title: Induction of macrophage TNF alpha, IL-1, IL-6, and PGE2 production by DTH-initiating factors.
abstract: The elicitation of delayed-type hypersensitivity (DTH) reactions in mice is due to the sequential action of two different, antigen-specific, Thy-1+ cells. We have previously cloned the early-acting DTH-initiating cell from nude mice that were immunized and boosted by contact sensitization with oxazolone (OX). This cell clone, WP-3.27, releases an antigen-specific factor (OX-F) that sensitizes mast cells such that specific antigen challenge will induce serotonin release which mediates the early phase of DTH. In normal mice contact sensitized with picryl chloride (PCl), a similar polyclonal factor (PCl-F) has a similar activity and is also known to bind to macrophages. Thus, we measured macrophage production of TNF alpha, IL-1, IL-6, and PGE2 in response to the hapten affinity-purified DTH-initiating factors OX-F and PCl-F. Both factors induced significant release of each cytokine and PGE2. The production of TNF alpha, IL-1, and IL-6 was measured by bioassays. Northern blot analysis showed rapid accumulation of cytokine mRNA (2-4 hr), while maximal production of PGE2 occurred at approximately 8 hr. These macrophage activating properties of OX-F and PCl-F were not due to contamination with LPS as determined by the low levels of LPS present in OX-F and PCl-F and by the failure of polymyxin B to inhibit factor-induced PGE2 and TNF alpha production. Also, macrophage activation was shown not to be due to the action of several lymphokines known to be produced by WP3.27. Separation of OX-F and PCl-F by preparative isoelectric focusing showed a similar pattern: there were two major peaks of PGE2-inducing activity observed for both factors (for PCl-F at pI of 2-3 and 5.0, and for OX-F at pI of 3.5-4 and 5.0), but not for a sham factor produced by WEHI-3 cells. The ability of DTH-initiating factors to rapidly induce macrophage cytokine release and PGE2 synthesis 4-6 hr later may suggest a role for these mediators during the respective early vascular and late cellular phases of inflammation in DTH.
authors.affiliation: Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510-8056.
authors.forename: N R
authors.initials: NR
authors.lastname: Ferreri
authors.forename: I
authors.initials: I
authors.lastname: Millet
authors.forename: V
authors.initials: V
authors.lastname: Paliwal
authors.forename: W
authors.initials: W
authors.lastname: Herzog
authors.forename: D
authors.initials: D
authors.lastname: Solomon
authors.forename: R
authors.initials: R
authors.lastname: Ramabhadran
authors.forename: P W
authors.initials: PW
authors.lastname: Askenase
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D016207:Cytokines / Q000502:physiology; D003877:Dermatitis, Contact / Q000276:immunology*; D015232:Dinoprostone / Q000096:biosynthesis*; D015870:Gene Expression; D006968:Hypersensitivity, Delayed / Q000276:immunology*; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D015850:Interleukin-6 / Q000096:biosynthesis* / Q000235:genetics; D008264:Macrophages / Q000276:immunology*; D008297:Male; D051379:Mice; D008808:Mice, Inbred CBA; D010081:Oxazolone; D010853:Picryl Chloride; D012333:RNA, Messenger / Q000235:genetics; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D016207:Cytokines; D007375:Interleukin-1; D015850:Interleukin-6; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D010081:Oxazolone; D015232:Dinoprostone; D010853:Picryl Chloride