferred substrates (e.g., Au, SiO2, and TiO2) but also on polymeric substrates such as poly(ethylene terephthalate) (PET), cyclic olefin copolymer (COC), and nylon.

Antifouling Surface Coating on Various Substrates by Inducing Tyrosinase-Mediated Oxidation of a Tyrosine-Conjugated Sulfobetaine Derivative.

Inspired by the melanogenesis occurring in nature, we report tyrosinase-mediated antifouling surface coating by synthesizing a tyrosine-conjugated sulfobetaine derivative (Tyr-SB). Synthetic Tyr-SB contains zwitterionic sulfobetaine and tyrosine, whose phenolic amine group acts as a dormant coating precursor. In contrast to catecholamine derivatives, tyrosine derivatives are stable against auto-oxidation and are enzymatically oxidized only in the presence of tyrosinase to initiate melanin-like oxidation. When the surface of interest was applied during the course of Tyr-SB oxidation, a superhydrophilic poly(Tyr-SB) film was coated on the surfaces, thereby showing antifouling performance against proteins or adherent cells. Because the oxidation of Tyr-SB occurred under mild aqueous conditions (pH 6-7) without the use of any chemical oxidants, such as sodium periodate or ammonium persulfate, we anticipate that the coating method described herein will serve as a biocompatible tool in the field of biosensors, cell surface engineering, and medical devices, whose interfaces differ in chemistry.

Development of Universal and Clickable Film by Mimicking Melanogenesis: On-Demand Oxidation of Tyrosine-Based Azido Derivative by Tyrosinase.

A tyrosine-based azido derivative (TBAD) that permits both substrate-independent surface coating and clickable film functionalization by mimicking natural melanogenesis is synthesized here. In contrast to catechol derivatives, which are generally susceptible to oxidation by air under ambient conditions, the monophenol-based TBAD remains stable under alkaline and neutral conditions and is activated to oxidized quinone in situ by tyrosinase to initiate melanin-like polymerization. The resulting poly(TBAD) film can be formed on various substrates including noble metals, metal oxides, and synthetic polymers, which can undergo click reaction with terminal alkyne moieties on the entire surface or a specific region through Cu(I)-catalyzed azide-alkyne cycloaddition. The enzyme-mediated coating can rapidly form thin films (≈10 nm) and produce a uniform film morphology, which are important aspects in surface chemistry. This on-demand, clickable coating may become a significant tool for bioconjugation, soft lithography, and labeling techniques.

S/Mo ratio and petal size controlled MoS2 nanoflowers with low temperature metal organic chemical vapor deposition and their application in solar cells.

Vertically aligned two-dimensional (2D) molybdenum disulfide nanoflowers (MoS2 NFs) have drawn considerable attention as a novel functional material with potential for next-generation applications owing to their inherently distinctive structure and extraordinary properties. We report a simple metal organic chemical vapor deposition (MOCVD) method that can grow high crystal quality, large-scale and highly homogeneous MoS2 NFs through precisely controlling the partial pressure ratio of H2S reaction gas, P SR, to Mo(CO)6 precursor, P MoP, at a substrate temperature of 250 °C. We investigate microscopically and spectroscopically that the S/Mo ratio, optical properties and orientation of the grown MoS2 NFs can be controlled by adjusting the partial pressure ratio, P SR/P MoP. It is also shown that the low temperature MOCVD (LT-MOCVD) growth method can regulate the petal size of MoS2 NFs through the growth time, thereby controlling photoluminescence intensity. More importantly, the MoS2 NFs/GaAs heterojunction flexible solar cell exhibiting a power conversion efficiency of ∼1.3% under air mass 1.5 G illumination demonstrates the utility of the LT-MOCVD method that enables the direct growth of MoS2 NFs on the flexible devices. Our work can pave the way for practical, easy-to-fabricate 2D materials integrated flexible devices in optical and photonic applications.

Feasibility of training using full immersion virtual reality video game in young stroke survivor: A case report.

BACKGROUND: With the recent developments in science, full-immersion virtual reality devices have been developed, which may have feasibility for stroke rehabilitation. OBJECTIVE: This case report investigated the feasibility of training using a full-immersion virtual reality video game for improving motor function, balance, and gait in a young stroke survivor. METHOD: The case was a 27-year-old woman with stroke. A training using full-immersion virtual reality video game (Sony PlayStation®VR) was performed for 30 minutes per session, 3 sessions per week, for 6 weeks, with a total of 18 sessions. Before training and at each training for 6 weeks, with a total of 19 times, the motor function, balance, and gait were assessed using the Motor Assessment Scale (MAS), Berg Balance Scale (BBS), Timed Up and Go (TUG) Test, and Tinetti Balance Assessment, 10 Meter Walk Test (10MWT), Tinetti Gait Assessment, and Dynamic Gait Index (DGI). RESULTS: During the training, there were no adverse events reported. The case achieved 14 points higher than the pre-assessment with 34 points on the MAS, 16 points higher than the pre-assessment with 48 points on BBS, 6.85 sec lower than the pre-assessment, with 13.58 sec on TUG, 5 points higher than the pre-assessment with 13 points on the Tinetti Balance Assessment, 5.36 sec lower than the pre-assessment, with 8.15 sec on the 10MWT, 4 points higher than the pre-assessment with 10 points on the Tinetti Gait Assessment, and 10 points higher than the pre-assessment with 21 points on the DGI. CONCLUSION: This case report suggests that training using a full-immersion virtual reality video game may be a safe and effective method to improve motor function, balance, and gait in a young stroke survivor.

Threonyl-tRNA Synthetase Promotes T Helper Type 1 Cell Responses by Inducing Dendritic Cell Maturation and IL-12 Production via an NF-κB Pathway.

Threonyl-tRNA synthetase (TRS) is an aminoacyl-tRNA synthetase that catalyzes the aminoacylation of tRNA by transferring threonine. In addition to an essential role in translation, TRS was extracellularly detected in autoimmune diseases and also exhibited pro-angiogenetic activity. TRS is reported to be secreted into the extracellular space when vascular endothelial cells encounter tumor necrosis factor-α. As T helper (Th) type 1 response and IFN-γ levels are associated with autoimmunity and angiogenesis, in this study, we investigated the effects of TRS on dendritic cell (DC) activation and CD4 T cell polarization. TRS-treated DCs exhibited up-regulated expression of activation-related cell-surface molecules, including CD40, CD80, CD86, and MHC class II. Treatment of DCs with TRS resulted in a significant increase of IL-12 production. TRS triggered nuclear translocation of the NF-κB p65 subunit along with the degradation of IκB proteins and the phosphorylation of MAPKs in DCs. Additionally, MAPK inhibitors markedly recovered the degradation of IκB proteins and the increased IL-12 production in TRS-treated DCs, suggesting the involvement of MAPKs as the upstream regulators of NF-κB in TRS-induced DC maturation and activation. Importantly, TRS-stimulated DCs significantly increased the populations of IFN-γ+CD4 T cells, and the levels of IFN-γ when co-cultured with CD4+ T cells. The addition of a neutralizing anti-IL-12 mAb to the cell cultures of TRS-treated DCs and CD4+ T cells resulted in decreased IFN-γ production, indicating that TRS-stimulated DCs may enhance the Th1 response through DC-derived IL-12. Injection of OT-II mice with OVA-pulsed, TRS-treated DCs also enhanced Ag-specific Th1 responses in vivo. Importantly, injection with TRS-treated DC exhibited increased populations of IFN-γ+-CD4+ and -CD8+ T cells as well as secretion level of IFN-γ, resulting in viral clearance and increased survival periods in mice infected with influenza A virus (IAV), as the Th1 response is associated with the enhanced cellular immunity, including anti-viral activity. Taken together, these results indicate that TRS promotes the maturation and activation of DCs, DC-mediated Th1 responses, and anti-viral effect on IAV infection.

Sulfonate Version of OHPAS Linker Has Two Distinct Pathways of Breakdown: Elimination Route Allows Para-Hydroxy-Protected Benzylsulfonate (PHP-BS) to Serve as an Alternative Self-Immolative Group.

Recently we have reported that the ortho-hydroxy-protected aryl sulfate (OHPAS) system can be exploited as a new self-immolative group (SIG) for phenolic payloads. We extended the system to nonphenolic payloads by simply introducing a para-hydroxy benzyl (PHB) spacer. As an additional variation of the system, we explored a benzylsulfonate version of the OHPAS system and found that it has two distinct breakdown pathways, cyclization and 1,4-elimination, the latter of which implies that para-hydroxy-protected (PHP) benzylsulfonate (BS) can also be used as an alternative SIG. The PHP-BS system was found to be stable chemically and in mouse and human plasma, having payload release rates comparable to those of the original OHPAS conjugates.

IL-33 changes CD25hi Tregs to Th17 cells through a dendritic cell-mediated pathway.

Interleukin (IL)-33 is an alarmin factor that is highly secreted in a variety of autoimmune diseases, induces maturation of dendritic cells (DCs) and differentiation of T helper 17 (Th17) cells. As the balance between Th17 cells and regulatory T cells (Tregs) is important to maintain immune homeostasis, in this study, we investigated the effects of IL-33 on Treg cell response. We observed that direct treatment with IL-33 had no effect on Treg differentiation, whereas IL-33-matured DCs (IL33-matDCs) inhibited the differentiation of CD4+ T cells to Tregs by decreasing the expression of Foxp3. Furthermore, co-culture with IL-33-matDCs changed stable Tregs (CD25hiCD4+ Tregs) to IL-17-producing cells, whereas IL-33-matDCs had little effects on unstable Tregs (CD25loCD4+ Tregs). The stable Tregs were demonstrated to express high levels of IL-6 receptors. Blocking of IL-6 secreted from IL-33-matDCs suppressed the conversion of Tregs to Th17 cells, indicating the greater propensity to convert stable Tregs to Th17 cells is due to IL-6 signaling. Taken together, these results demonstrate that IL-33 inhibits Treg differentiation and the conversion of stable Tregs to Th17 cells via DCs.

Differential effects of luteolin and its glycosides on invasion and apoptosis in MDA-MB-231 triple-negative breast cancer cells.

Luteolin is known to have anticancer activity in various cancers. Recent studies have shown that luteolin glycosides, such as luteolin-8-C-ß-fucopyranoside, 7-methoxy-luteolin-8-C-ß-(6- deoxyxylopyranos-3-uloside) and luteolin-8-C-ß-d-glucopyranoside, flavonoids that are present in Arthraxon hispidus, exert antimigratory and anti-invasive effects, but no cytotoxic effect in estrogen receptor-positive MCF7 breast cancer cells. In the present study, we further investigated and compared differential effects of luteolin and its glycosides in MDA-MB-231 triple-negative breast cancer cells. Luteolin suppressed the expression of matrix metalloproteinase-9 and inhibited migration and invasion in MDA-MB-231 cells treated with the tumor promotor 12-O-tetradecanoylphorbol-13-acetate at non-cytotoxic concentrations (0, 5, and 10 µM). Furthermore, at cytotoxic concentrations (20 and 40 µM), luteolin induced apoptosis via extrinsic and intrinsic pathways in MDA-MB-231 cells. However, luteolin glycosides did not exert any cytotoxic, antimigratory, or anti-invasive effect in MDA-MB-231 cells. In brief, luteolin had both antimetastatic and cytotoxic effects on MDA-MB-231 cells, whereas luteolin glycosides had no effect on this cell line. Taking together the present results and our previous findings on the differential effects of luteolin and its glycosides on MDA-MB-231 and MCF-7 breast cancer cells, luteolin and its glycosides can be suggested as a potential candidate for breast cancer therapy.

NLRP3 negatively regulates Treg differentiation through Kpna2-mediated nuclear translocation.

Naïve CD4+ T cells in the periphery differentiate into regulatory T cells (Tregs) in which Foxp3 is expressed for their suppressive function. NLRP3, a pro-inflammatory molecule, is known to be involved in inflammasome activation associated with several diseases. Recently, the expression of NLRP3 in CD4+ T cells, as well as in myeloid cells, has been described; however, a role of T cell-intrinsic NLRP3 in Treg differentiation remains unknown. Here, we report that NLRP3 impeded the expression of Foxp3 independent of inflammasome activation in Tregs. NLRP3-deficient mice elevate Treg generation in various organs in the de novo pathway. NLRP3 deficiency increased the amount and suppressive activity of Treg populations, whereas NLRP3 overexpression reduced Foxp3 expression and Treg abundance. Importantly, NLRP3 interacted with Kpna2 and translocated to the nucleus from the cytoplasm under Treg-polarizing conditions. Taken together, our results identify a novel role for NLRP3 as a new negative regulator of Treg differentiation, mediated via its interaction with Kpna2 for nuclear translocation.

Introduction of Para-Hydroxy Benzyl Spacer Greatly Expands the Utility of Ortho-Hydroxy-Protected Aryl Sulfate System: Application to Nonphenolic Payloads.

The ortho-hydroxy-protected aryl sulfate (OHPAS) linker is composed of a diaryl sulfate backbone equipped with a latent phenol moiety at the ortho position of one of the aryl units. The Ar-OH released when the ortho phenol undergoes intramolecular cyclization and displaces the second aryl unit can be viewed as a payload. We have shown in the preceding paper that the OHPAS linkers are highly stable chemically and in various plasmas, yet release payloads when exposed to suitable triggering conditions. As an extension of the OHPAS system, we employed a para-hydroxy benzyl (PHB) spacer for coupling to nonphenolic payloads; this tactic again provided a highly stable system capable of smooth release of appended payloads. The PHB modification works beautifully for tertiary amine and N-heterocycle payloads.

Aryl Sulfate is a Useful Motif for Conjugating and Releasing Phenolic Molecules: Sulfur Fluorine Exchange Click Chemistry Enables Discovery of Ortho-Hydroxy-Protected Aryl Sulfate Linker.

A new self-immolative linker motif, Ortho Hydroxy-Protected Aryl Sulfate (OHPAS), was devised, and OHPAS-containing antibody drug conjugates (ADC) were tested in vitro and in vivo. Conveniently synthesized using Sulfur Fluorine Exchange (SuFEx) chemistry, it is based structurally on diaryl sulfate, with one aryl acting as a payload and the other as a self-immolative sulfate unit having a latent phenol function at the ortho position. The chemically stable OHPAS linker was stable in plasma samples from 5 different species, yet it can release the payload molecule smoothly upon chemical or biological triggering. The payload release proceeds via intramolecular cyclization, producing a cyclic sulfate coproduct that eventually hydrolyzes to a catechol monosulfate. A set of OHPAS-containing ADCs based on Trastuzumab were prepared with a drug to antibody ratio of ∼2, and were shown to be cytotoxic in 5 different cancer cell lines in vitro and dose-dependently inhibited tumor growth in a NCI-N87 mouse xenograft model. We conclude that OHPAS conjugates will be of considerable use for delivering phenol-containing payloads to tissues targeted for medical intervention.

Development of a wheeled walker braking device using the four-bar mechanism.

The elderly population in many countries has been rising rapidly, and falls are a serious event many elderly people experience. Assistive equipment is actively used to reduce falls among elderly people. Popular types of assistive equipment include canes, electric wheelchairs, and wheeled walkers. Wheeled walkers support the body of elderly people, making their gait comfortable as they age or recover from injuries. Wheeled walkers may be equipped with hand brakes; however, frail older people may experience difficulty using such hand brakes, as they require force to operate. Thus, in the present study, a braking method using a wire connected to a user's belt or clothes was designed and implemented; if the tension of the wire connecting the safety device and the user exceeds a critical value, the wheeled walker brakes, which can prevent the rapid motion of walkers. Two feasibility tests of the wheeled walker with the braking device were conducted: one with 10 healthy adults in their 20s and the other with 10 elderly people over 65 years of age; the tests measured the braking time and speed control using a speed measuring device. The results of the first and second feasibility tests demonstrated that the average braking time of participants was 50.3 ms and 50.7 ms, respectively. All participants in the feasibility tests succeeded in the speed control test. Thus, based on the results, the braking device on the wheeled walker worked properly.

Effect of Horizontal Whole-Body Vibration Training on Trunk and Lower-Extremity Muscle Tone and Activation, Balance, and Gait in a Child with Cerebral Palsy.

BACKGROUND The aim of the present study was to investigate the effect of horizontal whole-body vibration (WBV) training on trunk and lower-extremity muscle tone and activation, balance, and gait in a child with spastic diplegia cerebral palsy. CASE REPORT A 10-year-old male with spastic diplegia cerebral palsy received horizontal WBV training followed by conventional physiotherapy (50 min per day, 12 days per month), but only conventional physiotherapy during followup. Muscle tone was assessed using the Modified Ashworth Scale (MAS) and muscle activation with surface electromyography. Balance was assessed using the Timed Up and Go test (TUG) and Pediatric Balance Scale (PBS), and gait parameters were assessed using the GAITRite system. Assessment was performed at 3 points: pre-intervention, post-intervention, and follow-up. Following the intervention, MAS decreased in both the hip extensor and right ankle plantar flexor. Muscle activation increased post-intervention in the bilateral erector spinae (ES), rectus abdominis (RA), rectus femoris (RF), and right tibialis anterior (TA) during standing, and in the left RA, bilateral RF, gastrocnemius (GCM), and left TA during squatting. At follow-up, activation increased in the right ES, left RA, and RF during standing. At post-intervention and follow-up, improvement was observed in PBS score, gait velocity, right step length, and right stride length, with decreased single-leg support time, and double support and toe deviation angle. CONCLUSIONS Horizontal WBV training can safely and effectively maintain and improve physical performance and can be considered for inclusion in rehabilitation programs.

IL-33-matured dendritic cells promote Th17 cell responses via IL-1ß and IL-6.

IL-33 is associated with a variety of autoimmune diseases, such as sclerosis, inflammatory bowel disease, and rheumatoid arthritis. Although IL-33 is mainly involved in the induction of Th2 cells, however, the relationship between IL-33 and Th17 cells is still largely unknown. In this study, we investigated the effects of IL-33 on DC-mediated CD4+ T cell activation and Th17 cell differentiation because DCs are essential cells for presenting self-antigens to CD4+ T cells in autoimmune disease conditions. OT-II mice were injected with IL-33-treated DCs or untreated DCs that were primed by OVA323-339 peptide, and their Th17 cell responses were compared. Th17 cell population and IL-17 expression levels were significantly increased in draining lymph nodes of mice injected with IL-33-treated DCs, compared with those in mice injected with untreated DCs. IL-33 treatment maturated DCs to present self-antigens and to increase production of proinflammatory cytokines such as IL-1ß and IL-6, which have a crucial role in Th17 cell differentiation. We found that the IL-33-matured DCs enhanced the expression of an early T cell activation marker (CD69) and the Th17 master transcription factor (RORγt), but IL-33 did not directly affect CD4+ T cell differentiation or increase Th17 polarization. Notably, neutralizing IL-1ß and/or IL-6 significantly decreased IL-17 expression levels and Th17 cell population which were increased by the coculture of CD4+ T cells with IL-33-matured DCs, indicating that IL-33 may induce Th17 cell responses via IL-1ß and IL-6 derived from IL-33-matured DCs.

l-Asparaginase-mediated downregulation of c-Myc promotes 1,25(OH)2 D3 -induced myeloid differentiation in acute myeloid leukemia cells.

Treatment of acute myeloid leukemia (AML) largely depends on chemotherapy, but current regimens have been unsatisfactory for long-term remission. Although differentiation induction therapy utilizing 1,25(OH)2 D3 (VD3) has shown great promise for the improvement of AML treatment efficacy, severe side effects caused by its supraphysiological dose limit its clinical application. Here we investigated the combinatorial effect of l-asparaginase (ASNase)-mediated amino acid depletion and the latent alternation of VD3 activity on the induction of myeloid differentiation. ASNase treatment enhanced VD3-driven phenotypic and functional differentiation of three-different AML cell lines into monocyte/macrophages, along with c-Myc downregulation. Using gene silencing with shRNA and a chemical blocker, we found that reduced c-Myc is a critical factor for improving VD3 efficacy. c-Myc-dependent inhibition of mTORC1 signaling and induction of autophagy were involved in the enhanced AML cell differentiation. In addition, in a postculture of AML cells after each treatment, ASNase supports the antileukemic effect of VD3 by inhibiting cell growth and inducing apoptosis. Finally, we confirmed that the administration of ASNase significantly improved VD3 efficacy in the prolongation of survival time in mice bearing tumor xenograft. Our results are the first to demonstrate the extended application of ASNase, which is currently used for acute lymphoid leukemia, in VD3-mediated differentiation induction therapy for AML, and suggest that this drug combination may be a promising novel strategy for curing AML.