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The priming effects of the products of stimulated mononuclear cells on the response of neutrophils to C5a des arg
Author(s) -
Crouch Sharon,
Fletcher John
Publication year - 1991
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1991.tb07971.x
Subject(s) - respiratory burst , lipopolysaccharide , tumor necrosis factor alpha , peripheral blood mononuclear cell , zymosan , priming (agriculture) , immunology , stimulation , cytokine , granulocyte , biology , endocrinology , chemistry , medicine , in vitro , biochemistry , botany , germination
Summary Certain recombinant human cytokines have been shown to enhance polymorphonuclear leucocyte (PMN) responses to subsequent stimulation. Mononuclear cells (MNC) from normal healthy individuals were stimulated for 5 h with 1 μg/ml bacterial lipopolysaccharide (LPS) in order to induce production and secretion of inflammatory cytokines into the surrounding medium. These mononuclear cell conditioned media (MNCM) were then used to prime PMN isolated from healthy volunteers. Preincubating the PMN with MNCM for 15 min at 4°C followed by washing and warming to 37°C caused a 344% increase ( n = 26) in the rate of superoxide anion production in response to zymosan‐activated serum (ZAS), a source of C5a des arg. This effect could not be reproduced with recombinant human forms of interleukin 1 beta (Il‐1 beta) or granulocyte‐macrophage‐colony stimulating factor (GM‐CSF), although, with the latter, there was some effect when the preincubation stage was carried out for 60 min at 37°C. Only recombinant human tumour necrosis factor‐alpha (rh‐TNF‐alpha) gave a similar PMN priming effect to that seen with MNCM. This effect could not be reversed by washing away either the MNCM or rh‐TNF‐alpha. The priming effect could be markedly reduced (74.8%, n = 6) by employing the use of polyclonal antibody to TNF‐alpha in the preincubation step; assaying for TNF‐alpha in these MNCMs showed that the degree of priming corresponded to the amount of TNF‐alpha present. Rh‐TNF‐alpha alone appeared to have very little direct stimulatory effect on respiratory burst activity. The results show that TNF‐alpha produced by LPS stimulated MNC after 5 h binds to a PMN surface receptor in the cold and warming of the cells to 37°C allows for an immediate and dramatic response to ZAS stimulation. This suggests that TNF‐alpha is the important cytokine upregulating PMN responses to other physiological mediators, including C 5 a des arg during the early phases of an inflammatory reaction.