title: Anti-human leukocyte antigen-DR (MHC class II) humanized monoclonal antibody, IMMU-114, suppresses human to bovine cellular responses.
abstract: The effect of an anti-human leukocyte antigen-DR (MHC class II) humanized monoclonal antibody, IMMU-114, against the human to bovine cellular response was investigated.
  
  
  METHODS
  Human peripheral mononuclear cells (PBMCs) were cocultured with inactivated self-PBMCs (Self), bovine PBMCs with control antibody (Xeno), or bovine PBMCs with IMMU-114 (IMMU-114). Cellular responses were investigated by thymidine incorporation assay, CFSE (carboxyfluorescein diacetate succinimidyl ester)-mixed lymphocyte reaction, and cytokine production in culture medium.
  
  
  RESULTS
  Thymidine incorporation rates at a 1:1 responder to stimulator ratio for Xeno + control antibody, Xeno + IMMU-114, Self + control antibody, and Self + IMMU-114 were 14201.3 ± 1968.4, 513.0 ± 49.5, 952.7 ± 128.7, and 423.3 ± 138.8 cpm, respectively (P = 0.032). Those at a 1:2 ratio were 6518.0 ± 690.1, 896.6 ± 92.9, 1051.0 ± 123.6, and 736.0 ± 35.6 cpm, respectively (P = 0.036). CFSE-mixed lymphocyte reaction demonstrated that the frequencies of CFSE-low, CD4(+), and CD25(+) activating T cells in Self, Xeno, and IMMU-114 were 0.27 ± 0.04%, 3.65 ± 0.53%, and 1.23 ± 0.15%, respectively (P = 0.027). Cytokine production in culture medium indicated that IMMU-114 decreased Th1-type cytokines, including interleukin-2, interferon-γ, and tumor necrosis factor-α.
  
  
  CONCLUSIONS
  IMMU-114 effectively suppresses human to bovine cellular responses. The mechanism involves direct inhibition of the interaction between class II human leukocyte antigen-DR-positive cells and CD4(+) T cells, and indirect suppression of Th1 cytokine production.
authors.affiliation: Second Department of Surgery, Dokkyo Medical University, Kitakobayashi 880, Mibu, Shimotsuga, Tochigi, Japan.
authors.forename: Kyung Hwa
authors.initials: KH
authors.lastname: Park
authors.forename: Tokihiko
authors.initials: T
authors.lastname: Sawada
authors.forename: Toru
authors.initials: T
authors.lastname: Murakami
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Ishii
authors.forename: Mineko
authors.initials: M
authors.lastname: Yasuo
authors.forename: Manami
authors.initials: M
authors.lastname: Urakawa
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Aoyagi
authors.forename: Shohei
authors.initials: S
authors.lastname: Fuchinoue
authors.forename: Keiichi
authors.initials: K
authors.lastname: Kubota
mesh_terms: D000818:Animals; D000910:Antibodies, Heterophile / Q000276:immunology; D061067:Antibodies, Monoclonal, Humanized / Q000276:immunology* / Q000494:pharmacology; D015478:Antigens, Heterophile / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D006684:HLA-DR Antigens / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D007165:Immunosuppression Therapy / Q000379:methods; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D013936:Thymidine / Q000493:pharmacokinetics; D014183:Transplantation, Heterologous / Q000276:immunology*
chemical_list: D000910:Antibodies, Heterophile; D061067:Antibodies, Monoclonal, Humanized; D015478:Antigens, Heterophile; D016207:Cytokines; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; C577236:IMMU-114; D013936:Thymidine