Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UNIPROT:P15088 (mast cell)
 14,925 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Keratinocytes are capable of releasing distinct immunomodulating cytokines such as epidermal cell-derived thymocyte activating factor (ETAF) and an epidermal cell-derived natural killer cell augmenting factor (ENKAF). The present study was performed to determine whether human keratinocytes also may secrete an interleukin 3 (IL-3)-like mediator and thereby participate in the regulation of mast cell activity in the skin. Supernatants of freshly isolated human epidermal cells (EC) and malignant keratinocyte cell lines (A 431, SCC) were tested for their capacity to induce the proliferation of IL-3-dependent cell lines 32 DCL and FDCP. Human epidermal cell interleukin 3 (EC IL-3) is spontaneously released by freshly isolated EC, A 431, and squamous cell carcinoma (SCC) cells. However, both normal EC and A 431 cells produced increased levels of EC IL-3 activity when cultured in the presence of different stimulants, such as phorbol myristate acetate and lipopolysaccharide. The EC IL-3 activity was not inhibited when treated with a monoclonal anti-IL-1 or anti-IL-2-antibody. Biochemical characterization showed that human EC IL-3 has a molecular weight of 17K, elutes of DEAE-ion exchange high-performance liquid chromatography (HPLC) as one major peak at 0.36 M NaCl, and upon HPLC-chromatofocusing exhibits 3 isoelectric points of 7.8, 7.5, and 5.6. Upon reversed-phase HPLC, EC IL-3 activity eluted at about 100% acetonitrile. When highly purified EC IL-3 was labeled with 125I and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, a single homogeneous band exhibiting a molecular weight of 17K was seen, which correlated with the IL-3 activity and was free of ETAF/IL-1, IL-2, and interferon activity. These data indicate that human EC synthesize an IL-3-like cytokine which is distinct from ETAF/IL-1, IL-2, and interferon and thereby may participate in the regulation of mast cell activity during inflammatory and fibrotic, as well as hypersensitivity reactions.
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PMID:Human keratinocytes and epidermoid carcinoma cell lines produce a cytokine with interleukin 3-like activity. 243 14

High-performance liquid chromatography (HPLC) is useful for the purification and separation of immunoregulatory cytokines, such as macrophage-derived interleukin 1 (IL 1). In addition to macrophages, epidermal cells also release a mediator, epidermal cell (EC) derived thymocyte-activating factor (ETAF), which cannot be separated from IL 1. Moreover, it has been shown recently that EC produce a distinct interleukin 3-like mast cell-activating factor (EC IL 3). This study was performed to investigate whether HPLC may be useful for the separation of EC-derived cytokines, such as ETAF and EC IL 3. For factor production, a murine EC line (Pam 212) was used. ETAF activity was measured using the thymocyte costimulator assay. EC IL 3 was was determined by induction of the proliferative activity of an IL 3-dependent cell line (32 DCL). Using a TSK 125 size-exclusion column and phosphate-buffered saline (pH 7.2) as the mobile phase, ETAF was eluted with an apparent molecular weight of 17 kD, and EC IL 3 with a molecular weight of 28 kD. When EC supernatants were chromatofocused on a Mono P column, ETAF activity was eluted with apparent pI values of 6.8, 6.2 and 5.3, and EC IL 3 activity with pI 7.8, 7.4 and 7.1. When reversed-phase HPLC (RP-HPLC) (equilibration with water and a 0-100% concave acetonitrile gradient) was applied ETAF exhibited four distinct peaks, whereas EC IL 3 was eluted as one major peak between 70 and 80% acetonitrile. Separation on a Bio-Gel HPHT column with a sodium phosphate gradient was not satisfactory, but the recovery was high.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:High-performance liquid chromatographic separation of distinct epidermal cell-derived cytokines. 389 54