No description available.

No description available.

Rosetting is an adhesion property of malaria parasites whereby infected erythrocytes bind to two or more uninfected erythrocytes, forming a so-called rosette. Rosetting of Plasmodium falciparum is associated with disease severity and high parasitaemia in sub-Saharan Africa, although currently the function of rosetting remains unknown. An early IFNg response elicited from the innate immune system is associated with resolution of malaria infection in mice. Published data suggests that optimal IFNg production may require contact between peripheral blood mononuclear cells and P. falciparum infected erythrocytes. The first part of this thesis investigates the hypothesis that rosetting is an immune evasion strategy to hide infected erythrocytes from detection by innate immune cells. Across five laboratory strains of P. falciparum rosetting was not associated with differential IFNg production when parasites were grown in group O blood. Reappraisal of the data with respect to blood group for one strain found that rosetting significantly reduced the IFNg response to parasites grown in group A blood (P=0.022, Wilcoxon signed-rank test), where it is known that rosettes are bigger and stronger. This is consistent with the hypothesis that rosetting is an immune evasion strategy and the first study to find evidence for a function of rosetting. Further work is needed in order to generalise this finding. The cytokine response to P. falciparum varies between people and this variation may be indicative of disease progression. In mice infected with malaria it is also apparent that parasite strain can determine the cytokine response of the host. It is unclear whether P. falciparum strains vary in their ability to induce cytokines. The second part of this thesis investigates variation in cytokine induction between P. falciparum strains. Across four laboratory strains of P. falciparum, IFNg production was significantly dependent on parasite strain (F3,178= 48.49, P<0.001). Production of GM-CSF, IL-1b, IL-6, IL-10 and TNFa significantly correlated with production of IFNg (P<0.001, Pearson correlation) and followed the same strain-dependent pattern. The ratio of pro-inflammatory cytokines to IL-10 was also dependent on parasite strain. These data provide strong evidence for P. falciparum strain-dependent cytokine responses which may be an important determinant of disease outcome. Phagocytosis by splenic macrophages is proposed to be the principle mechanism of parasitaemia control in malaria infection. CD36 mediated phagocytosis may by an important mechanism of non-opsonic parasite clearance. The final part of this thesis investigates the hypothesis that rosetting is an immune evasion strategy of P. falciparum in order to evade phagocytic clearance, in particular that mediated by CD36. Overall the data obtained were inconsistent. Phagocytosis was significantly reduced in rosetting versus non-rosetting parasites in some strains (e.g. R29; P=0.048, paired T test), whereas others showed no effect (e.g. Muz12; P=0.228, paired T test) or increased versus non-rosetting parasites (e.g. HB3, P=0.004, paired T test). The relationship between CD36 binding and phagocytosis was also unclear, and anti-CD36 antibody did not effectively block phagocytosis, suggesting the involvement of alternative mechanisms. Further experiments are needed to clarify these observations. Data presented in this thesis are suggestive that rosetting in non-group O blood may be an immune evasion strategy with regard to IFNg production by innate immune cells, mechanistically linking rosetting with enhanced parasitaemia and disease severity. Furthermore, parasite strain significantly affects cytokine production and may be a determinant of disease outcome. This thesis demonstrates the importance of continued research into the effect of parasite virulence on the immune response, with particular emphasis on rosetting.

The aim of this dissertation is to develop theoretical models for the motion of rigid and flexible particles through very tight spaces. The geometries of conduits which will be investigated are cylindrical tubes, parallel plane walls and rectangular channels. This work is motivated by an interest in the flow and deformation of single red blood cells in very narrow capillaries, in spleen and in bone marrow. Mammalian red cells are highly flexible, but their deformations satisfy two significant constraints. They must deform at constant volume, because the contents of the cell are incompressible, and also at nearly constant surface area, because the red cell membrane strongly resists dilation. Consequently, there exists a minimal tube diameter below which passage of intact cells is not possible. A cell in a tube with this diameter has its critical shape: a cylinder with hemispherical ends. The motion of red cells is analysed using lubrication theory. When the tube diameter is slightly larger than the minimal value, the cell shape is close to its shape in the critical case. However, the rear end of the cell becomes flattened and then concave with a relatively small further increase in the diameter. The changes in cell shape and the resulting rheological parameters are analysed using matched asymptotic expansions for the high-velocity limit and using numerical solutions. A rapid decrease in the apparent viscosity of red cell suspensions with increasing tube diameter is predicted over the range of diameters considered. The red cell velocity is found to exceed the mean bulk velocity by an amount which increases with increasing tube diameter. The same type of analysis is applied to the flow and deformation of red blood cells between two parallel plates with near-minimal spacings. First, the critical shape of the particle and the minimum gap width are determined using calculus of variations. In this case, it is a disk with a rounded edge. The flow in the plasma layers between the cell and the plates is described using lubrication theory. Approximate solutions can be obtained using a locally two-dimensional analysis at each point of the rim of the cell. Cell shapes, pressure distributions, membrane stresses and particle velocities are deduced as functions of geometrical parameters. One significant finding is that the gap width between the cell and the wall decreases with distance from the axis of symmetry parallel to the flow direction. The red cell velocity may be smaller or larger than the mean fluid velocity far from the cell, depending on the spacing of the plates, with equality when the width of the red cell is about ninety percent of the spacing between plates. The same procedure is also applied to rectangular channels.

IT2018

by Kin-wai Lee. / Includes bibliographies / Thesis (M.Phil.)--Chinese University of Hong Kong, 1982

Thesis. 1978. Ph.D.--Massachusetts Institute of Technology. Dept. of Mechanical Engineering. / MICROFICHE COPY AVAILABLE IN ARCHIVES AND ENGINEERING. / Vita. / Includes bibliographical references. / by Thomas H. Papanek. / Ph.D.

Red blood cell (RBC) transfusions are associated with adverse effects, such as an increased risk of bacterial infection. In preparing RBCs for transfusion, donor RBCs are refrigerator stored for extended periods of time, during which they undergo oxidative damage, ultimately leading to their rapid post-transfusion clearance from the circulation. Macrophages play important roles in recycling iron derived from the clearance of RBCs. They are also a critically important component of host defense, protecting against invading pathogens. However, the effects on macrophage biology of acutely ingesting large numbers of RBCs are not completely understood. To investigate this issue, we used a mouse model of RBC transfusion and clearance, which mimics the clinical setting. In this model, transfusions of refrigerator storage-damaged (i.e., “old”) RBCs led to increased erythrophagocytosis by splenic red pulp macrophages (RPMs). This robust erythrophagocytosis induced ferroptosis, an iron-dependent form of cell death, in RPMs. This was accompanied by increases in reactive oxygen species and lipid peroxidation in vivo, which were reduced by treatment in vitro with ferrostatin-1, a ferroptosis inhibitor. Old RBC transfusions also induced RPM-dependent chemokine expression by splenic Ly6Chi monocytes, which signaled Ly6Chi monocyte migration from bone marrow to spleen, where these cells subsequently differentiated into RPMs. The combination of cell division among remaining splenic RPMs, along with the influx of bone marrow-derived Ly6Chi monocytes, suggests that, following RPM depletion induced by robust erythrophagocytosis, there is a coordinated effort to restore homeostasis of the RPM population by local self-maintenance and contributions from circulating monocytes. However, the effects on the overall functioning of the splenic Ly6Chi monocytes and remaining RPMs are unclear, especially their responses to subsequent immune challenges. In a mouse model of RBC storage and transfusion, we found that, following a transfusion of old RBCs, macrophages were less capable of phagocytosing a subsequent particle stimulus, such as bacteria (i.e., Escherichia coli and Staphylococcus aureus) or additional old RBCs. However, splenic Ly6Chi monocytes became activated in a specific timeframe following the initial old RBC transfusion, thereby increasing their phagocytic capacity. Nonetheless, despite contributions from activated splenic Ly6Chi monocytes, RPM function was indispensable for clearing S. aureus; this functional impairment may make the transfusion recipient susceptible to S. aureus sepsis. In conclusion, these findings may be clinically relevant to pathological conditions that can arise as a result of increased erythrophagocytosis, such as transfusion-related immunomodulation and impaired host immunity.

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No description available.