Multiple Target Cells Research Articles

Network pharmacology-based study to investigate the mechanism of compound houttuynia mixture against influenza virus infection by suppressing TLR7/MyD88 signaling pathway

Aim of the studyCompound houttuynia mixture (CHM) has been approved to cure respiratory diseases in China. However, the anti-influenza virus effect and underlying mechanism of CHM remains to be confirmed. Materials and methodsThe in vitro activity of CHM against H3N2 virus was evaluated using MDCK, A549 and THP-1 cells as an in vitro model. The in vivo protective effect on H3N2 virus infection was investigated. Moreover, serum cytokines were measured with high throughput liquid phase protein chip. The anti-influenza mechanism of CHM was predicted by network pharmacology and further validated with immunoblotting. ResultsOur results indicated that CHM has significant inhibitory effect on the replication of influenza A H3N2 virus. Furthermore, CHM could effectively reduce the mortality caused by lethal H3N2 virus infection by prolonging the survival time, reducing lung pathological injury, and suppressing excessive cytokines storm. Network pharmacology revealed that the protective effect of CHM on influenza virus infection was involved in multiple targets and multiple pathways. Mechanistic validation indicated that inhibiting the excessive activation of TLR7/MyD88 signaling pathway may be the critical mechanism of CHM exerting the protective effect against influenza virus infection. ConclusionRegulating of TLR7/MyD88/NF-κB signaling pathway in multiple target cells might be one of key mechanisms of CHM by inhibiting virus replication and excessive inflammatory. Our findings indicated that CHM might be an effective treatment for influenza virus infection.

Development of a Novel, Allogeneic GPRC5D-Directed CAR for Treatment of Multiple Myeloma Patients

Despite the success of BCMA-targeted therapy in advanced multiple myeloma, the disease is still considered incurable. Relapse following BCMA-directed cellular and antibody-based therapies is commonly associated with decreased surface expression of BCMA on malignant plasma cells. The G protein-coupled receptor, class C group 5 member D (GPRC5D) represents an additional target whose expression is limited to bone marrow and hair follicles, with selectively increased expression on malignant plasma cells. Thus far, the use of GPRC5D-targeted CAR T cells has provided a strong clinical benefit in multiple myeloma with limited on-target, off-tumor effects. To this end, we have validated novel, fully human GPRC5D-specific CARs for use with our hypoimmune (HIP), allogeneic CAR T cell platform. This will allow rapid delivery of allogeneic GPRC5D CAR T cells from healthy donors that simultaneously circumvent “non-self” recognition through TCR, B2M and CIITA gene disruption and promote persistence through innate immune evasion by leveraging CD47 overexpression. For early selection of candidate GPRC5D binders, T cells from healthy donors were transduced with VSV-G pseudotyped lentivirus containing second-generation, fully-human GPRC5D CARs. A clinically validated GPRC5D CAR was produced in similar fashion and used as benchmark control for initial characterization experiments. Transduction efficiency of GPRC5D CARs was analyzed via flow cytometry. GPRC5D CAR efficacy was analyzed in vitro via 24-hour luciferase or long-term Incucyte cytotoxicity assays against RPMI-8226 and MM.1S multiple myeloma target cells. Systemic MM.1S in vivo tumor models were performed in NSG mice to further assess efficacy of lead CAR candidates. Here we show our candidate GPRC5D CARs demonstrate in vitro cytotoxicity and effector cytokine production that is comparable to clinically validated benchmark controls. In 5-day Incucyte cytotoxicity assays, GPRC5D CAR T cells expand up to 10-fold and reduce tumor cell counts at starting E:T ratios of 1:8. Furthermore, in MM.1S in vivo tumor challenge models, these GPRC5D CARs significantly reduce flux and prolonged survival relative to tumor only controls with tumor clearance that was in line with clinical benchmark controls. Using ex vivo VSV-G production, our GPRC5D CAR T cells control multiple myeloma tumor cells both in vitro and in vivo, demonstrating efficacy that is on par with GPRC5D clinical benchmark CAR T cells. Studies to further define the efficacy of HIP-modified GPRC5D CAR T cells are ongoing. Moving forward, further interrogating GPRC5D CAR activity against multiple myeloma tumor cells with varying levels of target antigen expression will be critical to understanding the benefit of deploying GPRC5D CAR T cells within the multiple myeloma clinical landscape.

Intrinsically bioactive and biomimetic nanoparticle-derived therapies alleviate asthma by regulating multiple pathological cells

Asthma is a serious global public health concern. Airway neutrophilic inflammation is closely related to severe asthma, for which effective and safe therapies remain to be developed. Here we report nanotherapies capable of simultaneously regulating multiple target cells relevant to the pathogenesis of neutrophilic asthma. A nanotherapy LaCD NP based on a cyclic oligosaccharide-derived bioactive material was engineered. LaCD NP effectively accumulated in the injured lungs of asthmatic mice and mainly distributed in neutrophils, macrophages, and airway epithelial cells after intravenous or inhalation delivery, thereby ameliorating asthmatic symptoms and attenuating pulmonary neutrophilic inflammation as well as reducing airway hyperresponsiveness, remodeling, and mucus production. Surface engineering via neutrophil cell membrane further enhanced targeting and therapeutic effects of LaCD NP. Mechanistically, LaCD NP can inhibit the recruitment and activation of neutrophils, especially reducing the neutrophil extracellular traps formation and NLRP3 inflammasome activation in neutrophils. Also, LaCD NP can suppress macrophage-mediated pro-inflammatory responses and prevent airway epithelial cell death and smooth muscle cell proliferation, by mitigating neutrophilic inflammation and its direct effects on relevant cells. Importantly, LaCD NP showed good safety performance. Consequently, LaCD-derived multi-bioactive nanotherapies are promising for effective treatment of neutrophilic asthma and other neutrophil-associated diseases.

The Molecular Basis of the Anti-Inflammatory Property of Astragaloside IV for the Treatment of Diabetes and Its Complications.

Astragali Radix is a significant traditional Chinese medication, and has a long history of clinical application in the treatment of diabetes mellitus (DM) and its complications. AS-IV is an active saponin isolated from it. Modern pharmacological study shows that AS-IV has anti-inflammatory, anti-oxidant and immunomodulatory activities. The popular inflammatory etiology of diabetes suggests that DM is a natural immune and low-grade inflammatory disease. Pharmacological intervention of the inflammatory response may provide promising and alternative approaches for the prevention and treatment of DM and its complications. Therefore, this article focuses on the potential of AS-IV in the treatment of DM from the perspective of an anti-inflammatory molecular basis. AS-IV plays a role by regulating a variety of anti-inflammatory pathways in multiple organs, tissues and target cells throughout the body. The blockade of the NF-κB inflammatory signaling pathway may be the central link of AS-IV's anti-inflammatory effect, resulting in a reduction in the tissue structure and function damage stimulated by inflammatory factors. In addition, AS-IV can delay the onset of DM and its complications by inhibiting inflammation-related oxidative stress, fibrosis and apoptosis signals. In conclusion, AS-IV has therapeutic prospects from the perspective of reducing the inflammation of DM and its complications. An in-depth study on the anti-inflammatory mechanism of AS-IV is of great significance for the effective use of Chinese herbal medicine and the promotion of its status and influence on the world.

In Vivo Generation of Memory-like NK Cells for the Treatment of AML and Myelodysplastic Syndrome; Early Clinical Applications of INKmuneTM

Natural killer (NK) cells can have potent anti-tumour responses and recently highlighted potential for memory-like functions, but the best approach to generate memory-like NK (mlNK) cells remains unclear. Priming of NK cells with the pharmaceutical-grade, replication incompetent tumour cell product, INKmuneTM generates tumour-induced mlNK (TIML-NK) cells with enhanced cytokine production and cytotoxicity against multiple NK-resistant tumour target cell lines in vitro, analogous to previously reported cytokine-primed memory-like NK cells. In addition, proteomic profiling of TIML-NK cells revealed differential abundance of proteins involved in promoting mitochondrial survival and function as well as critical nutrient receptors, which may provide a unique benefit of NK cell activation whilst sparing mitochondrial damage typically associated with cytokine-mediated activation. INKmune priming increased NK glycolysis and oxidative phosphorylation whilst enhancing mitochondrial respiratory capacity and maintaining glycolytic reserve. Four patients have been treated with a cycle of three, weekly infusions of 1x108 INKmuneTM cells. All patients received all three doses without incident and without side effects. Patient 1 was a 78yr old man with A 3yr history of refractory MLD MDS who was transfusion and platelet dependent and required G-CSF. Within 7 days of first infusion, over 50% of his peripheral blood NK cells activated and this increased with each dose. By day +29, 72% of his NK cells were activated and this remained above 68% at day +119 when monitoring ended. His ECOG status fell from 2 pre-treatment to 0 at D+119 and his RAD-1 ctDNA from 45 to 38. Analysis of systemic cytokines showed increases in MIP-1a/b, TNF-a and sIL2R which paralleled the changes in percentages of CD69+ NK cells. IL6 levels peaked after the second infusion but levels remained low and there was no evidence of CRS. At one year follow-up he remained well, platelet and G-CSF independent, with reduced transfusion requirements and had resumed playing sport with friends. Subjects 2, 3 and 4, were all multiply relapsed and refractory AML patients; two of whom had relapsed after HLA-mismatched HSCT. All showed rapid activation of peripheral NK cells post infusion with sustained activity throughout follow-up. In all cases these activated NK cells killed NK-resistant tumour cells in vitro without additional activation. Subject 2 was a neutropenic 21yr old female with 70% a mixed chimera and refractory AML (M2) post VUD HSCT at time of treatment. Within 1 month of initiation of INKmune treatment she was discharged with PMN >500 and her mixed chimera resolved to full donor chimera. At Day 140 post INKmune her bone marrow NK cells remained highly activated (>60% CD69+, K562 and Raji lysis of >70%). Her AML recurred 4 months post INKmune infusion and she died from relapsed disease 8 months after INKmune treatment. Subject 3 was a 21yr old male with refractory AML (M6) after two failed HSCT (haplo m/m and VUD). He showed the same rapid generation of activated NK cells in vivo but to a lesser degree (maximum 35% CD69+, sustained increased lysis of K562 but no lysis of Raji cells) and no evidence of clinical improvement. Subject 4, a multiply relapsed AML patient has just been treated and has shown peripheral NK proliferation and maturation at D+8. The subject experienced no adverse effects post 1st and 2nd doses and further follow-up will be presented. INKmuneTM infusions proved safe and led to rapid and sustained activation of peripheral NK cells with concomitant increases in relevant systemic cytokines. The TIML-NK have a distinct phenotypic, proteomic and metabolomic signature which mirrors the activity in vitro and in vivo. This is the first time memory-like NK cells have been generated in vivo without the need for cytokine induction or support and our findings inform the development of more optimal NK cell-based immunotherapeutic strategies for cancer. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Aptamer-Based Traceless Multiplexed Cell Isolation Systems.

In both biomedical research and clinical cell therapy manufacturing, there is a need for cell isolation systems that recover purified cells in the absence of any selection agent. Reported traceless cell isolation methods using engineered antigen-binding fragments or aptamers have been limited to processing a single cell type at a time. There remains an unmet need for cell isolation processes that rapidly sort multiple target cell types. Here, we utilized two aptamers along with their designated complementary strands (reversal agents) to tracelessly isolate two cell types from a mixed cell population with one aptamer-labeling step and two sequential cell elution steps with reversal agents. We engineered a CD71-binding aptamer (rvCD71apt) and a reversal agent pair to be used simultaneously with our previously reported traceless purification approach using the CD8 aptamer (rvCD8apt) and its reversal agent. We verified the compatibility of the two aptamer displacement mechanisms by flow cytometry and the feasibility of incorporating rvCD71apt with a magnetic solid state. We then combined rvCD71apt with rvCD8apt to isolate activated CD4+ T cells and resting CD8+ cells by eluting these target cells into separate fractions with orthogonal strand displacements. This is the first demonstration of isolating different cell types using two aptamers and reversal agents at the same time. Potentially, different or more aptamers can be included in this traceless multiplexed isolation system for diverse applications with a shortened operation time and a lower production cost.

Acute exercise mobilizes NKT-like cells with a cytotoxic transcriptomic profile but does not augment the potency of cytokine-induced killer (CIK) cells.

CD3+/CD56+ Natural killer (NK) cell-like T-cells (NKT-like cells) represent <5% of blood lymphocytes, display a cytotoxic phenotype, and can kill various cancers. NKT-like cells can be expanded ex vivo into cytokine-induced killer (CIK) cells, however this therapeutic cell product has had mixed results against hematological malignancies in clinical trials. The aim of this study was to determine if NKT-like cells mobilized during acute cycling exercise could be used to generate more potent anti-tumor CIK cells from healthy donors. An acute exercise bout increased NKT-like cell numbers in blood 2-fold. Single cell RNA sequencing revealed that exercise mobilized NKT-like cells have an upregulation of genes and transcriptomic programs associated with enhanced anti-tumor activity, including cytotoxicity, cytokine responsiveness, and migration. Exercise, however, did not augment the ex vivo expansion of CIK cells or alter their surface phenotypes after 21-days of culture. CIK cells expanded at rest, during exercise (at 60% and 80% VO2max) or after (1h post) were equally capable of killing leukemia, lymphoma, and multiple myeloma target cells with and without cytokine (IL-2) and antibody (OKT3) priming in vitro. We conclude that acute exercise in healthy donors mobilizes NKT-like cells with an upregulation of transcriptomic programs involved in anti-tumor activity, but does not augment the ex vivo expansion of CIK cells.

Novel Drug Delivery Technologies and Targets for Renal Disease.

The burden of acute and chronic kidney diseases to the health care system is exacerbated by the high mortality that this disease carries paired with the still limited availability of comprehensive therapies. A reason partially resides in the complexity of the kidney, with multiple potential target cell types and a complex structural environment that complicate strategies to protect and recover renal function after injury. Management of both acute and chronic renal disease, irrespective of the cause, are mainly focused on supportive treatments and renal replacement strategies when needed. Emerging preclinical evidence supports the feasibility of drug delivery technology for the kidney, and recent studies have contributed to building a robust catalog of peptides, proteins, nanoparticles, liposomes, extracellular vesicles, and other carriers that may be fused to therapeutic peptides, proteins, nucleic acids, or small molecule drugs. These fusions can display a precise renal uptake, an enhanced circulating time, and a directed intraorgan biodistribution while protecting their cargo to improve therapeutic efficacy. However, several hurdles that slow the transition towards clinical applications are still in the way, such as solubility, toxicity, and sub-optimal renal targeting. This review will discuss the feasibility and current limitations of drug delivery technologies for the treatment of renal disease, offering an update on their potential and the future directions of these promising strategies.

Trajectory planning for UAV navigation in dynamic environments with matrix alignment Dijkstra

The trajectory planning for Unmanned aerial vehicles (UAVs) in the dynamic environments is a challenging task. Many restrictions should be taken into consideration, including dynamic terrain collision, no-fly zone criteria, power and fuel criteria and so on. However, some methods treat dynamic restrictions as static in order to reduce cost and obtain efficient and acceptable paths. To achieve optimal, efficient and acceptable paths, we first use a high-dimension matrix, extended hierarchical graph, to model the unexplored dynamic grid environment and to convert weather conditions to passable coefficient. The original shortest path planning task is then translated to plan the safest path. Therefore, we propose a new forward exploration and backward navigation algorithm, matrix alignment Dijkstra (MAD), to pilot UAVs. We use matrix alignment operation to simulate the parallel exploration process of all cells from time t to \(t+1\). This exploration process can be accelerated on a GPU. Finally, we recall the optimal path according to the navigator matrix. In addition, we can achieve UAV path planning from one source cell to multiple target cells within one single run. We validate our method on a real dynamic weather dataset and get competitive results in both efficiency and accuracy. We analyse the performance of MAD on a grid-based benchmark dataset and artificial maze data. Simulated results show MAD is especially helpful when the grid-based environment is large-scale and dynamic.

Abstract 2889: Novel bispecific humanized BCMA-CD3 antibodies specifically target multiple myeloma cells

Abstract BCMA antigen is overexpressed in multiple myeloma cells and has been shown to be a promising target for novel cellular and antibody therapeutics. The humanized BCMA (clone 4C8A) antibody that effectively targeted multiple myeloma in a CAR (chimeric antigen receptor) format was used for designing several formats of bispecific BCMA-CD3 antibodies. Several different designs including univalent and bivalent BCMA-CD3 CrossMAB; triple chain BCMA-FAB-CD3 ScFv-Fc and tetravalent BCMA ScFv-CD3 ScFv-Fc antibodies were tested for affinity and binding with BCMA-positive cells and T cells. Real time cytotoxic activity (RTCA) and IFN-gamma secretion were also tested with multiple myeloma and CHO-BCMA target cells. All BCMA bispecific antibody designs demonstrated specific affinity with CHO-BCMA but not CHO cells in the nanoM range of 0.3-33 nM; all designs bound to T cells by FACS. The triple chain BCMA-FAB-CD3 ScFv-Fc designs had the highest Kd of binding with CHO-BCMA cells (Kd=0.3 nM) and with multiple myeloma MM1S, and H929 cells (0.1 and 0.3 nM, respectively). All antibodies with T cells specifically killed CHO-BCMA and multiple myeloma cells in a dose-dependent manner but not CHO or K562 cells, measured by RTCA assay. The bivalent BCMA-CD3 CrossMAB and triple chain BCMA FAB CD3scFv-Fc antibodies with T cells secreted IFN-gamma with EC50 =0.01-0.02 nM. In addition, the BCMA bispecific antibodies had high in vivo efficacy using a MM1S xenograft NSG mouse model. Significance: This data demonstrates high efficacy of novel hBCMA-CD3 antibodies with multiple myeloma cells and provides a basis for future pre-clinical and clinical development. Citation Format: Yanwei Huang, John Sienkiewicz, Jinying Sun, Liselle Guiang, Feng Li, Liming Yang, Lijun Wu, Vita M. Golubovskaya. Novel bispecific humanized BCMA-CD3 antibodies specifically target multiple myeloma cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2889.

Cytotoxic T lymphocytes targeting a conserved SARS-CoV-2 spike epitope are efficient serial killers.

Understanding immune response to infections and vaccines lags understanding humoral responses. While neutralizing antibody responses wane over time, T cells are instrumental in long-term immunity. We apply machine learning and time-lapse imaging microscopy in nanowell grids (TIMING) to study thousands of videos of T cells with specificity for SARS-CoV-2 eliminating targets bearing spike protein as a surrogate for viral infection. The data on effector functions, including cytokine secretion and cytotoxicity, provide the first direct evidence that cytotoxic T lymphocytes from a convalescent patient targeting an epitope conserved across all known variants of concern are serial killers capable of eliminating multiple infected target cells. These data have implications for vaccine development and for the recovery and monitoring of infected individuals.

Ultrastructure of Rat Rostral Nucleus of the Solitary Tract Terminals in the Parabrachial Nucleus and Medullary Reticular Formation.

Neurons in the rostral nucleus of the solitary tract (rNST) receive taste information from the tongue and relay it mainly to the parabrachial nucleus (PBN) and the medullary reticular formation (RF) through two functionally different neural circuits. To help understand how the information from the rNST neurons is transmitted within these brainstem relay nuclei in the taste pathway, we examined the terminals of the rNST neurons in the PBN and RF by use of anterograde horseradish peroxidase (HRP) labeling, postembedding immunogold staining for glutamate, serial section electron microscopy, and quantitative analysis. Most of the anterogradely labeled, glutamate-immunopositive axon terminals made a synaptic contact with only a single postsynaptic element in PBN and RF, suggesting that the sensory information from rNST neurons, at the individual terminal level, is not passed to multiple target cells. Labeled terminals were usually presynaptic to distal dendritic shafts in both target nuclei. However, the frequency of labeled terminals that contacted dendritic spines was significantly higher in the PBN than in the RF, and the frequency of labeled terminals that contacted somata or proximal dendrites was significantly higher in the RF than in the PBN. Labeled terminals receiving axoaxonic synapses, which are a morphological substrate for presynaptic modulation frequently found in primary sensory afferents, were not observed. These findings suggest that the sensory information from rNST neurons is processed in a relatively simple manner in both PBN and RF, but in a distinctly different manner in the PBN as opposed to the RF.

Dynamical Analysis of Secondary Dengue Viral Infection with Multiple Target Cells and Diffusion by Mathematical Model

Dengue is an epidemic disease rapidly spreading throughout many parts of the world, which is a serious public health concern. Understanding disease mechanisms through mathematical modeling is one of the most effective tools for this purpose. The aim of this manuscript is to develop and analyze a dynamical system of PDEs that describes the secondary infection caused by DENV, considering (i) the diffusion due to spatial mobility of cells and DENV particles, (ii) the interactions between multiple target cells, DENV, and antibodies of two types (heterologous and homologous). Global existence, positivity, and boundedness are proved for the system with homogeneous Neumann boundary conditions. Three threshold parameters are computed to characterize the existence and stability conditions of the model’s four steady states. Via means of Lyapunov functional, the global stability of all steady states is carried out. Our results show that the uninfected steady state is globally asymptotically stable if the basic reproduction number is less than or equal to unity, which leads to the disappearance of the disease from the body. When the basic reproduction number is greater than unity, the disease persists in the body with an active or inactive immune antibody response. To demonstrate such theoretical results, numerical simulations are presented.

Endogenous and Therapeutic Estrogens: Maestro Conductors of the Microenvironment of ER+ Breast Cancers.

Simple SummaryBreast cancers that express estrogen receptor alpha (ER+) are the most common subtype of breast cancers. Although surgery and anti-estrogen therapies are successful for most of these patients, treatment-resistant ER+ metastatic cancers account for the majority of breast-cancer-related deaths. Immunotherapies have shown promise for other cancer types, but these have been much less effective for ER+ breast cancers. In this review, we update progress in our understanding of the immune microenvironment and the community of other cells that surround ER+ cancer cells at the primary and metastatic sites, the responses of these different cell types to various anti-estrogen therapies, and the net outcomes in experimental and clinical studies. We highlight evolving technologies that will provide greater insight into the biology of ER+ breast cancer and the foundation for new treatment and prevention strategies, in order to reduce mortality of this disease.Estrogen receptor alpha (ERα) marks heterogeneous breast cancers which display a repertoire of somatic genomic mutations and an immune environment that differs from other breast cancer subtypes. These cancers also exhibit distinct biological behaviors; despite an overall better prognosis than HER2+ or triple negative breast cancers, disseminated dormant cells can lead to disease recurrence decades after the initial diagnosis and treatment. Estrogen is the best studied driver of these cancers, and antagonism or reduction of estrogen activity is the cornerstone of therapeutic approaches. In addition to reducing proliferation of ERα+ cancer cells, these treatments also alter signals to multiple other target cells in the environment, including immune cell subpopulations, cancer-associated fibroblasts, and endothelial cells via several distinct estrogen receptors. In this review, we update progress in our understanding of the stromal cells populating the microenvironments of primary and metastatic ER+ tumors, the effects of estrogen on tumor and stromal cells to modulate immune activity and the extracellular matrix, and net outcomes in experimental and clinical studies. We highlight new approaches that will illuminate the unique biology of these cancers, provide the foundation for developing new treatment and prevention strategies, and reduce mortality of this disease.

Mathematical analysis of multi-target cells and multi-strain age-structured model with two HIV infection routes

In this paper, we propose detailed and reasonable viral dynamics by using a multi-compartment model that incorporating the age since the infection of multiple infected cells, multiple target cells (Langerhans-cells and CD4[Formula: see text] T-cells), multiple viral strains (CCR5 and CXR4 HIV) and multiple infection routes (cell-to-cell and cell-to-virus). The basic reproduction number, [Formula: see text], of the whole model is derived from two transmission mechanisms: one is the potential trigger from the infection routes for a single target cell and other is the joint effect of multiple viral infections for multi-target cells. Accordingly, we study the global stability of the steady states for the single target model. For the whole model, we prove that the infection-free steady state is globally asymptotically stable if [Formula: see text], whereas viruses persist uniformly if [Formula: see text]. Numerical simulations are carried out to illustrate the theoretical results. Sensitive analyses expound the effect of model parameters on the comprehensive reproduction number. It is remarkable to find that simultaneous control of HIV infection for two target cells can effectively reduce the viral loads within-host. Finally, our work suggests that the synergetic mechanism of multi-target cells and multi-strain cannot be ignored during treatment.

Extracellular Vesicles in neural cell interaction and CNS homeostasis.

Central nervous system (CNS) homeostasis critically depends on the interaction between neurons and glia cells. Extracellular vesicles (EVs) recently emerged as versatile messengers in CNS cell communication. EVs are released by neurons and glia in activity‐dependent manner and address multiple target cells within and outside the nervous system. Here, we summarize the recent advances in understanding the physiological roles of EVs in the nervous system and their ability to deliver signals across the CNS barriers. In addition to the disposal of cellular components via EVs and clearance by phagocytic cells, EVs are involved in plasticity‐associated processes, mediate trophic support and neuroprotection, promote axonal maintenance, and modulate neuroinflammation. While individual functional components of the EV cargo are becoming progressively identified, the role of neural EVs as compound multimodal signaling entities remains to be elucidated. Novel transgenic models and imaging technologies allow EV tracking in vivo and provide further insight into EV targeting and their mode of action. Overall, EVs represent key players in the maintenance of CNS homeostasis essential for the lifelong performance of neural networks and thus provide a wide spectrum of biomedical applications.

Antibody Afucosylation Augments CD16-Mediated Serial Killing and IFNγ Secretion by Human Natural Killer Cells.

One mechanism by which monoclonal antibodies (mAb) help treat cancer or autoimmune disease is through triggering antibody-dependent cellular cytotoxicity (ADCC) via CD16 on Natural Killer (NK) cells. Afucosylation is known to increase the affinity of mAbs for CD16 on NK cells and here, we set out to assess how mAb afucosylation affects the dynamics of NK cell interactions, receptor expression and effector functions. An IgG1 version of a clinically important anti-CD20 mAb was compared to its afucosylated counterpart (anti-CD20-AF). Opsonization of CD20-expressing target cells, 721.221 or Daudi, with anti-CD20-AF increased NK cell cytotoxicity and IFNγ secretion, compared to anti-CD20. The afucosylated mAb also caused a more rapid and greater loss of CD16 from NK cell surfaces. Loss of CD16 has recently been shown to be important for NK cell detachment and sequential engagement of multiple target cells. Here, live-cell time-lapse microscopy of individual cell-cell interactions in an aqueous environment and a three-dimensional matrix, revealed that anti-CD20-AF induced more rapid killing of opsonized target cells. In addition, NK cells detached more quickly from target cells opsonized with anti-CD20-AF compared to anti-CD20, which increased engagement of multiple targets and enabled a greater proportion of NK cells to perform serial killing. Inhibition of CD16 shedding with TAPI-0 led to reduced detachment and serial killing. Thus, disassembly of the immune synapse caused by loss of cell surface CD16 is a factor determining the efficiency of ADCC and antibody afucosylation alters the dynamics of intercellular interactions to boost serial killing.

Dynamics of a delayed integro-differential HIV infection model with multiple target cells and nonlocal dispersal

In this paper, we formulate an age-space-structured HIV infection model that incorporating infection age, multiple target cells, and nonlocal dispersal. Applying the characteristic line method, we reduce the infection-age model to a delayed integro-differential system. The global well-posedness and boundedness of the semiflow for the system are established. The principal eigenvalue of the nonlocal dispersal problem is formulated, and it plays the same role as the basic reproduction number $$R_0$$ (the spectral radius of the next generation operator), which determines the global behavior of the steady states of the system. More precisely, the infection-free steady state is globally asymptotically stable (g.a.s) when $$R_0<1$$ , the virus is always present and the infected steady state is g.a.s when $$R_0>1$$ . Numerical simulations are carried out reinforcing these analytical results. In particular, three different kernel functions are given out to study the impact of dispersal form on the HIV infection within the host. Finally, our simulation works show that (i) increasing the dispersal rate and decreasing the intracellular delay will be increasing the final viral loads; (ii) the dispersal kernel function affects the value of $$R_0$$ and the final viral loads, and it is revealed that the dispersal form plays a crucial role in the process of HIV infection within the host.

Mathematical modelling of multiple target cells with delay

Target cells contagion has sparked the necessity of a multiple target cell model with chronic infection and intracellular delay (MTC-CI-ID). This paper seeks to advance a viral dynamical model with multiple target cells and corroborate the global stability of their steady states. Hence, the affiliation of the virus with two classes of target cells, CD4 + Tcells and macrophage. The time delay amidst viral entry and the output of virions were intimated and the basic reproductive number was procured, as the long term behavior of the model and less than unity. The contagious free equilibrium E0 was unveiled as locally and globally asymptotically stable. The conditions of an infected CD4 + Tcell and macrophage on viral production were engaged. It was ascertained that CD4 + Tcell accounts for larger amounts of virions in the blood as per macrophage cells. Hence, varying time delay has no impact on peak viral levels, but only shelves the viral peaking time. The adopted model has unrestrained bearing on HIV-1 therapy.

Droplet-Based Cytotoxicity Assay: Implementation of Time-Efficient Screening of Antitumor Activity of Natural Killer Cells

Natural killer (NK) cells are key players of the innate immune system. Due to their rapid cytotoxicity against infectious pathogens, hematologic malignancies, and solid tumors, NK cells represent solid candidates for cell-based immunotherapy. Despite the progress made in recent years, the heterogeneity in their cytotoxic behavior represents a drawback. With the goal of screening the intrinsic diversity of NK cells, droplet-based microfluidic technology is exploited to develop a single-cell time-efficient cytotoxicity assay. Toward this end, NK-92 cells are coencapsulated with hematological tumor cell lines in water-in-oil droplets of different sizes and their cytotoxic activity is evaluated. The effect of droplet-based confinement on NK cytotoxicity is investigated by controlling the droplet volume. The successful optimization of the droplet size allows for time efficiency compared to cytotoxicity assays based on flow cytometry. Additionally, the ability of individual NK-92 cells to kill multiple target cells in series is explored, expanding the knowledge about the serial killing process dynamics. The developed droplet-based microfluidic assay does not require the labeling of NK cells and represents a step toward developing of a forthcoming process for the selection of NK cells with the highest cytotoxicity against specific targets.