title: Human rotavirus-specific IgM Memory B cells have differential cloning efficiencies and switch capacities and play a role in antiviral immunity in vivo.
abstract: Protective immunity to rotavirus (RV) is primarily mediated by antibodies produced by RV-specific memory B cells (RV-mBc). Of note, most of these cells express IgM, but the function of this subset is poorly understood. Here, using limiting dilution assays of highly sort-purified human IgM(+) mBc, we found that 62% and 21% of total (non-antigen-specific) IgM(+) and RV-IgM(+) mBc, respectively, switched in vitro to IgG production after polyclonal stimulation. Moreover, in these assays, the median cloning efficiencies of total IgM(+) (17%) and RV-IgM(+) (7%) mBc were lower than those of the corresponding switched (IgG(+) IgA(+)) total (34%) and RV-mBc (17%), leading to an underestimate of their actual frequency. In order to evaluate the in vivo role of IgM(+) RV-mBc in antiviral immunity, NOD/Shi-scid interleukin-2 receptor-deficient (IL-2Rγ(null)) immunodeficient mice were adoptively transferred highly purified human IgM(+) mBc and infected with virulent murine rotavirus. These mice developed high titers of serum human RV-IgM and IgG and had significantly lower levels than control mice of both antigenemia and viremia. Finally, we determined that human RV-IgM(+) mBc are phenotypically diverse and significantly enriched in the IgM(hi) IgD(low) subset. Thus, RV-IgM(+) mBc are heterogeneous, occur more frequently than estimated by traditional limiting dilution analysis, have the capacity to switch Ig class in vitro as well as in vivo, and can mediate systemic antiviral immunity.
authors.affiliation: Instituto de Genética Humana, Facultad de Medicina, Pontificia Universidad Javeriana, Bogotá, Colombia.
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Narváez
authors.forename: Ningguo
authors.initials: N
authors.lastname: Feng
authors.forename: Camilo
authors.initials: C
authors.lastname: Vásquez
authors.forename: Adrish
authors.initials: A
authors.lastname: Sen
authors.forename: Juana
authors.initials: J
authors.lastname: Angel
authors.forename: Harry B
authors.initials: HB
authors.lastname: Greenberg
authors.forename: Manuel A
authors.initials: MA
authors.lastname: Franco
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology; D002469:Cell Separation; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry; D006801:Humans; D007070:Immunoglobulin A / Q000737:chemistry; D007072:Immunoglobulin D / Q000737:chemistry; D007074:Immunoglobulin G; D007075:Immunoglobulin M / Q000737:chemistry* / Q000378:metabolism; D007156:Immunologic Memory; D053631:Interleukin Receptor Common gamma Subunit / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D008822:Mice, Transgenic; D010641:Phenotype; D012401:Rotavirus / Q000378:metabolism*; D012400:Rotavirus Infections / Q000378:metabolism
chemical_list: D007070:Immunoglobulin A; D007072:Immunoglobulin D; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D053631:Interleukin Receptor Common gamma Subunit