Clusters of red blood cells in microcapillary flow: hydrodynamic versus macromolecule induced interaction.

We present experiments on RBCs that flow through micro-capillaries under physiological conditions. The strong flow-shape coupling of these deformable objects leads to a rich variety of cluster formation. We show that the RBC clusters form as a subtle imbrication between hydrodynamic interactions and adhesion forces because of plasma proteins, mimicked by the polymer dextran. Clusters form along the capillaries and macromolecule-induced adhesion contributes to their stability. However, at high yet physiological flow velocities, shear stresses overcome part of the adhesion forces, and cluster stabilization due to hydrodynamics becomes stronger. For the case of pure hydrodynamic interaction, cell-to-cell distances have a pronounced bimodal distribution. Our 2D-numerical simulations on vesicles capture the transition between adhesive and non-adhesive clusters at different flow velocities.

Amoeboid swimming in a channel.

Several micro-organisms, such as bacteria, algae, or spermatozoa, use flagellar or ciliary activity to swim in a fluid, while many other micro-organisms instead use ample shape deformation, described as amoeboid, to propel themselves either by crawling on a substrate or swimming. Many eukaryotic cells were believed to require an underlying substratum to migrate (crawl) by using membrane deformation (like blebbing or generation of lamellipodia) but there is now increasing evidence that a large variety of cells (including those of the immune system) can migrate without the assistance of focal adhesion, allowing them to swim as efficiently as they can crawl. This paper details the analysis of amoeboid swimming in a confined fluid by modeling the swimmer as an inextensible membrane deploying local active forces (with zero total force and torque). The swimmer displays a rich behavior: it may settle into a straight trajectory in the channel or navigate from one wall to the other depending on its confinement. The nature of the swimmer is also found to be affected by confinement: the swimmer can behave, on average over one swimming cycle, as a pusher at low confinement, and becomes a puller at higher confinement, or vice versa. The swimmer's nature is thus not an intrinsic property. The scaling of the swimmer velocity V with the force amplitude A is analyzed in detail showing that at small enough A, V∼A(2)/η(2) (where η is the viscosity of the ambient fluid), whereas at large enough A, V is independent of the force and is determined solely by the stroke cycle frequency and the swimmer size. This finding starkly contrasts with models where motion is based on ciliary and flagellar activity, where V∼A/η. To conclude, two definitions of efficiency as put forward in the literature are analyzed with distinct outcomes. We find that one type of efficiency has an optimum as a function of confinement while the other does not. Future perspectives are outlined.

Prediction of anomalous blood viscosity in confined shear flow.

Red blood cells play a major role in body metabolism by supplying oxygen from the microvasculature to different organs and tissues. Understanding blood flow properties in microcirculation is an essential step towards elucidating fundamental and practical issues. Numerical simulations of a blood model under a confined linear shear flow reveal that confinement markedly modifies the properties of blood flow. A nontrivial spatiotemporal organization of blood elements is shown to trigger hitherto unrevealed flow properties regarding the viscosity η, namely ample oscillations of its normalized value [η] = (η-η(0))/(η(0)ϕ) as a function of hematocrit ϕ (η(0) = solvent viscosity). A scaling law for the viscosity as a function of hematocrit and confinement is proposed. This finding can contribute to the conception of new strategies to efficiently detect blood disorders, via in vitro diagnosis based on confined blood rheology. It also constitutes a contribution for a fundamental understanding of rheology of confined complex fluids.

Vesicle dynamics in a confined Poiseuille flow: from steady state to chaos.

Red blood cells (RBCs) are the major component of blood, and the flow of blood is dictated by that of RBCs. We employ vesicles, which consist of closed bilayer membranes enclosing a fluid, as a model system to study the behavior of RBCs under a confined Poiseuille flow. We extensively explore two main parameters: (i) the degree of confinement of vesicles within the channel and (ii) the flow strength. Rich and complex dynamics for vesicles are revealed, ranging from steady-state shapes (in the form of parachute and slipper shapes) to chaotic dynamics of shape. Chaos occurs through a cascade of multiple periodic oscillations of the vesicle shape. We summarize our results in a phase diagram in the parameter plane (degree of confinement and flow strength). This finding highlights the level of complexity of a flowing vesicle in the small Reynolds number where the flow is laminar in the absence of vesicles and can be rendered turbulent due to elasticity of vesicles.

Rheology of a vesicle suspension with finite concentration: a numerical study.

Vesicles, closed membranes made of a bilayer of phospholipids, are considered as a biomimetic system for the mechanics of red blood cells. The understanding of their dynamics under flow and their rheology is expected to help the understanding of the behavior of blood flow. We conduct numerical simulations of a suspension of vesicles in two dimensions at a finite concentration in a shear flow imposed by countertranslating rigid bounding walls by using an appropriate Green's function. We study the dynamics of vesicles, their spatial configurations, and their rheology, namely, the effective viscosity η(eff). A key parameter is the viscosity contrast λ (the ratio between the viscosity of the encapsulated fluid over that of the suspending fluid). For small enough λ, vesicles are known to exhibit tank treading (TT), while at higher λ they exhibit tumbling (TB). We find that η(eff) decreases in the TT regime, passes a minimum at a critical λ=λ(c), and increases in the TB regime. This result confirms previous theoretical and numerical works performed in the extremely dilute regime, pointing to the robustness of the picture even in the presence of hydrodynamic interactions. Our results agree also with very recent numerical simulations performed in three dimensions both in the dilute and more concentrated regime. This points to the fact that dimensionality does not alter the qualitative features of η(eff). However, they disagree with recent simulations in two dimensions. We provide arguments about the possible sources of this disagreement.

Mucosal imprinting of vaccine-induced CD8⁺ T cells is crucial to inhibit the growth of mucosal tumors.

Although many human cancers are located in mucosal sites, most cancer vaccines are tested against subcutaneous tumors in preclinical models. We therefore wondered whether mucosa-specific homing instructions to the immune system might influence mucosal tumor outgrowth. We showed that the growth of orthotopic head and neck or lung cancers was inhibited when a cancer vaccine was delivered by the intranasal mucosal route but not the intramuscular route. This antitumor effect was dependent on CD8⁺ T cells. Indeed, only intranasal vaccination elicited mucosal-specific CD8⁺ T cells expressing the mucosal integrin CD49a. Blockade of CD49a decreased intratumoral CD8⁺ T cell infiltration and the efficacy of cancer vaccine on mucosal tumor. We then showed that after intranasal vaccination, dendritic cells from lung parenchyma, but not those from spleen, induced the expression of CD49a on cocultured specific CD8⁺ T cells. Tumor-infiltrating lymphocytes from human mucosal lung cancer also expressed CD49a, which supports the relevance and possible extrapolation of these results in humans. We thus identified a link between the route of vaccination and the induction of a mucosal homing program on induced CD8⁺ T cells that controlled their trafficking. Immunization route directly affected the efficacy of the cancer vaccine to control mucosal tumors.

Tumor-specific targeting of pancreatic cancer with Shiga toxin B-subunit.

Pancreatic carcinoma is one of the most aggressive tumor entities, and standard chemotherapy provides only modest benefit. Therefore, specific targeting of pancreatic cancer for early diagnosis and therapeutic intervention is of great interest. We have previously shown that the cellular receptor for Shiga toxin B (STxB), the glycosphingolipid globotriaosylceramide (Gb(3) or CD77) is strongly increased in colorectal adenocarcinoma and their metastases. Here, we report an upregulation of Gb(3) in pancreatic adenocarcinoma (21 of 27 cases) as compared with matched normal tissue (n = 27). The mean expression was highly significantly increased from 30 ± 16 ng Gb(3)/mg tissue in normal pancreas to 61 ± 41 ng Gb(3)/mg tissue (mean ± SD, P = 0.0006), as evidenced by thin layer chromatography. Upregulation of Gb(3) levels did not depend on tumor stage or grading and showed no correlation with clinical outcome. Tumor cells and endothelial cells were identified as the source of increased Gb(3) expression by immunocytochemistry. Pancreatic cancer cell lines showed rapid intracellular uptake of STxB to the Golgi apparatus, following the retrograde pathway. The therapeutic application of STxB was tested by specific delivery of covalently coupled SN38, an active metabolite of the topoisomerase I inhibitor irinotecan. The cytotoxic effect of the STxB-SN38 compound in pancreatic cancer cell lines was increased more than 100-fold compared with irinotecan. Moreover, this effect was effectively blocked by competing incubation with nonlabeled STxB, showing the specificity of the targeting. Thus, STxB constitutes a promising new tool for specific targeting of pancreatic cancer.

Nonequilibrium fluctuations of an interface under shear.

The steady-state properties of an interface in a stationary Couette flow are addressed within the framework of fluctuating hydrodynamics. Our study reveals that thermal fluctuations are driven out of equilibrium by an effective shear rate that differs from the applied one. In agreement with experiments, we find that the mean-square displacement of the interface is reduced by the flow. We also show that nonequilibrium fluctuations present a certain degree of universality in the sense that all features of the fluids can be factorized into a single control parameter. Finally, the results are discussed in the light of recent experimental and numerical studies.