title: IFN-gamma is the only anti-rotavirus cytokine found after in vitro stimulation of memory CD4+ T cells from mice immunized with a chimeric VP6 protein.
abstract: CD4+ T cells are the only lymphocytes required for protection of mice against rotavirus shedding after mucosal immunization with chimeric VP6 (MBP::VP6) and the adjuvant LT(R192G). One possible effector of protection is CD4+ T-cell cytokines. To determine if memory CD4+ T cells of immunized mice produce cytokines with direct anti-rotavirus activity, an in vitro infection model was developed using mouse CMT-93 cells and rhesus rotavirus (RRV). Spleen and lamina propria (LP) cells, as well as purified splenic CD4T cells obtained after intranasal immunization of BALB/c mice with MBP::VP6/LT(R192G) released large quantities of two cytokines (IL-17 and IFN-gamma) into cell supernatants when stimulated with MBP::VP6. Production of these same cytokines is rapidly upregulated in intestinal lymphocytes after rotavirus inoculation of immunized mice. IL-17 pretreatment of CMT-93 cells had no effect on subsequent RRV replication, but IFN-gamma was the most potent inhibitor within a panel of nine cytokines tested. Supernatants obtained after in vitro stimulation of splenic CD4+ T cells of immunized mice had high levels of anti-RRV activity and their pretreatment with mAb against IFN-gamma caused essentially complete loss of activity. Thus, IFN-gamma was the only cytokine identified in stimulated CD4+ T cells from immunized mice that directly inhibited rotavirus replication.
authors.affiliation: Division of Infectious Diseases, Children's Hospital Medical Center, Cincinnati, Ohio 45229, USA. monica.mcneal@cchmc.org
authors.forename: Monica M
authors.initials: MM
authors.lastname: McNeal
authors.forename: Susan C
authors.initials: SC
authors.lastname: Stone
authors.forename: Mitali
authors.initials: M
authors.lastname: Basu
authors.forename: John D
authors.initials: JD
authors.lastname: Clements
authors.forename: Anthony H-C
authors.initials: AH
authors.lastname: Choi
authors.forename: Richard L
authors.initials: RL
authors.lastname: Ward
mesh_terms: D000281:Administration, Intranasal; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D001427:Bacterial Toxins / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D036022:Capsid Proteins / Q000276:immunology*; D002352:Carrier Proteins / Q000276:immunology; D002460:Cell Line; D016207:Cytokines / Q000032:analysis / Q000502:physiology; D004768:Enterotoxins / Q000276:immunology; D004847:Epithelial Cells / Q000821:virology; D029968:Escherichia coli Proteins / Q000276:immunology; D007114:Immunization; D007156:Immunologic Memory; D007371:Interferon-gamma / Q000032:analysis / Q000502:physiology*; D058619:Maltose-Binding Proteins; D051379:Mice / Q000276:immunology / Q000821:virology; D008807:Mice, Inbred BALB C; D009092:Mucous Membrane / Q000276:immunology; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology; D012401:Rotavirus / Q000276:immunology / Q000502:physiology; D012400:Rotavirus Infections / Q000517:prevention & control; D022243:Rotavirus Vaccines / Q000008:administration & dosage / Q000276:immunology; D013154:Spleen / Q000166:cytology / Q000276:immunology; D019444:Vaccines, DNA / Q000276:immunology*; D014779:Virus Replication
chemical_list: D000956:Antigens, Viral; D001427:Bacterial Toxins; D036022:Capsid Proteins; D002352:Carrier Proteins; D016207:Cytokines; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins; D022243:Rotavirus Vaccines; C061332:VP6 protein, Rotavirus; D019444:Vaccines, DNA; D007371:Interferon-gamma; C027538:heat-labile enterotoxin, E coli