title: Construction and Functional Characterization of a Fusion Protein Interleukin-21/Immunoglobulin for Long-Term In Vivo Biodisponibility.
abstract: Interleukin (IL)-21 has been intensively studied for use in therapy of autoimmune diseases, cancers, and chronic viruses due to its immunomodulatory properties, especially on CD4(+) and CD8(+) T cells and natural killer (NK) cells. The objective of this study was to produce an optimized form of IL-21 with improved stability. Plasmids encoding the murine IL-21 alone (pIL-21) or IL-21 genetically fused to portions from mouse IgG3 (pIL-21/Ig) were constructed, and the efficiency of expression, protein kinetics, biodisponibility, and function were analyzed. The genetic constructions of pIL-21 and pIL-21/Ig were transfected into HEK 293 cells, and significant levels of functional IL-21 were obtained. The amino acid of murine IL-21 and IgG3 cloned showed 100% identity with correspondent published sequences. At 24 h of incubation, increased levels of IL-21 were detected in the supernatants of pIL-21. At 72 h of culture, the levels of IL-21 in the supernatant of cells transfected with pIL-21/Ig were significantly higher than those secreted by pIL-21-transfected cells. Furthermore, the data showed that our chimeric IL-21/Ig present improved systemic disponibility in BALB/c mice and conserved the intrinsic ability to increase the frequency of CD4(+) T cells, NKT cells, and CD8(+) T cells.
authors.affiliation: 1 Laboratório de Biologia Molecular e Cultivo de Células, Centro Universitário Franciscano (UNIFRA) , Santa Maria, Brazil .|2 Laboratório de Imunologia Celular e Molecular, Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), Porto Alegre, Brazil .
authors.forename: Rodrigo Benedetti
authors.initials: RB
authors.lastname: Gassen
authors.affiliation: 3 Laboratório de Imunologia Celular e Molecular, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA) , Porto Alegre, Brazil .
authors.forename: Pedro Roosevelt T
authors.initials: PR
authors.lastname: Romão
authors.affiliation: 1 Laboratório de Biologia Molecular e Cultivo de Células, Centro Universitário Franciscano (UNIFRA) , Santa Maria, Brazil .|2 Laboratório de Imunologia Celular e Molecular, Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), Porto Alegre, Brazil .
authors.forename: Deise Nascimentode
authors.initials: DN
authors.lastname: Freitas
authors.affiliation: 1 Laboratório de Biologia Molecular e Cultivo de Células, Centro Universitário Franciscano (UNIFRA) , Santa Maria, Brazil .|3 Laboratório de Imunologia Celular e Molecular, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA) , Porto Alegre, Brazil .
authors.forename: Luiz Carlos
authors.initials: LC
authors.lastname: Rodrigues Junior
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects; D005260:Female; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism*; D007267:Injections; D007694:Killer Cells, Natural / Q000187:drug effects; D008807:Mice, Inbred BALB C; D015202:Protein Engineering / Q000379:methods*; D055550:Protein Stability; D018123:Receptors, Interleukin / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*
chemical_list: D007074:Immunoglobulin G; D018123:Receptors, Interleukin; D011993:Recombinant Fusion Proteins; C415369:interleukin-22 receptor