Biology subjects

Ciminski, K.

Publications and source records attributed to Ciminski, K..

5 recordsLinked to original sources

bioRxiv2026

Bat-borne influenza A H9N2 virus shows species-specific replication and transmission

Bat-borne H9N2 IAV (bat H9N2), initially detected in bat feces in Egypt in 2017, has recently attracted attention because of its unexpected pre-pandemic properties, including airborne transmission between ferrets and efficient replication in human lung explant cultures. However, its host range and potential reservoir hosts remain poorly defined, particularly with respect to additional bat hosts and mammalian or avian mixing-vessel species. Here, we compare the susceptibility of two pteropodid bat species, Rousettus aegyptiacus and Eidolon helvum, and additionally assessed pigs and quails following intranasal inoculation with bat H9N2. Juvenile R. aegyptiacus showed only low-level viral replication and shedding, largely restricted to the upper respiratory tract, and did not show transmission to contact bats. In contrast, both juvenile and adult E. helvum, despite showing no clinical signs, showed high-level viral replication and shedding in the upper respiratory tract, with efficient and early transmission to contact bats. Viral RNA was most abundant in the respiratory turbinates, the nasopharyngeal meatus and the tracheal epithelium, where it was associated with moderate to severe necrotizing rhinitis and tracheitis. The lower respiratory tract was less involved, with viral RNA confined to scattered bronchioles and associated with focal necrotizing bronchiolitis. Other organs, including the gastrointestinal tract, lacked histologic lesions and viral RNA labeling. In pigs and quails, intranasal inoculation did not result in detectable replication, transmission, or seroconversion. Our findings demonstrate marked differences in the susceptibility to bat H9N2 between the two fruit bat species tested and identify E. helvum as a highly susceptible host capable of rapid and efficient transmission. Conversely, pigs and quails were not susceptible, suggesting that additional viral adaptations may be required to allow for a productive infection in these species.

microbiology↗

bioRxiv2026

MHC-II acts as a fusion-triggering receptor for bat influenza virus

Influenza A virus hemagglutinin is a prototypical class I viral fusion protein that binds sialylated glycans and is activated by low pH in endosomes. In contrast, bat-derived IAV subtypes H17N10 and H18N11 use major histocompatibility complex class II (MHC-II) as an entry receptor, but how this receptor contributes to membrane fusion remains unknown. We find that MHC-II-dependent hemagglutinin subtypes H17, H18, and H19 possess an increased negative net charge relative to canonical HAs. Using cryo-electron tomography, we demonstrate that H18N11 morphology remains stable and H18 is in prefusion conformation at strongly acidic pH. Remarkably, H18 undergoes fusion-relevant conformational changes only when both MHC-II binding and low pH are present. By reconstitution of H18N11 fusion with liposomes and purified MHC-II, we show that receptor engagement is required to trigger the fusion activity of H18. These findings identify MHC-II as a receptor that directly triggers membrane fusion and reveal a previously unrecognized receptor-dependent mechanism of influenza virus entry. SignificanceBat-derived influenza A viruses (IAV) challenged established paradigms because they use major histocompatibility complex class II (MHC-II) molecules instead of sialylated glycans for cell entry. Recent studies suggest that MHC-II usage extends beyond bats, with avian, swine, and human strains showing dual receptor specificity. However, how MHC-II contributes to membrane fusion has remained unclear. We show that, unlike conventional HAs that are activated by acidity alone, the H18 fusion protein requires receptor engagement and low pH to undergo fusion-relevant conformational changes leading to membrane fusion. Hence, we reveal a novel receptor-dependent mechanism of membrane fusion for the MHC-II dependent entry of H18N11.

microbiology↗

bioRxiv2024

The bat Influenza A virus subtype H18N11 induces nanoscale MHCII clustering upon host cell attachment

Prior to the discovery of the bat influenza A virus (IAV) subtypes H17N10 and H18N11, it was believed that all IAVs bind via the viral hemagglutinin (HA) to sialic acid residues to mediate attachment and subsequent viral entry. Both HA subtypes, H17 and H18, however, do not bind to sialic acids but instead engage a proteinaceous receptor: major histocompatibility complex class II (MHCII). The mechanistic details of this hitherto unknown protein-mediated entry are not understood. Since conventional IAVs require attachment to clusters of sialylated glycans to overcome the low affinity of the HA-sialic acid interaction, we hypothesized that bat HA would likewise interact with multiple MHCII molecules. Here, we used photoactivated localization microscopy (PALM) on fixed and live cells expressing MHCII fused to appropriate fluorescent reporters. We show that bat IAV particles attach to pre-existing MHCII clusters present on susceptible cells and that the local HA-MHCII interaction results in an increased cluster size. To measure the impact of viral attachment on the dynamics of MHCII, we utilized an experimental setup designated "inverse infection" where intact viral particles were immobilized on coverslips before live MHCII-expressing cells were seeded on top. Recording the trajectories of single MHCII molecules, this approach revealed that the mobility of MHCII was indeed slowed down in viral proximity leading to a local enrichment of MHCII molecules beneath the viral particle. Taken together, these data suggest that attachment of viral particles leads to clustering of MHCII, a process similar to the MHCII dynamics observed during the formation of an immunological synapse.

microbiology↗

bioRxiv2021

Engineered chimeric T cell receptor fusion construct (TRuC)-expressing T cells prevent translational shutdown in SARS-CoV-2-infected cells

SARS-CoV-2, the causative agent of Covid-19, is known to evade the immune system by several mechanisms. This includes the shutdown of the host cellular protein synthesis, which abrogates the induction of antiviral interferon responses. The virus initiates the infection of susceptible cells by binding with its spike protein (S) to the host angiotensin-converting enzyme 2 (ACE2). Here we applied the T cell receptor fusion construct (TRuC) technology to engineer T cells against such infected cells. In our TRuCs an S-binding domain is fused to the CD3{varepsilon} component of the T cell receptor (TCR) complex, enabling recognition of S-containing cells in an HLA independent manner. This domain either consists of the S-binding part of ACE2 or a single-chain variable fragment of an anti-S antibody. We show that the TRuC T cells are activated by and kill cells that express S of SARS-CoV-2 and its alpha (B.1.1.7) and beta (B.1.351) variants at the cell surface. Treatment of SARS-CoV-2 infected cells with our engineered T cells did not lead to massive cytotoxicity towards the infected cells, but resulted in a complete rescue of the translational shutdown despite ongoing viral replication. Our data show that engineered TRuC T cell products might be used against SARS-CoV-2 by exposing infected cells to the host innate immune system.

bioengineering↗

bioRxiv2021

Enhanced FcγRIII/CD16 activation by discrete ligands as independent correlates of disease severity in COVID-19 patients

A dysregulated immune response with high levels of SARS-CoV-2 specific IgG antibodies characterizes patients with severe or critical COVID-19. Although a robust IgG response is traditionally considered to be protective, excessive triggering of activating Fc-gamma-receptors (Fc{gamma}Rs) could be detrimental and cause immunopathology. Here, we document that patients who develop soluble circulating IgG immune complexes (sICs) during infection are subject to enhanced immunopathology driven by Fc{gamma}R activation. Utilizing cell-based reporter systems we provide evidence that sICs are predominantly formed prior to a specific humoral response against SARS-CoV-2. sIC formation, together with increased afucosylation of SARS-CoV-2 specific IgG eventually leads to an enhanced CD16 (Fc{gamma}RIII) activation of immune cells reaching activation levels comparable active systemic lupus erythematosus (SLE) disease. Our data suggest a vicious cycle of escalating immunopathology driven by an early formation of sICs in predisposed patients. These findings reconcile the seemingly paradoxical findings of high antiviral IgG responses and systemic immune dysregulation in severe COVID-19. Clinical implicationsThe identification of sICs as drivers of an escalating immunopathology in predisposed patients opens new avenues regarding intervention strategies to alleviate critical COVID-19 progression. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=78 SRC="FIGDIR/small/449893v4_ufig1.gif" ALT="Figure 1"> View larger version (17K): org.highwire.dtl.DTLVardef@b4616corg.highwire.dtl.DTLVardef@682d1aorg.highwire.dtl.DTLVardef@16946cborg.highwire.dtl.DTLVardef@a6ef7d_HPS_FORMAT_FIGEXP M_FIG C_FIG A vicious cycle of immunopathology in COVID-19 patients is driven by soluble multimeric immune complexes (sICs). SARS-CoV-2 infection triggers sIC formation in prone individuals. Activation of Fc{gamma}RIII/CD16 expressing immune cells by sICs precedes a humoral response to SARS-CoV2 infection. sICs and infection add to IgG afucosylation, further enhancing Fc{gamma}RIII/CD16 activation by opsonized targets. High inflammation induces further sIC mediated immune cell activation ultimately leading to an escalating immunopathology.

immunology↗