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Biomedical subjects

M Cumberbatch

Publications and source records attributed to M Cumberbatch.

At least 37 records · Page 2Linked to original sources
PubMed1999

Inhibition by dexamethasone of Langerhans cell migration: influence of epidermal cytokine signals.

The influence of dexamethasone (DEX), a synthetic glucocorticoid, on the induction in mice of Langerhans cell (LC) migration has been investigated. Systemic treatment of mice with DEX was found to inhibit significantly the ability of a topically applied contact allergen (oxazolone) to induce the migration of LC from the epidermis and their subsequent accumulation as dendritic cells (DC) in draining lymph nodes. The stimulation of LC migration during skin sensitization is dependent upon signals provided by the epidermal cytokines tumour necrosis factor alpha (TNF-alpha) and interleukin 1beta (IL-1beta). It was found that treatment with DEX was unable to inhibit either LC migration or DC accumulation induced by the intradermal injection of TNF-alpha. In contrast, LC migration provoked by similar exposure of mice to IL-1beta (the action of which is dependent upon the de novo synthesis of TNF-alpha) was inhibited by DEX as was the arrival of DC in draining lymph nodes induced by this cytokine. Taken together, the data reported here indicate that DEX is able to inhibit very markedly the stimulation of LC migration during skin sensitization and it is proposed that such inhibition may represent an important aspect of the immunosuppressive properties of glucocorticoids and of their proven utility in the treatment of cutaneous inflammatory disorders. The results also indicate that DEX does not inhibit LC migration secondary to direct effects on cell motility. The proposal is that impaired LC migration results from the regulation by DEX of the de novo synthesis and/or release of TNF-alpha, an inducible epidermal cytokine that provides one important signal for LC to traffic from the skin.

Animals↗
PubMed1999

Tumour necrosis factor-alpha induces Langerhans cell migration in humans.

The role of tumour necrosis factor (TNF)-alpha in the mobilization and migration of human epidermal Langerhans cells (LC) has been investigated. Intradermal injection of normal human volunteers with homologous recombinant TNF-alpha was found to cause a dose-dependent reduction in the frequency of LC within epidermal sheets 2 h later. Equivalent results were obtained when epidermal LC were identified on the basis of either CD1a or HLA-DR expression. At the dose of TNF-alpha used routinely (500 U), treatment resulted in an average reduction in LC density of approximately 24%. Treatment with TNF-alpha was associated with a perivascular polymorphonuclear infiltration at 2 h, but the epidermis appeared normal with neither fibrinoid necrosis nor vasculitis, and LC morphology was not affected significantly. These results demonstrate that TNF-alpha provides an important signal for LC migration in humans and is likely therefore to play a crucial part in the induction of cutaneous immune responses.

Adult↗
PubMed1999

Possible antimigraine mechanisms of action of the 5HT1F receptor agonist LY334370.

This study investigated whether the selective 5HT1F receptor agonist LY334370 has other possible antimigraine mechanisms in addition to the proposed inhibition of dural plasma extravasation. LY334370 (up to 10(-5) M) had no vasoconstrictor effects on human cerebral arteries in vitro. It had no effect (up to 10 mg kg-1, i.v.) on neurogenic vasodilation of dural blood vessels produced by electrical stimulation of the dura mater in anesthetized rats. Nor had it any effect (at 3 mg kg-1, i.v.) on the hyperalgesia produced by injection of carrageenan into the paw of conscious rats or on nociceptive reflex responses in the spinalized, decerebrate rabbit (up to 3 mg kg-1, i.v.), indicating that it has no general analgesic properties. However, it significantly inhibited activation of second-order neurons in the trigeminal nucleus caudalis produced by electrical stimulation of the dura mater in anesthetised rats at 3 mg kg-1, i.v. These results provide evidence to suggest that LY334370 has a central mechanism of action in blocking the transmission of nociceptive impulses within the trigeminal nucleus caudalis and that this may represent a mechanism through which it has its antimigraine effect.

Animals↗
PubMed1998

Characteristics and regulation of the expression on interleukin 1 receptors by murine Langerhans cells and keratinocytes.

There is increasing evidence that interleukin 1beta (IL-1beta), a product in murine epidermis of Langerhans cells (LC) exclusively, contributes to the initiation and regulation of LC migration in response to skin sensitization. The hypothesis is that IL-1beta induces the production by keratinocytes of tumour necrosis factor alpha (TNF-alpha) which acts in a paracrine fashion on LC to provide one signal for migration. In addition, it is believed that IL-1beta acts in an autocrine fashion on LC to provide a second, TNF-alpha-independent, signal for the initiation of this response. The viability of this hypothesis is dependent upon the availability of appropriate membrane receptors. We describe therefore experiments designed to investigate the IL-1 receptor (IL-1R) status of keratinocytes, LC and lymph node dendritic cells (DC). Flow cytometric analyses of epidermal cell suspensions revealed at least 60% of LC positive for the type I IL-1R (IL-1RI). In contrast, only a small proportion of keratinocytes displayed surface IL-1RI, although high intracellular expression of this receptor could be detected either by flow cytometric analysis of cells permeabilized with saponin or by immunofluorescence microscopy. Expression of the type II IL-1R (IL-1RII) was detected at relatively low levels on both LC and keratinocytes. Interestingly, DC isolated from the lymph nodes of sensitized mice displayed upregulated expression of IL-1RI and lower levels of IL-1RII compared to LC. The conclusion drawn is that the IL-1R phenotype of LC and keratinocytes under resting conditions is consistent with the proposed contribution of IL-1beta to LC migration. Furthermore, the regulation of IL-1R expression by epidermal cells and DC will undoubtedly influence the development of cutaneous immune responses.

Animals↗
PubMed1998

Langerhans cells and chemical allergy.

Epidermal Langerhans cells (LCs) play a pivotal role in the induction of cutaneous immune responses, including those provoked by chemical allergens. The delivery by LCs of allergen to draining lymph nodes requires cell migration from the skin, a process that is dependent upon the availability of epidermal cytokines -particularly TNF-alpha and IL-1beta. Here we consider the ways in which these cytokines interact with LCs to both induce and regulate their mobilization in response to skin sensitization. In addition, the effects of these cytokines on both the selectivity of LC migration from the skin and protection of LCs from cell death are considered. Finally, the possible counter-regulatory activity of other cutaneous cytokines and the influence of LCs on the development of selective T lymphocyte responses are explored.

Allergens↗
PubMed1997

Alpha 6 integrins are required for Langerhans cell migration from the epidermis.

Topical exposure of mice to chemical allergens results in the migration of epidermal Langerhans cells (LCs) from the skin and their accumulation as immunostimulatory dendritic cells (DCs) in draining lymph nodes. Epidermal cell-derived cytokines have been implicated in the maturation and migration of LCs, but the adhesion molecules that regulate LC migration have not been studied. We hypothesized that integrin-mediated interactions with extracellular matrix components of the skin and lymph node may regulate LC/DC migration. We found that alpha 6 integrins and alpha 4 integrins were differentially expressed by epidermal LCs and lymph node DCs. A majority of LCs (70%) expressed the alpha 6 integrin subunit, whereas DCs did not express alpha 6 integrins. In contrast, the alpha 4 integrin subunit was expressed at high levels on DCs but at much lower levels on LCs. The anti-alpha 6 integrin antibody, GoH3, which blocks binding to laminin, completely prevented the spontaneous migration of LCs from skin explants in vitro and the rapid migration of LCs from mouse ear skin induced after intradermal administration of TNF-alpha in vivo. GoH3 also reduced the accumulation of DCs in draining lymph nodes by a maximum of 70% after topical administration of the chemical allergen oxazolone. LCs remaining in the epidermis in the presence of GoH3 adopted a rounded morphology, rather than the interdigitating appearance typical of LCs in naive skin, suggesting that the cells had detached from neighboring keratinocytes and withdrawn cellular processes in preparation for migration, but were unable to leave the epidermis. The anti-alpha 4 integrin antibody PS/2, which blocks binding to fibronectin, had no effect on LC migration from the epidermis either in vitro or in vivo, or on the accumulation of DCs in draining lymph nodes after oxazolone application. RGD-containing peptides were also without effect on LC migration from skin explants. These results identify an important role for alpha 6 integrins in the migration of LC from the epidermis to the draining lymph node by regulating access across the epidermal basement membrane. In contrast, alpha 4 integrins, or other integrin-dependent interactions with fibronectin that are mediated by the RGD recognition sequence, did not influence LC migration from the epidermis. In addition, alpha 4 integrins did not affect the accumulation of LCs as DCs in draining lymph nodes.

Animals↗
PubMed1997

Interleukin 1 beta and the stimulation of Langerhans cell migration: comparisons with tumour necrosis factor alpha.

Epidermal Langerhans cells (LC) and the cells into which they mature are believed to play a pivotal role in cutaneous immune function. The induction phase of contact sensitization is associated with the migration of LC from the skin and their accumulation as dendritic cells (DC) in lymph nodes draining the site of exposure. We have demonstrated previously that tumour necrosis factor alpha (TNF-alpha), an epidermal cytokine produced by keratinocytes, provides one signal for LC migration. We describe here experiments designed to evaluate the influence of interleukin 1 beta (IL-1 beta), a product exclusively of LC in murine epidermis, on LC migration, LC morphology and DC accumulation, and to compare the effects of this cytokine with those of TNF-alpha. Both cytokines induced a significant reduction in the frequency of epidermal LC and the arrival of DC in draining lymph nodes. Changes in both parameters were induced more rapidly following intradermal administration of TNF-alpha than were observed after treatment with IL-1 beta. However, the reduction in LC frequency was more persistent with IL-1 beta. Both cytokines caused the activation of LC, characterized by the acquisition of a more dendritic morphology and the increased expression of Ia molecules. These results demonstrate that IL-1 beta and TNF-alpha can each stimulate the migration of epidermal LC, but that the changes induced by these cytokines are not identical.

Animals↗
PubMed1997

A re-appraisal of the skin-sensitizing activity of 2,4-dinitrothiocyanobenzene.

A debate continues regarding the immunological properties of 2,4-dinitrothiocyanobenzene (DNTB). In some investigations this chemical was shown not to cause skin sensitization when applied topically but to induce instead hyporesponsiveness or immunological tolerance. In other studies DNTB was found to cause skin sensitization, but not tolerance. However, this chemical continues to be used to discriminate between the properties of skin sensitizing and non-sensitizing chemicals. This study demonstrates that topical exposure of mice to DNTB induces skin sensitization in mice and that this is associated with the accumulation of dendritic cells in draining lymph nodes and the stimulation of lymph node cell proliferation; the latter responses being of equivalent magnitude to those stimulated by 2,4-dinitrochlorobenzene (DNCB), a chemical known to cause contact sensitization. Moreover, exposure of mice to DNTB, as with exposure to DNCB, resulted in the development of a cytokine secretion pattern by draining lymph node cells (LNC) characteristic of contact allergens. Thus, DNTB and DNCB each induced the production by LNC of high levels of interferon-gamma, but little or no interleukin 4 or interleukin 10. Finally, DNTB was shown in the guinea pig maximization test to behave as an extreme skin sensitizer. These results confirm that DNTB should not be regarded as a universal tolerogen and that it possesses a significant potential to induce contact sensitization. The use of this chemical as a presumptive non-sensitizer and/or tolerogen for the evaluation of the selectivity of new predictive test methods for the identification of contact allergens is therefore considered to be inappropriate.

Administration, Topical↗
PubMed1997

Langerhans cells require signals from both tumour necrosis factor-alpha and interleukin-1 beta for migration.

The induction phase of contact sensitization is associated with the movement of epidermal Langerhans cells (LC) from the skin and their migration, via afferent lymphatics, to draining lymph nodes where they accumulate as immunostimulatory dendritic cells (DC). It has been demonstrated previously that tumour necrosis factor-alpha (TNF-alpha) provides an important signal for LC migration and that in the absence of this cytokine, movement of LC from the epidermis to regional lymph nodes is inhibited. Recent evidence indicates that interleukin-1 beta (IL-1 beta), a cytokine produced in murine epidermis exclusively by LC, may also play a role in LC migration. The purpose of the investigations described here was to clarify, using relevant neutralizing anti-cytokine antibodies, the contributions made by TNF-alpha and IL-1 beta to the migration of LC from the epidermis. It was found that like anti-TNF-alpha, anti-IL-1 beta administered systemically to mice (by intraperitoneal injection), prior to skin sensitization with the contact allergen oxazolone, resulted in a marked inhibition of DC accumulation in draining lymph nodes. It was shown also that anti-IL-1 beta inhibited TNF-alpha-induced LC migration and DC accumulation and that; in similar fashion, the stimulation of LC migration and DC accumulation induced by IL-1 beta was compromised by prior treatment with anti-TNF-alpha. Based upon these data it is proposed that the stimulation of LC migration in response to skin sensitization requires the receipt by LC of two independent signals, one provided by TNF-alpha and the other by IL-1 beta. Morphological analyses of LC in epidermal sheets prepared from animals exposed to these cytokines with or without prior systemic treatment with anti-cytokine antibody suggested that the changes induced in LC by TNF-alpha and IL-1 beta may include the altered expression of adhesion molecules and acquisition of the ability to interact with and pass through the basement membrane.

Animals↗
PubMed1996

Sensitization to 2,4-dinitrochlorobenzene: influence of vehicle on absorption and lymph node activation.

Effective skin sensitization is dependent upon immune activation of lymph nodes draining the site of exposure. The influence of vehicle formulation on the vigour of lymph node cell proliferative responses to 2,4-dinitrochlorobenzene (DNCB) has been examined. Mice (BALB/c strain) were exposed topically to 0.5% DNCB dissolved in either acetone or propylene glycol (PG). A significantly greater lymph node cell proliferative response was induced by DNCB in acetone. The observed differences were not attributable to variations in the numbers of immunostimulatory dendritic cells accumulating in the draining nodes following sensitization. In parallel studies, the absorption and cutaneous disposition of DNCB dissolved in acetone or PG were measured in vitro using static diffusion cells and full thickness mouse skin. Although flux of DNCB through the skin was comparable with both vehicles over 24 h, the absorption of the allergen during the first 4 h of exposure was significantly faster when acetone was used as the vehicle. Localization of DNCB demonstrated that much less of the chemical allergen was present in the skin at 4 h when applied in PG vehicle. However, there were no measurable vehicle effects on skin disposition of DNCB at 24 h. These data indicate that the sensitization potential of DNCB is influenced significantly by the nature of the vehicle used, possibly due to consequential effects on chemical absorption and disposition. The studies described in this paper reveal that the application vehicle may have a significant influence on the ability of DNCB to induce immune activation of draining lymph nodes and hence skin sensitization and that this may in turn be associated with important changes in the absorption and/or disposition of the chemical within the skin.

Acetone↗
PubMed1996

Influence of dibutyl phthalate on dermal sensitization to fluorescein isothiocyanate.

What limited evidence there is indicates that the formulation in which a chemical allergen is encountered on the skin can have a marked impact upon the induction of cutaneous immune responses and the subsequent development of contact sensitization. The purpose of the present investigations was to examine further this phenomenon by analysis of the influence of dibutyl phthalate (DBP) on dermal sensitization to fluorescein isothiocyanate (FITC), a skin sensitizing fluorochrome. Addition of DBP augmented very substantially, in a dose-dependent fashion, the ability of topically applied FITC to stimulate proliferative responses in mice by draining lymph node cells (LNC), a correlate of skin sensitizing potential. Under these conditions, exposure of mice to DBP alone failed to elicit significant LNC responses. The influence of DBP on the accumulation of dendritic cells (DC) induced by FITC was examined also. Although 10% DBP had little effect on the numbers of DC found within draining nodes 18 hr following exposure of mice to FITC, the phthalate did result in a very substantial increase in the frequency of lymph node DC bearing detectable antigen (FITC+ DC). Furthermore, in the presence of DBP the median amount of FITC associated with antigen-bearing DC was higher. In vitro skin absorption studies indicated that DBP was associated with a small increase in percutaneous absorption of FITC. Collectively these data demonstrate that the vehicle formulation can exert a marked influence on dermal sensitization and that one mechanism which may be relevant is the increased acquisition of antigen by DC, associated possibly with altered penetration of the allergen into or through the skin.

Administration, Topical↗
PubMed1996

Adhesion molecule expression by epidermal Langerhans cells and lymph node dendritic cells: a comparison.

The migration of epidermal Langerhans cells (LC) and their transport of antigen from the skin to draining lymph nodes are of considerable importance in the induction of cutaneous immune responses, including contact sensitization. While in transit to the lymph nodes, LC are subject to a number of phenotypic changes required for their movement from the skin and acquisition of the capacity for antigen presentation. Among these are alterations in the expression of adhesion molecules that regulate interactions with the surrounding tissue matrix and with T lymphocytes. In the study described here, we investigated, using a combination of immunocytochemistry and flow cytometry, the expression by LC, and the lymph node dendritic cells into which they mature, of the three adhesion molecules, E-cadherin, intercellular adhesion molecule-1 (ICAM-1) and the membrane glycoprotein CD44. The migration of LC was associated with a marked reduction in the expression of E-cadherin, but a parallel upregulation of ICAM-1. No change in the expression of CD44 was detectable. The significance of these changes and their relevance for the functional maturation of LC are discussed.

Animals↗
PubMed1996

Constitutive and inducible expression of interleukin-6 by Langerhans cells and lymph node dendritic cells.

During the induction phase of contact sensitization and other cutaneous immune responses a proportion of epidermal Langerhans cells (LC) is induced to leave the skin and migrate via afferent lymphatics to lymph nodes draining the site of exposure. The cells that accumulate in draining nodes have acquired the characteristics of immunostimulatory dendritic cells and effectively present antigen to responsive T lymphocytes. In the present study we have questioned whether LC in the epidermis and the lymph node dendritic cells into which they develop express interleukin-6 (IL-6), a cytokine that has been shown to serve as an important costimulator of T lymphocyte activation. In situ immunocytochemical analyses using a biotin-streptavidin staining technique revealed that dendritic cells resident in the epidermis of untreated mice constitutively express this cytokine. Keratinocytes expressed detectable IL-6 only following local exposure to the contact allergen oxazolone. Such treatment also appeared to enhance the expression by epidermal dendritic cells of this cytokine. Analyses of unfractionated and LC-enriched and -depleted populations of epidermal cells revealed a close correlation between major histocompatibility complex (MHC) class II (Ia) antigen expression and staining for IL-6, implicating LC as the sole or major source of this cytokine in unstimulated epidermis. Finally, compared with tissue isolated from mice treated with vehicle alone, draining lymph nodes prepared from animals 18 hr following sensitization with oxazolone displayed a substantial increase in both the frequency of dendritic cells and the number of IL-6+ cells within the paracortex. These data demonstrate that resident epidermal LC and the dendritic cells into which they develop are important sources of IL-6. Their constitutive and inducible expression of this cytokine will facilitate the induction of cutaneous immune responses.

Animals↗