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Extensive studies with subtype A BG505-derived HIV envelope glycoprotein (Env) immunogens have revealed that the dominant autologous neutralizing epitope in rabbits is located in an exposed region of the heavily glycosylated trimer that lacks potential N-linked glycosylation sites at positions 230, 241, and 289. The Env derived from B41, a subtype B virus, shares a glycan hole centered on positions 230 and 289. To test whether broader neutralization to the common glycan hole can be achieved, we immunized rabbits with B41 SOSIP (gp120-gp41 disulfide [SOS] with an isoleucine-to-proline mutation [IP] in gp41) alone, as well as B41 and BG505 coimmunization. We isolated autologous neutralizing antibodies (nAbs) and described their structure in complex with the B41 Env. Our data suggest that distinct autologous nAb lineages are induced by BG505 and B41 immunogens, even when both were administered together. In contrast to previously described BG505 glycan hole antibodies, the B41-specific nAbs accommodate the >97% conserved N241 glycan, which is present in B41. Single-particle cryo-electron microscopy studies confirmed that B41- and BG505-specific nAbs bind to overlapping glycan hole epitopes. We then used our high-resolution data to guide mutations in the BG505 glycan hole epitope in an attempt to broaden the reactivity of a B41-specific nAb, but we recovered only partial binding. Our data demonstrate that the lack of cross-reactivity in glycan hole antibodies is due to amino acid differences within the epitope, and our attempts to rationally design cross-reactive trimers resulted in only limited success. Thus, even for the immunodominant glycan hole shared between BG505 and B41, the prospect of designing prime-boost immunogens remains difficult. IMPORTANCE A glycan hole is one of the most dominant autologous neutralizing epitopes targeted on BG505 and B41 SOSIP trimer-immunized rabbits. Our high-resolution cryo-electron microscopy (cryoEM) studies of B41 in complex with a B41-specific antibody complex elucidate the molecular basis of this strain-specific glycan hole response. We conclude that even for the immunodominant glycan hole shared between BG505 and B41, the prospect of designing prime-boost immunogens remains difficult., (Copyright © 2020 Yang et al.)

Influenza A virus infection is a persistent threat to public health worldwide due to its ability to evade immune surveillance through rapid genetic drift and shift. Current vaccines against influenza A virus provide immunity to viral isolates that are similar to vaccine strains. High-affinity neutralizing antibodies against conserved epitopes could provide immunity to diverse influenza virus strains and protection against future pandemic viruses. In this study, by using a highly sensitive H5N1 pseudotype-based neutralization assay to screen human monoclonal antibodies produced by memory B cells from an H5N1-infected individual and molecular cloning techniques, we developed three fully human monoclonal antibodies. Among them, antibody 65C6 exhibited potent neutralization activity against all H5 clades and subclades except for subclade 7.2 and prophylactic and therapeutic efficacy against highly pathogenic avian influenza H5N1 viruses in mice. Studies on hemagglutinin (HA)-antibody complexes by electron microscopy and epitope mapping indicate that antibody 65C6 binds to a conformational epitope comprising amino acid residues at positions 118, 121, 161, 164, and 167 (according to mature H5 numbering) on the tip of the membrane-distal globular domain of HA. Thus, we conclude that antibody 65C6 recognizes a neutralization epitope in the globular head of HA that is conserved among almost all divergent H5N1 influenza stains.

The rate of isolation of organisms resembling Haemophilus vaginalis (Corynebacterium vaginale) from vaginal specimens was not significantly affected by anaerobic versus carbon dioxide incubation atmospheres or whether specimens were inoculated on isolation media immediately after collection or after a delay of 6 h. Forty-one clinically isolated strains were provisionally divided into 30 H. vaginalis strains and 11 H. vaginalis-like (HVL) strains based on morphological and growth characteristics. The H. vaginalis strains were less reactive in API-20A identification test strips, (Analytab Products, Inc.) using Lombard-Dowell broth, than in a modified basal medium that contained proteose peptone no. 3 (Difco). The numbers and kinds of substrates fermented by 30 clinical and 2 reference strains of H. vaginalis varied among conventional, API, Minitek (Baltimore Biological Laboratory), and rapid buffered substrate fermentation systems. A greater number and variety of carbohydrates were fermented by the 11 HVL strains more consistently in all four test systems. Analysis of volatile and nonvolatile fermentation end products by gas-liquid chromatography did not reveal significant differences between the H. vaginalis and HVL strains. However, the latter group grew in peptone-yeast extract-glucose broth, whereas the H. vaginalis strains did not grow without the addition of starch to peptone-yeast extract-glucose. All of the reference and clinical strains were similar in their susceptibilities to a variety of antimicrobial compounds except sulfonamides, which inhibited the HVL strains and bifidobacteria but not the H. vaginalis strains. Sulfonamide susceptibility or resistance corresponded in part to the H. vaginalis and HVL-bifidobacteria strain reactions on selected conventional fermentation substrates. Susceptibility or resistance to sulfonamides and metronidazole in conjunction with fermentation tests is described to aid in the separation of H. vaginalis from other possibly unrecognized biotypes of H. vaginalis or other vaginal bacteria that presumptively resemble the organism. A human blood medium known as V agar was also of considerable value in distinguishing H. vaginalis from HVL strains, because only the H. vaginalis strains produced diffuse beta-hemolysis on V agar.