Isolation of cross‐reactive human monoclonal antibodies that prevent binding of human noroviruses to histo‐blood group antigens

Abstract

In order to identify the repertoire of antibodies generated on natural infection of norovirus (NoV) in humans, and to characterize the human monoclonal antibodies against NoV, three phage-displayed antibody libraries originating from healthy person(s) were screened using purified virus-like particles (VLPs) of strain Narita 104 (r104, genogroup II, genotype 4) or strain Chiba 407 (rCV, genogroup I, genotype 4) as antigens. On screening with r104, 62 clones were isolated. Among these antibodies, two clones, 12A11 and 12B10, showed intra-genogroup cross-reactivity to genotypes 1, 3-7, 12, and 14, and genotypes 1, 4, 6, and 7 of genogroup II, respectively. In addition, antibodies belonging to the same group were isolated from two different libraries. On screening with rCV, five clones were isolated, two of which were cross-reactive. One, CV-2F5, reacted to genotypes 1-4, and 8 of genogroup I, and the other, CV-1A5, showed inter-genogroup cross-reactivity to all the VLPs employed in this study. The blocking activities of the monoclonal antibodies against the interaction of homotypic VLPs (VLPs used in the panning procedure) with histo-blood group antigens were also assessed as an alternative to neutralization assay. Although the blocking activity of 12A11 was partially limited 12B10 prevented the binding of r104 to histo-blood group antigens that had been reported to bind r104. The blocking activity of CV-2F5 against the attachment of rCV to suitable histo-blood group antigens was weak, but the blocking activity of CV-1A5 was well recognized. Thus, 12B10 and CV-1A5 were suggested to be cross-reactive monoclonal antibodies with neutralizing activity.

Experimental Human Infection with Norwalk Virus Elicits a Surrogate Neutralizing Antibody Response with Cross-Genogroup Activity

“…Furthermore, our study suggests that experimental infection of healthy adults with Norwalk virus (a GI NoV) can elicit blocking antibody against heterotypic NoV antigens both within the GI genogroup as well as to the more divergent GII NoVs. This observation extends previous descriptions of broadly cross-reactive serum activity (20,(23)(24)(25)(26) and is the first description, to our knowledge, of the development of a serum-blocking antibody response with cross-genogroup specificity in the context of experimental human challenge with an NoV.…”

“…The impact of NoV diversity on the breadth and duration of protective immunity remains poorly understood. Some previous experimental NoV challenge studies and epidemiologic studies suggested that NoVs elicit antibodies with intragenogroup cross-reactivity (15)(16)(17)(18)(19)(20)(21)(22), while other studies reported evidence for intergenogroup cross-reactive serum antibody (23)(24)(25)(26)(27)(28).…”

Advances in Norovirus Biology

“…Analysis of monoclonal antibodies raised against HuNoV VLPs has demonstrated the development of a wide spectrum of antibodies with strain-, genotype-, genogroup-, or intergenogroup-specific reactivities. While strain-specific epitopes are located in the hypervariable P2 domain of VP1 [e.g., (Allen et al, 2009; Debbink et al, 2012)], more broadly cross-reactive epitopes are located in conserved regions of the S and P1 domains [e.g., (Higo-Moriguchi et al, 2014)]. Although classical HuNoV neutralization assays cannot be performed due to the lack of a cell culture system, a widely used surrogate assay (‘blockade assay’) measures the ability of antibodies to block VLP binding to histo-blood group antigens (HBGAs), which are viral attachment factors on the intestinal epithelium (Harrington et al, 2002).…”

Mapping Broadly Reactive Norovirus Genogroup I and II Monoclonal Antibodies

“…The P domain is further subdivided into the P1 subdomain (residues 226 to 278 and 406 to 520 for GI.1 Norwalk virus [NV]) and the P2 subdomain (residues 279 to 405 for GI.1 NV) (23). P2 represents the most exposed surface of the viral particle and is involved in cellular histo-blood group antigen (HBGA) binding (26)(27)(28).…”

“…Most information on the antigenic characteristics of NoVs comes from the study of monoclonal antibodies (MAbs) generated against VLPs from both GI and GII viruses (27,(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40). The majority of these MAbs are genogroup specific and recognize only viruses closely related to the immunogen used to generate the MAb.…”