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Skin patches (SPs) have rapidly advanced to rehabilitation, health monitoring, self-powered and integrated systems. Accordingly, design of nanomaterials, flexible substrates, hydrogels and nanofibers can facilitate the therapeutic application of SPs.

Triple-negative breast cancer (TNBC) is one of the most fatal subtype of breast cancer. Tumor immune microenvironments (TIMEs) have critical influences on TNBC therapies. However, syngeneic mouse tumor models are insufficient to evaluate immunotherapeutic efficacy, because the different immune responses between mice and human. Therefore, human xenograft tumor mouse models with humanized immune systems (HIS) are more suitable for evaluation of immunotherapeutic efficacy. In these study, two immune humanized mouse models transferred with human hematopoietic stem cells (hHSC-HIS) or human peripheral mononuclear cells (hPBMC-HIS) in our advanced severe immune deficiency (ASID) mice were generated. Both systemic immune profiling and TIMEs of TNBC MDA-MB-231 tumors from hHSC- and hPBMC-HIS ASID models were compared. Generation of the comprehensive human immune system and large amounts of human macrophages/bone marrow-derived stromal cells around the TIMEs were noted in hHSC-HIS ASID mice, but the growth of TNBC was not affected. In contrast, T cells dominated the systemic immune system and TIMEs in hPBMC-HIS ASID mice, and inhibited the tumor growth. In summary, the TIMEs of hHSC-HIS ASID model is suitable for validations of immunotherapies reversing immune suppression. The TIMEs of hPBMC-HIS ASID can be applied for cytotoxicity test of tumor specific T cells.

A series of Ni complexes supported by SNO Schiff-base derivatives were synthesized in this study. Complex synthesis and characterization data are reported herein. Treatment of the pro-ligands [L

Long-lasting immunological memory is the ultimate goal of vaccination. Homeostatic maintenance of memory CD8We engineered a genetically engineered mouse in which IL-15R complementary DNA (cDNA) had been inserted in-frame with lecithin-retinol acyltransferase gene and bred onto an IL-15R-KO (15R-KO) genetic background (L15R) that expressed IL-15R in HSCs at normal levels, but not in other liver cells. Outside of the liver of L15R mice, IL-15R expression was found in a number of organs, but not in dendritic cells and macrophages. The low IL-15R expression in the bone marrow (BM) of L15R mice was eliminated by the reconstitution of lethally-irradiated L15R mice with 15R-KO BM to generate L15RC mice. Because MemCD8TC maintenance is mediated by 15HD, not empty IL-15R, 15HD content in L15R mice was determined and found for liver, lung, kidney, and heart. L15R and L15RC mice developed and maintained long-lasting, systemic antigen-specific MemCD8TCs that were efficacious against tumor growth and Listeria monocytogenes infection in an antigen-specific manner. Among the four organs with 15HD content, liver-associated MemCD8TCs were different from those found in the lung, kidney, and heart in two ways: (1) they were quantitatively the most numerous, and (2) they appeared uniquely in the form of clusters in a specialized structure, sinusoidal niches of the liver.The liver, the largest organ of the body, is endowed with the capability of effectuating long-lasting functional cytotoxic T cell memory.

We reported for the first time the utilization of hexadentate benzothiazole-based diamine-bisphenolate ligands to synthesize structurally well-characterized dinickel dicarboxylate complexes and studied their catalysis for copolymerization of carbon dioxide with epoxides. Dinickel carboxylate complexes having a 1,3-diamine-bridged backbone were demonstrated to be high-performance catalysts for alternating copolymerization of CO

The highlight of in vivo measurement to monitor abdominal compartment syndrome (ACS) caused by surgery has emerged since ACS without care will lead to death and organ failure if ignored in advance. Typically, it is intuitive to conventionally implant a monitoring electronic device or swallow an electronics device (which is an emerging technology) such as pressure sensor in body for a long time to continuously detect the abdominal compartment abnormal pressure; however, the pressure sensor in the body may have risk. In this letter, we design and fabricate a flexible capacitance sensor to detect the tiny capacitance change due to the acidity (pH value) in the saliva caused by the ACS. The devised circular-type sensor senses the 2-fF capacitance change in air, 8-fF change in deionized water, 12-fF change in glucose liquid, and 14 fF in 0.5% hydrochloric acid. The proposed highly sensitive capacitance flexible sensor is not only used to detect saliva pH change caused by ACS but can also extend to a wearable device to monitor human biomarkers, such as pH and Na+ in sweat, by integrating the microfluid device.

Functionalized polycarbonates from carbon dioxide have become an emerging topic owing to a large number of potential applications recently. In this contribution, we have developed efficient catalysis strategies along with dinuclear nickel catalysts to synthesize functional CO2-based polycarbonates and investigated their unique applications in the field of biomaterials. Novel bimetallic nickel catalysts based on bis(benzotriazole iminophenolate) (BiIBTP) ligands have been prepared for copolymerization of carbon dioxide (CO2) with epoxides to produce CO2-derived polycarbonates. Nickel complexes [(C83CBiIBTP)Ni2(DNP)2(MeOH)2] (1), [(C83CBiIBTP)Ni2(DNP)(H2O)3][DNP] (2) and [(C83CBiIBTP)Ni2(NP)(MeOH)2][NP] (3) (DNP = 2,4-dinitrophenolate; NP = 2-nitrophenolate) from the facile one-pot synthesis have been structurally characterized by single crystal X-ray crystallography, and their molecular structures belong to dinuclear Ni(II)/Ni(II) species containing a hexadentate BiIBTP ligand and two nitrophenolate coligands. We found that alternating copolymerization of CO2 with cyclohexene oxide (CHO) could proceed efficiently via catalysis using dinickel complex 1 or 3 to afford CO2-based poly(cyclohexene carbonate)s (PCHCs) with >99% carbonate linkages under optimized conditions. Beyond CO2/CHO copolymerization, copolymerization of 4-vinyl-1,2-cyclohexene oxide (VCHO), CHO and CO2 to obtain the corresponding polycarbonates with the vinyl functionalities (PCHC-co-PVCHC) was also studied. Utilizing the vinyl functionalities, one copolymer PCHC-co-PVCHC was successfully converted into charged PCHCs (CPCHCs) with different levels of charge density through thiol–ene click functionalization. Having an amphiphilic structure, these CPCHCs have been used as a biodegradable polymeric surfactant to prepare an oil-in-water (O/W) mini-emulsion. CPCHCs were further converted into degradable nanoparticles (NPs) through our developed mini-emulsion interfacial cross-linking technique and biomedical functions of the NPs such as drug loading capacity, drug release profile and cytotoxicity have been demonstrated.

Hierarchically porous carbon materials with interconnected frameworks of macro- and mesopores are desirable for electrochemical applications in biosensors, electrocatalysis, and supercapacitors. In this study, we report a facile synthetic route to fabricate hierarchically porous carbon materials by controlled macro- and mesophase separation of a mixture of polystyrene

We reported a series of well-characterized di-nuclear nickel catalysts containing 1,3-diamine-bis(benzotriazole phenolate) derivatives for high-performance copolymerization of CO2 and epoxides. The di-nickel di-acetate complex 1 was found to be a highly active catalyst to alternatingly copolymerize CO2 with cyclohexene oxide (CHO), producing poly(cyclohexene carbonate)s (PCHCs) with a high turnover frequency (TOF) up to 7800 h−1 under optimized conditions. Interestingly, complex 1 also appears to be the first example of nickel-catalyzed CO2-copolymerization with CHO that could proceed at 1 atm CO2 pressure. Not only has the excellent catalysis of CO2/CHO copolymerization by di-Ni 1 been enabled, but also 1 has been further applied to effectively catalyze the copolymerization of CO2 with 4-vinyl-1,2-cyclohexene oxide or cyclopentene oxide (CPO) to obtain the corresponding polycarbonates. Particularly, 1 was also reported for the first time to be highly efficient for nickel-catalyzed CO2/CPO copolymerization and enabled the formation of poly(cyclopentene carbonate)s with a TOF > 700 h−1 and excellent carbonate linkage contents (>99%). Kinetic studies of the Ni-catalyzed CO2-copolymerization of CHO were performed and the activation energy for PCHC formation by 1 is 60.6 kJ mol−1.

Four novel di-nuclear yttrium catalysts coordinated by bis(benzotriazole iminophenolate) (BiIBTP) ligands have been synthesized and structurally characterized. The one-pot procedure of BiIBTP pro-ligand with [Y(NO3)3.6H2O] (1.0 equiv.) and 2-nitrophenol derivative (1.0 equiv.) in the presence of triethylamine (5.0 equiv.) under refluxing MeOH solution gave bimetallic yttrium nitrophenolate complexes 1–4, [(μ-BiIBTP)Y(L)]2 (BiIBTP = C13CBiIBTP or C83CBiIBTP; L = 2-nitrophenolate or 2,4-dinitrophenolate) with good to high yields. The solid-state structure of 1 and 3–4 exhibits a di-yttrium(III) di-nitrophenolate species with the pentadentate BiIBTP ligand chelating two metal atoms. Versatile catalysis towards ring-opening polymerization of cyclic ester and copolymerization of internal epoxides with carbon dioxide (CO2) or phthalic anhydride (PA) catalyzed by these di-yttrium nitrophenolate complexes was systematically studied. Experimental results revealed that air-stable complexes 3 and 4 were shown to effectively catalyze not only lactide polymerization but also copolymerization of cyclohexene oxide (CHO) with CO2 or PA. Among these compounds, di-Y complex 4 was the most active catalyst for the ring-opening polymerization of l -lactide in combination with 9-anthracenemethanol, providing poly( l -lactide)s having the expected molecular weight and narrow molecular weight distribution. Furthermore, catalyst 3 was capable of copolymerizing CO2 and CHO in the presence of co-catalysts under mild conditions, producing perfectly alternating poly(cyclohexene carbonate)s with the controllable character. In addition, the molecular weight controllable poly(PA-co-CHO)s with high ester linkage contents could be afforded on copolymerizing PA and CHO utilizing di-yttrium complex 3 as the catalyst. This work provides the facilely synthesized di-nuclear yttrium complexes as versatile catalysts for three different types of polymerization catalysis.

A novel series of well-defined dicarboxylate dinuclear nickel complexes containing benzotriazole based 1,3-diamine-bisphenolate (1,3-DiBTP) ligands were readily synthesized through a one-pot procedure, which were highly active single-component catalysts for copolymerization of CO

A family of di-nuclear bis(benzotriazole iminophenolate) (BiIBTP) cobalt complexes containing diverse ancillary carboxylate derivatives have been synthesized and structurally characterized. The one-pot synthesis of the BiIBTP ligand precursor with cobalt perchlorate salt (2.0 equiv.) and carboxylic acid derivatives (2.0 or 5.0 equiv.) in the presence of triethylamine (5.0 equiv.) under refluxing methanolic solution generated bimetallic di-carboxylate Co(II)/Co(II) complexes [(C83CBiIBTP)Co2(O2CR)2] (R = C6H5 (1), C6F5 (2), 4-CF3–C6H4 (3), 4-OMe–C6H4 (4), CF3 (5)) in ≧65% yields. Interestingly, the Co(II)/Co(III) mixed-valence complex 6 resulted from the treatment of 1 with silver perchlorate (1.0 equiv.) as the oxidizing agent under an O2-atmosphere in 50% yield. The crystal structure of 6 reveals an ionic and di-nuclear benzoate species composed of a cationic moiety formulated as [(C83CBiIBTP)Co2(O2CC6H5)2]+ and a counterbalanced perchlorate anion, and both metal atoms are attributed to hexa-coordinated cobalt ions with varied coordination environments. Catalysis results of CO2/epoxide copolymerization indicated that complex 1 was more efficient than 2–6 where compound 6 was shown to be the least active. Co complex 1 incorporating benzoate coligands was demonstrated to effectively catalyze the CO2-copolymerization of cyclohexene oxide (CHO), 4-vinyl-1,2-cyclohexene oxide or cyclopentene oxide, producing the associated CO2-based polycarbonates with >99% carbonate repeated units under optimal conditions. Not only the controllable character of complex 1 for CO2/CHO copolymerization is enabled, but also 1 has been shown to catalyze such a copolymerization in the “immortal” manner. Using the same di-cobalt catalyst in combination with excess ratios of neopentyl glycol (up to 150 equiv.) as the chain transfer agent could give low molecular weight poly(cyclohexene carbonate) polyols with monomodal molecular weight distributions. This work offers the facilely prepared di-nuclear cobalt complexes as catalysts for the efficient catalysis of CO2-copolymerization.

A series of di-nuclear metal acetate complexes 1–6 incorporated by nitrogen heterocycle-containing salen-type ligands have been synthesized, structurally characterized and performed as catalysts to prepare biodegradable polycarbonates and polyesters. Their catalytic performances for copolymerization of carbon dioxide-epoxides or cyclic anhydride-epoxides were systematically examined. Bimetallic nickel(II) complexes 1, 2 and 5 were active catalysts for the alternating copolymerization of cyclohexene oxide (CHO) with CO2; di-nickel complex 1 was shown to be the most effective and selective, leading to obtaining poly(cyclohexene carbonate)s with the best efficiency among them. Moreover, complex 1 was also found to be versatile for the ring-opening copolymerization of CO2 with different cyclic epoxides to give the corresponding polycarbonates. Additionally, di-cobalt(II) analogs 3, 4 and 6 were efficient catalysts for the alternating copolymerization of CHO and phthalic anhydride (PA) under mild conditions. Based on the results of catalytic studies, complex 3 was demonstrated to be the most active one CHO-PA copolymerization, producing the polymeric products with a “controlled” manner involving controllable molecular weights and narrow polydispersity. Interestingly, Co complex 3 was also able to catalyze the copolymerization of PA with 4-vinyl-1,2-cyclohexene oxide to obtain the associated polyester with the vinyl functionality on the side chains, which was further functionalized with tertiary amine moieties via thiol-ene click functionalization and converted to nanofibers through electrospinning. Due to the incorporation of polar groups, the resulting tertiary amine-modified polyester nanofibers that exhibit an improved hydrophilic property relative to their un-modified counterpart have been considered to have high potential to be utilized as a new functional fiber material.

Four new 2D indium metal-organic frameworks (MOFs) (Me2NH2)[In(SBA)2] (1), (Me2NH2)[In(SBA)(BDC)] (2), (Me2NH2)[In(SBA)(BDC-NH2)] (3), and (NH4)3[In3Cl2(BPDC)5] (4), (H2SBA = 4,4'-sulfonyldibenzoic acid; H2BDC = 1,4-benzenedicarboxylic acid; H2BDC-NH2 = 2-amino-1,4-benzenedicarboxylic acid; H2BPDC = 4,4'-biphenyldicarboxylic acid) have been synthesized under solvothermal reaction conditions for compounds 1 to 3 and the DES (deep eutectic solvent) reaction has been attempted for compound 4. The structure of these MOFs has been determined by using single crystal X-ray diffraction study and all of theses four 2D monolayer framework with porous properties. The N2 gas sorption measurements indicated that Brunauer-Emmer-Teller (BET) and Langmuir surface areas of compound 1 are 207 and 301 m2 g-1, respectively, which is probably the first one having substantial gas uptake properties in the entire 2D In-MOF family to date. Furthermore, these new indium MOFs on the addition of n-Bu4NBr were active for the cycloaddition of CO2 and propylene oxide, generating propylene carbonates in high conversions under mild conditions. Particularly, the most active MOF 4 was found to efficiently couple CO2 with a series of terminal epoxides to give the corresponding cyclic organic carbonates with high selectivities.

The development and activation of MHC class II (MHC-II)–restricted CD4+ T cells are distinct immunological processes that are strictly MHC-II–dependent. To address their relative dependence on MHC-II, we established a novel ENU-induced mutant mouse on the C57BL/6 background, named I-A12%, with ∼8-fold reduced I-A expression on the surface of B cells, dendritic cells, cortical thymic epithelial cells, and medullary thymic epithelial cells. I-A100% and I-A12% mice are highly similar with respect to the numbers of double-positive thymocytes, CD4+CD8− T cells, regulatory T cells, CD4+ T cell marker expression, lifespan, and Th/regulatory T cell function. Despite the demonstration of functional intrathymic negative selection in I-A12% mice, transfer of I-A12% CD25−CD4+ T cells into RAG-knockout hosts revealed increased autoaggression activity against the liver. Compared to I-A100% mice, infection of I-A12% mice with graded doses of Listeria monotcytogenes or influenza virus revealed comparable and significantly reduced generation of Ag-specific CD4+ T cells at high and low infection doses, respectively. A significantly weakened Ag-specific recall cytokine production response was also found for I-A12% mice previously infected with a relative low dose of L. monocytogenes. CD44hiCD4+ T cells from I-A100% and I-A12% mice previously infected with a relatively high L. monocytogenes dose displayed highly similar Ag-specific multicytokine production profiles. In contrast, polyclonal activation of endogenous memory-like I-A12% CD44hiCD4+ T cells revealed highly elevated production of multiple cytokines. Our results demonstrate that there exist distinct thresholds for different MHC-II–dependent immunological processes. The I-A12% mutant mouse model we describe in the present study is a valuable tool for investigations on the quantitative cause–effect relationship in MHC-II–dependent normal and autoimmune responses.

The acute phase response (APR) is a systemic first-line defense against challenges including infection, trauma, stress, and neoplasia. Alteration of acute phase protein (APP) levels in plasma is the most important change during acute phase response. C-reactive protein (CRP), which increases dramatically during inflammation onset, is an indicator of inflammation. To monitor the process of APR, we generated human CRP promoter-driven luciferase transgenic (hCRP-Luc) mice to quantify the hCRP promoter activation in vivo. The naïve female hCRP-Luc mice express low basal levels of liver bioluminescence, but the naïve male hCRP-Luc mice do not. Thus, female hCRP-Luc mice are suitable for monitoring the process of APR. The liver bioluminescence of female hCRP-Luc mice can be induced by several toll-like receptor (TLR) ligands. The expression of liver bioluminescence was highly sensitive to endotoxin stimulation in a dose-dependent manner. On-off-on bioluminescence response was noted in female hCRP-Luc mice upon two endotoxin stimulations one month apart. The LPS-induced bioluminescence of the female hCRP-Luc mice was IL-6-mediated and associated with APP alpha-1-acid glycoprotein expression. In conclusion, the female hCRP-Luc mouse is a non-invasive, sensitive and reusable reporter tool for APR.

This study reports that a series of structurally well-defined mono- and di-nuclear cobalt complexes containing the amine-bis(benzotriazole phenolate) ligand (amine-BiBTP) have been synthesized and fully characterized. The one-pot reaction of the amine-BiBTP ligand precursor with cobalt nitrate salt in the presence of triethylamine under refluxing methanolic solution generated di-nuclear cobalt(II) complexes [(C1NNBiBTP)2Co2] (1). The mono-Co(II) complex [(C1NNBiBTP)CoR] (R = DMAP (2), tBuPy (3)) could be prepared by 1 with a stoichiometric amount of 4-(dimethylamino)pyridine (DMAP) or 4-tert-butylpyridine (tBuPy) to yield a five-coordinated mononuclear cobalt species. Interestingly, the Co(III) complexes 4–7 resulted from the treatment of 2 or 3 with silver nitrate/trifluoroacetate as the oxidizing agent, and an ionic and mononuclear species formulated as [(C1NNBiBTP)CoR2]X (R = DMAP, X = NO3 (4), R = DMAP, X = CF3COO (5), R = tBuPy, X = NO3 (6), R = tBuPy, X = CF3COO (7)) could be formed. Versatile catalysis towards copolymerization of internal epoxides with anhydride catalyzed by these Co(II)/Co(III) was systematically studied under mild conditions. Experimental results revealed that complex 4 was shown to effectively catalyze copolymerization of cyclohexene oxide (CHO) with phthalic anhydride (PA), resulting in copolymers with a high alternating microstructure possessing ≥ 99% ester-linkage contents.

A unique cluster of lithium isopropoxide assisted tantalum isopropoxide derivative incorporating bidentate ketiminate ligand is reported and structurally characterized. Reaction between Li(OCMeCHCMeNAr) and TaCl5 in toluene at room temperature generates compound 1, (OCMeCHCMeNAr)TaCl4, in relatively high yield. Further reacting of 1 with excess amount of LiiOPr in diethylether affords the tantalum-lithium bimetallic cluster {[Li(OCMeCHCMeNAr)]Ta(OiPr)5(LiOiPr)(LiCl)}2 (2). The molecular geometry of 2 reveals as an edge-sharing bis(open-cube) that it consists two Li-Cl edge-sharing bis(open-cube)TaLi3O5Cl units in its core structure.

This study reports a two-step method to synthesize spermidine-capped fluorescent carbon quantum dots (Spd-CQDs) and their potential application as an antibacterial agent. Fluorescent carbon quantum dots (CQDs) are synthesized by pyrolysis of ammonium citrate in the solid state and then modified with spermidine by a simple heating treatment without a coupling agent. Spermidine, a naturally occurring polyamine, binds with DNA, lipids, and proteins involved in many important processes within organisms such as DNA stability, and cell growth, proliferation, and death. The antimicrobial activity of the as-synthesized Spd-CQDs (size ≈4.6 nm) has been tested against non-multidrug-resistant E. coli, S. aureus, B. subtilis, and P. aeruginosa bacteria and also multidrug-resistant bacteria, methicillin-resistant S. aureus (MRSA). The minimal inhibitory concentration value of Spd-CQDs is much lower (>25 000-fold) than that of spermidine, indicating their promising antibacterial characteristics. The mechanism of antibacterial activity is investigated, and the results indicate that Spd-CQDs cause significant damage to the bacterial membrane. In vitro cytotoxicity and hemolysis analyses reveal the high biocompatibility of Spd-CQDs. To demonstrate its practical application, in vitro MRSA-infected wound healing studies in rats have been conducted, which show faster healing, better epithelialization, and formation of collagen fibers when Spd-CQDs are used as a dressing material.

The synthesis of structurally well-characterized dinuclear nickel complexes bearing a new type of pyrazolyl based diamine-bisphenolate ligand was reported for the first time. Catalysis towards nickel-catalyzed copolymerization of CO2 with cyclohexene oxide (CHO) by these newly prepared bimetallic complexes was systematically investigated. Particularly, dinickel complex 4 supporting 2-nitrophenoxide coligands was demonstrated to effectively catalyze CO2-copolymerization of CHO, 4-vinyl-1,2-cyclohexene oxide or cyclopentene oxide, generating the corresponding CO2-based polycarbonates with >99% carbonate linkages under optimal conditions. Kinetic studies of CO2/CHO copolymerization revealed that such copolymerization catalysis is first-order in dinickel complex 4 and second-order in CHO concentration.

Four new 2D indium metal-organic frameworks (MOFs) (Me2NH2)[In(SBA)2] (1), (Me2NH2)[In(SBA)(BDC)] (2), (Me2NH2)[In(SBA)(BDC-NH2)] (3), and (NH4)3[In3Cl2(BPDC)5] (4), (H2SBA = 4,4'-sulfonyldibenzoic acid; H2BDC = 1,4-benzenedicarboxylic acid; H2BDC-NH2 = 2-amino-1,4-benzenedicarboxylic acid; H2BPDC = 4,4'-biphenyldicarboxylic acid) have been synthesized under solvothermal reaction conditions for compounds 1 to 3 and the DES (deep eutectic solvent) reaction has been attempted for compound 4. The structure of these MOFs has been determined by using single crystal X-ray diffraction study and all of theses four 2D monolayer framework with porous properties. The N2 gas sorption measurements indicated that Brunauer-Emmer-Teller (BET) and Langmuir surface areas of compound 1 are 207 and 301 m2 g-1, respectively, which is probably the first one having substantial gas uptake properties in the entire 2D In-MOF family to date. Furthermore, these new indium MOFs on the addition of n-Bu4NBr were active for the cycloaddition of CO2 and propylene oxide, generating propylene carbonates in high conversions under mild conditions. Particularly, the most active MOF 4 was found to efficiently couple CO2 with a series of terminal epoxides to give the corresponding cyclic organic carbonates with high selectivities.

BCR engagement leads to activation and clonal expansion of B cells. The I-A12% mutant mouse possesses a branch site point mutation in the H2-Aa gene that causes highly reduced I-Aa protein expression. As I-A is a heterodimer made up of I-Aa and I-Ab, reduced I-Aa results not only in reduced surface I-A expression but also in an excess of unpaired I-Ab. B cells that develop in I-A12% mice proliferated in response to LPS stimulation but failed to do so upon BCR stimulation. Developing I-A12% B cells were engaged in unfolded protein response due to an excess of unpaired I-Ab. BCR responsiveness was restored by transduced I-Aa expression and by BiP, the unfolded protein response sensor. Reducing the load of unpaired I-Ab also restored BCR responsiveness of I-A12% B cells. Mef2c protein, a transcription factor required for BCR-stimulated proliferation, was missing in I-A12% B cells, and that transduced Mef2c expression restored BCR responsiveness. Mef2c protein appeared in I-A12% B cells after addition of proteasome inhibitors. Mef2c degradation was mediated by Skp2 E3 ligase, and that knockdown of Skp2 mRNA in I-A12% B cells restored BCR responsiveness. Our results point to a generalized incompatibility between BCR responsiveness and increased Skp2 stability. They also imply the existence of regulatory mechanisms other than Ig gene rearrangement that govern Mef2c turnover in a specific, exquisite, and dynamic fashion.

We report the synthesis of structurally well-characterized di-nuclear nickel bis(benzotriazole iminophenolate) (BiIBTP) complexes containing diverse mono- or bi-dentate monoanionic coligands for the utilization of copolymerization of carbon dioxide (CO2) with internal epoxides. The molecular structures of complexes 1–3 can be ascribed to bimetallic nickel(II) species containing a hexadentate BiIBTP main ligand and two homo-coligands of pentafluorobenzoate, pentafluorophenolate or chloride, whereas 4 and 5 are BiIBTP-ligated di-nickel chloride analogues mixed with carboxylate coligands. Di-nickel complexes 1 and 3 supported by O2CC6F5 and Cl coligands were demonstrated to be efficient catalysts for alternating copolymerization of CO2 with cyclohexene oxide (CHO) to afford CO2-based poly(cyclohexene carbonate)s (PCHCs) with >99% carbonate repeated units. Particularly, di-Ni di-chloride complex 3 was capable of giving high activity with a turnover frequency of 688 h−1 for such copolymerization under the optimal conditions. Interestingly, di-Ni catalyst 1 enables us to produce the low molecular weight di-hydroxyl end-capped PCHCs with unimodal molecular weight distributions while commercially available CHO without further purification was used as the epoxide monomer. In addition to CO2-CHO copolymerization, bimetallic complex 1 was also found to effectively copolymerize CO2 with cyclopentene oxide, generating highly alternating poly(cyclopentene carbonate)s with molecular weights in a broad range. Kinetic studies of CO2/CHO copolymerization catalyzed by 1 were investigated using 1H NMR spectroscopy and revealed the first-order dependence for both catalyst 1 and CHO concentrations.

Glycine N-methyltransferase (GNMT) is known for its function as a tumor suppressor gene. Since 100% of female Gnmt(-/-) mice developed hepatocellular carcinoma, we hypothesized that Gnmt(-/-) mice may have defective immune surveillance. In this study, we examined the immune modulation of GNMT in T-cell responses using experimental autoimmune encephalomyelitis (EAE). The results showed that EAE severity was reduced significantly in Gnmt(-/-) mice. Pathological examination of the spinal cords revealed that Gnmt(-/-) mice had significantly lower levels of mononuclear cell infiltration and demyelination than the wild-type mice. In addition, quantitative real-time PCR showed that expression levels of proinflammatory cytokines, including interferon (IFN)-γ and interleukin (IL)-17A, were much lower in the spinal cord of Gnmt(-/-) than in that of wild-type mice. Accordingly, myelin oligodendrocyte glycoprotein (MOG)-specific T-cell proliferation and induction of T-helper (Th)1 and Th17 cells were markedly suppressed in MOG(35-55)-induced Gnmt(-/-) mice. Moreover, the number of regulatory T (Treg) cells was increased significantly in these mice. When the T-cell receptor was stimulated, the proliferative capacity and the activation status of mTOR-associated downstream signaling were decreased significantly in Gnmt(-/-) CD4(+) T cells via an IL-2- and CD25-independent manner. Moreover, GNMT deficiency enhanced the differentiation of Treg cells without affecting the differentiation of Th1 and Th17 cells. Furthermore, the severity of EAE in mice adoptive transferred with GNMT-deficient CD4(+) T cells was much milder than in those with wild-type CD4(+) T cells. In summary, our findings suggest that GNMT is involved in the pathogenesis of EAE and plays a crucial role in the regulation of CD4(+) T-cell functions.

P14 TCR transgenic CD8+ T cells (LCMV gp33-specific) were activated by antigen in the presence of either IL-2 or IL-2 + IL-4 to generate effector cytotoxic T lymphocytes (CTLs). The therapeutic effectiveness of such IL-2- or IL-2 + IL-4-grown CTLs was tested in mice that had received intravenous inoculations of B16.gp33 melanoma cells 7 days previously. Administration of P14 CTLs activated by antigen + IL-2 + IL-4 was significantly more effective at reducing melanoma colony formation in the lung than those grown in the presence of antigen + IL-2. Highly significant improvement in survival was observed with 80% of B16.gp33-inoculated mice showing long-term survival after therapy with 10 x 10(6) antigen + IL-2 + IL-4-activated P14 CTLs. Similar therapeutic effectiveness of antigen + IL-2 + IL-4-activated P14 CTLs against subcutaneously inoculated B16.gp33 melanoma cells was also found. There was significant reduction in P14 CD8+ T cells in the peripheral blood of B16.gp33-inoculated mice than in mice that did not receive B16.gp33 melanoma cells, indicating possible homing of P14 CD8+ T cells to the site of tumor growth or antigen-induced apoptotic cell death. These results may have implications in tumor therapy using CTLs grown ex vivo, especially during early stages of tumor formation. They also support the concept that the therapeutic effectiveness of CTLs can be governed by the cytokine context in which they are activated.

Immunological memory provides faster and stronger immune responses to protect host from infectious pathogens and tumor cells. T cell memory is a major part of immunological memory, and plays an important role to eliminate tumor cells or cells infected by intracellular pathogen. Cytokines including IL-15, IL-7 and IL-4 have been reported to enhance the generation or maintenance of memory T cells. IL-15 enhances homeostatic proliferation and survival of memory CD8+ T cells. An ENU-affected mouse pedigree carried a Tyr301stop point mutation of Runx3, which resulted in a truncated form of Runx3. These mice started to show spontaneous diarrhea after weaning (spontaneous colitis, SC). SC mice also expressed CD4-derepression in the CD8+ T cells, and loss of memory phenotype CD8+ T cells. SC CD8+ T cells showed proliferation deficiency and decreased memory CD8+ T cell precursors after activation and also had poor response to IL-15-induced proliferation. However, SC host mice enhanced in vivo long-term survival of activated WT CD8+ cells through improvement of homeostatic proliferation. This improvement of homeostatic proliferation was due to elevated serum IL-15 levels. In summary, C-terminal truncated Runx3 in mice results in upregulation of IL-15 expression and provides enhanced homeostatic environment for memory CD8+ T cells; however, CD8+ T cells have intrinsic memory defect. Thus, the SC mouse model is a valuable tool for investigations on the effect of immune system upon spontaneous colitis.

Inflammatory bowel disease (IBD) is a common, chronic, progressive inflammatory intestinal disease in the world. The etiology of IBD is due to the imbalance of host immune responses to intestinal flora. Many human genes play the important role to induce improper immune responses, including IL-10, STAT3, IRGM, AGT16L1, NOD2 and RUNX3. It has been reported that homozygous Runx3-ko mice develop colitis spontaneously. However, homozygous Runx3-ko mice show very high mortality rate in the first 2 week of life, and are hard to study the role of Runx3 in the spontaneous colitis. A mouse pedigree (spontaneous colitis, SC) carried an ENU-induced Tyr301stop point mutation of Runx3, which resulted in a short form truncated form of Runx3. SC mice started to show spontaneous diarrhea after weaning. SC mice exhibited prominant enlargement of colon that revealed large amounts of inflammatory cells infiltration from mucosal to muscular layers. Immunofluorescent staining showed large amounts of CD4+ T cells involved in the inflammatory colon. Moreover, inflammatory colons of SC mice expressed higher mRNA levels of IL-17A, and serum IL-17A level were also upregulated in SC mice. In addition, SC CD4+ T cells expressed stronger IL-17A than WT CD4+ T cells after in vitro activation. Transfer of total SC CD4+ T cells into Rag1-ko host mice resulted in autoimmune colitis. In summary, C-terminal truncated Runx3 results in autoimmune colitis through elevated Th17 immune responses. Thus, the SC mouse model is a valuable tool for investigations on the effect of immune system upon spontaneous colitis.

Immunotherapy aiming to rescue or boost antitumor immunity is an emerging strategy for treatment of cancers. The efficacy of immunotherapy is strongly controlled by the immunological milieu of cancer patients. Myeloid-derived suppressor cells (MDSCs) are heterogeneous immature myeloid cell populations with immunosuppressive functions accumulating in individuals during tumor progression. The signaling mechanisms of MDSC activation have been well studied. However, there is little known about the metabolic status of MDSCs and the physiological role of their metabolic reprogramming. In this study, we discovered that myeloid cells upregulated their glycolytic genes when encountered with tumor-derived factors. MDSCs exhibited higher glycolytic rate than their normal cell compartment did, which contributed to the accumulation of the MDSCs in tumor-bearing hosts. Upregulation of glycolysis prevented excess reactive oxygen species (ROS) production by MDSCs, which protected MDSCs from apoptosis. Most importantly, we identified the glycolytic metabolite, phosphoenolpyruvate (PEP), as a vital antioxidant agent able to prevent excess ROS production and therefore contributed to the survival of MDSCs. These findings suggest that glycolytic metabolites have important roles in the modulation of fitness of MDSCs and could be potential targets for anti-MDSC strategy. Targeting MDSCs with analogs of specific glycolytic metabolites, for example, 2-phosphoglycerate or PEP may diminish the accumulation of MDSCs and reverse the immunosuppressive milieu in tumor-bearing individuals.

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Upon adoptive transfer into histocompatible mice, naive CD8+ T cells stimulated ex vivo by TCR+IL-4 turn into long-lived functional memory cells. The liver contains a large number of so formed memory CD8+ T cells, referred to as liver memory T cells (Tlm) in the form of cell clusters. The CD62Llow expression and nonlymphoid tissue distribution of Tlm cells are similar to effector memory (Tem) cells, yet their deficient cytotoxicity and IFN-γ inducibility are unlike Tem cells. Adoptive transfer of admixtures of TCR+IL-4–activated Vβ8+ and Vβ5+ CD8+ T cells into congenic hosts reveals Tlm clusters that are composed of all Vβ5+ or Vβ8+, not mixed Vβ5+/Vβ8+ cells, indicating that Tlm clusters are formed by clonal expansion. Clonally expanded CD8+ T cell clusters are also seen in the liver of Listeria monocytogenes-immune mice. Tlm clusters closely associate with hepatic stellate cells and their formation is IL-15/IL-15R–dependent. CD62Llow TLM cells can home to the liver and secondary lymphoid tissues, remain CD62Llow, or acquire central memory (Tcm)-characteristic CD62Lhi expression. Our findings show the liver as a major site of CD8+ memory T cell growth and that Tlm cells contribute to the pool of peripheral memory cells. These previously unappreciated Tlm characteristics indicate the inadequacy of the current Tem/Tcm classification scheme and help ongoing efforts aimed at establishing a unifying memory T cell development pathway. Lastly, our finding of Tlm clusters suggests caution against interpreting focal lymphocyte infiltration in clinical settings as pathology and not normal physiology.

Anti-PD1 therapy has shown promising anti-tumor activities in multiple cancer types, including hepatocellular carcinoma (HCC), and PD-L1 expression in tumor cells was considered a potential predictive marker for treatment efficacy. The present study explored whether PD-L1/L2 expression may affect treatment efficacy of anti-PD1 therapy for HCC. PD-L1 (MR203953; Origene Technologies, Rockville, MD) or PD-L2 (MR222499; Origene Technologies) was transfected into the BNL-MEA murine liver cancer cell lines and stable clones were selected. Effects of PDL1/L2 over-expression were confirmed by Western blotting and immunofluorescence assay. Effects of PDL1/L2 over-expression on in vitro growth characteristics were compared by MTT, colony formation, and 3-D culture assays. Orthotopic liver cancer models were established by sub-capsular injection of parental or transfected cancer cells into the left lobe of mouse liver. The anti-tumor efficacy of anti-mouse PD1 therapy (clone 4H2, Ono Pharmaceutical Co), 0.2 mg by intraperitoneal injection on day 5, 8, 11, 14, and 17 after tumor implantation, was measured by change in tumor volume using the orthotopic liver cancer models. Distribution of T-cell sub-populations in tumor-infiltrating lymphocytes (TILs) after treatment were measured by flow cytometry. The growth characteristics of parental and PD-L1/L2-expressing BNL-MEA cells did not differ significantly, as measured by MTT, colony formation, and 3-D culture assays. Survival of animals bearing the orthotopic liver tumor with or without PD-L1/L2 over-expression was also similar. Anti-PD1 therapy appeared more efficacious in PD-L1-over-expressing liver cancer models than the parental or PD-L2-over-expressing models. Distinctive TILs distribution patterns between parental and PD-L1/L2 over-expressing liver cancer models were found after anti-PD1 therapy. Conclusions: The PD-L1/L2 expressing liver cancer models may help evaluate the efficacy and mechanisms of action of anti-PD1 therapy for HCC. Citation Format: Da-Liang Ou, Shu-Ching Chen, Yu-Chia Su, Ann-Lii Cheng, Chiun Hsu. Development of PD-L1/L2 over-expressing mouse liver cancer models to test the efficacy of anti-PD1 therapy for hepatocellular carcinoma. [abstract]. In: Proceedings of the CRI-CIMT-EATI-AACR Inaugural International Cancer Immunotherapy Conference: Translating Science into Survival; September 16-19, 2015; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(1 Suppl):Abstract nr B136.

Glycine N-methyltransferase (GNMT) is known for its function as a tumor suppressor gene. Since 100% of female Gnmt−/− mice developed hepatocellular carcinoma, we hypothesized that Gnmt−/− mice may have defective immune surveillance. In this study, we examined the immune modulation of GNMT in T cell responses using experimental autoimmune encephalomyelitis (EAE). The results showed that EAE severity was significantly reduced in Gnmt−/− mice. Pathological examination of the spinal cords revealed that Gnmt−/- mice had significantly lower level of mononuclear cell infiltration and demyelination than the wild-type mice. In addition, quantitative real-time PCR showed that expression levels of pro-inflammatory cytokines, including IFN-gamma and IL-17A, were much lower in the spinal cord of Gnmt−/- than in that of wild-type mice. Accordingly, myelin oligodendrocyte glycoprotein (MOG)-specific T cell proliferation and induction of T helper (Th) 1 and Th17 cells were markedly suppressed in MOG35-55 induced Gnmt−/- mice. Moreover, the amount of regulatory T cells was significantly increased in these mice. When the T cell receptor was stimulated, the proliferative capacity and the activation status of mTOR-associated downstream signaling were significantly decreased in GNMT−/- CD4+ T cells via an IL-2 and CD25-independent manner. Moreover, GNMT deficiency enhanced the differentiation of regulatory T cells without affecting the differentiation of Th1 and Th17 cells. Furthermore, the severity of EAE in mice adoptive transferred with GNMT-deficient CD4+ T cells was much milder than in those with wild-type CD4+ T cells. In summary, our findings suggest that GNMT is involved in the pathogenesis of EAE and plays a crucial role in the regulation of CD4+ T cell functions. Citation Format: Chung-Hsien Li, Ming-Hong Lin, Yen-Fu Chen, Shih-Han Chu, Pang-Hsien Tu, Cheng-Chieh Fang, Chia-Hung Yen, Peir-In Liang, Jason C. Huang, Yu-Chia Su, Huey-Kang Sytwu, Yi-Ming A. Chen. Role of glycine N-methyltransferase in the regulation of T cell responses in experimental autoimmune encephalomyelitis. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1295. doi:10.1158/1538-7445.AM2015-1295

Objective The major aim of the study was to establish a murine model of chronic lymphocytic leukemia with B-1 cells derived from a New Zealand white mouse. Material and Methods Malignant B-1 cells (named CLL-RT cells) derived from a New Zealand white mouse were injected into the peritoneal cavity of severe combined immunodeficiency mice. Upon follow-up of recipient mice, the lymphomas showed characteristics similar to chronic lymphocytic leukemia (CLL) with Ritcher's transformation. Results Blood samples from the recipient mice showed that CLL-RT cells increased rapidly in peripheral blood after 5 weeks. Serum interleukin-10 also increased significantly in recipient mice, as in human chronic lymphocytic leukemia patients. These CLL-RT cells showed a high nucleus-to-cytoplasm ratio. These cells could metastasize via circulation in the recipients and form diffuse lymphomas in various tissues. These aggressive and diffuse lymphomas were similar to Ritcher's transformation of human CLL. The cell surface antigens of the spleen and peritoneal resident cells were analyzed by flow cytometry. The CLL-RT cells constantly expressed surface immunoglobulins M and G, and CD5, CD19, B220, and CD40 molecules. They did not express any CD11b, CD3, MAC-3, CD23, NK1.1, or H-2K d molecules. Conclusions The characteristics of our animal model are very similar to human CLL. This animal system could be an ideal model for the human disease. We believe the animal model would be valuable in therapeutic studies and aid in the identification of the specific genetic alleles associated with the disease.