Roles of serum in innate immune responses of human leukocytes to synthetic lipopeptide

Abstract

Tripalmitoyl-S-glyceryl-l-Cys-Ser-(Lys)₄ (Pam₃CSK₄) is a highly conserved molecular motif found in various classes of lipoproteins. The requirement for leukocyte to respond to synthetic Pam₃CSK₄ were studied. Pam₃CSK₄ primed neutrophils for a respiratory burst in a serum-dependent manner. Pam₃CSK₄ upregulated CD11b, CD14, and cytochrome b₅₅₈, and downregulated Leu-8. Treatment of neutrophils with anti-CD14 antibodies and treatment of serum with anti-LPS binding protein (LBP) antibodies resulted in the inhibition of priming for respiratory burst by Pam₃CSK₄. It should be noted that LBP could not replicate the effects of serum in priming of neutrophils for respiratory burst by Pam₃CSK₄. Serum LBP bound to immobilized Pam₃CSK₄. Pam₃CSK₄ induced the interleukin-8 (IL-8) production by leukocytes in a serum-dependent manner. Further, Pam₃CSK₄-induced priming of neutrophils for respiratory burst was not inhibited by the LPS antagonists LA-14-PP, Rhodobacter sphaeroides LPS, or E5531, and Pam₃CSK₄-induced IL-8 production by leukocytes was not affected by LPS antagonist, E5531, indicating that Pam₃CSK₄ was recognized by a different receptor than LPS. Thus, Pam₃CSK₄ and LPS had similar biological activities and similar requirement to act on leukocytes, but were recognized by different receptors. Serum in the action of Pam₃CSK₄ on leukocytes was not replicated by LBP, suggesting that Pam₃CSK₄ might be disaggregated by serum to result in the activation of leukocytes.

Keywords: LPS binding protein; Neutrophil; Pam(3)CSK(4); Priming; Respiratory burst; Serum.