title: Dynamics of cytokines and immunoglobulins serum profiles in primary and secondary Cryptosporidium parvum infection: usefulness of Luminex® xMAP technology.
abstract: Infection by Cryptosporidium parvum triggers a complex array of innate and adaptive cell mediated immune response, playing an important role in controlling the infection. To date, there are no studies applying the Luminex® xMAP technology to determine profiles of cytokines and immunoglobulins in the context of an infection by C. parvum. In this study, we analyzed these immune mediators in the serum of immunocompetent mice inoculated with C. parvum oocysts, using Luminex, to understand how the immune system responds to an infection by this parasite. Animal sera were also analyzed by ELISA to determine the expressed immunoglobulin isotype profile, and compare the obtained trend with data obtained by Luminex. Specific-pathogen-free BALB/C mice were inoculated with oocysts of C. parvum at days 0 and 22. Peripheral blood was aseptically collected from sacrificed mice on several time points, and immune mediators were evaluated in serum samples. Infection was confirmed by the presence of C. parvum DNA in feces by a nested-PCR assay (60-kDa glycoprotein). Luminex results showed predominance in the secretion of IgG1 and IgG2a, confirmed by ELISA, which also showed that IgG1 is the major immunoglobulin isotype produced during the infection. The analysis of cytokines suggests a preferential Th(1) over the Th(2) response, with increased production of TNF-α, IFN-γ and GM-CSF. This work contributed to a better understanding of the immune response to the infection by C. parvum, as well as demonstrated the advantage of Luminex® xMAP technology to study immune mediators, using small sample volumes.
authors.affiliation: Unidade de Parasitologia Médica, Grupo de Protozoários Oportunistas/VIH e Outros Protozoários, CMDT, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Lisboa 1349-008, Portugal.
authors.forename: Vera
authors.initials: V
authors.lastname: Codices
authors.forename: Catarina
authors.initials: C
authors.lastname: Martins
authors.forename: Carlos
authors.initials: C
authors.lastname: Novo
authors.forename: Bruno
authors.initials: B
authors.lastname: de Sousa
authors.forename: Ângela
authors.initials: Â
authors.lastname: Lopes
authors.forename: Miguel
authors.initials: M
authors.lastname: Borrego
authors.forename: Olga
authors.initials: O
authors.lastname: Matos
mesh_terms: D000818:Animals; D002417:Cattle; D003457:Cryptosporidiosis / Q000276:immunology* / Q000469:parasitology; D016785:Cryptosporidium parvum / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D016207:Cytokines / Q000097:blood*; D016054:DNA, Protozoan / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005243:Feces / Q000469:parasitology; D005260:Female; D005434:Flow Cytometry; D007121:Immunocompetence; D007136:Immunoglobulins / Q000097:blood*; D051379:Mice; D008807:Mice, Inbred BALB C; D008863:Microspheres; D016133:Polymerase Chain Reaction; D013047:Specific Pathogen-Free Organisms
chemical_list: D016207:Cytokines; D016054:DNA, Protozoan; D007136:Immunoglobulins