Targeting age‐specific changes in CD4+ T cell metabolism ameliorates alloimmune responses and prolongs graft survival

Age impacts alloimmunity. Effects of aging on T‐cell metabolism and the potential to interfere with immunosuppressants have not been explored yet. Here, we dissected metabolic pathways of CD4⁺ and CD8⁺ T cells in aging and offer novel immunosuppressive targets. Upon activation, CD4⁺ T cells from old mice failed to exhibit adequate metabolic reprogramming resulting into compromised metabolic pathways, including oxidative phosphorylation (OXPHOS) and glycolysis. Comparable results were also observed in elderly human patients. Although glutaminolysis remained the dominant and age‐independent source of mitochondria for activated CD4⁺ T cells, old but not young CD4⁺ T cells relied heavily on glutaminolysis. Treating young and old murine and human CD4⁺ T cells with 6‐diazo‐5‐oxo‐l‐norleucine (DON), a glutaminolysis inhibitor resulted in significantly reduced IFN‐γ production and compromised proliferative capacities specifically of old CD4⁺ T cells. Of translational relevance, old and young mice that had been transplanted with fully mismatched skin grafts and treated with DON demonstrated dampened Th1‐ and Th17‐driven alloimmune responses. Moreover, DON diminished cytokine production and proliferation of old CD4⁺ T cells in vivo leading to a significantly prolonged allograft survival specifically in old recipients. Graft prolongation in young animals, in contrast, was only achieved when DON was applied in combination with an inhibition of glycolysis (2‐deoxy‐d‐glucose, 2‐DG) and OXPHOS (metformin), two alternative metabolic pathways. Notably, metabolic treatment had not been linked to toxicities. Remarkably, immunosuppressive capacities of DON were specific to CD4⁺ T cells as adoptively transferred young CD4⁺ T cells prevented immunosuppressive capacities of DON on allograft survival in old recipients. Depletion of CD8⁺ T cells did not alter transplant outcomes in either young or old recipients. Taken together, our data introduce an age‐specific metabolic reprogramming of CD4⁺ T cells. Targeting those pathways offers novel and age‐specific approaches for immunosuppression.     

Genome-wide CRISPR Screens Reveal Host Factors Critical for SARS-CoV-2 Infection

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Functional lung imaging with synchrotron radiation: Methods and preclinical applications

Many lung disease processes are characterized by structural and functional heterogeneity that is not directly appreciable with traditional physiological measurements. Experimental methods and lung function modeling to study regional lung function are crucial for better understanding of disease mechanisms and for targeting treatment. Synchrotron radiation offers useful properties to this end: coherence, utilized in phase-contrast imaging, and high flux and a wide energy spectrum which allow the selection of very narrow energy bands of radiation, thus allowing imaging at very specific energies. K-edge subtraction imaging (KES) has thus been developed at synchrotrons for both human and small animal imaging. The unique properties of synchrotron radiation extend X-ray computed tomography (CT) capabilities to quantitatively assess lung morphology, and also to map regional lung ventilation, perfusion, inflammation and biomechanical properties, with microscopic spatial resolution. Four-dimensional imaging, allows the investigation of the dynamics of regional lung functional parameters simultaneously with structural deformation of the lung as a function of time. This review summarizes synchrotron radiation imaging methods and overviews examples of its application in the study of disease mechanisms in preclinical animal models, as well as the potential for clinical translation both through the knowledge gained using these techniques and transfer of imaging technology to laboratory X-ray sources.     

Respiratory enzymes in muscle: Interaction between environmental temperature, nutrition and growth

1. 1.In young pigs living at 35 or 10°C on a high or low energy intake, respiratory enzyme activities in longissimus dorsi muscle were greater both in the cold and on low intake. The elevated activities in the cold were unlikely to be related entirely to shivering since they were also found in muscle from the diaphragm.

2. 2.In a second study, pigs were kept close to thermal neutrality (26°C) on different levels of food intake and for different periods of time. For all animals, as body weight increased there was a decline in respiratory enzyme activity and the number of dark fibres in skeletal muscle. For those of the same weight, but different age and food intake, there was no difference in enzyme activity or number of dark fibres per unit area.

3. 3.At least part of the difference in respiratory enzyme activities related to energy intake must therefore be due to differences in body size. However, size is not the sole determinant of enzyme activity in skeletal muscle, since in animals of similar size those living at 10°C have greater enzyme activities than those at 35°C.

Conformationally altered aortic myosin light chains

Aorta smooth myosin contains two types of light chain, LC20 and LC17, which fold together with the N-terminal region of each heavy chain to form the globular head region of myosin. We demonstrate an altered conformation of LC20 after its separation from heavy chain by high concentrations of urea, on the basis of the following evidende: 1) A polyclonal antibody against LC20 was not able to recognize this conformationally altered form; 2) Myosin reconstituted from heavy chains and urea-dissociated light chains exhibited extremely low ATPase activity. Circular dichroism unfolding profiles showed that light chains dissociated from heavy chains by SDS appeared to be more stable than those generated by urea dissociation.     

Evidence for a branched respiratory system with two cytochrome oxidases in autotrophically grown Alcaligenes latus

The respiratory system of chemolithoautotrophically-grown Alcaligenes latus contains a, b, and c type cytochromes. Two cytochrome oxidases were identified by their carbon monoxide difference spectra and their differing sensitivities to cyanide and carbon monoxide. The oxidases were cytochrome o and an a-type cytochrome. Ubiquinone was present in A. latus membranes and could be reduced by H₂. The quinone analogue, 2-heptyl-4-hydroxy-quinoline-N-oxide (HQNO), was a strong inhibitor of the H₂ oxidase reaction, but did not prevent the reduction of either ubiquinone or the cytochromes.Abbreviations HQNO 2-heptyl-4-hydroxy-quinoline-N-oxide - TMPD N,N,N,N-tetramethyl-p-phenylenediamine     

Kinetic measurement of bacterial adherence to eukaryotic cells

We developed a kinetic assay using a monolayer of differentiated respiratory epithelium in culture to assess bacterial adherence. Mean residence time of bacteria in the tissue culture chamber was estimated from a model-independent (moment) analysis of the rate of bacterial washout from perfused Rose chambers. Results with this method compared favorably with visual assessment of adherence and double radiolabel method with H. influenzae. Adherence was assessed with low inoculae of H. influenzae, P. cepacia and P. aeruginosa avoiding cytotoxic effects seen when large inoculae are added to eukaryotic cells. This method will provide a means of assessing adherence of pathogenic respiratory bacteria to their cellular target at low inoculae.     

Convergence to stationary distributions in two-species stochastic competition models

Two sets of sufficient conditions are given for convergence to stationary distributions, for some general models of two species competing in a randomly varying environment. The models are nonlinear stochastic difference equations which define Markov chains. One set of sufficient conditions involves strong continuity and -irreducibility of the transition probability for the chain. The second set has a much weaker irreducibility condition, but is only applicable to monotonic models. The results are applied to a stochastic two-species Ricker model, and to Chesson's lottery model with vacant space, to illustrate how the assumptions can be checked in specific models.     

我国沙漠中部地区主要不同生态类型植物脯氨酸的累积、光合、呼吸和叶绿素含量

中生植物脯氨酸含量为0．42mg/g.dw, 低于少浆旱生植物（1．73mg/g.dw)。多浆旱生植物脯氨酸含量最高（7．22mg/g.dw),为前两者的17倍和4倍,两类旱生植物在干旱条件下（非灌溉）脯氨酸含量均高于灌水处理。少浆与多浆旱生植物的光合强度（16．74,14．04CO2mg/g.dw.h)差异不大,而中生植物（37．57mg/g.dW.h)略高于多浆旱生植物（4．73CO2Mg/g.dw.h),与中生植物（7.60Co2mg/g.dw.h)接近,光合／呼吸值,少浆,多浆与中生植物分别为2．50,3．09和4．59,说明中生植物的合成明显大于消耗,季节动态中,中生植物显著高于两类旱生植物,叶绿素总量三类植物差异甚微。