Such a limitation might also apply to the N-terminal RGD neutralization target on the KSHV gB (Akula em et al

Such a limitation might also apply to the N-terminal RGD neutralization target on the KSHV gB (Akula em et al. /em , 2002). Alternatively, antibody-bound gBs may still retain Trifloxystrobin an appreciable Rabbit polyclonal to ZBED5 chance of participating in fusion, given the right cellular setting. infection completely and blocked the infection of NMuMG epithelial cells hardly at all. Virions saturated with antibody also remained infectious to mice. Thus, the MHV-68 gB presents at best a very difficult target for antibody-mediated neutralization. INTRODUCTION Herpesviruses are among the most successful of all vertebrate parasites. They robustly establish persistent, productive infections of immunocompetent hosts. This is achieved without generalized immunosuppression, tolerance of viral gene products or significant viral antigenic variation. For example, the murine cytomegalovirus glycoprotein B (gB) is immunogenic, yet the region associated with neutralization is antigenically conserved in field isolates (Xu (Fenner significance of this neutralization. Partial neutralization of Kaposi’s sarcoma-associated herpesvirus (KSHV) has been reported for rabbit sera raised against recombinant gH, gL (Naranatt neutralizing activity of human immune sera (Dialyna and by the limited opportunities for manipulation BL21 and induction with IPTG (Boname (Fig.?2d). Their epitope(s) were therefore independent of neutralization with single or combined neutralizing mAbs by adding saturating amounts of gB-specific and gH/gL-specific neutralizing mAbs to MHV-68 virions and then infecting mice intranasally with the equivalent of 1?p.f.u. each (Fig.?6f). All of the mice became infected. Thus, at least some cells in the respiratory tract remained susceptible to infection by antibody-coated virions, i.e. NMuMG cells represented neutralization more faithfully than BHK-21 cells. High antibody avidity is crucial for gB-directed neutralization Secreted, pentameric IgM has a valency five times that of IgG. This high avidity helps to preserve suboptimal binding interactions in early antibody responses. However, MHV-68 does not induce sizeable long-term IgM responses C the virus is essentially a T cell-dependent antigen that elicits IgG (Stevenson & Doherty, 1999; Sangster context, we have begun to analyse the glycoprotein-specific antibody response of MHV-68-infected mice. The MHV-68 gB is an obvious candidate for neutralization: it is a highly conserved virion protein that is essential for infectivity and is equivalent to a reported neutralization target on other herpesviruses. However, although gB-specific mAbs could readily be derived, gB-specific neutralizing mAbs were rare. Even the most effective mAb reduced infectivity to a threshold value rather than to zero, was cell-type specific in its potency and was critically dependent on high avidity for even modest neutralization. Most tellingly, mAb combinations that appeared to neutralize quite well failed to prevent infection may be distinct qualitatively as well as quantitatively from those for reducing infectivity (Spiekermann neutralization tests therefore provide a crucial reality check for infectivity reductions. As gB-directed neutralization acted close to the obligate infection step of viral membrane fusion, an alternative entry pathway seemed unlikely. The infectivity preserved with gB-directed neutralization probably reflected an appreciable chance of fusion occurring, even when every mAb binding site was occupied. One possibility is that not every gB molecule on virions was accessible. The MHV-68 gB Trifloxystrobin N terminus C like that of several other gammaherpesviruses C is predicted to be heavily showed at least as much resistance to gB-directed neutralization as did that propagated in BHK-21 cells (Fig.?6b). Such a limitation might also apply to the N-terminal RGD neutralization target on the KSHV gB (Akula em et al. /em , 2002). Alternatively, antibody-bound gBs may still retain an appreciable chance of participating in fusion, given Trifloxystrobin the right cellular setting. MHV-68 infects both fibroblasts (Gill em et al. /em , 2006) and epithelial cells (data not shown) via endocytosis, so any antibody blocking membrane fusion must remain attached in endosomes. Analogy with other viruses would suggest Trifloxystrobin that MHV-68 membrane fusion results from energetically favourable conformational changes in viral glycoproteins at endosomal pH C essentially, viruses tap the cellular energy invested in endosomal acidification to drive membrane fusion. If the more mobile regions of viral fusion proteins are inaccessible, it may be difficult for antibodies to bind their targets strongly enough to block pH-driven conformation changes completely. The high avidity of IgM pentamers C which should be maintained in the oxidizing redox potential of endosomes (Austin em et al. /em , 2005) C would allow them to compete more effectively, and this may explain why the best gB-specific neutralizing mAb was an IgM. Competition between antibody binding and glycoprotein conformation switching could also explain the differences in neutralization between cell types C NMuMG cells providing an environment more conducive to conformation changes than BHK-21 cells. Whatever the mechanistic explanation, the key practical point was that an NMuMG-like, susceptible target was available for incoming, antibody-bound virions to infect em in vivo /em . One way to overcome the difficulty of holding on to viral glycoproteins in.