Epidermal Cell-derived Thymocyte-activating Factor Research Articles

EGFR and IL-1 Signaling Synergistically Promote Keratinocyte Antimicrobial Defenses in a Differentiation-Dependent Manner

Ligands of the EGF family regulate autocrine keratinocyte proliferation, and IL-1 family cytokines orchestrate epithelial defense responses. Although members of both families are overexpressed in wound healing and psoriasis, their roles in regulating the innate immune functions of keratinocytes remain incompletely explored. Using sensitive assays, we found significant increases of heparin-binding EGF-like growth factor, transforming growth factor-α, and amphiregulin mRNA and protein in lesional psoriasis compared with uninvolved or control skin. In normal human keratinocyte (NHK) monolayers, EGFR ligands were ineffective in inducing DEFB4, S100A7, and CCL20 mRNAs and human β-defensin (hBD)-2 peptide. Combined with IL-1α, however, EGFR ligands provoked 250 × more DEFB4 and CCL20 and a 9-fold rise in S100A7 mRNA relative to the EGFR ligand alone. This synergy was also reflected in secreted hBD-2 protein, both from NHK and reconstituted human epidermis. Keratinocyte differentiation was critical for these responses, as postconfluent NHK yielded mRNA and protein levels an order of magnitude greater than subconfluent cells. Differentiation also influenced signal transduction, with subconfluent cells using NF-κB and postconfluent cells using EGFR, MEK1/2, and p38. We propose that EGFR ligands are important modifiers of IL-1 activity, synergizing with IL-1 to stimulate epidermal production of hBD-2, S100A7, and CCL20, three of the most upregulated transcripts in psoriatic plaques.

The Interleukin-1 Axis and Cutaneous Inflammation.

Since the discovery that epidermal cell-derived thymocyte-activating factor was identical to interleukin (IL)-1 alpha and -beta in 1986, these molecules have been implicated in the pathogenesis of skin diseases. In 1995, it has become clear that a group of gene products function to regulate the activity of IL-1. IL-1 alpha and mature 17-kD IL-1 beta (cleaved from precursor by IL-1 beta-converting enzyme) bind to the type 1 IL-1 receptor to transduce a signal. This process can be antagonized at the level of the receptor by two distinct forms of the IL-1 receptor antagonist, which bind to the type I receptor but do not transduce a signal. The process can also be antagonized at the level of the ligand by either cell-bound or soluble type 2 IL-1 receptor. This type 2 IL-1 receptor binds ligand but does not transduce a signal. Keratinocytes can make each of these variables in vitro, and the balance between agonists and antagonists dictates the biologic outcome of a putative IL-1-mediated event. Transgenic mice that overexpress each of these factors individually in epidermis will be useful for enhancing our understanding of the cutaneous biology of IL-1.

The Interleukin-1 Axis and Cutaneous Inflammation

Since the discovery that epidermal cell-derived thymocyte-activating factor was identical to interleukin (IL)-1 alpha and -beta in 1986, these molecules have been implicated in the pathogenesis of skin diseases. In 1995, it has become clear that a group of gene products function to regulate the activity of IL-1. IL-1 alpha and mature 17-kD IL-1 beta (cleaved from precursor by IL-1 beta-converting enzyme) bind to the type 1 IL-1 receptor to transduce a signal. This process can be antagonized at the level of the receptor by two distinct forms of the IL-1 receptor antagonist, which bind to the type I receptor but do not transduce a signal. The process can also be antagonized at the level of the ligand by either cell-bound or soluble type 2 IL-1 receptor. This type 2 IL-1 receptor binds ligand but does not transduce a signal. Keratinocytes can make each of these variables in vitro, and the balance between agonists and antagonists dictates the biologic outcome of a putative IL-1-mediated event. Transgenic mice that overexpress each of these factors individually in epidermis will be useful for enhancing our understanding of the cutaneous biology of IL-1.

INHIBITORY EFFECT OF 8-METHOXYPSORALEN PLUS ULTRAVIOLET-A ON INTERLEUKIN-1 PRODUCTION BY MURINE KERATINOCYTES

We report the effects of 8-methoxypsoralen (8-MOP) plus ultraviolet-A (UV-A) irradiation on interleukin-1 (IL-1) production by murine epidermal keratinocytes, correlating its effect on IL-1 with cell viability, DNA synthesis, and 8-MOP-DNA photoadduct formation. Freshly isolated murine keratinocytes were treated with various doses of 8-MOP (5-100 ng/mL; incubation time, 30 min) plus 1 J/cm2 UV-A and cultured for 1-3 days. The IL-1/epidermal cell-derived thymocyte-activating factor (ETAF) activity in both supernatant and cell extract was reduced proportionately with increasing doses of 8-MOP/UV-A. Interleukin-1 inhibitors induced by 8-MOP plus UV-A were not detected in either supernatant or cell extract. A clear reduction of the IL-1 production was induced by the treatment as low as 15 ng/mL 8-MOP plus 1 J/cm2 UV-A, which led to the formation of 0.52 8-MOP photoadducts per million DNa bases and affected neither cell viability nor DNA synthesis of the treated cells. Cells treated with 100 ng/mL 8-MOP and 1 J/cm2 UV-A exhibited 57% suppression of IL-1 production in both 2- and 3-day culture samples. This treatment resulted in the formation of 3.8 photoadducts per million bases as well as significant abrogation of DNA synthesis although cell viability was unchanged. These observations provide some insights into the phototoxicity mechanisms of 8-MOP and the effect of PUVA therapy on the cytokine regulation in keratinocytes.

Production of Interleukin 1 Alpha by a Trichilemmoma Cell Line

In order to obtain a good source of human epidermal cell derived thymocyte activating factor (ETAF) for biological study, several cell lines of epidermal origin were cultured. One of the trichilemmoma cell lines, KTL-1, was found to produce ETAF satisfactorily in its culture supernatant. The culture supernatant was collected and analyzed to determine the nature of the ETAF activity. The ETAF activity was completely inhibited by anti-IL 1 alpha antibody, but not by anti-IL 1 beta antibody. Messenger RNA expression by KTL-1 cells was analyzed by using cDNA of IL 1 alpha as a probe. Messenger RNA was not detected in KTL-1 cells by either dot blot analysis or northern blot analysis. However, it was expressed weakly in the cytoplasm by in situ hybridization technique. Messenger RNA expression was easily detected in KTL-1 cells stimulated by lipopolysaccharide (LPS). Messenger RNA expression reached a peak at 3 hours after stimulation of LPS. A trichilemmoma cell line, KTL-1, can be a good source of human ETAF for biological study.

Normal human epidermal keratinocyte-derived neutrophil chemotactic factor.

Human epidermal keratinocytes constitutively produce a variety of cytokines, including neutrophil chemotactic peptide named epidermal cell-derived thymocyte-activating factor, which has been later confirmed to be interleukin 1 (IL-1). Because recombinant IL-1 lacks chemotactic activity, in the present study, we examined the exact nature of the neutrophil chemotactic peptide in the culture supernatant of normal human epidermal keratinocytes. Normal human epidermal keratinocytes produced a neutrophil chemotactic factor, which was also chemotactic for T lymphocytes. Molecular sieve chromatography revealed an approximate molecular size of 11,000 daltons. The activity was retained after heating at 100 degrees C for 10 min, and at a pH between 4 and 11, but was partially inactivated at pH 3, or by trypsin treatment. The chemotactic activity was not inhibited by the treatment with anti-IL-1 antibody. Its production by keratinocytes was stimulated by IL-1 and lipopolysaccharide but not by UV irradiation, tumor necrosis factor-alpha or by interferon-gamma. The neutrophil chemotactic activity in vivo was confirmed by the intradermal injection of the factor into guinea pigs. Blocking study with monoclonal antibodies against NAP-1/IL-8 confirmed that the neutrophil chemotactic factor is IL-8.

Lidocaine-Induced Inhibition of the Production of interleukin 1-Like Epidermal Cell-Derived Thymocyte Activating Factor

For the purpose of investigating the influence of the cationic anesthetic, lidocaine, on the production of epidermal cell-derived thymocyte activating factor (ETAF), murine epidermal cells were incubated for 1 h with 0.02-6 mg/ml lidocaine, washed, and incubated for an additional 23 h. The ETAF activity was assayed as the co-mitogenic activity of the crude epidermal cell supernatants on phytohemagglutinin-stimulated murine thymocytes. Lidocaine reduced the ETAF activity significantly, most markedly at a concentration of 2 mg/ml. The reduction was not caused by cytotoxicity, by co-production of inhibitory factors, or by modification of the ETAF molecule. Although the murine thymocyte assay was highly sensitive to lidocaine, the reduction of ETAF activity was not the result of carryover of lidocaine to the thymocyte assay. Our results indicate that lidocaine inhibits ETAF production in vitro, and suggest that conventional procedures, such as lidocaine anesthesia, which are acceptable for morphologic techniques, might not be suitable for functional studies of the cellular components of the skin.

A new role for epidermal cell-derived thymocyte activating factor/IL-1 as an antagonist for distinct epidermal cell function.

It is known that many immunologic responses to IL-1 are antagonized by the neuropeptide alpha-melanocyte stimulating hormone (alpha-MSH). This led us to investigate the possible reciprocal effects of IL-1 and the functionally related epidermal cytokines, epidermal cell-derived thymocyte activating factor (ETAF) and IL-6, on the melanogenic effect of alpha-MSH on murine Cloudman melanoma cells. When these cells were treated with ETAF in combination with alpha-MSH or its potent analog [Nle4,D-Phe7]-alpha-MSH, the melanotropin induced increase in tyrosinase activity, and thus melanin synthesis, was abrogated. This inhibitory effect of ETAF was not mediated by competitive binding to the melanotropin receptor, because ETAF also blocked the melanogenic response of melanoma cells to isobutyl methylxanthine (IBMX) and to PGE1 and PGE2. ETAF had no effect on cellular proliferation. Inhibition of the stimulated tyrosinase activity by ETAF was not due to diminished cAMP synthesis or increased cAMP degradation. Cells treated concomitantly with ETAF and alpha-MSH, IBMX, or PGE1 had the same cAMP levels as cells treated with alpha-MSH, IBMX, or PGE1 alone. In contrast to ETAF, human rIL-1 alpha or IL-1 beta alone or in combination did not have an inhibitory effect on melanogenesis. IL-6 significantly inhibited the basal level of tyrosinase and partially abrogated the alpha-MSH-induced tyrosinase activity. IL-6 also stimulated cellular proliferation when added alone or in combination with alpha-MSH. Granulocyte-macrophage colony stimulating factor (GM-CSF) did not alter either the tyrosinase activity or cellular replication at the concentrations tested. IL-1 alpha, GM-CSF, and IL-6 or IL-1 alpha and GM-CSF added together did not significantly affect the MSH-induced tyrosinase activity. These results ascribe a new potential function for ETAF and IL-6 as modulators of the melanogenic response of pigment cells.

Lichenoid Tissue Reaction Induced by Local Transfer of Ia-Reactive T-Cell Clones. III. Role of Ia+ Keratinocytes in the Epidermotropic Migration of the T Cells

The role of Ia+ keratinocytes in epidermotropism of T cells was analyzed by using self-IA specific autoreactive cloned T cells with epidermotropic nature (termed BB5) and those without it (termed C10). These T-cell clones were injected into the footpads of syngeneic nude mice whose keratinocytes had been induced to express Ia by the iv injection of normal mouse serum. Ia expression by keratinocytes was associated with the increased epidermal invasion of BB5 cells, but did not render C10 cells capable of migrating into the epidermis. The migration of the T cells to epidermis was also studied in vitro using a migration assay under agar. Ia expression by keratinocytes significantly enhanced the in vitro migration of BB5 cells to the epidermis, but had no effect on the migration of C10 cells and freshly isolated unstimulated lymphocytes. We surmise from these results that Ia+ keratinocytes may facilitate the epidermal invasion of the T cells with epidermotropic nature, but not those without it. However, the possibility certainly exists that the observed preferential migration of BB5 cells to Ia+ keratinocytes may be secondary to the alteration of other factors associated with the Ia expression. The injection of normal mouse serum was accompanied by an increase in the production of epidermal cell-derived thymocyte activating factor (ETAF) indicating that the increased ETAF production may have contributed to some of the observed preferential migration of BB5 cells.

Effects of topical corticosteroid therapy on Langerhans cell antigen presenting function in human skin.

We have investigated the mechanisms by which topical corticosteroids modulate cutaneous immune reactions in man. Volunteers applied clobetasone butyrate 0.05% (Eumovate; EV), betamethasone valerate 0.1% (Betnovate; BV), clobetasol propionate 0.05% (Dermovate; DV), and control vehicles twice daily to forearm skin for 7 days. Steroid therapy significantly decreased the number of HLA-DR/T6 (CD1a) positive Langerhans cells (LCs) per mm2 in suction blister-derived epidermal sheets, expressed as a mean percentage of controls, as follows: EV 69.2%; BV 67.3%; DV 37.8%. LC antigen presenting capacity was determined in the allogeneic and autologous epidermal cell-lymphocyte reactions. The LC-dependent allostimulatory capacity of epidermal cells, expressed as a mean percentage of controls, was also significantly reduced by steroid therapy: EV 45.1%; BV 41.9%; DV 23.4%. Following therapy with clobetasol propionate 0.05%, the capacity of epidermal cells to present tetanus toxoid to, and to augment concanavalin A mediated lymphocyte stimulation of, autologous lymphocytes was reduced to 33.6% and 19.7% respectively of controls. Depression of epidermal cell allostimulatory capacity was not the result of a steroid-induced decrease in the production of epidermal cell-derived thymocyte activating factor (ETAF)/interleukin 1 by keratinocytes, since it could not be reversed by addition of exogenous interleukin 1. Indomethacin, added to block any potential prostaglandin synthesis during the culture period, did not restore the allostimulatory capacity of epidermal cells from steroid-treated sites. Addition of epidermal cells from DV-treated sites depressed the capacity of control epidermal cells to stimulate lymphocytes in the allogeneic epidermal-lymphocyte reaction. Our results demonstrate that the anti-inflammatory action of topical corticosteroids in man is associated not only with a reduction in the number of HLA-DR/T6 positive LCs, but also with a marked decrease in Langerhans cell-dependent T lymphocyte activation. The effects of the different steroids on both of these parameters correlated with their potency as determined in the standard occlusive vasoconstrictor assay. Topical corticosteroids are widely used for the treatment of inflammatory skin disorders, and inhibit not only the elicitation phase, but also the induction phase, of allergic contact dermatitis reactions.(ABSTRACT TRUNCATED AT 400 WORDS)

Autocrine growth stimulation of human keratinocytes by epidermal cell-derived thymocyte-activating factor: implications for skin aging

The monocyte-derived cytokine interleukin-1 (IL-1) has growth-promoting activity for a variety of cell types, including lymphocytes and fibroblasts. We have previously shown that the epidermal cell-derived thymocyte-activating factor (ETAF) strongly resembles IL-1 in terms of biological, biochemical, and molecular biological properties. Because some lymphokines are known ot alter epidermal cell growth and differentiation and because cultured epidermal keratinocytes are capable of autocrine growth stimulation in vitro through "conditioning" of their culture medium, we sought to evaluate the effect of ETAF on keratinocyte growth. While there was marked donor variability in the responsiveness of keratinocytes to ETAF, partially purified preparations of ETAF showed substantial ability to stimulate the growth of keratinocytes, particularly those of newborn donors. In addition, in conditioned media there appeared to be activities distinct from ETAF that also promoted keratinocyte growth. Keratinocytes in serum-free medium secreted large amounts of ETAF, as reported previously, and keratinocyte cultures derived from newborn donors secreted significantly more than did those derived from adult donors. These results are consistent with an autocrine growth regulatory role of ETAF in human epidermis and with an age-associated loss of this phenomenon.

Role of epidermal cell thymocyte-activating factor in the proliferation and differentiation of murine B cells.

The role of antigen nonspecific cytokines in T- and B-lymphocyte responses is now well established. Interleukin-1 (IL-1) has been shown to augment B-cell maturation and proliferation. While the major source of IL-1 is from monocytes or macrophages, other cell types have been shown to produce IL-1-like cytokines. Epidermal cells produce a cytokine termed "epidermal cell-derived thymocyte-activating factor" (ETAF) which is similar if not identical with monocyte-derived IL-1. In this report we show that ETAF induces polyclonal stimulation of murine B cells. We show that ETAF augments B cell proliferation and differentiation in the absence of any added antigens or mitogens. This activity can be partially inhibited by anti-IL-1 antibodies. ETAF appears to activate B cells directly, although its activity is increased in the presence of T cells. Thus, ETAF may be involved in local polyclonal antibody responses occurring in the skin.

Expression of mRNA Homologous to Interleukin 1 in Human Epidermal Cells

Epidermal cells produce an interleukin 1 (IL-1)-related molecule known as epidermal cell-derived thymocyte activating factor (ETAF). Given their similar physical and biologic properties, it was presumed that ETAF would share homology to known IL-1 sequences. A murine IL-1 alpha complimentary DNA (cDNA) probe hybridized with a 2.1 kb RNA from the murine keratinocyte cell line PAMM 212. Further, a 24-base synthetic IL-1 beta probe complementary to a region 700 bases from the putative 5'-end of human IL-1 beta hybridized with 2.7- and 1.6-kb RNAs in human keratinocytes, monocytes, and promyelocytic leukemia cells. Reverse transcripts primed with the IL-1 beta oligonucleotide revealed major first-strand cDNAs from human keratinocyte COLO-16 cell mRNA and human monocyte mRNA of about 700 bases, consistent with the expected cDNA from IL-1 beta. A partial library constructed from this cDNA contained clones that hybridized with the IL-1 beta oligonucleotide. The 750 bp-IL-1 beta-like cDNA clones were not identical to IL-1 beta as judged by restriction mapping and Northern analysis. These cDNA clones hybridized with the 1.6-kb keratinocyte mRNA but not to the 1.6-kb monocyte nor to the 2.7-kb mRNA. The 1.6-kb keratinocyte mRNA may encode a new IL-1-like species.

Human Keratinocytes and Epidermoid Carcinoma Cell Lines Produce a Cytokine with Interleukin 3-Like Activity

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.

In vitro modulation of T6 expression on gingival Langerhans cells by interleukin-1 inhibitors and ETAF.

T6 antigen is a highly specific marker for human Langerhans cells (LC). Previous studies have demonstrated that Interleukin-1 (IL-1) and an IL-1 inhibitor (ILS) modulate LC T6 expression (T6E) in explant culture. The present study examined the in vitro modulation of T6E by two molecules: epidermal-cell-derived thymocyte-activating factor (ETAF), and a bone-derived protein (BP) implicated in the control of bone homeostasis. The effect of purified ILS was also examined. ETAF mimicked the stimulatory effect of IL-1 on LC T6E, while BP depressed T6E in a manner resembling that seen with ILS. No agents altered Class II (DR and DQ) expression by LC. BP was a specific IL-1 inhibitor, and did not inhibit thymocyte proliferation in the standard IL-1 bioassay in the absence of IL-1. These results demonstrate that molecules resembling IL-1 or ILS can modulate T6E, and implicate locally produced ETAF and ILS in the regulation of T6E in the oral mucosa in vivo.

Epidermal Cell-Derived Lymphocyte Differentiating Factor (ELDIF) Inhibits In Vitro Lymphoproliferative Responses and Interleukin 2 Production

We have examined the biologic characteristics and immunologic properties of epidermal cell-derived lymphocyte differentiating factor (ELDIF), a lymphocyte differentiating factor produced by cultured human keratinocytes. The ELDIF was semipurified by a gel filtration procedure. This factor, which is distinct from prostaglandins, epidermal cell-derived thymocyte activating factor (ETAF), and the well-known thymic hormones (thymulin, thymopoietin, and thymosin alpha 1) did not exhibit any interleukin (IL)-1, IL-2, or IL-3 activity. It strongly inhibited in vitro lymphoproliferative responses of normal mouse spleen cells to phytohemagglutinin, concanavalin A, and lipopolysaccharide. This dose-dependent phenomenon was associated with a suppression of IL-2 production rather than any toxic effect. It can be concluded that ELDIF, a product of human epidermal cells, which displays in vitro T-cell differentiation and regulatory activities, could be of major importance in vivo in the control of cutaneous inflammatory reactions.

Formation of Epidermal Cell Thymocyte-Activating Factor (ETAF) From Cultured Human Keratinocytes From Normal and Uninvolved Psoriatic Skin

Cultures of keratinocytes from uninvolved psoriatic skin have been shown to exhibit increased DNA synthesis. Since epidermal cell-derived thymocyte-activating factor (ETAF) can stimulate the proliferation of human keratinocytes in vitro, we have compared the formation of ETAF in cultured keratinocytes from normal and uninvolved psoriatic skin. Trypsinized epidermal cells were plated at nonconfluent concentrations in dishes coated with a collagen type I gel. Normal keratinocyte cultures spontaneously released ETAF into the supernatant from day 1, reaching a maximum level (mean 124 +/- 4 units/micrograms DNA) on day 7, just before the cultures became completely confluent (days 9-12). The ETAF release then gradually decreased until a plateau (mean 16 +/- 7 units/micrograms DNA) was reached on day 18. In normal keratinocyte cultures the incorporation of [3H]thymidine into DNA changed in parallel with the ETAF release. Psoriatic keratinocytes had normal rates of ETAF release and of DNA synthesis until the time of culture confluency. However, when complete confluency was obtained, psoriatic keratinocytes continued to synthesize DNA at high levels (increased by 146% on day 15) while their ETAF release declined. Analysis of intracellular ETAF showed a significant correlation with the extracellular ETAF in cultures from both normal (r = 0.77, n = 10, p less than 0.02) and uninvolved psoriatic skin (r = 0.76, n = 12, p less than 0.01). These results indicate a relation between ETAF formation and DNA synthesis of normal keratinocyte cultures. However, factors other than ETAF appear to be responsible for the elevated DNA synthesis of confluent keratinocyte cultures from uninvolved psoriatic skin.

Keratinocyte Membrane-Associated Epidermal Cell-Derived Thymocyte-Activating Factor (ETAF)

This study describes the association between secreted keratinocyte interleukin 1 (IL-1) and its presence on the keratinocyte cell surface. These properties were studied in normal and transformed human keratinocytes as well as in transformed murine keratinocytes. We will present evidence that the secretion of IL-1 by human and murine keratinocytes is associated with the presence of IL-1 on the keratinocyte membrane. In addition, although transformed murine keratinocytes secrete other cytokines, namely keratinocyte T-cell growth factor (KTGF) and IL-3, no KTGF or IL-3 activity can be demonstrated on the cell surface.

Stimulation of Muscle Protein Degradation by Murine and Human Epidermal Cytokines: Relationship to Thermal Injury

Accelerated muscle proteolysis is a characteristic of systemic reaction following trauma, sepsis, or extensive thermal injury. The factors involved in this accelerated muscle breakdown have not been fully described. However, recently leukocytic pyrogen or interleukin 1 (IL-1) have been implicated in the induction of muscle protein degradation in septicemia or trauma. The epidermal cytokine epidermal cell-derived thymocyte activating factor (ETAF) is biochemically and functionally similar to IL-1. Injury to skin can augment ETAF activity. Using a murine model, we found that thermal injury can significantly enhance ETAF/IL-1 activity in a dose-dependent fashion. In addition, ETAF can cause net muscle protein breakdown in vitro. Thus, increased amounts of ETAF produced by thermally injured skin may contribute to the accelerated muscle breakdown in extensive thermal injury.

Human keratinocytes contain mRNA indistinguishable from monocyte interleukin 1 alpha and beta mRNA. Keratinocyte epidermal cell-derived thymocyte-activating factor is identical to interleukin 1.

Keratinocytes produce an IL-1 like factor termed epidermal cell-derived thymocyte-activating factor (ETAF). In this study, we show that ETAF and IL-1 are identical by the following criteria: Both normal and malignant human keratinocytes contain mRNAs identical to monocytic IL-1 alpha and IL-1 beta mRNA, as determined by an S1 nuclease protection assay; and IL-1 activity in medium conditioned by these cells can be neutralized by antibodies specific for human IL-1. The IL-1 alpha and IL-1 beta mRNAs can be identified in cultured human keratinocytes in the absence of identifiable stimulation; this basal level of mRNA can be further induced to accumulate with certain defined stimuli. Cultured normal human keratinocytes (HFKs) contain 2-4 times more IL-1 alpha than IL-1 beta mRNA; in contrast, human peripheral blood monocytes contain 10-20 times more IL-1 beta than IL-1 alpha mRNA. The IL-1 activity released by these HFK can be neutralized by an antibody that neutralizes both alpha and beta IL-1, but not by an antibody that neutralizes only IL-1 beta. While human monocytes produce a large excess of IL-1 beta after appropriate stimulation, these data suggest that IL-1 alpha is a major (and may be the predominant) form of IL-1 produced by human keratinocytes.