title: Inhibition of JAK3 and PKC via Immunosuppressive Drugs Tofacitinib and Sotrastaurin Inhibits Proliferation of Human B Lymphocytes In Vitro.
abstract: BACKGROUND
  Antibody-mediated response in solid organ transplantation is critical for graft dysfunction and loss. The use of immunosuppressive agents partially inhibits the B-lymphocyte response leading to a risk of acute and chronic antibody-mediated rejection. This study evaluated the impact of JAK3 and PKC inhibitors tofacitinib (Tofa) and sotrastaurin (STN), respectively, on B-cell proliferation, apoptosis, and activation in vitro.
  
  
  METHODS
  Human B cells isolated from peripheral blood of healthy volunteers were cocultured with CD40 ligand-transfected fibroblasts as feeder cells in the presence of interleukin (IL) 2, IL-10, and IL-21. The cocultures were treated with immunosuppressants Tofa, STN, and rapamycin (as a control), to analyze the proliferation and apoptosis of B cells by means of Cyquant and flow cytometry, respectively. CD27 and IgG staining were applied to evaluate whether treatments modified the activation of B cells.
  
  
  RESULTS
  Tofa and STN were able to inhibit B-cell proliferation to the same extent as rapamycin, without inducing cell apoptosis. After 6 days in coculture with feeder cells, all B cells showed CD27 memory B-cell phenotype. None of the immunosuppressive treatments modified the proportion between class-switched and non-class-switched memory B cells observed in nontreated cultures. The high predominance of CD27+CD24+phenotype was not modified by any immunosuppressive treatment.
  
  
  CONCLUSIONS
  Our results show that Tofa and STN can suppress B-cell antibody responses to an extent similar to rapamycin, in vitro; therefore these compounds may be a useful therapy against antibody-mediated rejection in transplantation.
authors.affiliation: Departamento de Nefrología y Trasplante Renal, Hospital Clínic de Barcelona, Barcelona, Spain; Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: M N
authors.initials: MN
authors.lastname: Martina
authors.affiliation: Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: M J
authors.initials: MJ
authors.lastname: Ramirez Bajo
authors.affiliation: Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Bañon-Maneus
authors.affiliation: Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Moya Rull
authors.affiliation: Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: N
authors.initials: N
authors.lastname: Hierro-Garcia
authors.affiliation: Departamento de Nefrología y Trasplante Renal, Hospital Clínic de Barcelona, Barcelona, Spain; Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Revuelta
authors.affiliation: Departamento de Nefrología y Trasplante Renal, Hospital Clínic de Barcelona, Barcelona, Spain; Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: J M
authors.initials: JM
authors.lastname: Campistol
authors.affiliation: Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain.
authors.forename: J
authors.initials: J
authors.lastname: Rovira
authors.affiliation: Departamento de Nefrología y Trasplante Renal, Hospital Clínic de Barcelona, Barcelona, Spain; Laboratori Experimental de Nefrologia i Trasplantament (LENIT) Fundació; Clínic IDIBAPS, Barcelona, Spain. Electronic address: fdiekman@clinic.ub.es.
authors.forename: F
authors.initials: F
authors.lastname: Diekmann
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D001402:B-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D023201:CD40 Ligand / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D016753:Interleukin-10 / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D007378:Interleukins / Q000493:pharmacokinetics; D053616:Janus Kinase 3 / Q000037:antagonists & inhibitors*; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D008213:Lymphocyte Activation / Q000187:drug effects; D010880:Piperidines / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D011758:Pyrroles / Q000494:pharmacology*; D011799:Quinazolines / Q000494:pharmacology*; D020123:Sirolimus / Q000494:pharmacology; D000098546:Interleukin-21
chemical_list: C508609:IL10 protein, human; D007166:Immunosuppressive Agents; D007376:Interleukin-2; D007378:Interleukins; D010880:Piperidines; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D011758:Pyrroles; D011799:Quinazolines; D016753:Interleukin-10; D023201:CD40 Ligand; C543528:sotrastaurin; C479163:tofacitinib; C507933:JAK3 protein, human; D053616:Janus Kinase 3; D011493:Protein Kinase C; D000098546:Interleukin-21; D020123:Sirolimus