A novel CSP C-terminal epitope targeted by an antibody with protective activity against Plasmodium falciparum

Potent and durable vaccine responses will be required for control of malaria caused by Plasmodium falciparum (Pf). RTS,S/AS01 is the first, and to date, the only vaccine that has demonstrated significant reduction of clinical and severe malaria in endemic cohorts in Phase 3 trials. Although the vaccine is protective, efficacy declines over time with kinetics paralleling the decline in antibody responses to the Pf circumsporozoite protein (PfCSP). Although most attention has focused on antibodies to repeat motifs on PfCSP, antibodies to other regions may play a role in protection. Here, we expressed and characterized seven monoclonal antibodies to the C-terminal domain of CSP (ctCSP) from volunteers immunized with RTS,S/AS01. Competition and crystal structure studies indicated that the antibodies target two different sites on opposite faces of ctCSP. One site contains a polymorphic region (denoted α-ctCSP) and has been previously characterized, whereas the second is a previously undescribed site on the conserved β-sheet face of the ctCSP (denoted β-ctCSP). Antibodies to the β-ctCSP site exhibited broad reactivity with a diverse panel of ctCSP peptides whose sequences were derived from field isolates of P. falciparum whereas antibodies to the α-ctCSP site showed very limited cross reactivity. Importantly, an antibody to the β-site demonstrated inhibition activity against malaria infection in a murine model. This study identifies a previously unidentified conserved epitope on CSP that could be targeted by prophylactic antibodies and exploited in structure-based vaccine design.     

A cross-neutralizing antibody between HIV-1 and influenza virus

Incessant antigenic evolution enables the persistence and spread of influenza virus in the human population. As the principal target of the immune response, the hemagglutinin (HA) surface antigen on influenza viruses continuously acquires and replaces N-linked glycosylation sites to shield immunogenic protein epitopes using host-derived glycans. Anti-glycan antibodies, such as 2G12, target the HIV-1 envelope protein (Env), which is even more extensively glycosylated and contains under-processed oligomannose-type clusters on its dense glycan shield. Here, we illustrate that 2G12 can also neutralize human seasonal influenza A H3N2 viruses that have evolved to present similar oligomannose-type clusters on their HAs from around 20 years after the 1968 pandemic. Using structural biology and mass spectrometric approaches, we find that two N-glycosylation sites close to the receptor binding site (RBS) on influenza hemagglutinin represent the oligomannose cluster recognized by 2G12. One of these glycan sites is highly conserved in all human H3N2 strains and the other emerged during virus evolution. These two N-glycosylation sites have also become crucial for fitness of recent H3N2 strains. These findings shed light on the evolution of the glycan shield on influenza virus and suggest 2G12-like antibodies can potentially act as broad neutralizers to target human enveloped viruses.     

Vacuolar H+-ATPase is involved in preventing heavy metal-induced oxidative stress in Saccharomyces cerevisiae

In Saccharomyces cerevisiae, vacuolar H⁺-ATPase (V-ATPase) involved in the regulation of intracellular pH homeostasis has been shown to be important for tolerances to cadmium, cobalt and nickel. However, the molecular mechanism underlying the protective role of V-ATPase against these metals remains unclear. In this study, we show that cadmium, cobalt and nickel disturbed intracellular pH balance by triggering cytosolic acidification and vacuolar alkalinization, likely via their membrane permeabilizing effects. Since V-ATPase plays a crucial role in pumping excessive cytosolic protons into the vacuole, the metal-sensitive phenotypes of the Δvma2 and Δvma3 mutants lacking V-ATPase activity were supposed to result from highly acidified cytosol. However, we found that the metal-sensitive phenotypes of these mutants were caused by increased production of reactive oxygen species, likely as a result of decreased expression and activities of manganese superoxide dismutase and catalase. In addition, the loss of V-ATPase function led to aberrant vacuolar morphology and defective endocytic trafficking. Furthermore, the sensitivities of the Δvma mutants to other chemical compounds (i.e. acetic acid, H₂O₂, menadione, tunicamycin and cycloheximide) were a consequence of increased endogenous oxidative stress. These findings, therefore, suggest the important role of V-ATPase in preventing endogenous oxidative stress induced by metals and other chemical compounds.     

Uterine artery flow velocity waveform,arterial flow indices,follicular dynamics,and sex hormones during preovulatory period in synchronized ovulatory cycle of Bos indicus beef cows

A greater understanding of the uterine artery’s (UtA) biology is essential to the increase in female reproductive abilities. The UtA flow velocity waveform, blood flow volume (BFV), pulsatility and resistance indices (PI and RI), blood flow velocities, dynamics of the dominant follicle (DF), and estradiol (E2) and progesterone (P4) levels in an induced ovulatory cycle were evaluated in Thai native cattle. Twenty cows were induced with synchronized ovulation through a P4-releasing device, from Day ?9 to Day ?4, concurrent with the administration of two doses of a gonadotropin-releasing hormone on Day ?9 and Day ?1, and two doses of prostaglandin F_2α on Day ?4 and 8?h later. Day 0 was designated as the day of ovulation. The cows underwent Doppler sonographic determination and blood collection from Day ?4 to Day 0. The cows were classified in the non-ovulating (n?=?5) and ovulating groups (n?=?15). The ovulating cows presented higher BFV values, blood flow velocities, DF growth rates, and E2 levels; yet lower PI values and P4 concentrations, than those of the non-ovulating cows. The BFV values and the blood flow velocities were greater, but the RI and PI values were lower in the ovulatory side UtA than in the contraovulatory side UtA. The BFV values were positively correlated with blood flow velocities, DF growth rates and E2 concentrations in the ovulating cows; confirming the importance of UtA blood flow, follicular growth, and E2-vasodilation during preovulatory phase in the induced ovulatory cycle of Bos indicus beef cows.