Immunoglobulin E plus antigen challenge induces a novel intercrine/chemokine in mouse mast cells.

Abstract

SummaryIn an attempt to characterize genes participating in the allergic late phase reaction, we have isolated a novel intercrine/chemokine (called MARC) from a cDNA library of the stimulated mouse mast cell line, CPlI. As measured by Northern blotting, it is strongly upregulated at the mRNA level after the physiological challenge of the cells with immunoglobulin (Ig)E plus antigen. Unstimulated cells completely lack significant, stable expression, as do a number of other, different cell lines (uninduced and induced) and mouse tissues. In contrast to the Northern blot analysis, a polymerase chain reaction (PCR) analysis, performed on CPII ceUs and on Perco11 gradient purified mouse peritoneal mast cells, revealed a basal level of transcription in the uninduced stage. After 2 h of IgE plus antigen challenge, a quantitative reverse transcriptase-PCR, using a spiked in MIMIC, showed a level of transcripts more than 100-fold higher in the CPII cells and 5-20-fold higher in purified mouse peritoneal cavity mast cells. This rapid induction after the Fc+RI challenge, the identification of the gene as a member of the chemokine family, and its upregulated expression in peritoneal mast cells, all suggest an involvement in certain acute and chronic pathological mast cell-driven diseases.I ntercrines/r are small soluble proteins that regulate the physiological tra~cking and the partial activation ofleukocytes (1). In contrast to most other known cytokines and lymphokines, chemokines show a considerable homology (identity and similarity) at the amino acid level Additionally, they are characterized by a common protein structure of two loops formed via disulfide bridges of four highly conserved cysteines. Based on the location of the two NH2-terminal cysteines, this superfamily is subdivided into a CXC (IL-8 or PF4 family) and CC (RANTES/sis family) branch (2, 3). In the last, six members in the human system and five corresponding mouse genes are currently isolated (4, 5).Spedfic sites of production in the body and the low amount produced in vivo make the direct isolation and characterization of these proteins from healthy individuals nearly impossible. A few members, like macrophage inflammatory protein (MIP)-la, MIP-IB and monocyte chemotactic protein (MCP)I, have been purified at protein level from overexpressing tumor cell lines by functional monitoring, and the corresponding genes were subsequently identified (6, 7). However, the majority in the CC branch were isolated by inductionspecific differential hybridization of cDNA libraries (see reference 4). This reflects the fact that most members are strongly upregulated at the transcriptional level after cell activation (4). Supernatants from transiently transfected cell lines were used for a detailed functional analysis afterwards (8).Type I allergic reactions are characterized at the level of mast cells by a biphasic response. In an immediate reaction (up to several minutes) preformed low molecular weight substances, like histamine and serotonin, are released after IgE plu...

Gene expression of CC chemokines in experimental crescentic glomerulonephritis (CGN)

“…Since the cDNA sequences of rat CC chemokines, with the exception of MCP-1 [15], had not been published, we designed primers for PCR on the basis of the published sequences of murine MCP-3 [16], MIP-1a [17], MIP-1b [18], RANTES [19] and TCA3 [20]. RT-PCR with samples of rat RNA yielded a single band for each chemokine apart from RANTES (see below), and the size of each cDNA was identical to that of the corresponding mouse homologue.…”

“…5 mM MgCl 2 , 100 mg/ml bovine serum albumin (BSA), 1 mM each of dATP, dCTP, dGTP and dTTP, 0 . 5 mg oligo(dT) [12][13][14][15][16][17][18] and 200 U M-MLV Reverse Transcriptase (GIBCO BRL, Gaithersburg, MD). Reactions were stopped by heat inactivation at 90ЊC for 5 min and chilled on ice.…”

STAT3 and MITF cooperatively induce cellular transformation through upregulation of c-fos expression

“…JE and A7 are chemokines that are induced by immunoglobulin (Ig) E plus antigen stimulation in mast cells. (Burd et al, 1989;Kulmburg et al, 1992;Ong et al, 1993). Therefore, STAT3/MITF target genes are strongly linked to the function of mast cells and melanocytes, which constitutively express MITF.…”

Suppression by intradermal administration of heparin of eosinophil accumulation but not oedema formation in inflammatory reactions in guinea‐pig skin

“…Mast cells appear to play an important role in allergic diseases such as asthma and rhinitis (Wasserman, 1987;Holgate & Church, 1992). Thus, mast cells are directly responsible for the acute clinical manifestations of allergic diseases (Holgate & Church, 1992) but may also influence the chronic states through the release of cytokines (Plaut et al, 1989;Bradding et al, 1992;Kulmburg et al, 1992). Another feature of allergic inflammation is the presence of eosinophils which are potent effector cells capable of causing tissue destruction and dysfunction (Djukanovic et al, 1990;Kay, 1991).…”

“…Another feature of allergic inflammation is the presence of eosinophils which are potent effector cells capable of causing tissue destruction and dysfunction (Djukanovic et al, 1990;Kay, 1991). Mast cells are thought to contribute to the recruitment of eosinophils through the release of mediators including phospholipids and chemotactic cytokines (Kulmburg et al, 1992) and this may be relevant to allergic disorders such as asthma. ' Author for correspondence.…”