SARS-CoV-2 mutations in MHC-I-restricted epitopes evade CD8+ T cell responses.

CD8+ T cell immunity to SARS-CoV-2 has been implicated in COVID-19 severity and virus control. Here, we identified nonsynonymous mutations in MHC-I-restricted CD8+ T cell epitopes after deep sequencing of 747 SARS-CoV-2 virus isolates. Mutant peptides exhibited diminished or abrogated MHC-I binding in a cell-free in vitro assay. Reduced MHC-I binding of mutant peptides was associated with decreased proliferation, IFN-γ production and cytotoxic activity of CD8+ T cells isolated from HLA-matched COVID-19 patients. Single cell RNA sequencing of ex vivo expanded, tetramer-sorted CD8+ T cells from COVID-19 patients further revealed qualitative differences in the transcriptional response to mutant peptides. Our findings highlight the capacity of SARS-CoV-2 to subvert CD8+ T cell surveillance through point mutations in MHC-I-restricted viral epitopes.

Profiling antibody responses to infections by Chlamydia abortus enables identification of potential virulence factors and candidates for serodiagnosis.

Enzootic abortion of ewes (EAE) due to infection with the obligate intracellular pathogen Chlamydia (C.) abortus is an important zoonosis leading to considerable economic loss to agriculture worldwide. The pathogen can be transmitted to humans and may lead to serious infection in pregnant women. Knowledge about epidemiology, clinical course and transmission to humans is hampered by the lack of reliable diagnostic tools. Immunoreactive proteins, which are expressed in infected animals and humans, may serve as novel candidates for diagnostic marker proteins and represent putative virulence factors. In order to broaden the spectrum of immunogenic C. abortus proteins we applied 2D immunoblot analysis and screening of an expression library using human and animal sera. We have identified 48 immunoreactive proteins representing potential diagnostic markers and also putative virulence factors, such as CAB080 (homologue of the "macrophage infectivity potentiator", MIP), CAB167 (homologue of the "translocated actin recruitment protein", TARP), CAB712 (homologue of the "chlamydial protease-like activity factor", CPAF), CAB776 (homologue of the "Polymorphic membrane protein D", PmpD), and the "hypothetical proteins" CAB063, CAB408 and CAB821, which are predicted to be type III secreted. We selected two putative virulence factors for further characterization, i.e. CAB080 (cMIP) and CAB063, and studied their expression profiles at transcript and protein levels. Analysis of the subcellular localization of both proteins throughout the developmental cycle revealed CAB063 being the first C. abortus protein shown to be translocated to the host cell nucleus.

A synthetic peptide ELISA for the screening of rubella virus neutralizing antibodies in order to ascertain immunity.

One hundred and fifty-one human sera from patients exposed to rubella virus (RV) and shown to be negative for IgM antibodies were tested for total RV-IgG, hemagglutination inhibition (HAI) and for virus neutralizing (VN) antibodies using a peptide enzyme-linked immunosorbent assay (ELISA) based on BCH-178, a peptide representing one of several known neutralizing epitopes on RV hemagglutinin (E1). The data showed that, among 39 and 51 sera with HAI and RV-IgG titres of 1/128 and >150 IU/ml, respectively, neutralizing antibody readings using the BCH-178 ELISA were above cut-off values. However, 13% of HAI positive sera (titre > or =1/16) and 16% of RV-IgG ELISA positive sera (> or =20 IU/ml) were below the cut-off value of the BCH-178 ELISA. This may explain why several cases of congenital rubella syndrome (CRS) have been observed in spite of positive titres. We suggest that a diagnosis of sufficient immunity against RV infection or reinfection may be safer if an additional test detecting antibodies against VN RV epitopes is positive as well.