The cytokines NAP-1 (IL-8), MCP-1, IL-1 beta, and GRO in rabbit inflammatory skin lesions produced by the chemical irritant sulfur mustard

Abstract

Developing and healing dermal inflammatory lesions were produced in rabbits by the topical application of dilute sulfur mustard (SM), the military vesicant. In tissue sections of such lesions, cells containing the mRNA of important cytokines were identified with in situ hybridization techniques. These cytokines were neutrophil attractant/activation protein-1 (NAP-1 (also called IL-8), monocyte chemoattractant (activating) protein 1 (MCP-1), interleukin 1 (beta) (IL-1 (beta)), and GRO (a growth factor and chemokine). Mononuclear cells (mainly macrophages and activated fibroblasts) contained the mRNA of all four of these cytokines. A higher percentage of cytokine-producing mononuclear cells (macrophages and activated fibroblasts) was present in lesions at 2 days (their peak size) than at 6 days, when they were almost healed. Granulocytes emigrated from the bloodstream, passed through the lesions, and were the major constituent of the protective crust. This sequence correlated with the distribution of cells able to produce NAP-1: At 2 days and 6 days, the mononuclears that contained messenger RNA for this granulocyte chemoattractant were found mainly in the upper part of the dermis. At 2 days and 6 days, cells containing the mRNA of IL-1, a primary cytokine, were also found predominantly in the upper dermis, i.e., nearest the site of injury. In contrast, mononuclears containing the mRNA of MCP-1 (a monocyte chemoattractant), and the mRNA of GRO (a granulocyte chemoattractant) were more equally distributed throughout the dermis. SM stimulated hair follicle epithelial cells to up-regulate GRO mRNA and, to a lesser degree, NAP-1 mRNA. Apparently, the irritation produced by SM directly or indirectly induces such epithelial cells to manufacture these growth factors. In the rabbit, hair follicles are known to be the main source of new epithelial cells after the covering epithelium has been destroyed. Therefore, GRO is probably a major autocrine-paracrine stimulus for such repair. A brief review of the role of cytokines in dermal inflammation is presented.

Nonspecific and immune-specific up-regulation of cytokines in rabbit dermal tuberculous (BCG) lesions

“…This procedure is essentially that of Gillitzer et al [36,37] with the slight modifications described prevously [3]. The in situ hybridizations for cytokine mRNAs were performed with 35 S-antisense RNA probes (complementary to cellular mRNA).…”

“…Preparation and sources of 35 S-antisense and 35 S-sense IL-1␤, TNF-␣, MCP-1, and IL-8 (NAP-1) RNAs for in situ hybridization 35 S-antisense and 35 S-sense RNA probes were made, as previously described [3], from plasmids containing cDNA inserts of rabbit IL-1␤, MCP-1, IL-8, and TNF-␣. The T3/T7 Riboprobe Synthesis System (Promega Corp., Madison, WI) with 35 S-UTP supplied by Dupont-New England Nuclear Products (Boston, MA) was used.…”

“…IL-1␤ and TNF-␣ are primary cytokines that up-regulate the production of other cytokines [1][2][3]. These primary cytokines also up-regulate adhesion molecules in the microvasculature that enable the diapedesis of mononuclear cells (MN) and polymorphonuclear neutrophils (PMN) into inflammatory sites [4][5][6].…”

MCP-1 is selectively expressed in the late phase by cytokine-stimulated human neutrophils: TNF-α plays a role in maximal MCP-1 mRNA expression

“…The main immunoreactivity for MCP-1 in the early phase of delayed-type hypersensitivity (DTH) reaction induced in the rat skin was associated with infiltrating PMN [26]. In contrast, in an acute inflammation caused in rabbit skin with sulfur mustard, MCP-1 mRNA was detected in macrophages by in situ hybridization, but not in PMN [27]. Mori et al investigated the genes expressed in PMN infiltrating into rabbit peritoneal cavities after casein injection.…”

Sulfur mustard and respiratory diseases: Revisit with special reference to the “Comments on ‘Sulfur Mustard and Respiratory Diseases’, Tang and Loke () and a prepared Integrated Mechanism for Chronic Pulmonary Disease from Exposure to Sulfur Mustard” by Saburi and Ghanei ()

“…However, results from Emad's group, and some of Ghanei's group (such as Aghanouri et al, 2004;Attaran et al 2010;Zarin et al, 2010) are more relevant to pulmonary diseases as they focused on localized tissue changes of cytokines and chemokines in bronchoalveolar lavage fluid or lung biopsies. To link changes of cytokines and chemokines at system level (in serum) to pulmonary diseases may be too speculative as it has been well known that mustard could also induce inflammatory (cytokine and chemokine) changes in other organs such as skin (Arroyo et al, 2000(Arroyo et al, , 2003(Arroyo et al, , 2004Blaha et al, 2000a,b;Cowan et al, 2002;Kehe et al, 2009;Ricketts et al, 2000;Sabourin et al, 2002;Tanaka et al, 1997;Tsuruta et al, 1996;Wormser et al, 2005), eye (Ghasemi et al, 2012), liver (Anand et al, 2009). …”