Spike-specific antibodies are central to effective COVID19 immunity. Research efforts have focused on antibodies that neutralize the ACE2-Spike interaction but not on non-neutralizing antibodies. Antibody-dependent phagocytosis is an immune mechanism enhanced by opsonization, where typically, more bound antibodies trigger a stronger phagocyte response. Here, we show that Spike-specific antibodies, dependent on concentration, can either enhance or reduce Spike-bead phagocytosis by monocytes independently of the antibody neutralization potential. Surprisingly, we find that both convalescent patient plasma and patient-derived monoclonal antibodies lead to maximum opsonization already at low levels of bound antibodies and is reduced as antibody binding to Spike protein increases. Moreover, we show that this Spike-dependent modulation of opsonization correlate with the outcome in an experimental SARS-CoV-2 infection model. These results suggest that the levels of anti-Spike antibodies could influence monocyte-mediated immune functions and propose that non-neutralizing antibodies could confer protection to SARS-CoV-2 infection by mediating phagocytosis.

Group A streptococci have evolved multiple strategies to evade human antibodies, making it challenging to create effective vaccines or antibody treatments. Here, we have generated antibodies derived from the memory B cells of an individual who had successfully cleared a group A streptococcal infection. The antibodies bind with high affinity in the central region of the surface‐bound M protein. Such antibodies are typically non‐opsonic. However, one antibody could effectively promote vital immune functions, including phagocytosis and in vivo protection. Remarkably, this antibody primarily interacts through a bivalent dual‐Fab cis mode, where the Fabs bind to two distinct epitopes in the M protein. The dual‐Fab cis‐binding phenomenon is conserved across different groups of M types. In contrast, other antibodies binding with normal single‐Fab mode to the same region cannot bypass the M protein's virulent effects. A broadly binding, protective monoclonal antibody could be a candidate for anti‐streptococcal therapy. Our findings highlight the concept of dual‐Fab cis binding as a means to access conserved, and normally non‐opsonic regions, regions for protective antibody targeting.

Antibodies play a central role in the immune defense against SARS-CoV-2. Emerging evidence has shown that nonneutralizing antibodies are important for immune defense through Fc-mediated effector functions. Antibody subclass is known to affect downstream Fc function. However, whether the antibody subclass plays a role in anti-SARS-CoV-2 immunity remains unclear. Here, we subclass-switched eight human IgG1 anti-spike monoclonal antibodies (mAbs) to the IgG3 subclass by exchanging their constant domains. The IgG3 mAbs exhibited altered avidities to the spike protein and more potent Fc-mediated phagocytosis and complement activation than their IgG1 counterparts. Moreover, combining mAbs into oligoclonal cocktails led to enhanced Fc- and complement receptor-mediated phagocytosis, superior to even the most potent single IgG3 mAb when compared at equivalent concentrations. Finally, in an in vivo model, we show that opsonic mAbs of both subclasses can be protective against a SARS-CoV-2 infection, despite the antibodies being nonneutralizing. Our results suggest that opsonic IgG3 oligoclonal cocktails are a promising idea to explore for therapy against SARS-CoV-2, its emerging variants, and potentially other viruses.

One hundred and three Streptococcus pyogenes isolates recovered mainly from streptococcal throat infections in Lebanon were characterized by emm and PFGE typing. Thirty-three emm types and subtypes were detected among the isolates. PFGE was more discriminatory as a typing method. The prevalent emm types were emm1 (12.6 %), emm22 (8.7 %), emm28 (7.7 %), emm88 (7.7 %) and emm4 (6.8 %) and all isolates were susceptible to vancomycin and penicillin G. Ten per cent of the isolates were resistant to erythromycin and 3 % were resistant to erythromycin and clindamycin, showing the macrolide-lincosamide-streptogramin B phenotype. The emm sequences and PFGE pattern database that were generated in this study will serve as a basis for information for long-term evolutionary and epidemiological studies of local S. pyogenes recovered not only in Lebanon, but also in neighbouring countries. INTRODUCTIONStreptococcus pyogenes is one of the most clinically important Gram-positive pathogens. S. pyogenes causes disease states that vary between mild sore throat to the flesh-eating disease necrotizing fasciitis. S. pyogenes causes, other than the acute diseases, a set of severe post-disease sequelae such as rheumatic fever and post-streptococcal glomerulonephritis (Yoonim et al., 2005).The successful nature of S. pyogenes as a pathogen relates to its wide array of virulence factors, be they secreted or cell bound. S. pyogenes strains express a multitude of genes coding for DNases, antibiotic resistance and the highly virulent superantigens that cause streptococcal toxic shock syndrome in addition to other nonsuppurative sequelae (Aziz & Kotb, 2008). S. pyogenes also exhibits C5a peptidase, streptolysin O and streptolysin S activity, which aid in streptococcal cytotoxicity and virulence (Sagar et al., 2008). The S. pyogenes M protein is probably the most notorious virulence factor that this micro-organism possesses. It is a fibrillar cell wall protein that aids in adherence to human cells and prevention of opsonophagocytosis. This protein, due to molecular mimicry within protein sequences found in human tissues, is implicated in rheumatic fever (Guilherme et al., 2006). S. pyogenes emm typing has become one of the best approaches used in molecular typing of this microorganism. Many methods have been exploited to best type S. pyogenes, such as 16S rRNA gene sequencing (Stanley et al., 1995), restriction fragment length polymorphism analysis (Desai et al., 1999), random amplified polymorphic DNA analysis (Seppälä et al., 1994) and PFGE (Chiou et al., 2004). However, sequencing-based emm typing by the use of oligonucleotides that target the Nterminus of the M-protein coding gene remains the most practical method of S. pyogenes typing (Beall et al., 2000). The reason behind this is that the gene coding for the group A Streptococcus M protein contains a hypervariable 59 region that is subject to many single nucleotide polymorphisms, which serves as the basis for emm typing S. pyogenes isolates (Yoonim et al., 2005).To our knowledge, this ...

The fungal pathogen Candida albicans is a leading causative agent of death in immunocompromised individuals. Many factors have been implicated in virulence including filamentation-inducing transcription factors, adhesins, lipases and proteases. Many of these factors are glycosylphosphatidylinositol-anchored cell surface antigenic determinant proteins. Pga1 is one such protein shown to be upregulated during cell wall regeneration. The purpose of this study was to characterise the role Pga1 plays by creating a homozygous pga1 null strain and comparing the phenotype with the parental strain. It was observed that the mutant strain showed less oxidative stress tolerance and an increased susceptibility to calcofluor white, a cell surface disrupting agent that inhibits chitin fibre assembly which translated as a 40% decrease in cell wall chitin content. Furthermore, the mutant exhibited a 50% reduction in adhesion and a 33% reduction in biofilm formation compared with the parental strain, which was reflected as a slight reduction in virulence. Our data suggest that Pga1 plays an important role in cell wall rigidity and stability. It was also observed that the pga1 null was over filamentous on both liquid and solid media and exhibited increased resistance to SDS suggesting upregulation of filamentation-inducing genes and cell surface components to partially compensate for the deletion.