title: Mucosal and systemic isotype-specific antibody responses and protection in conventional pigs exposed to virulent or attenuated porcine epidemic diarrhoea virus.
abstract: Eleven-day-old conventionally reared piglets were inoculated orally with two different doses of the cell-culture adapted strain CV-777 of the porcine epidemic diarrhoea virus (PEDV) or the virulent isolate of the same strain and challenged with the same virulent PEDV 3 weeks later. Pigs inoculated with the two doses of the attenuated virus did not show any typical sign of the disease, and virus shedding was not frequent. In contrast, 31% of pigs exposed to the virulent PEDV developed diarrhoea and virus shedding was demonstrated in 100%. At different postinoculation day (PID) and postchallenge day (PCD) virus-specific antibody-secreting cells (ASC) in gut associated lymphoid tissues (duodenum and ileum lamina propria and mesenteric lymph nodes) and systemic locations (blood and spleen) were assessed by enzyme-linked immunospot (ELISPOT). Only a small response was detected in the groups inoculated with attenuated PEDV, whereas in the group previously exposed to the virulent virus on PID 21 a large number of IgG and IgA ASC was detected. Isotype-specific antibody responses in serum were investigated by ELISA. IgG responses were detected in all groups, although the highest response corresponded to the group inoculated with virulent virus and only this group showed an IgA response. The pigs exposed to virulent PEDV were completely protected against the challenge with a higher dose of the same virulent virus on PID 21 and none of them shed the virus. The pigs inoculated with the attenuated strain were partially protected against the challenge, and 25% of the low dose- and 50% of the high dose-exposed pigs did not shed virus after challenge. All the pigs from a control group, not previously exposed to the virus, excreted the virus in faeces. A strong positive correlation was established between protection and the ASC responses detected in gut associated lymphoid tissues and blood at the challenge day and also between protection and serum isotype-specific antibody titers on that day. In addition, the IgA and IgG ASC responses detected in the blood on PID 21 also correlated with the responses found in the gut associated lymphoid tissues. The ASC and serum antibody responses after the challenge corresponded to a secondary immune response in the groups inoculated with attenuated virus, whereas a primary response was evident in the control group. No increase was seen in any of the parameters studied in the pigs inoculated with virulent PEDV.
authors.affiliation: Departamento de Sanidad Animal (Enfermedades Infecciosas y Epidemiología), Facultad de Veterinaria, Universidad de León, E-24071, León, Spain.
authors.forename: M L
authors.initials: ML
authors.lastname: de Arriba
authors.forename: A
authors.initials: A
authors.lastname: Carvajal
authors.forename: J
authors.initials: J
authors.lastname: Pozo
authors.forename: P
authors.initials: P
authors.lastname: Rubio
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis*; D000921:Antibody-Producing Cells / Q000276:immunology; D002522:Chlorocebus aethiops; D003332:Coronaviridae / Q000276:immunology*; D018352:Coronavirus Infections / Q000276:immunology / Q000662:veterinary*; D003967:Diarrhea / Q000276:immunology / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay; D018928:Immunity, Mucosal; D007070:Immunoglobulin A / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D007132:Immunoglobulin Isotypes / Q000096:biosynthesis*; D013552:Swine; D013553:Swine Diseases / Q000276:immunology*; D014709:Vero Cells; D014774:Virulence; D017201:Virus Shedding
chemical_list: D000914:Antibodies, Viral; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes