Reliable quantification of the antibody response to SARS-CoV-2 is highly relevant, e.g., for identifying possible vaccine failure and estimating the time of protection. We compared the performance of five CE marked tests that quantify antibodies against the viral spike protein.

The human pathogen Streptococcus pyogenes (group A streptococcus [GAS]) pilus components, suggested to play a role in pathogenesis, are encoded in the variable FCT (fibronectin-and collagen-binding T-antigen) region. We investigated the functions of sortase A (SrtA), sortase C2 (SrtC2), and the FctA protein of the most prevalent type 3 FCT region from a serotype M49 strain. Although it is considered a housekeeping sortase, SrtA's activity is involved in pilus formation in addition to its essentiality for GAS extracellular matrix protein binding, host cell adherence/internalization, survival in human blood, and biofilm formation. SrtC2 activity is crucial for pilus formation but dispensable for the other phenotypes tested in vitro. FctA is the major pilus backbone protein, simultaneously acting as the M49 T antigen, and requires SrtC2 and LepA, a signal peptidase I homologue, for monomeric surface expression and polymerization, respectively. Collagen-binding protein Cpa expression supports pilus formation at the pilus base. Immunofluorescence microscopy and fluorescence-activated cell sorting analysis revealed several unexpected expression patterns, as follows: (i) the monomeric pilus protein FctA was found exclusively at the old poles of GAS cells, (ii) FctA protein expression increased with lower temperatures, and (iii) FctA protein expression was restricted to 20 to 50% of a given GAS M49 population, suggesting regulation by a bistability mode. Notably, disruption of pilus assembly by sortase deletion rendered GAS serotype M49 significantly more aggressive in a dermonecrotic mouse infection model, indicating that sortase activity and, consequently, pilus expression allow a subpopulation of this GAS serotype to be less aggressive. Thus, pilus expression may not be a virulence attribute of GAS per se.Streptococcus pyogenes (group A streptococcus [GAS]) is a bacterial pathogen that is perfectly adapted to colonization, infection, and persistence in its human host (8,10,14,22). Many of the associated virulence factors expressed by this bacterium are encoded in discrete regions of the GAS genome (28). Two of these pathogenicity regions (Mga and FCT [see below]) each comprise genes for secreted and surface-exposed virulence factors and at least one stand-alone transcriptional regulator. These genes have been shown to act together in a growth phase-dependent regulatory network to coordinate GAS host cell adherence, internalization, and intracellular persistence (summarized in references 28 and 29).The FCT region (fibronectin-and collagen-binding T-antigen region) (6) is present in all GAS genomes specifically tested (26). The FCT region always contains a RALP transcriptional regulator whose type correlates with the type of emm pathogenicity region and the preferred infection site of the GAS strain, i.e., the throat or the skin. Since a similar association was also shown for the fibronectin-and collagenbinding proteins encoded by this region, the FCT region gene products could contribute to tissue-specific GAS infecti...

The diffusion dynamics of lipids and GPI-anchored proteins is investigated using superresolution STED microscopy combined with single-molecule fluorescence correlation spectroscopy in the cellular membranes. The actin cytoskeleton is shown to play an essential role in the diffusion characteristics of molecules.

Diffusion and interaction dynamics of molecules at the plasma membrane play an important role in cellular signalling. These have been suggested to be strongly associated with the actin cytoskeleton. Here, we utilise super-resolution STED microscopy combined with fluorescence correlation spectroscopy (STED-FCS) to access the sub-diffraction diffusion regime of different fluorescent lipid analogues and GPI-anchored proteins (GPI-APs) in the cellular plasma membrane, and compare it to the diffusion regime of these molecules in cell-derived actin-free giant plasma membrane vesicles (GPMVs). We show that phospholipids and sphingomyelin, which undergo hindered diffusion in the live cell membrane, diffuse freely in the GPMVs. In contrast to sphingomyelin, which is transiently trapped on molecular-scale complexes in intact cells, diffusion of the ganglioside lipid GM1 suggests transient incorporation into nanodomains, which is less influenced by the actin cortex. Finally, our data on GPI-APs indicate two molecular pools in living cells, one pool showing high mobility with trapped and compartmentalized diffusion, and the other forming immobile clusters both of which disappear in GPMVs. Our data underlines the crucial role of the actin cortex in maintaining hindered diffusion modes of most but not all membrane molecules.