Characterising plasmacytoid and myeloid AXL+ SIGLEC-6+ dendritic cell functions and their interactions with HIV.

AXL+ Siglec-6+ dendritic cells (ASDC) are novel myeloid DCs which can be subdivided into CD11c+ and CD123+ expressing subsets. We showed for the first time that these two ASDC subsets are present in inflamed human anogenital tissues where HIV transmission occurs. Their presence in inflamed tissues was supported by single cell RNA analysis of public databases of such tissues including psoriasis diseased skin and colorectal cancer. Almost all previous studies have examined ASDCs as a combined population. Our data revealed that the two ASDC subsets differ markedly in their functions when compared with each other and to pDCs. Relative to their cell functions, both subsets of blood ASDCs but not pDCs expressed co-stimulatory and maturation markers which were more prevalent on CD11c+ ASDCs, thus inducing more T cell proliferation and activation than their CD123+ counterparts. There was also a significant polarisation of naïve T cells by both ASDC subsets toward Th2, Th9, Th22, Th17 and Treg but less toward a Th1 phenotype. Furthermore, we investigated the expression of chemokine receptors that facilitate ASDCs and pDCs migration from blood to inflamed tissues, their HIV binding receptors, and their interactions with HIV and CD4 T cells. For HIV infection, within 2 hours of HIV exposure, CD11c+ ASDCs showed a trend in more viral transfer to T cells than CD123+ ASDCs and pDCs for first phase transfer. However, for second phase transfer, CD123+ ASDCs showed a trend in transferring more HIV than CD11c+ ASDCs and there was no viral transfer from pDCs. As anogenital inflammation is a prerequisite for HIV transmission, strategies to inhibit ASDC recruitment into inflamed tissues and their ability to transmit HIV to CD4 T cells should be considered.

Myeloperoxidase Gene Deletion Causes Drastic Microbiome Shifts in Mice and Does Not Mitigate Dextran Sodium Sulphate-Induced Colitis.

Neutrophil-myeloperoxidase (MPO) is a heme-containing peroxidase which produces excess amounts of hypochlorous acid during inflammation. While pharmacological MPO inhibition mitigates all indices of experimental colitis, no studies have corroborated the role of MPO using knockout (KO) models. Therefore, we investigated MPO deficient mice in a murine model of colitis. Wild type (Wt) and MPO-deficient mice were treated with dextran sodium sulphate (DSS) in a chronic model of experimental colitis with three acute cycles of DSS-induced colitis over 63 days, emulating IBD relapse and remission cycles. Mice were immunologically profiled at the gut muscoa and the faecal microbiome was assessed via 16S rRNA amplicon sequencing. Contrary to previous pharmacological antagonist studies targeting MPO, MPO-deficient mice showed no protection from experimental colitis during cyclical DSS-challenge. We are the first to report drastic faecal microbiota shifts in MPO-deficient mice, showing a significantly different microbiome profile on Day 1 of treatment, with a similar shift and distinction on Day 29 (half-way point), via qualitative and quantitative descriptions of phylogenetic distances. Herein, we provide the first evidence of substantial microbiome shifts in MPO-deficiency, which may influence disease progression. Our findings have significant implications for the utility of MPO-KO mice in investigating disease models.

OMIP-096: A 24-color flow cytometry panel to identify and characterize CD4+ and CD8+ tissue-resident T cells in human skin, intestinal, and type II mucosal tissue.

There is a great need to understand human immune cells within tissue, where disease manifests and infection occurs. Tissue-resident memory T cells (TRMs) were discovered over a decade ago, there is a great need to understand their role in human disease. We developed a 24-color flow cytometry panel to comprehensively interrogate CD4+ and CD8+ TRMs isolated from human tissues. When interrogating cells within human tissue, enzymatic methods used to liberate cells from within the tissue can cause cleavage of cell surface markers needed to phenotype these cells. Here we carefully select antibody clones and evaluate the effect of enzymatic digestion on the expression of markers relevant to the identification of T cell residency, as well as markers relevant to the activation and immunoregulation status of these cells. We have designed this panel to be applicable across a range of human tissues including skin, intestine, and type II mucosae such as the vagina.

Models for resonant acoustic metasurfaces with application to moth wing ultrasound absorption.

Taking as bioinspiration the remarkable acoustic absorption properties of moth wings, we develop a simple analytical model that describes the interaction between acoustic pressure fields, and thin elastic plates incorporating resonant sub-structures. The moth wing is an exemplar of a natural acoustic metamaterial; the wings are deeply subwavelength in thickness at the frequencies of interest, the absorption is broadband and the tiny scales resonate on the moth wing acting in concert. The simplified model incorporates only the essential physics and the scales are idealized to flat rigid rectangular plates coupled via a spring to an elastic plate that forms the wing; all the components are deep-subwavelength at desired frequencies. Based on Fourier analysis, complemented by phenomenological modelling, our theory shows excellent agreement with simulation mimicking the moth-wing structure. Moth wings operate as broadband sound absorbers employing a range of scale sizes. We demonstrate that a random distribution of scale sizes generates a broadband absorption spectrum. To further illustrate the potential of the model, we design a deeply sub-wavelength acoustic counterpart of electromagnetically induced reflectance. This article is part of the theme issue 'Wave generation and transmission in multi-scale complex media and structured metamaterials (part 2)'.

Moth wings as sound absorber metasurface.

In noise control applications, a perfect metasurface absorber would have the desirable traits of not only mitigating unwanted sound, but also being much thinner than the wavelengths of interest. Such deep-subwavelength performance is difficult to achieve technologically, yet moth wings, as natural metamaterials, offer functionality as efficient sound absorbers through the action of the numerous resonant scales that decorate their wing membrane. Here, we quantify the potential for moth wings to act as a sound-absorbing metasurface coating for acoustically reflective substrates. Moth wings were found to be efficient sound absorbers, reducing reflection from an acoustically hard surface by up to 87% at the lowest frequency tested (20 kHz), despite a thickness to wavelength ratio of up to 1/50. Remarkably, after the removal of the scales from the dorsal surface the wing's orientation on the surface changed its absorptive performance: absorption remains high when the bald wing membrane faces the sound but breaks down almost completely in the reverse orientation. Numerical simulations confirm the strong influence of the air gap below the wing membrane but only when it is adorned with scales. The finding that moth wings act as deep-subwavelength sound-absorbing metasurfaces opens the door to bioinspired, high-performance sound mitigation solutions.

Wingtip folds and ripples on saturniid moths create decoy echoes against bat biosonar.

Sensory coevolution has equipped certain moth species with passive acoustic defenses to counter predation by echolocating bats.1,2 Some large silkmoths (Saturniidae) possess curved and twisted biosonar decoys at the tip of elongated hindwing tails.3,4 These are thought to create strong echoes that deflect biosonar-guided bat attacks away from the moth's body to less essential parts of their anatomy. We found that closely related silkmoths lacking such hindwing decoys instead often possess intriguing ripples and folds on the conspicuously lobed tips of their forewings. The striking analogy of twisted shapes displayed far from the body suggests these forewing structures might function as alternative acoustic decoys. Here we reveal that acoustic reflectivity and hence detectability of such wingtips is higher than that of the body at ultrasonic frequencies used by hunting bats. Wingtip reflectivity is higher the more elaborate the structure and the further from the body. Importantly, wingtip reflectivity is often considerably higher than in a well-studied functional hindwing decoy. Such increased reflectivity would misdirect the bat's sonar-guided attack toward the wingtip, resulting in similar fitness benefits to hindwing acoustic decoys. Structurally, folded wingtips present echo-generating surfaces to many directions, and folds and ripples can act as retroreflectors that together create conspicuous targets. Phylogenetically, folds and ripples at wingtips have evolved multiple times independently within silkmoths and always as alternatives to hindwing decoys. We conclude that they function as acoustic wingtip decoys against bat biosonar. VIDEO ABSTRACT.

Plasmacytoid dendritic cells have divergent effects on HIV infection of initial target cells and induce a pro-retention phenotype.

Although HIV infection inhibits interferon responses in its target cells in vitro, interferon signatures can be detected in vivo soon after sexual transmission, mainly attributed to plasmacytoid dendritic cells (pDCs). In this study, we examined the physiological contributions of pDCs to early HIV acquisition using coculture models of pDCs with myeloid DCs, macrophages and the resting central, transitional and effector memory CD4 T cell subsets. pDCs impacted infection in a cell-specific manner. In myeloid cells, HIV infection was decreased via antiviral effects, cell maturation and downregulation of CCR5 expression. In contrast, in resting memory CD4 T cells, pDCs induced a subset-specific increase in intracellular HIV p24 protein expression without any activation or increase in CCR5 expression, as measured by flow cytometry. This increase was due to reactivation rather than enhanced viral spread, as blocking HIV entry via CCR5 did not alter the increased intracellular p24 expression. Furthermore, the load and proportion of cells expressing HIV DNA were restricted in the presence of pDCs while reverse transcriptase and p24 ELISA assays showed no increase in particle associated reverse transcriptase or extracellular p24 production. In addition, pDCs also markedly induced the expression of CD69 on infected CD4 T cells and other markers of CD4 T cell tissue retention. These phenotypic changes showed marked parallels with resident memory CD4 T cells isolated from anogenital tissue using enzymatic digestion. Production of IFNα by pDCs was the main driving factor for all these results. Thus, pDCs may reduce HIV spread during initial mucosal acquisition by inhibiting replication in myeloid cells while reactivating latent virus in resting memory CD4 T cells and retaining them for immune clearance.

Human anogenital monocyte-derived dendritic cells and langerin+cDC2 are major HIV target cells.

Tissue mononuclear phagocytes (MNP) are specialised in pathogen detection and antigen presentation. As such they deliver HIV to its primary target cells; CD4 T cells. Most MNP HIV transmission studies have focused on epithelial MNPs. However, as mucosal trauma and inflammation are now known to be strongly associated with HIV transmission, here we examine the role of sub-epithelial MNPs which are present in a diverse array of subsets. We show that HIV can penetrate the epithelial surface to interact with sub-epithelial resident MNPs in anogenital explants and define the full array of subsets that are present in the human anogenital and colorectal tissues that HIV may encounter during sexual transmission. In doing so we identify two subsets that preferentially take up HIV, become infected and transmit the virus to CD4 T cells; CD14+CD1c+ monocyte-derived dendritic cells and langerin-expressing conventional dendritic cells 2 (cDC2).

The Role of Tissue Resident Memory CD4 T Cells in Herpes Simplex Viral and HIV Infection.

Tissue-resident memory T cells (TRM) were first described in 2009. While initially the major focus was on CD8+ TRM, there has recently been increased interest in defining the phenotype and the role of CD4+ TRM in diseases. Circulating CD4+ T cells seed CD4+ TRM, but there also appears to be an equilibrium between CD4+ TRM and blood CD4+ T cells. CD4+ TRM are more mobile than CD8+ TRM, usually localized deeper within the dermis/lamina propria and yet may exhibit synergy with CD8+ TRM in disease control. This has been demonstrated in herpes simplex infections in mice. In human recurrent herpes infections, both CD4+ and CD8+ TRM persisting between lesions may control asymptomatic shedding through interferon-gamma secretion, although this has been more clearly shown for CD8+ T cells. The exact role of the CD4+/CD8+ TRM axis in the trigeminal ganglia and/or cornea in controlling recurrent herpetic keratitis is unknown. In HIV, CD4+ TRM have now been shown to be a major target for productive and latent infection in the cervix. In HSV and HIV co-infections, CD4+ TRM persisting in the dermis support HIV replication. Further understanding of the role of CD4+ TRM and their induction by vaccines may help control sexual transmission by both viruses.

Moth wings are acoustic metamaterials.

Metamaterials assemble multiple subwavelength elements to create structures with extraordinary physical properties (1-4). Optical metamaterials are rare in nature and no natural acoustic metamaterials are known. Here, we reveal that the intricate scale layer on moth wings forms a metamaterial ultrasound absorber (peak absorption = 72% of sound intensity at 78 kHz) that is 111 times thinner than the longest absorbed wavelength. Individual scales act as resonant (5) unit cells that are linked via a shared wing membrane to form this metamaterial, and collectively they generate hard-to-attain broadband deep-subwavelength absorption. Their collective absorption exceeds the sum of their individual contributions. This sound absorber provides moth wings with acoustic camouflage (6) against echolocating bats. It combines broadband absorption of all frequencies used by bats with light and ultrathin structures that meet aerodynamic constraints on wing weight and thickness. The morphological implementation seen in this evolved acoustic metamaterial reveals enticing ways to design high-performance noise mitigation devices.

Thoracic scales of moths as a stealth coating against bat biosonar.

Many moths are endowed with ultrasound-sensitive ears that serve the detection and evasion of echolocating bats. Moths lacking such ears could still gain protection from bat biosonar by using stealth acoustic camouflage, absorbing sound waves rather than reflecting them back as echoes. The thorax of a moth is bulky and hence acoustically highly reflective. This renders it an obvious target for any bat. Much of the thorax of moths is covered in hair-like scales, the layout of which is remarkably similar in structure and arrangement to natural fibrous materials commonly used in sound insulation. Despite this structural similarity, the effect of thorax scales on moth echoes has never been characterized. Here, we test whether and how moth thorax scales function as an acoustic absorber. From tomographic echo images, we find that the thin layer of thoracic scales of diurnal butterflies affects the strength of ultrasound echoes from the thorax very little, while the thorax scales of earless moths absorbs an average of 67 ± 9% of impinging ultrasonic sound energy. We show that the thorax scales of moths provide acoustic camouflage by acting as broadband (20-160 kHz) stealth coating. Modelling results suggest the scales are acting as a porous sound absorber; however, the thorax scales of moths achieve a considerably higher absorption than technical fibrous porous absorbers with the same structural parameters. Such scales, despite being thin and lightweight, constitute a broadband, multidirectional and efficient ultrasound absorber that reduces the moths' detectability to hunting bats and gives them a survival advantage.

Deaf moths employ acoustic Müllerian mimicry against bats using wingbeat-powered tymbals.

Emitting ultrasound upon hearing an attacking bat is an effective defence strategy used by several moth taxa. Here we reveal how Yponomeuta moths acquire sophisticated acoustic protection despite being deaf themselves and hence unable to respond to bat attacks. Instead, flying Yponomeuta produce bursts of ultrasonic clicks perpetually; a striated patch in their hind wing clicks as the beating wing rotates and bends. This wing structure is strikingly similar to the thorax tymbals with which arctiine moths produce their anti-bat sounds. And indeed, Yponomeuta sounds closely mimic such arctiine signals, revealing convergence in form and function. Because both moth taxa contain noxious compounds, we conclude they are mutual Müllerian acoustic mimics. Yponomeuta's perpetual clicking would however also attract bat predators. In response, their click amplitude is reduced and affords acoustic protection just as far as required, matching the distance over which bat biosonar would pick up Yponomeuta echoes anyway - advanced acoustic defences for a deaf moth.

Biomechanics of a moth scale at ultrasonic frequencies.

The wings of moths and butterflies are densely covered in scales that exhibit intricate shapes and sculptured nanostructures. While certain butterfly scales create nanoscale photonic effects, moth scales show different nanostructures suggesting different functionality. Here we investigate moth-scale vibrodynamics to understand their role in creating acoustic camouflage against bat echolocation, where scales on wings provide ultrasound absorber functionality. For this, individual scales can be considered as building blocks with adapted biomechanical properties at ultrasonic frequencies. The 3D nanostructure of a full Bunaea alcinoe moth forewing scale was characterized using confocal microscopy. Structurally, this scale is double layered and endowed with different perforation rates on the upper and lower laminae, which are interconnected by trabeculae pillars. From these observations a parameterized model of the scale's nanostructure was formed and its effective elastic stiffness matrix extracted. Macroscale numerical modeling of scale vibrodynamics showed close qualitative and quantitative agreement with scanning laser Doppler vibrometry measurement of this scale's oscillations, suggesting that the governing biomechanics have been captured accurately. Importantly, this scale of B. alcinoe exhibits its first three resonances in the typical echolocation frequency range of bats, suggesting it has evolved as a resonant absorber. Damping coefficients of the moth-scale resonator and ultrasonic absorption of a scaled wing were estimated using numerical modeling. The calculated absorption coefficient of 0.50 agrees with the published maximum acoustic effect of wing scaling. Understanding scale vibroacoustic behavior helps create macroscopic structures with the capacity for broadband acoustic camouflage.

Jet-paddling jellies: swimming performance in the Rhizostomeae jellyfish Catostylus mosaicus.

Jellyfish are a successful and diverse class of animals that swim via jet propulsion, with swimming performance and propulsive efficiency being related to the animal's feeding ecology and body morphology. The Rhizostomeae jellyfish lack tentacles but possess four oral lobes and eight trailing arms at the centre of their bell, giving them a body morphology quite unlike that of other free-swimming medusae. The implications of this body morphology on the mechanisms by which thrust is produced are unknown. Here, we determined the wake structure and propulsive efficiency in the blue-blubber jellyfish Catostylus mosaicus (order: Rhizostomeae). The animal is propelled during both bell contraction and bell relaxation by different thrust-generating mechanisms. During bell contraction, a jet of fluid is expelled from the subumbrellar cavity, which results from the interaction between the counter-rotating stopping (from the preceding contraction cycle) and starting vortices, creating a vortex superstructure and propulsion. This species is also able to utilise passive energy recapture, which increases the animal's swimming velocity towards the end of the bell expansion phase when the bell diameter is constant. The thrust produced during this phase is the result of the flexible bell margin manoeuvring the stopping vortex into the subumbrellar cavity during bell relaxation, enhancing its circulation, and creating a region of high pressure on the inner surface of the bell and, consequently, thrust. These mechanisms of thrust generation result in C. mosaicus having a relatively high propulsive efficiency compared with other swimmers, indicating that economical locomotion could be a contributing factor in the ecological success of these medusan swimmers.

Swimming mechanics and propulsive efficiency in the chambered nautilus.

The chambered nautilus (Nautilus pompilius) encounters severe environmental hypoxia during diurnal vertical movements in the ocean. The metabolic cost of locomotion (Cmet) and swimming performance depend on how efficiently momentum is imparted to the water and how long on-board oxygen stores last. While propulsive efficiency is generally thought to be relatively low in jet propelled animals, the low Cmet in Nautilus indicates that this is not the case. We measured the wake structure in Nautilus during jet propulsion swimming, to determine their propulsive efficiency. Animals swam with either an anterior-first or posterior-first orientation. With increasing swimming speed, whole cycle propulsive efficiency increased during posterior-first swimming but decreased during anterior-first swimming, reaching a maximum of 0.76. The highest propulsive efficiencies were achieved by using an asymmetrical contractile cycle in which the fluid ejection phase was relatively longer than the refilling phase, reducing the volume flow rate of the ejected fluid. Our results demonstrate that a relatively high whole cycle propulsive efficiency underlies the low Cmet in Nautilus, representing a strategy to reduce the metabolic demands in an animal that spends a significant part of its daily life in a hypoxic environment.