Multifunctional cytokine production marks influenza A virus-specific CD4 T cells with high expression of survival molecules.

Cytokine production by memory T cells is a key mechanism of T cell mediated protection. However, we have limited understanding of the persistence of cytokine producing T cells during memory cell maintenance and secondary responses. We interrogated antigen-specific CD4 T cells using a mouse influenza A virus infection model. Although CD4 T cells detected using MHCII tetramers declined in lymphoid and non-lymphoid organs, we found similar numbers of cytokine+ CD4 T cells at days 9 and 30 in the lymphoid organs. CD4 T cells with the capacity to produce cytokines expressed higher levels of pro-survival molecules, CD127 and Bcl2, than non-cytokine+ cells. Transcriptomic analysis revealed a heterogeneous population of memory CD4 T cells with three clusters of cytokine+ cells. These clusters match flow cytometry data and reveal an enhanced survival signature in cells capable of producing multiple cytokines. Following re-infection, multifunctional T cells expressed low levels of the proliferation marker, Ki67, whereas cells that only produce the anti-viral cytokine, interferon-γ, were more likely to be Ki67+ . Despite this, multifunctional memory T cells formed a substantial fraction of the secondary memory pool. Together these data indicate that survival rather than proliferation may dictate which populations persist within the memory pool.

Plasma Alarin Level and Its Influencing Factors in Obese Newly Diagnosed Type 2 Diabetes Patients.

OBJECTIVE: To investigate the plasma alarin level in newly diagnosed obese type 2 diabetes mellitus (T2DM) and its correlation with glucose and lipid metabolism and insulin resistance. METHODS: From October 2018 to June 2020, 239 newly diagnosed T2DM patients were collected. According to obesity, patients were divided into T2DM obese group (n=135) and T2DM non-obese group (n =104). Gender, age, body mass index (BMI), blood lipids, blood glucose, glycosylated hemoglobin A1c (HbA1c), fasting insulin (FINS), plasma alarin concentration, homeostasis model assessment for insulin resistance (HOMA-IR), homeostasis model assessment for ß-cell function (HOMA-ß) and other clinical data were collected and analyzed. RESULTS: BMI, triacylglycerol (TG), total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C), fasting blood glucose (FPG), HbA1c, FINS, plasma alarin levels and HOMA-IR in the control group, T2DM non-obese group and T2DM obese group increased sequentially, and high-density lipoprotein-cholesterol (HDL-L) and HOMA-ß decreased sequentially (P<0.05). Correlation analysis results showed that plasma alarin levels in T2DM patients were positively correlated with waistline, BMI, TC, LDL-C, FPG, HbA1c, FINS and HOMA-IR (P<0.05), and negatively correlated with HDL-C and HOMA-ß (P <0.05), and the correlation coefficient of T2DM obese group was significantly higher than that of T2DM non-obese group (P<0.05). Multiple linear stepwise regression analysis showed that BMI, FPG, HbA1c, HOMA-ß, and HOMA-IR were independent factors related to plasma alarin levels in T2DM non-obese and T2DM obese patients, and the correlation coefficient of the T2DM obese group was significantly higher than that of the T2DM non-obese group (P <0.05). CONCLUSION: Plasma alarin levels increase in newly diagnosed T2DM and obese T2DM patients, which are affected by TC, BMI, FPG, HbA1c, HOMA-ß and HOMA-IR, and may be involved in development of T2DM.

High Molecular Weight Silk Fibroin Prepared by Papain Degumming.

A major challenge for the silk textile industry and for the process of silk-based biomaterials is to find a degumming method that can completely remove sericin while avoiding obvious hydrolysis damage to the silk fibroin. In this study, papain was used to degum Bombyx mori silk fibers under nearly neutral conditions based on the specificity of papain to sericin. The degumming efficiency was investigated, as well as the mechanical properties and molecular weight of the sericin-free silk fibroin. The results indicated that increasing the papain concentration aided in sericin removal, as the concentration increased to 3.0 g/L, the degummed fibers showed a clean, smooth surface morphology and exhibited a yellow color when stained by picric acid and carmine, confirming the complete removal of sericin from silk fibroin. Furthermore, an analysis of the amino acid composition indicated that the silk fibroin suffered less damage because papain specifically cleaved the binding sites between L-arginine or L-lysine residue and another amino acid residue in sericin, leading to a significantly higher molecular weight and improved tensile strength compared to traditional sodium carbonate degumming. This study provides a novel degumming method which cannot only completely remove sericin, but also maintain the original strong mechanical properties and high molecular weight of silk fibroin.

The Interactions of Quantum Dot-Labeled Silk Fibroin Micro/Nanoparticles with Cells.

When silk fibroin particles are used for controlled drug delivery, particle size plays a key role in the location of the carrier on the cells as well as the transport pathway, utilization efficiency, and therapeutic effect of the drugs. In this study, the interactions of different-sized silk fibroin particles and cell lines were investigated. Silk fibroin microparticles with dry size of 1.9 ± 0.4 µm (2.7 ± 0.3 µm in wet state) and silk fibroin nanoparticles with dry size of 51.5 ± 11.0 nm (174.8 ± 12.5 nm in wet state) were prepared by salting-out method and high-voltage electrospray method, respectively. CdSe/ZnS quantum dots were coupled to the surface of the micro/nanoparticles. Photostability observations indicated that the fluorescence stability of the quantum dots was much higher than that of fluorescein isothiocyanate. In vitro, microparticles and nanoparticles were co-cultured with human umbilical vein endothelial cells EA.hy 926 and cervical cancer cells HeLa, respectively. The fluorescence test and cell viability showed that the EA.hy926 cells tended to be adhered to the microparticle surfaces and the cell proliferation was significantly promoted, while the nanoparticles were more likely to be internalized in HeLa cells and the cell proliferation was notably inhibited. Our findings might provide useful information concerning effective drug delivery that microparticles may be preferred if the drugs need to be delivered to normal cell surface, while nanoparticles may be preferred if the drugs need to be transmitted in tumor cells.