Topical cyclosporine A inhibits the phorbol ester induced hyperplastic inflammatory response but not protein kinase C activation in mouse epidermis.

Cyclosporine A (CsA) is efficacious in the treatment of psoriasis. Although CsA is known to inhibit T-lymphocyte proliferation in vitro, whether this is its mode of action in psoriasis is uncertain. 12-0-tetradecanoylphorbol-13-acetate (TPA) induces an inflammatory, hyperplastic response in mouse skin, with many of the biochemical and histologic aberrations that occur in psoriatic epidermis. Protein kinase C is the major cellular phorbol ester receptor, and most responses of cells to TPA are mediated by PK-C, which is directly activated by TPA. We therefore have investigated the effects of CsA on pleiotypic responses induced by TPA and whether CsA acts in vivo as a direct inhibitor of PK-C. Simultaneous application of CsA (1.7 mumol) and TPA (10 nmol) to mouse skin significantly reduced inflammatory cell infiltration and increased epidermal thickness induced by TPA treatment alone. CsA had to be applied within 30 min of TPA application in order to have a significant inhibitory effect. Optimal doses of CsA inhibited TPA-induced ODC activity, TGase activity, arachidonic acid release, and interleukin-1 beta (IL-1 beta) mRNA to the same degree (approximately 80%), despite measurement at widely different times (30 min-12 h) required to obtain maximal induction by TPA. CsA did not, however, directly inhibit activation of PK-C by TPA. These data demonstrate that CsA blocks the pleiotypic responses of mouse skin to TPA treatment involving biochemical events, inflammatory cell infiltration, and epidermal hyperplasia. The molecular site(s) of action of CsA appears to be distal to the initial activation of PK-C by TPA and clearly inhibits PK-C inducible events. Furthermore, the above data suggest that CsA may directly affect keratinocytes in vivo.

Chloroquine and onchocerciasis.

My original elective proposal was to study the prevalence and nature of onchocerciasis or 'river blindness' along the Canandé River in Esmeraldas province of Ecuador (Cooper, 1986). However, unknown to me, this study was completed and published before I arrived (Guderian et al, 1987). Fortunately, on arrival I was able to join the same group on a study of the efficacy of chloroquine in the control of onchocerciasis.

Sphingosine inhibits phorbol ester-induced inflammation, ornithine decarboxylase activity, and activation of protein kinase C in mouse skin.

Tumor promoting phorbol esters, such as 12-0-tetradecanoylphorbol-13-acetate (TPA), when applied topically to mouse skin cause inflammation and hyperplasia. The major cellular phorbol ester receptor is a calcium and phospholipid dependent protein kinase, protein kinase C (PK-C). PK-C is directly activated by TPA and most of the responses of cells to TPA appear to be mediated by PK-C. This suggests that PK-C may play a key role as a mediator of inflammation and growth in TPA treated mouse skin. Sphingosine has been reported to be a potent inhibitor of PK-C in vitro and in intact leukocytes. We therefore have investigated the effects of sphingosine upon TPA-induced inflammation, hyperplasia, induction of ornithine decarboxylase (ODC) activity and ODC mRNA, and activation of PK-C in mouse skin. The results demonstrate that sphingosine is a potent inhibitor of all of the TPA-induced responses examined. These data are compatible with the hypothesis that PK-C is a major mediator of the phorbol ester response in mouse skin. Furthermore, PK-C inhibitors may have therapeutic potential in inflammatory skin diseases such as psoriasis.

Induction of dermal and subcutaneous inflammation by recombinant cachectin/tumor necrosis factor (TNF alpha) in the mouse.

The ability of cachectin/tumor necrosis factor (TNF alpha) to induce acute dermal and subcutaneous inflammation was examined in a murine model. A number of other proteins, and diluent alone were examined as controls. After subcutaneous injection into the mouse footpad, recombinant human TNF alpha (rHuTNF alpha) induced acute inflammation with an initial marked dermal and subcutaneous neutrophil infiltrate by approximately 3 h, with a peak between 4 and 24 h and resolution by 79 h. Recombinant interleukin-2, cytochrome c, and heat-inactivated rHuTNF alpha induced negligible inflammation. Recombinant human lymphotoxin (TNF beta), another control protein, also induced acute inflammation in our system. Because TNF alpha and TFN beta are partially homologous, they may be acting through a similar mechanism. This pro-inflammatory effect of TNF alpha may result from chemotactic activity as well as by induction of secondary mediators. Inflammation induced by TNF alpha was partially suppressed by indomethacin treatment, suggesting that products of the cyclo-oxyganase pathway may mediate a portion of the inflammation involved. Five daily injections of rHuTNF alpha into the mouse footpad resulted in a predominantly mononuclear infiltrate and focal fibrosis. These results suggest that TNF alpha may be an important mediator of acute inflammation in vivo and might provide a signal for the production of collagen.

Actions of platelet activating factor (PAF) homologues and their combinations on neutrophil chemokinesis and cutaneous inflammatory responses in man.

The inflammatory actions of synthetic C16:0 and C18:0 platelet activating factor (PAF) homologues, both alone and in combination, have been compared in an in vitro human neutrophil chemokinesis assay and by intradermal injection in human skin. In the chemokinesis assay, the maximum distance moved by neutrophils in the presence of C18:0 PAF was significantly greater than that seen with the C16:0 compound. A mixture of C16:0 and C18:0 PAFs in a ratio of 1:9 appeared to be more active than in a ratio of 3:1. Intradermal injection of the C16:0 and C18:0 PAF homologues induced dose-dependent increases in weal volume and flare area responses which were not significantly different. Combination of these phospholipids in a ratio of 3:1 or 1:9 of C16:0:C18:0 did not significantly alter the dose response curves. Thus, changes in the chain length of the alkyl substituent of synthetic PAF homologues and combination of these homologues, in ratios found in vivo or formed by leukocytes in vitro, did not alter the cutaneous inflammatory responses to PAF in man. The C18:0 homologue was, however, more active as a human neutrophil chemoattractant in vitro.

Mast cell-dependent amplification of an immunologically nonspecific inflammatory response. Mast cells are required for the full expression of cutaneous acute inflammation induced by phorbol 12-myristate 13-acetate.

Mast cells clearly are critical for the expression of some IgE-dependent responses, but their roles in other forms of inflammation are uncertain. We previously described a new model system for defining the unique contribution of mast cells to biologic responses in vivo, genetically mast cell-deficient WBB6F1-W/Wv mice that have undergone selective local repair of their mast cell deficiency by the injection of IL-3-dependent cultured mast cells derived from the congenic normal (WBB6F1-+/+) mice. Using this approach, we analyzed the contribution of mast cells to the acute inflammation induced by the epicutaneous application of PMA. Even though PMA can activate a wide variety of cell types that may contribute to acute inflammation, we found that mast cells were required for the full expression of the tissue swelling and leukocyte infiltration associated with the response to the agent in vivo. Thus, in WBB6F1-W/Wv mice selectively reconstituted with dermal mast cells by intradermal injection of cultured WBB6F1-+/+ mast cells into the left ear only, PMA induced approximately twice the tissue swelling and neutrophil infiltration in the mast cell-reconstituted left ears as in the contralateral control ears. This represents the first use of W/Wv mice locally reconstituted with mast cells to confirm the hypothesis that mast cells can represent an important amplification mechanism in acute inflammatory responses of nonimmunologic origin. It also defines a model system that may be generally useful for investigating mast cell-dependent and -independent aspects of acute inflammatory responses.

A direct in vivo comparison of the inflammatory properties of human C5a and C5a des Arg in human skin.

C5a is an 11,000-Da complement-derived inflammatory glycoprotein that has been shown to mediate inflammatory reactions in vitro as well as in vivo in human skin. The C5a degradation product, C5a des Arg, is rapidly formed after exposure of C5a to serum carboxypeptidase N and may represent the relevant C5-derived inflammatory peptide in vivo. To examine the biologic activity of human C5a des Arg in vivo and to compare it with that seen with human C5a, we purified and characterized homogeneous preparations of human C5a and C5a des Arg and injected them intradermally into seven normal volunteers. C5a des Arg exhibited biochemical and biologic properties in vitro that were different from those of C5a. When injected into human skin, C5a des Arg was less potent than C5a, in respect to both minimal dose eliciting wheal and flare reactions and maximal wheal and flare elicited at a given dose, but C5a des Arg still elicited cutaneous wheal and flare reactions at physiologically relevant concentrations. Histologically, C5a des Arg skin test sites showed dense polymorphonuclear neutrophil-rich infiltrates associated with leukocytoclasis, dermal mast cell degranulation, and endothelial cell swelling. These were virtually indistinguishable from reactions elicited by C5a and occurred with concentrations attainable in vivo. Cutaneous wheal and flare reactions elicited by either C5a or C5a des Arg were partially inhibited by H1 antihistamines but were unaffected by selected nonsteroidal anti-inflammatory agents.

Sources of extracellular lysosomal enzymes released in organ-culture by developing and healing inflammatory lesions.

Developing and healing inflammatory lesions were topically produced in the skin of rabbits by sulfur mustard (SM). After the rabbits were sacrificed, the various lesions were removed and organ-cultured. The organ-culture fluids extracted the extracellular lysosomal enzymes (acid phosphatase, beta-glucuronidase, beta-galactosidase, and lysozyme), so that they could be measured biochemically along with lactic dehydrogenase (LDH), an enzyme marker for cell death. In tissue sections, the number and types of cells were counted, and their lysosomal enzyme content evaluated histochemically. The culture fluids from peak lesions contained much lower levels of all five enzymes than did culture fluids from healing lesions. When histological-histochemical-biochemical correlations were made, serum, macrophages (MN), and activated fibroblasts (but not tissue PMN) appeared to be major sources of extracellular lysosomal enzymes in peak lesions; and the dead PMN in the crusts and the activated fibroblasts in the tissues appeared to be major sources in healing lesions. The high lysosomal enzyme content of the crusts covering the lesions suggests that this passive barrier may also play an active role in promoting healing and in protecting against invasion by microorganisms.

Association between clinical symptoms and lymphocyte abnormalities in a population with chronic domestic exposure to industrial solvent-contaminated domestic water supply and a high incidence of leukaemia.

An unusually high incidence of leukaemia and recurrent infections was noted in children exposed in utero to domestic water supply contaminated with industrial solvents including trichloroethylene, perchloroethylene and 1,2-transdichloroethylene. Medical and laboratory investigations were carried out on 28 family members of the patients with leukaemia with particular emphasis on the immunological system to determine if they displayed symptoms associated with acute or chronic exposure to these chlorinated hydrocarbons. The principal organ systems affected were neurological, immunological and cardiological. Damage to these systems was found in all subjects by history, physical and laboratory parameters. Damage to the immunological system was manifest by altered ratios of T lymphocyte subpopulations, increased incidence of auto-antibodies, increased infections and recurrent rashes.

In vitro damage of basal lamina-associated anionic sites by hydroxyl radical.

We examined the effect of chemically generated hydroxyl radical on basal lamina-associated anionic sites. Cytochemical studies showed that hydroxyl radical produced a loss of cationic tracer-positive, anionic particles, and this effect was inhibited by a specific scavenger, thiourea. These data might suggest that anionic sites were degraded by hydroxyl radical which was derived, for example, from the activated leukocytes in close contact with the dermal-epidermal junction during acute inflammation resulting in the disturbance of the charge-selective permeability barrier.

Recent investigations of mechanisms of chemically induced skin irritation in laboratory mice.

The time course, dose response, components of inflammation, and involvement of putative mediators of inflammation in irritation induced by different chemicals was compared using a mouse ear swelling technique. Differences in time courses of inflammation produced by the irritants were not solely due to differences in rates of penetration. Changes in blood flow and permeability of vessels were phasic with different numbers of phases induced by different irritants. Effects of antagonist, synthesis, inhibitors, and depleting agents of putative inflammatory mediators on intensity of inflammation varied for different irritants. These studies demonstrate that all chemicals do not produce skin irritation by a common inflammatory pathway.

Reduction of anthralin inflammation by potassium hydroxide and Teepol.

Application of 1% potassium hydroxide (KOH) reduced subsequent development of anthralin inflammation without loss of its therapeutic effect on psoriasis. Teepol had a similar but smaller effect on subsequent development of inflammation. The action of KOH appears to have resulted from enhanced oxidation of anthralin to inactive products and the action of Teepol to have increased anthralin solubility and removal. The effect of KOH and Teepol decreased with time after anthralin application and both were ineffective by 24 h, indicating that anthralin persists on the skin in an active form for up to 24 h after a single application. The reduction of anthralin inflammation without loss of therapeutic effect is potentially useful in short contact anthralin therapy.

Dapsone syndrome--a case report.

A 31-year-old male patient with lepromatous leprosy developed fever, malaise, nausea, anorexia, lymphadenopathy, hepatitis, exfoliative dermatitis and ainhum like lesions while on multidrug therapy comprised of dapsone, clofazimine and rifampicin. The provocation tests confirmed the dapsone to be cause of this event.

Inflammatory mediators and modulators released in organ culture from rabbit skin lesions produced in vivo by sulfur mustard. III. Electrophoretic protein fractions, trypsin-inhibitory capacity, alpha 1-proteinase inhibitor, and alpha 1- and alpha 2-macroglobulin proteinase inhibitors of culture fluids and serum.

This is the third report in a series on the inflammatory mediators and modulators released in organ culture from skin lesions of various ages, which were produced in vivo in rabbits by the military vesicant, sulfur mustard (SM). It describes the electrophoretic protein fractions and trypsin-inhibitory capacities of the various culture fluids and the amounts of alpha 1-proteinase inhibitor and alpha-macroglobulin proteinase inhibitors in these fluids. With one-dimensional electrophoresis, the albumin and beta-globulin fractions of protein in culture fluids varied little with the development and healing of the SM lesions. These fractions proportionally resembled the corresponding fractions found in serum. The alpha 1-globulin fraction was proportionally smaller than the corresponding fractions of serum as the lesions healed. The alpha 2-globulin fraction was proportionally smaller than the corresponding fractions of serum at all stages of lesion development and healing. The gamma-globulin fraction was proportionally larger as the lesions healed. With two-dimensional electrophoresis, about 68%, 46%, and 35% of the protein spots in culture fluids from representative 1-day and 6-day SM lesions and normal skin, respectively, matched those from serum. In each case, the large, diffuse, serum albumin spot represented about two-thirds of the protein present. Thus, gravimetrically, in normal skin and in both developing and healing lesions, the extracellular proteins were 80-90% of serum origin. The trypsin-inhibitory capacity (TIC) per milligram protein in the culture fluids of healing lesions was markedly less than the TIC per milligram protein in the fluids of peak lesions. This decrease correlates well with the decrease found in the alpha 1-globulin fraction, which contains alpha 1-antiproteinase (alpha 1-PI) (and alpha 1-macroglobulin [alpha 1M] in rabbits). The alpha 1PI and the alpha 1M-alpha 2M proteinase inhibitors were identified in the culture fluids by means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western blots, specific antibodies, and the immuno-peroxidase technique. The levels of both free and proteinase-complexed alpha 1PI and alpha M inhibitors in the culture fluids decreased as the lesions healed. In both developing and healing lesions, at least half of the alpha 1PI and alpha M inhibitors seemed to be complexed with proteinases. Thus, serum seems to be a major source of unbounded extracellular protein within acute inflammatory lesions, and serum proteinase inhibitors seem to be the host's major defense against local damage by proteinases from serum, infiltrating leukocytes, and activated fibroblasts.

The inflammatory response of rabbit skin to topical arachidonic acid and its pharmacological modulation.

The inflammatory reaction induced by the intradermal injection of arachidonic acid into the rabbit dermis has been investigated. Plasma extravasation was measured by the leakage of 125I-albumin into the tissues and polymorphonuclear leukocyte (PMNL) accumulation was assessed histologically. Arachidonic, 5,8,11,14,17-eicosapentaenoic and 8,11,14-eicosatrienoic acids, but not oleic, linoleic or linolenic acids, caused a concentration-related plasma extravasation following their intra-dermal injection. The plasma extravasation induced by arachidonic acid was dependent on PMNLs. PMNL infiltration and plasma extravasation into arachidonic acid-injected skin sites was inhibited by the mixed cyclo-oxygenase-lipoxygenase inhibitor, BW755C. Arachidonic acid-induced plasma extravasation was inhibited by cyclo-oxygenase and 5-lipoxygenase inhibitors but not by the Paf antagonist, kadsurenone. The inflammation induced by arachidonic acid in the rabbit dermis may be a useful model for evaluating 5-lipoxygenase inhibitors which could be potentially useful anti-inflammatory agents for the treatment of psoriasis and other inflammatory diseases.

Inflammatory effect of intradermal administration of soluble phospholipase A2 in rabbits.

Extracellular phospholipase A2 (PLA2) has been found in association with inflamed sites in experimental animals and in humans. The tissue effects of soluble PLA2 have not been defined. We studied the development of inflammatory changes in rabbit skin subsequent to intradermal injection of active and inactivated venom and pancreatic PLA2, over a broad concentration range. PLA2, at concentrations encountered in human disease, caused acute inflammatory changes characterized grossly by erythema and induration, and histologically by inflammatory cell infiltration, vascular and tissue damage, and abscess formation. Extracellular PLA2 may be considered as one of the pathogenic factors in inflammatory reaction.

Rheumatoid rigor: gold induced myokymia. A report and review of the literature.

Myokymia is a spontaneous movement disorder characterized by slow, undulating, persistent movement of the involved muscles. It is an unusual manifestation of gold neurotoxicity, rarely reported in the rheumatologic literature. We report a case of gold induced myokymia associated with gold-induced dermatitis and nephropathy.

Toxicity due to remission inducing drugs in rheumatoid arthritis. Association with HLA-B35 and Cw4 antigens.

Twenty-five patients with rheumatoid arthritis (RA) who developed toxicity while taking remission inducing drugs and 30 without toxicity were studied for possible associations with class I and II HLA antigens. A strong association has been found between nephritis and dermatitis due to Tiopronin (a D-Penicillamine like compound) and class I antigens B35-Cw4, and between dermatitis due to gold thiosulphate and B35. Compared to healthy controls a lower DR5 frequency was observed in patients with RA except for the Tiopronin related nephritis group.

D-penicillamine-induced acute hypersensitivity pneumonitis and cholestatic hepatitis in a patient with rheumatoid arthritis.

Acute hypersensitivity pneumonitis is not a known side effect of D-penicillamine. Also, there are only a few reports of cholestatic hepatitis caused by this drug. The present report describes a case of rheumatoid arthritis where these manifestations appeared in the same patient ten days after the institution of D-penicillamine therapy. The etiological relationship with the drug was established, inadvertantly, by its accidental intake by the patient 5 months later with the reproduction of the same manifestations.