Complement Factor H Family Proteins Modulate Monocyte and Neutrophil Granulocyte Functions.

Besides being a key effector arm of innate immunity, a plethora of non-canonical functions of complement has recently been emerging. Factor H (FH), the main regulator of the alternative pathway of complement activation, has been reported to bind to various immune cells and regulate their functions, beyond its role in modulating complement activation. In this study we investigated the effect of FH, its alternative splice product FH-like protein 1 (FHL-1), the FH-related (FHR) proteins FHR-1 and FHR-5, and the recently developed artificial complement inhibitor mini-FH, on two key innate immune cells, monocytes and neutrophilic granulocytes. We found that, similar to FH, the other factor H family proteins FHL-1, FHR-1 and FHR-5, as well as the recombinant mini-FH, are able to bind to both monocytes and neutrophils. As a functional outcome, immobilized FH and FHR-1 inhibited PMA-induced NET formation, but increased the adherence and IL-8 production of neutrophils. FHL-1 increased only the adherence of the cells, while FHR-5 was ineffective in altering these functions. The adherence of monocytes was increased on FH, recombinant mini-FH and FHL-1 covered surfaces and, except for FHL-1, the same molecules also enhanced secretion of the inflammatory cytokines IL-1ß and TNFα. When monocytes were stimulated with LPS in the presence of immobilized FH family proteins, FH, FHL-1 and mini-FH enhanced whereas FHR-1 and FHR-5 decreased the secretion of TNFα; FHL-1 and mini-FH also enhanced IL-10 release compared to the effect of LPS alone. Our results reveal heterogeneous effects of FH and FH family members on monocytes and neutrophils, altering key features involved in pathogen killing, and also demonstrate that FH-based complement inhibitors, such as mini-FH, may have effects beyond their function of inhibiting complement activation. Thus, our data provide new insight into the non-canonical functions of FH, FHL-1, FHR-1 and FHR-5 that might be exploited during protection against infections and in vaccine development.

Interaction of the Factor H Family Proteins FHR-1 and FHR-5 With DNA and Dead Cells: Implications for the Regulation of Complement Activation and Opsonization.

Complement plays an essential role in the opsonophagocytic clearance of apoptotic/necrotic cells. Dysregulation of this process may lead to inflammatory and autoimmune diseases. Factor H (FH), a major soluble complement inhibitor, binds to dead cells and inhibits excessive complement activation on their surface, preventing lysis, and the release of intracellular material, including DNA. The FH-related (FHR) proteins share common ligands with FH, due to their homology with this complement regulator, but they lack the domains that mediate the complement inhibitory activity of FH. Because their roles in complement regulation is controversial and incompletely understood, we studied the interaction of FHR-1 and FHR-5 with DNA and dead cells and investigated whether they influence the regulatory role of FH and the complement activation on DNA and dead cells. FH, FHR-1, and FHR-5 bound to both plasmid DNA and human genomic DNA, where both FHR proteins inhibited FH-DNA interaction. The FH cofactor activity was inhibited by FHR-1 and FHR-5 due to the reduced binding of FH to DNA in the presence of the FHRs. Both FHRs caused increased complement activation on DNA. FHR-1 and FHR-5 bound to late apoptotic and necrotic cells and recruited monomeric C-reactive protein and pentraxin 3, and vice versa. Interactions of the FHRs with pentraxins resulted in enhanced activation of both the classical and the alternative complement pathways on dead cells when exposed to human serum. Altogether, our results demonstrate that FHR-1 and FHR-5 are competitive inhibitors of FH on DNA; moreover, FHR-pentraxin interactions promote opsonization of dead cells.

Strain Pattern in Supercooled Liquids.

Investigations of strain correlations at the glass transition reveal unexpected phenomena. The shear strain fluctuations show an Eshelby-strain pattern [∼cos(4θ)/r^{2}], characteristic of elastic response, even in liquids, at long times. We address this using a mode-coupling theory for the strain fluctuations in supercooled liquids and data from both video microscopy of a two-dimensional colloidal glass former and simulations of Brownian hard disks. We show that the long-ranged and long-lived strain signatures follow a scaling law valid close to the glass transition. For large enough viscosities, the Eshelby-strain pattern is visible even on time scales longer than the structural relaxation time τ and after the shear modulus has relaxed to zero.

Tests of mode-coupling theory in two dimensions.

We analyze the glassy dynamics of binary mixtures of hard disks in two dimensions. Predictions of the mode-coupling theory (MCT) are tested with extensive Brownian dynamics simulations. Measuring the collective particle density correlation functions in the vicinity of the glass transition, we verify four predicted mixing effects. For instance, for large size disparities, adding a small amount of small particles at a fixed packing fraction leads to a speedup in the long-time dynamics, while for small size disparities it leads to a slowing-down. Qualitative features of the nonergodicity parameters and the ß relaxation, which both depend in a nontrivial way on the mixing ratio, are found in the simulated correlators. Studying one system in detail, we are able to determine its ideal MCT glass transition point as φ(c)=0.7948 and test MCT predictions quantitatively.

Glass transition in confined geometry.

Extending mode-coupling theory, we elaborate a microscopic theory for the glass transition of liquids confined between two parallel flat hard walls. The theory contains the standard mode-coupling theory equations in bulk and in two dimensions as limiting cases and requires as input solely the equilibrium density profile and the structure factors of the fluid in confinement. We evaluate the phase diagram as a function of the distance of the plates for the case of a hard sphere fluid and obtain an oscillatory behavior of the glass transition line as a result of the structural changes related to layering.

Effect of mixing and spatial dimension on the glass transition.

We study the influence of composition changes on the glass transition of binary hard disk and hard sphere mixtures in the framework of mode coupling theory. We derive a general expression for the slope of a glass transition line. Applied to the binary mixture in the low concentration limits, this method allows a fast prediction of some properties of the glass transition lines. The glass transition diagram we find for binary hard disks strongly resembles the random close packing diagram. Compared to three dimensions from previous studies, the extension of the glass regime due to mixing is much more pronounced in two dimensions where plasticization only sets in at larger size disparities. For small size disparities we find a stabilization of the glass phase quadratic in the deviation of the size ratio from unity.

Delocalization-localization transition due to anharmonicity.

Analytical and numerical calculations for a reduced Fermi-Pasta-Ulam chain demonstrate that energy localization does not require more than one conserved quantity. Clear evidence for the existence of a sharp delocalization-localization transition at a critical amplitude A_c is given. Approaching A_c from above and below, diverging time scales occur. Above A_c, the energy packet converges towards a discrete breather. Nevertheless, ballistic energy transportation is present, demonstrating that its existence does not necessarily imply delocalization.