Journal: International immunology

Article Title: The contribution of Fc effector mechanisms in the efficacy of anti-CD154 immunotherapy depends on the nature of the immune challenge.

doi: 10.1093/intimm/dxh162

Figure Lengend Snippet: Fig. 1. Binding of hu5C8 mAbs to CD154, FccR and C1q. (A) Aglycosyl hu5C8 and glycosylated hu5C8 bound to human CD154 expressed on the surface of D1.1 cells with a relative affinity similar to that in a competitive FACS assay with biotinylated hu5C8. Aglycosyl hu5C8 has impaired FccR- binding capabilities; the ability of the anti-CD154 mAbs to form a bridge between rsCD154 and FccRI cells (B) or between CD154+ CHO and FccRIIb (C), FccRIIa (D) and FccRIIIa (E) cells was measured. (F) Aglycosyl hu5C8 has impaired complement-binding capabilities compared with glycosylated hu5C8 in a human C1q ELISA.

Article Snippet: The membrane was then probed for 2 h with 0.5 lg ml 1 of biotinylated hu5C8 (Biogen Idec, Cambridge, MA, USA) or biotinylated rsCD40L (Biogen Idec) to bind anti-hu5C8 or hu5C8, respectively.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay

Journal: International immunology

Article Title: The contribution of Fc effector mechanisms in the efficacy of anti-CD154 immunotherapy depends on the nature of the immune challenge.

doi: 10.1093/intimm/dxh162

Figure Lengend Snippet: Fig. 4. A membrane transfer assay capable of detecting the components of immune complexes was utilized to further analyze the serum from two aglycosyl hu5C8-treated rhesus monkeys that underwent renal transplant. Denatured immune complexes were immobilized on duplicate membranes and probed with biotinylated rsCD40L to measure hu5C8 or with biotinylated hu5C8 to detect anti-hu5C8. This method was validated by immobilized hu5C8 and rabbit anti-hu5C8 included as controls for each detection method. Serum samples obtained pre- or post-dose are as indicated. Anti-hu5C8 antibodies were detected in AT5D at days 12 and 16 and while hu5C8 was undetected by ELISA at day 12, it was readily detected here. Anti-hu5C8 antibodies were also detected in serum from AW34 on day 9.

Article Snippet: The membrane was then probed for 2 h with 0.5 lg ml 1 of biotinylated hu5C8 (Biogen Idec, Cambridge, MA, USA) or biotinylated rsCD40L (Biogen Idec) to bind anti-hu5C8 or hu5C8, respectively.

Techniques: Membrane, Enzyme-linked Immunosorbent Assay