dinitrobenzenes and Dermatitis--Contact

The most common reaction to fragrance materials seen by practicing dermatologists is allergic contact dermatitis. Photodermatitis is occasionally seen, as is contact urticaria, irritation, and depigmentation. Fragrances are the leading cause of allergic contact dermatitis due to cosmetics. The fragrance mixture can cause false-positive reactions; therefore, it is more desirable to test with a separate series of fragrance materials.

Topics: 1-Propanol; Acrolein; Balsams; Cosmetics; Coumarins; Dermatitis, Contact; Dinitrobenzenes; Eugenol; Humans; Perfume; Photosensitivity Disorders; Propanols; Resins, Plant; Skin Tests; Terpenes; Tetrahydronaphthalenes; Urticaria

Contact allergy to environmental xenobiotics is a common and important problem, but it is unclear why some chemicals are potent sensitizers and others weak/nonsensitizers. We explored this by investigating why similar chemicals, 2,4-dinitrochlorobenzene (DNCB) and 2,4-dinitrothiocyanobenzene (DNTB), differ in their ability to induce contact hypersensitivity (CHS). DNCB induced CHS in humans, whereas at similar doses DNTB did not. However, following DNCB sensitization, DNTB elicited CHS in vivo and stimulated DNCB-responsive T cells in vitro, suggesting that differences in response to these compounds lie in the sensitization phase. In contrast to DNCB, DNTB failed to induce emigration of epidermal Langerhans cells in naive individuals. Examination for protein dinitrophenylation in skin revealed that DNCB penetrated into the epidermis, whereas DNTB remained bound to a thiol-rich band within the stratum corneum. DNTB reacted rapidly with reduced glutathione in vitro and was associated with a decrease in the free thiol layer in the stratum corneum, but not in the nucleated epidermis. By contrast, DNCB required GST facilitation to react with gluthathione and, following penetration through the stratum corneum, depleted thiols in the viable epidermis. Chemical depletion of the thiol-rich band or removing it by tape stripping allowed increased penetration of DNTB into the epidermis. Our results suggest that the dissimilar sensitizing potencies of DNCB and DNTB in humans are determined by a previously undescribed outer epidermal biochemical redox barrier, a chemical component of the innate immune defense mechanisms that defend against sensitization by highly reactive environmental chemicals.

Topics: Adult; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Environmental Exposure; Female; Humans; Immunity, Innate; Irritants; Keratinocytes; Male; Oxidation-Reduction; Reverse Transcriptase Polymerase Chain Reaction; Skin; Skin Tests; Xenobiotics

Epicutaneous application of dinitrothiocyanobenzene (DNTB) induces tolerance against its related compound dinitrofluorobenzene (DNFB), because DNTB-pretreated mice cannot be sensitized against the potent hapten DNFB. This tolerance is hapten-specific and transferable. In this study, we demonstrate that IL-12 can break DNTB-mediated tolerance. Furthermore, naive mice treated with IL-12 before DNTB application responded to DNFB challenge with a pronounced ear swelling response without previous sensitization to DNFB, showing that IL-12 can convert the tolerogen DNTB into an immunogen. No differences in numbers or regulatory activity were observed between CD4+CD25+ regulatory T cells isolated from mice treated with DNFB, DNTB, or IL-12 followed by DNTB. However, the number of CD207+ Langerhans cells in regional lymph nodes of DNTB-treated mice was significantly lower than in animals treated with DNFB or IL-12 plus DNTB. Additionally, CD11c+ dendritic cells (DC) isolated from regional lymph nodes of DNTB-treated mice had a significantly lower ability to stimulate T cell proliferation and produced reduced amounts of inflammatory cytokines. Application of both DNFB and DNTB induced apoptotic cell death of DC in the epidermis and the regional lymph nodes. However, the number of apoptotic DC in regional lymph nodes was significantly higher in DNTB-treated animals compared with mice treated with DNFB or IL-12 plus DNTB. Therefore, we conclude that DNTB-mediated tolerance is secondary to inefficient Ag presentation as a result of apoptotic cell death of DC and that IL-12 converts the tolerogen DNTB into an immunogen by preventing DNTB-induced apoptosis of DC.

Topics: Animals; Apoptosis; Dendritic Cells; Dermatitis, Contact; Dinitrobenzenes; Epidermis; Haptens; Immune Tolerance; Interleukin-12; Lymph Nodes; Mice; Mice, Inbred BALB C; T-Lymphocytes, Regulatory

An important property of dendritic cells (DC), which contributes crucially to their strong immunogenic function, is their capacity to migrate from sites of antigen capture to the draining lymphoid organs. Here we studied in detail the migratory pathway and the differentiation of DC during migration in a skin organ culture model and, for comparison, in the conventional contact hypersensitivity system. We report several observations on the capacity of cutaneous DC to migrate in mouse ear skin. (i) Upon application of contact allergens in vivo the density of Langerhans cells in epidermal sheets decreased, as determined by immunostaining for major histocompatibility complex class II, ADPase, F4/80, CD11b, CD32, NLDC-145/DEC-205, and the cytoskeleton protein vimentin. Evaluation was performed by computer assisted morphometry. (ii) Chemically related nonsensitizing or tolerizing compounds left the density of Langerhans cells unchanged. (iii) Immunohistochemical double-staining of dermal sheets from skin organ cultures for major histocompatibility complex class II and CD54 excluded blood vessels as a cutaneous pathway of DC migration. (iv) Electron microscopy of organ cultures revealed dermal accumulations of DC (including Birbeck granule containing Langerhans cells) within typical lymphatic vessels. (v) Populations of migrating DC in organ cultures upregulated markers of maturity (the antigen recognized by monoclonal antibody 2A1, CD86), but retained indicators of immaturity (invariant chain, residual antigen processing function). These data provide additional evidence that during both the induction of contact hypersensitivity and in skin organ culture, Langerhans cells physically leave the epidermis. Both Langerhans cells and dermal DC enter lymphatic vessels. DC mature while they migrate through the skin.

Topics: Animals; Cell Count; Cell Movement; Cellular Senescence; Dendritic Cells; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Ear; Haptens; Immunohistochemistry; Lymphatic System; Mice; Mice, Inbred Strains; Microscopy, Electron; Organ Culture Techniques; Picryl Chloride; Skin

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.

Topics: Administration, Topical; Animals; Cytokines; Dermatitis, Contact; Dinitrobenzenes; Enzyme-Linked Immunosorbent Assay; Guinea Pigs; Immunization; Lymph Nodes; Mice; Mice, Inbred BALB C; Skin

Topics: Dermatitis, Contact; Dinitrobenzenes; Facial Dermatoses; Female; Humans; Perfume; Spouses

A population of CD8+ T cells from dinitrobenzene sulfonate-primed mice produce soluble effector molecules that down-regulate the magnitude of dinitrophenol-specific contact hypersensitivity reactions. These soluble molecules express the binding specificity and serologic determinants of alpha/beta TCR. To examine the requirement for the TCR-alpha chain in the production of these molecules, we have cloned the alpha-chain gene used to encode the surface TCR of MTs 79.1, a T cell hybridoma producing a DNP/Kd-specific soluble suppressive molecule, and tested the ability of this gene to reconstitute the production of the regulatory molecule in TCR alpha-chain gene deletion mutants. Transfection and expression of the alpha-chain construct into an alpha-chain deletion mutant of the parental hybridoma that expressed the parental beta-chain gene resulted in reconstitution of both surface TCR expression and production of the soluble suppressive molecule. As with the molecule produced by the MTs 79.1 parental cells, the inhibitory activity produced by these alpha-chain gene transfectants was DNP-specific and expressed determinants bound by anti-V beta 8 Abs. Transfection of the alpha-chain gene construct into an alpha-/beta- chain gene deletion mutant did not restore the production of the soluble regulatory molecule. These results indicate that in addition to the TCR beta-chain gene, expression of the TCR alpha-chain gene is also required for the production of these molecules. Our results strongly support the hypothesis that some forms of immunosuppression are mediated by soluble forms of the TCR.

Topics: Animals; Base Sequence; Dermatitis, Contact; Dinitrobenzenes; Female; Gene Expression Regulation; H-2 Antigens; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Receptors, Antigen, T-Cell, alpha-beta; Recombinant Fusion Proteins; Sequence Deletion; Suppressor Factors, Immunologic; T-Lymphocytes, Regulatory; Transfection

In murine contact sensitivity (CS) models, cutaneous immune responses are caused by the activity of two different Ag-specific Thy-1+ CD5+ cells. These two different cell subsets act in an obligate sequence to mediate separate early and late components of CS that are accompanied respectively by skin swelling responses at 2 and 24 h after local challenge with the hapten. The early-acting CS-initiating cells are not conventional T cells inasmuch as they are surface negative for CD4, CD8, and CD3. In contrast to this non-MHC-restricted, CS-initiating cell, the classical late-acting CS effector T cell that is recruited locally is CD3+, CD4+, CD8-, and is MHC-restricted. In our study, we have conducted experiments in which a mixture of immune CS-initiating and CS-effector cells were desensitized by incubation in vitro at 37 degrees C with various hapten-amino acid and hapten-carrier conjugates, before i.v. transfer and subsequent ear challenge of normal recipient mice. Desensitization was achieved with multivalent complexes of the relevant hapten conjugated to a variety of unrelated nonmurine carrier proteins, and, in some instances, with monovalent hapten conjugated to amino acid. Because the CS-mediating cells were desensitized in a hapten-specific manner, but these same cells transferred CS reactivity that was hapten/MHC-class II specific, it was suggested previously that these results argued in favor of a two-receptor model for T cell recognition of Ag, one receptor for hapten and the other for MHC. Our study suggests that these findings need to be reinterpreted because CS reactions are mediated by two different, Thy-1+ Ag-specific cells that act in an obligate sequence. Our data suggest that the late-acting Ag/MHC-specific CS-effector T cells are not hapten-specific. In fact, desensitization with hapten alone has a locus of action solely on the Ag receptor of the first-acting Thy-1+ CS-initiating cells--which are therefore truly hapten-specific and which are required for local recruitment of the Ag/MHC-specific late CS-effector T cells.

Topics: Animals; Antigens, Surface; CD3 Complex; Dermatitis, Contact; Dinitrobenzenes; Female; Haptens; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Picryl Chloride; Receptors, Antigen, T-Cell; Serum Albumin, Bovine; T-Lymphocyte Subsets; Temperature; Thy-1 Antigens; Time Factors; Trinitrobenzenes

We determined the lymphokines involved in the immune response to epicutaneously applied dinitrofluorobenzene (DNFB), a sensitizer, and dinitrothiocyanobenzene (DNTB), a tolerogen. Hapten-dependent T-cell proliferation and production of interleukin-2, interleukin-3 and interleukin-4 by lymph node cells (LNC) in mice painted with these cross-reactive haptens were measured by specific lymphokine assays. Proliferation of LNC in tolerized animals was lower than in sensitized mice. LNC from DNTB-treated mice produced lower amounts of interleukin-2, interleukin-3 and interleukin-4 than cells from DNFB-painted mice. These results may explain hapten-specific tolerance induced by DNTB which results in deficient production of both type 1 T-helper cell (Th1) and type 2 T-helper cell (Th2) lymphokines in response to hapten re-exposure. Deficient interleukin-4 production by cells from tolerized mice was corrected by the addition of exogenous interleukin-2. The suppressor function of adoptively transferred T cells from animals tolerized with dinitrothiocyanobenzene may be related to a shift in the balance of Th1 and Th2 lymphokines in favour of the latter, since recipient T cells might provide the source of interleukin-2 that induces interleukin-4 production by donor T cells.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Female; Haptens; Immune Tolerance; Interleukin-2; Interleukin-3; Interleukin-4; Lymphokines; Mice; Mice, Inbred BALB C

A 57-year-old man suffering from persistent light reaction with photocontact allergy to musk ambrette and contact allergy to fragrance mix was evaluated. A lowered minimal erythema dose to UV-B (MED-UV-B) was seen. Reactions to long-wave UV-A and visible radiation were normal. A skin biopsy from one MED-UV-B, taken 24 h after irradiation, showed acute spongiotic dermatitis.

Topics: Dermatitis, Contact; Dinitrobenzenes; Humans; Keratosis; Male; Middle Aged; Perfume; Photosensitivity Disorders; Ultraviolet Rays

Prostaglandins have been implicated in the immune suppression associated with the development of some tumours. Application of the prostaglandin synthetase inhibitor indomethacin, to murine skin prior to treatment with the chemical carcinogens benzo(a)pyrene (BP) or 7,12 dimethylbenz(a)anthracene (DMBA), restored contact sensitivity responses to 2,4-dinitrofluorobenzene in BP- but not DMBA-treated mice. However, indomethacin failed to restore antibody responses in either group of mice. Prolonged treatment with BP or DMBA led to cutaneous tumour formation. Indomethacin was found to delay the onset and reduce the size of tumours in BP- but not DMBA-treated mice. It is proposed that prostaglandin-induced suppression of cellular cutaneous immunity may play a role in BP- but not DMBA-induced cutaneous carcinogenesis.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antigen-Presenting Cells; Benzo(a)pyrene; Carcinoma, Squamous Cell; Dermatitis, Contact; Dinitrobenzenes; Immunity, Cellular; Indomethacin; Male; Mice; Mice, Inbred BALB C; Skin; Skin Neoplasms

In an attempt to define an immunological correlate of contact allergic potential it was investigated whether the magnitude of the lymphocyte proliferative response provoked following primary exposure to skin allergens influences the degree to which sensitization is induced. Previous studies have provided circumstantial evidence that there is a causal relationship between proliferation and sensitization. Thus, inhibition of the primary proliferative response to a sensitizing chemical, induced by prior exposure to an unrelated allergen, is associated with a significant depression of contact hypersensitivity. The results of comparative studies performed using a series of antigenically cross-reactive dinitrobenzene derivatives, 2,4-dinitrofluorobenzene, 2,4-dinitrochlorobenzene and 2,4-dinitrothiocyanobenzene are now reported. Evidence is presented that there is a direct correlation between cumulative proliferative activity in the draining lymph nodes and the severity of elicitation reactions. The tentative conclusion which can be drawn is that measurement of lymphocyte proliferative activity may form the basis of a method for comparative evaluation of the contact allergic potential of chemicals.

Topics: Animals; Antigens; Cell Division; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Dinitrofluorobenzene; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Mice; Mice, Inbred BALB C

By means of epicutaneous tests under standard conditions we studied the cell-mediated contact sensitivity to dinitrophenyl (DNP) conjugates of the stereoisomeric amino acids L-alpha-alanine and D-alpha-alanine, respectively, in patients sensitized with dinitrochlorobenzene (DNCB). There was no difference in the number of positive skin responses with the two enantiomeric compounds DNP-L-alpha-alanine (16/26) and DNP-D-alpha-alanine (17/26). However, as in earlier studies, no patient has shown a positive reaction to the position-isomeric DNP-beta-alanine.

Topics: Alanine; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Female; Haptens; Humans; Male; Patch Tests; Structure-Activity Relationship

When cultured with DNP-labeled I-A+ cells, Lyt 2+ T suppressor cells (Ts) from 2,4,-dinitrobenzene sulfonate (DNBS)-tolerized mice are activated to synthesize and release a suppressor factor (SSF) which suppresses the transfer of contact sensitivity to DNFB. The signals required to activate the DNBS-primed Ts to produce SSF were studied in greater detail. As previously observed with fixed DNP-labeled spleen cell stimulators, the supernatants from cultures of DNBS-primed spleen cells and glutaraldehyde-fixed DNP-labeled P388D1 cell monolayers did not contain SSF. When the tolerant cells were harvested from these monolayers and were treated with IL-1, the Ts released the synthesized SSF. Synthesis and release of SSF required Ts recognition of DNP/class I MHC on the hapten-presenting cells followed by interaction with the costimulator IL-1. When the tolerant cells were cultured with fixed DNP-labeled I-A+ or I-A- stimulators to induce SSF synthesis, release was induced by adding either unlabeled or TNP-labeled unprimed spleen cells to the cultures. The release of SSF was blocked when the second stimulators were pretreated with anti-I-A antibody but not with anti-DNP or anti-class I MHC antibodies. These results indicate that the release of SSF by DNBS-primed Lyt 2+ Ts is regulated by the activity of a self-I-A-reactive (i.e., autoreactive) T cell in the tolerant spleen cell population.

Topics: Animals; Antigens, Differentiation, T-Lymphocyte; Benzenesulfonates; Dermatitis, Contact; Dinitrobenzenes; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Immune Tolerance; Mice; Mice, Inbred BALB C; Suppressor Factors, Immunologic; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

We reported 2 patients with pigmented contact dermatitis caused by occupational airborne contactants, whitening dyes in clothes and formaldehyde in packing adhesive tapes. A women developed airborne pigmented contact dermatitis due to musk ambrette in incense. Patch testing confirmed the diagnosis. Since olden times, people in Japan have burnt incense when they worshipped their ancestors. Recently, it has been in fashion to enjoy perfumes and people may burn incense all day long every day. Our patient burnt 2 kinds of incense every day for about 5 years. We assumed musk ambrette was volatilized when incense was burnt, and contact on her face being dissolved in sebum, thus inducing allergic pigmented contact dermatitis.

Topics: Adolescent; Adult; Aged; Dermatitis, Contact; Dinitrobenzenes; Female; Humans; Middle Aged; Nitrobenzenes; Perfume

A 63-year-old male school teacher with itchy depigmented macules on his left dorsum manus, left shoulder and abdomen presented at our clinic on 8 July 1986. He had practiced the incense ceremony for about 15 years, and had burnt several incenses and sandalwood. 48 h closed patch testing revealed perfume in the incenses was the cause. We assumed that perfume in the incense was volatized in air when incense was burnt; skin surface contact occurred with airborne particle, which dissolved in sebum; thus allergic contact dermatitis accompanied by depigmentation might arise.

Topics: Dermatitis, Contact; Dinitrobenzenes; Humans; Male; Middle Aged; Perfume; Pigmentation Disorders; Polycyclic Sesquiterpenes; Sesquiterpenes; Skin Tests

Herein we described the direct suppressive effects of a monoclonal T cell hybridoma-derived, DNP-specific suppressor T cell factor (26.10.2 TsF) on the local transfer of delayed-type hypersensitivity (DTH) by a DNP-specific BALB/c T cell clone (dD1.9). The L3T4+, Lyt-2- dD1.9 T cell clone proliferated in response to DNP-OVA and DNBS, but not TNP-OVA or TNBS, in association with I-Ed determinants present on antigen-presenting cells. Similarly, local injection of histopaque-purified dD1.9 cell blasts resulted in DNP-specific, radioresistant, I-Ed-restricted, mononuclear cell-rich ear swelling responses. Incubation in 26.10.2 TsF specifically suppressed local transfer of DNP-specific DTH by dD1.9, but not local DTH responses transferred by BALB/c T cell clones specific for TNP or GAT. The suppressive effect of 26.10.2 TsF correlated with targeting on DNP-major histocompatibility complex determinants associated with the DTH T cell (TDH) targets. 26.10.2 TsF-mediated suppression was most pronounced after exposure of dD1.9 target cells to antigen (after the stimulation phase of the T cell clone maintenance procedure), and greatly reduced when dD1.9 was cultured for long periods in the absence of DNP (after the rest phase of clone maintenance). In additional support of this hypothesis, GAT-specific TDH, normally resistant to 26.10.2 TsF-mediated suppression, were rendered susceptible to suppression after surface DNPylation. The results demonstrate a direct, antigen-specific, effector phase regulatory effect of a monoclonal TsF on a cloned, antigen-specific T cell target, and strongly suggest that suppression is mediated via targeting on DNP determinants associated with the TDH target. Simplification of complex Ts circuitry operating in suppression of the efferent limb of DTH by the use of monoclonal TsF and cloned T cell targets should provide a basis for the future study of the molecular mechanisms of immune suppression.

Topics: Animals; Antibodies, Monoclonal; Antigens, Surface; Clone Cells; Dermatitis, Contact; Dinitrobenzenes; Epitopes; Female; H-2 Antigens; Hybridomas; Immunization, Passive; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Nitrobenzenes; Peptides; Phenotype; Polymers; Suppressor Factors, Immunologic; T-Lymphocytes, Regulatory

Topics: Acrylates; Animals; Dermatitis, Contact; Dermatitis, Occupational; Desensitization, Immunologic; Dinitrobenzenes; Guinea Pigs; Humans; Immune Tolerance; Nitrobenzenes

Topics: Animals; Antigens; Cells, Cultured; Dermatitis, Contact; Dimethylformamide; Dinitrobenzenes; Dinitrochlorobenzene; Disease Models, Animal; Guinea Pigs; Haptens; Immunization; Interleukin-1; Lymph Nodes; Mice; Mice, Inbred BALB C; T-Lymphocytes

The potential of 4 related nitro musk substances (musk tibetene, musk ketone, musk xylol, and musk moskene) to cause photoallergy, phototoxicity, and/or contact sensitivity was compared to that of musk ambrette, a known photoallergen. Musk ambrette caused a high incidence of photoallergy as indicated by the severity of the skin grades as compared to a control group. Musk tibetene and musk moskene were negative for phototoxicity, photoallergenicity and contact sensitivity under the test conditions. Musk xylol was shown to be a weak contact sensitizer. Musk ketone gave challenge responses suggestive of a weak phototoxin and a weak contact sensitizer. The latter was not affected by light exposure. These data suggest that except for musk ambrette, the nitro musks as a group do not have the potential to produce photoallergy. Some members of this type of perfume raw material could be classified as weak sensitizers (musk xylol, musk ketone) or weakly phototoxic (musk ketone). These latter biological qualities have not been expressed clinically.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Fatty Acids, Monounsaturated; Fatty Acids, Unsaturated; Guinea Pigs; Nitrobenzenes; Perfume; Photosensitivity Disorders

Musk ambrette, a contact and photocontact allergen, is a synthetic fragrance present in aftershave lotions and many toiletries. Thirty-four patients with contact and photocontact allergy to musk ambrette were studied. All had a facial eczema. Four distinctive clinical pictures were seen. These were plaques of eczema, jawline (mandibular) dermatitis, acute contact dermatitis and chronic actinic dermatitis. Twenty-six patients were light sensitive by history and 10 were diagnosed by clinical criteria as having chronic actinic dermatitis. Patch and photopatch testing to musk ambrette showed that seven patients had pure photocontact dermatitis to musk ambrette. Eight patients had contact dermatitis to musk ambrette but this was exacerbated by irradiation. Cross reaction with moskene occurred. Scrupulous avoidance of musk ambrette has resulted in clinical resolution in most patients.

Topics: Acute Disease; Adult; Aged; Chronic Disease; Dermatitis, Contact; Dinitrobenzenes; Facial Dermatoses; Humans; Male; Middle Aged; Nitrobenzenes; Patch Tests; Perfume; Photosensitivity Disorders; Ultraviolet Rays

Topics: Amino Acids; Animals; Dermatitis, Contact; Dinitrobenzenes; Guinea Pigs; Hypersensitivity, Delayed; Nitrobenzenes; Patch Tests; Skin; Skin Tests

Epicutaneous application of 2,4-dinitrothiocyanobenzene (DNTB) is said to result in specific immunological hyporesponsiveness and fails to induce contact sensitization. However, we demonstrate that topical exposure to DNTB causes activation of the draining lymph node in mice and the induction of contact sensitization in both rodents and a single human volunteer. In mice and rats, pre-exposure to DNTB failed to impair subsequent responsiveness to the cross-reactive allergen 2,4-dinitrochlorobenzene. These data provide evidence that DNTB cannot be regarded as an exclusive tolerogen when applied epicutaneously and indicate that attempts to define the characteristics of tolerising chemicals from analysis of this agent may be misleading.

Topics: Administration, Topical; Animals; Cross Reactions; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Female; Humans; Immune Tolerance; Lymph Nodes; Male; Mice; Mice, Inbred BALB C; Nitrobenzenes; Rats

Contact sensitivity (CS), anti-DNP (dinitrophenyl) IgG and IgE type antibodies were induced by skin applications of the contact sensitizer dinitrofluorobenzene. CS response was suppressed by treatment with the monovalent hapten dinitrobenzene sulfonate (DNBS) but was not affected by treatment with the DNP-conjugates (DNP22-Ficoll, DNP17-polyethylene glycol and DNP2-polyvinylpyrrolidone). On the other hand, IgE type response was depressed by treatment with the various DNP conjugates but was not affected by DNBS. The IgG type response was not appreciably affected by any of the treatments.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Dose-Response Relationship, Immunologic; Hypersensitivity; Immunity, Cellular; Immunoglobulin E; Immunoglobulin G; Immunosuppression Therapy; Mice; Mice, Inbred BALB C; Nitrobenzenes; Skin

Musk ambrette has become one of the commonest photosensitizers in many countries and is the cause of persistent light reactions in many patients. A simple method is presented for qualitative analysis for musk ambrette and other nitromusks in colognes using thin-layer chromatography with toluene as carrier solvent and visualization under UVL. It is useful for identifying such colognes so that proper recommendations can be given to patients sensitive to musk ambrette. 14 of 32 men's colognes, analysed by TLC and confirmed on GC/MS, were found to contain musk ambrette, the concentration varying from 0.02 to 0.39% w/v.

Topics: Chromatography, Gas; Chromatography, Thin Layer; Dermatitis, Contact; Dinitrobenzenes; Gas Chromatography-Mass Spectrometry; Humans; Male; Nitrobenzenes; Perfume; Xylenes

The most frequently used source of indoor lighting is the fluorescent tube. Although there are major variations in phosphors, the majority of these lamps are safe, efficient, and economical illuminators. These fluorescent light sources are currently our primary source of visible light; however, they emit small amounts of ultraviolet A light (UVA) as well as a somewhat larger percentage of infrared radiation. Photosensitivity diseases have been reported in each of these three broad wavelength bands. Specific examples include heat urticaria from infrared exposure, contact photosensitivity of the phototoxic type following exposure to dyes and visible light, and two relatively rare but disabling conditions from ultraviolet A exposure--solar urticaria and contact photosensitivity of the photoallergic type (persistent light reaction). During the past five years, eight patients with photosensitivity induced by musk ambrette and UVA have been treated at Columbia-Presbyterian Medical Center; six of these have been severely disabled and satisfy the criteria for persistent light reactors. Fifteen patients with solar urticaria have also been observed. Ten of these had reactions in the UVA range. The clinical and laboratory findings of these two groups of patients were presented.

Topics: Aged; Animals; Dermatitis, Contact; Dinitrobenzenes; Female; Humans; Lighting; Male; Middle Aged; Photosensitivity Disorders; Ultraviolet Rays; Urticaria

An assessment of T-lymphocyte proliferation and lymph node weight is proposed as a predictive test for contact sensitizers of industrial origin. Data are presented showing increased T-lymphocyte proliferation following epicutaneous application of a variety of industrially important acrylate-like chemicals which appear to correlate well with their ability to sensitize in the guinea pig. These data were compared with those obtained after application of 2,4-dinitro-1-fluorobenzene (DNFB) a strong sensitizer, and 2,4-dinitrothiocyanatebenzene (DNTB) a nonsensitizer when given epicutaneously. It is suggested that this quantitative approach, in parallel with a simple one-dose immunization, may provide a better picture of sensitization potential than the longer multidose immunizations currently in use.

Topics: Acrylates; Administration, Topical; Animals; Dermatitis, Contact; Dinitrobenzenes; Evaluation Studies as Topic; Female; Guinea Pigs; Lymph Nodes; Lymphocyte Activation; Male; Organ Size; T-Lymphocytes

For qualitative and quantitative analysis of musk ambrette and 4 other nitromusk compounds (musk ketone, moskene, musk tibetine, musk xylene) thin layer chromatography (TLC) and high pressure liquid chromatography (HPLC) techniques were developed. By TLC a reasonable separation was obtained and the limit of detection was 2-5 x 10(-7) g. By HPLC the separation was even better and the limit of detection for musk ambrette was 2 x 10(-9) g. The correlation between the amount of musk ambrette/ketone and the HPLC peak was linear. The TLC and HPLC techniques were used to demonstrate the presence of nitromusks in several commercial products, mainly aftershave lotions and eau-de-toilettes preparations. By ultraviolet spectrophotometry, absorption spectra were studied for the nitromusk compounds. The absorption maximum for musk ambrette was at 264 nm, that for moskene at 253 nm. Photopatch testing was carried out in 13 patients photoallergic to musk ambrette. Only 3 patients also reacted to other nitromusks. Photoallergey to musk ketone and musk tibetine is reported for the first time.

Topics: Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Dermatitis, Contact; Dinitrobenzenes; Fatty Acids, Monounsaturated; Fatty Acids, Unsaturated; Humans; Nitrobenzenes

A patient with photocontact allergy to musk ambrette was investigated with patch and photopatch testing with dilutions of musk ambrette and 4 other nitro-musk compounds (musk ketone, moskene, musk tibetine, musk xylene) as well as with in vitro ultraviolet-irradiated solutions of musk ambrette. Besides a strong photoallergy to musk ambrette, a plain contact allergy to photoproducts of musk ambrette was demonstrated. An attempt to isolate the photoproducts causing the plain contact allergy was performed by using a preparative thin layer chromatography technique. These isolated photoproducts were not, however, responsible for the contact allergy to the photodecomposed musk ambrette.

Topics: Adult; Dermatitis, Contact; Dinitrobenzenes; Humans; Male; Nitrobenzenes; Photochemistry; Skin Tests; Structure-Activity Relationship; Ultraviolet Rays

A patient with persistent photosensitivity and positive photopatch tests to musk ambrette and an after-shave lotion is reported. Phototests showed extreme sensitivity to UV radiation, especially UVB. Patch tests with the European Standard Series and some plant allergens were negative. Histology showed a granulomatous reaction with epithelioid and giant cells in the dermis.

Topics: Aged; Dermatitis, Contact; Dinitrobenzenes; Humans; Male; Nitrobenzenes; Patch Tests; Perfume; Photosensitivity Disorders; Skin

The immediate effects and mechanisms of desensitization of contact sensitivity to 2,4-dinitro-1-fluorobenzene (DNFB) were investigated. Mice were sensitized with DNFB, desensitized with antigen 2 weeks later, and challenged 1 day after desensitization. Significant inhibition (approximately 50%) of contact sensitivity was observed after iv injections of large doses of dinitrobenzene sulfonic acid (DNBS) or dinitropenol (DNP)-labeled spleen cells. Haptenated red blood cells (RBC) did not induce any significant immediate desensitization but produced significant inhibition of an anamnestic response 2 weeks later. The immediate desensitization induced by DNBS was antigen nonspecific. Although the contact sensitivity response itself could be inhibited by afferent- or efferent-acting suppressor cells, such cells were not demonstrated in desensitized animals. DNBS appears to desensitize by inactivating effector cells for contact sensitivity, although it appears that suppressor mechanisms could be activated by other physiochemical forms of the desensitizing antigen.

Topics: Animals; Dermatitis, Contact; Desensitization, Immunologic; Dinitrobenzenes; Dinitrofluorobenzene; Female; Haptens; Hypersensitivity, Delayed; Immune Tolerance; Immunologic Memory; Mice; Mice, Inbred BALB C; Nitrobenzenes; T-Lymphocytes, Regulatory

Of several dinitrobenzenes tested, 2,4-dinitrothiocyanatebenzene (DNTB) was found to be the only one that did not induce contact sensitivity when applied to the guinea pig ear epicutaneously, but when applied epicutaneously it induced tolerance to 2,4-dinitrofluorobenzene (DNFB). The manner in which DNFB, DNTB, and other dinitrobenzene compounds conjugated in vitro to soluble proteins, at physiologic pH, was examined. By measuring the free amino and sulfydryl radicals in the protein before and after conjugation, it was possible to determine to which groups the hapten was bound. It was found that although all the haptens bound to the free sulfydryl groups, DNTB was the only one that did not bind to amino groups. It is suggested that to be an epicutaenous tolerizer, as opposed to sensitizer, a hapten should bind to sulfydryl groups exclusively. It is hoped that a search for agents binding in a similar manner will reveal epicutaneous tolerizers for important industrial sensitizers.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Female; Free Radicals; Guinea Pigs; Male; Nitrobenzenes; Proteins; Skin

This manuscript describes the preparation and characterization of a syngeneic antiserum raised in BALB/c mice against nylon wool-purified lymph node (LN) T cells from 2,4-dinitrofluorobenzene- (DNFB) sensitized BALB/c mice. This antiserum (anti-T DH-DNP) plus complement blocks the ability of LN or purified T cells from DNFB-sensitized donors to transfer immunity to naive recipients. The inhibitory activity of the serum is due to antibodies; these antibodies have specificity for idiotype (Id) determinants as shown by their ability to bind to purified mouse anti-DNP antibodies but not to rabbit anti-DNP or normal mouse Ig. Inhibition of transfer of immunity by the anti-Id serum is not restricted by either H-2 gene products or Igh-1 allotypes. The antiserum is antigen specific when tested on LN cells from oxazolone-sensitized mice, but has a low level of cross-reactivity against T cells from 2,4,6-trinitrochlorobenzene- (TNCB) sensitized mice. In addition, it was shown that naturally occurring anti-Id antibodies, epsilon-DNP-L-lysine, or DNP-protein (but not NIP-protein) and the anti-Id in syngeneic anti-T DH-DNP serum compete for similar binding sites on DNFB-immune T cells. We interpret these findings to indicate that immunization of mice with T cells from DNFB-sensitized syngeneic mice induces the production of anti-Id antibodies. These anti-Id antibodies are specific for determinants expressed on the T cell receptor or receptor subsite that is specific for the hapten DNP.

Topics: Absorption; Animals; Binding, Competitive; Chromatography, Affinity; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Epitopes; Female; Hypersensitivity, Delayed; Immune Sera; Immunization, Passive; Immunoglobulin Idiotypes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred CBA; Nitrobenzenes; Receptors, Antigen, T-Cell; Species Specificity; T-Lymphocytes

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Epitopes; Glucans; Immune Tolerance; Immunity, Cellular; Immunoglobulin Idiotypes; Kinetics; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Nitrophenols; Phenylacetates; Receptors, Immunologic; T-Lymphocytes, Regulatory

Suppressor T cells induced by the i.v. injection of syngeneic DNP-LC were studied at varying times after tolerization. It was found that 7 days after tolerization both the spleen and lymph nodes of tolerant mice contained Ts active in suppressing the efferent (effector) phase of the contact sensitivity response (Ts-eff). At 14 days after tolerization, however, only the spleen showed significant suppressive activity that was active on the afferent (induction) phase of the contact sensitivity response. This second wave of afferent suppression was shown to be mediated by an antigen-specific, Ia-positive T cell (Ts-aff). Ts-aff could also be induced in normal, but not in CY-pretreated mice transfused with lymphoid cells containing primed Ts-eff, provided an appropriate induction period was allowed. Also the activity of Ts-aff, but not of Ts-eff, could be eliminated by treatment with anti-DNP antibody (i.e., idiotype) and C. Thus, Ts-aff appears not to be a maturation stage of Ts-eff. It was also shown that Ts-eff and Ts-aff interacted in an antagonistic fashion; cotransfer of these populations along with DNFB-immune TDH cells neutralized the efferent suppressive activity of Ts-eff for the immune cells, allowing successful passive transfer of immunity. The exact role of this antagonistic interaction is not clear, but it may play a part in maintaining the balance of suppression in the intact animal. The data are discussed in terms of a cellular network in immune suppression and are compared and contrasted to other suppressor systems in which network interactions have been described.

Topics: Animals; Cell Survival; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Female; Guinea Pigs; Immune Tolerance; Immunization, Passive; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred CBA; Spleen; T-Lymphocytes, Regulatory

Cutaneous basophil hypersensitivity (CBH)-inducing factor was demonstrated in immune sera obtained from dinitrofluorobenzene (DNFB)-sensitized animals 2 weeks after sensitization (DNP-GPS-2W). It showed hapten specificity and worked dose dependently. It was fractionated into a non-gamma-globulin fraction by Sephadex G-150 gel filtration following ammonium sulfate desalting and CM-cellulose chromatography. The factor was eluted into a fraction of a little smaller molecular weight than bovine serum albumin on Sephadex G-150 gel filtration. It passed through an antiguinea pig IgG1 column and was absorbed to a DNP-BGG column. On SDS-PAGE it failed to show any staining band because of low protein concentration. From these results CBH factor appearing in circulation in contact-sensitized animals was thought to be a somewhat different molecule from that of Askenase's factor, i.e. IgG1 antibody.

Topics: Animals; Basophils; Dermatitis, Contact; Dinitrobenzenes; Female; Guinea Pigs; Hypersensitivity, Delayed; Immune Sera; Immunoglobulin G; Skin

Topics: Animals; Benzenesulfonates; Binding, Competitive; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Immune Tolerance; Isoantibodies; Lymphocytes; Lymphokines; Mice; Mice, Inbred A; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred CBA; Nitrobenzenes; Phenotype; Rabbits; Suppressor Factors, Immunologic

Experiments were done to investigate the nature of the antigen receptor on lymph node(LN) T cells from mice sensitized to 2,4-dinitrofluorobenzene (DNFB). LN cells or purified T cells were treated in vitro with monovalent or different multivalent 2,4-dinitrophenol (DNP) ligands. The effect of this treatment was measured by testing the ability of the cells to transfer contact sensitivity (CS) to DNFB into naive recipients. We found that treatment of the T cells in vitro with either epsilon-DNP-L-lysine or DNP-protein conjugates inhibits the transfer of CS in a dose-dependent way. The inhibition is hapten specific and is not mediated by activation of suppressor cells. Inhibition of the T cells by hapten in vitro is rapid (15-30 min) and temperature independent but requires divalent cations in the treatment medium. In addition, it was found that hapten-treated T cells are unable to adsorb specific anti-idiotype antibody, and this inhibition of adsorption is hapten specific. Collectively, these data indicate that DNFB-immune T cells express a receptor specific for the hapten DNP and provide evidence that supports a two-receptor model for T cell recognition of antigen.

Topics: Animals; Binding, Competitive; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Dose-Response Relationship, Immunologic; Female; Guinea Pigs; Histocompatibility Antigens Class II; Immunization, Passive; Lymphocyte Activation; Lysine; Male; Mice; Mice, Inbred A; Mice, Inbred BALB C; Mice, Inbred CBA; Nitrobenzenes; Receptors, Antigen, T-Cell; T-Lymphocytes, Regulatory; Temperature; Time Factors

Studies concerning a patient with contact photosensitivity to musk ambrette, a commonly used fragrance, are reported. The patient had a persistent light reaction. The patient's history, clinical appearance, and phototest and photopatch test results are described. The observed patterns of these were identical to those previously noted in patients who had persistent light reactions after exposure to other photosensitizers such as the halogenated salicylanilides. The histopathologic findings in the phototest and photopatch test sites were those of an acute spongiotic dermatitis. No abnormal change was seen with ultraviolet A radiation exposure alone.

Topics: Aged; Dermatitis, Contact; Dinitrobenzenes; Humans; Male; Nitrobenzenes; Patch Tests; Perfume; Photosensitivity Disorders; Skin

Unresponsiveness to DNFB contact sensitivity induced by DNP-modified lymphoid cells (DNP-LC) is mediated by two separable pathways: a rapidly induced, long lasting inhibition of reactive T cell clones (donor tolerance), and a transient period of suppressor T cell (Ts) activity. The present report has examined the nature of the hapten-modified determinants responsible for the induction of these pathways by utilizing soluble DNP-LC cell lysate preparations as tolerogens. The results indicate that both DNP-modified MHC and non-MHC encoded determinants can mediate donor tolerance 7 days after tolerization. On the other hand, the induction of Ts requires DNP-modified MHC determinants, since DNP-LC lysates passed over lentil lectin or specific anti-H-2 immunoabsorbent columns lost their ability to induce Ts. Additional experiments showed that the injection of DNP-LC lysate compatible with the recipient strain at the H-2K and H-2D region of the MHC was sufficient for the induction of Ts. We propose that Ts induction involves the direct presentation of DNP-H-2 determinants to Ts precursors, whereas the induction of donor tolerance may involve host processing and presentation of DNP-modified membrane determinants in conjunction with host MHC structures.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Epitopes; Female; H-2 Antigens; Haptens; Immune Tolerance; Lectins; Lymphocytes; Male; Mice; Mice, Inbred BALB C; Mice, Inbred CBA; Rabbits; T-Lymphocytes

Injection of TNP-, DNP- or oxazolone-substituted syngeneic cells into mice causes the development of hapten-specific T suppressor cells which prevent the animals from being activity sensitized with homologous hapten. These cells injected together with immunized cells abrogate the latter's ability to transfer passively the contact sensitivity (CS) reaction into normal recipients. T lymphocytes from animals made unresponsive and sensitized with homologous hapten synthesize in vitro antigen-specific suppressor factors (SF) which when incubated with immune lymphocytes prevent them transferring adoptively the CS reaction. The type of cell used to induce suppression or production of suppressor factor (haptenated erythrocytes, thymocytes or macrophages) is not critical suggesting that a hapten-substituted common membrane structure is recognized as a tolerogen. The present work demonstrates that while the specific unresponsiveness induced by cell-bound hapten in vivo is long lasting, cells from tolerized animals are able to suppress the immunized cells in passive transfer or produce in vitro antigen-specific suppressor factors only when tested several days after tolerization.

Topics: Animals; Binding Sites; Carrier Proteins; Dermatitis, Contact; Dinitrobenzenes; Epitopes; Female; Haptens; Immune Tolerance; Immunization, Passive; Male; Mice; Mice, Inbred CBA; T-Lymphocytes; T-Lymphocytes, Regulatory; Time Factors; Trinitrobenzenes

We investigated genetic restrictions in the induction of immediate tolerance to DNFB contact sensitivity in mice. Using spleen cells from various donor strains haptenated at 500 micro M DNFB, we were unable to detect any restrictions in tolerance induction in recipients that were either syngeneic or allogeneic to the donor strain. However, if the concentration of hapten used in the in vitro labeling was decreased (from 500 micro M to 2.5 to 5 micro M DNFB), differences in tolerogenesis between the various donor strain haplotypes were found. Haptenated spleen cells labeled with 5 micro M DNFB produced a profound level of unresponsiveness in allogeneic recipients but produced minimal tolerance in syngeneic animals. This tolerant state was shown to be antigen-specific and was not produced by unmodified allogeneic cells alone. Further genetic analysis demonstrated that an efficient tolerant state was produced when the donor of the tolerogen and recipient differed at the MLS locus rather than at either the MHC or minor regions. This phenomenon required viable, Thy 1-bearing cells in the haptenated donor population for efficient tolerogenesis to DNFB contact sensitivity.

Topics: Animals; Cell Adhesion; Cell Survival; Chromosome Mapping; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Epitopes; Haploidy; Haptens; Immune Tolerance; Lymphocytes; Major Histocompatibility Complex; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred CBA; Mice, Inbred DBA; Nitrobenzenes; Rabbits

Several inbred strains of Syrian hamsters have been immunized by skin painting with highly reactive haptens. Classical contact hypersensitivity has been assayed by the ear swelling response: the response is hapten-specific, exaggerated after re-immunization with the original hapten, and can be transferred adoptively to naive syngeneic hamsters with viable lymphoid cells, suggesting that contact hypersensitivity is mediated in hamsters by T lymphocytes. Moreover, skin painting with hapten induces significant serum titers of anti-hapten antibodies, indicating that antigen-specific B lymphocytes are also activated. Hamsters can be rendered unresponsive to these same haptens by conventional methods: 1) i.v. inoculation of the soluble hapten sulfonate or 2) inoculation of hapten-derivatized syngeneic lymphoid cells. Hamsters treated with these "tolerizing" maneuvers develop profound hapten-specific unresponsiveness that can be adoptively transferred to naive recipients with living lymphoid cells. "Unresponsive" animals, however, make strong anti-hapten antibody responses that rival the humoral immune responses found after skin sensitization. The data suggest that an active process is involved in the induction and maintenance of the unresponsive state, but responsibility can not be assigned firmly to putative suppressor T cells or to an antibody-mediated B suppressor modality.

Topics: Animals; Antibody Formation; Cricetinae; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Ear; Female; Haptens; Immune Tolerance; Immunization, Passive; Lymphocytes; Male; Trinitrobenzenes

Contact sensitization was achieved with dinitrophenylated autologous lymph node cells killed by hypotonic lysis or sonication as well as live cells. The sensitizing ability of the sonicated cells disappeared after freeze-thawing or heating. The significance of these findings is discussed.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Guinea Pigs; Lymph Nodes; Male; Nitrobenzenes

Two male patients with photoallergic contact dermatitis to musk ambrette, 2-methoxy-3,5-dinitro-4-methyl-t-butylbenzene, are reported. Musk ambrette is a synthetic fragrance material commonly used in foods and cosmetics. The clinical distribution of lesions presented by these patients strongly suggested an adverse reaction to light. Over a period of several years no etiology had been determined in these cases despite extensive testing and hospitalization. Photopatch tests to musk ambrette were positive. No other etiology for photosensitivity was found. This report emphasizes that exposure to household or cosmetic products represents a potential source of contact photosensitivity. Patients presenting with dermatoses of unknown origin confined to the light-exposed areas should have fragrances considered as possible etiologic agents.

Topics: Aged; Cosmetics; Dermatitis, Contact; Dinitrobenzenes; Humans; Male; Nitrobenzenes; Patch Tests; Perfume; Photosensitivity Disorders; Skin

Previous reports have shown that picryl sulfonic acid (PSA) induces suppressor T cells that inhibit the effector phase of contact sensitivity, whereeas its DNP counterpart, dinitrobenzenesulfonate (DNBS) induces cells that inhibit the afferent phase of sensitization. Accordingly, cells from mice injected with DNBS, but not PSA, could be shown to inhibit the DNA synthesis in the lymph nodes that occurs during sensitization. It is now shown that PSA does induce T cells that suppress DNA synthesis but this can only be detected with enriched T cells or by using a regimen of PSA injection different frm previously used to induce suppressor cells for the effector phase. The T cells did not affect responses to oxazolone or dinitrofluorobenzene (DNFB) and were distinguishable from suppressors of the efferent phase in that they could be produced in adult thymectomized but not cyclophosphamide-treated mice. T cells from mice injected with DNBS that inhibited DNA synthesis to DNFB had the same properties.

Topics: Animals; Antilymphocyte Serum; Cyclophosphamide; Dermatitis, Contact; Dinitrobenzenes; DNA; Epitopes; Filtration; Idoxuridine; Male; Mice; Mice, Inbred CBA; Nitrobenzenes; Picryl Chloride; T-Lymphocytes; Thymectomy; Trinitrobenzenesulfonic Acid

Human lymphocytes from dinitrochlorobenzene (DNCB)-sensitized human subjects primed in first culture with dinitrophenylated-antigens yield a population of cells which respond in an accelerated manner to the same or similar antigen in second culture. Using this "clonal priming" approach, we have demonstrated that such a primed population showed maximal proliferative response to dinitrophenylated autologous cells. These DNP primed clones also showed responses to some dinitrophenylated allogeneic leukocytes. The magnitude of the accelerated blastogenic response with allogeneic leukocytes varied in most instances in relation to the degree of sharing of HLA-A, B, and DRw antigens with the original autologous stimulator. These findings show that the self-specific factors recognized in conjunction with the dinitrophenyl antigen are closely but not invariably associated with established major histocompatibility (MHC)-associated serologic typing results. While DNP primed clones fail to respond to unmodified autologous leukocytes, they often show significant responses to unmodified allogeneic leukocytes. If such accelerated responses to unmodified allogeneic leukocytes are not the result of nonspecific activation of allogeneic responding lymphocyte clones, these findings further indicate that DNP modified self can resemble some alloantigens.

Topics: Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Epitopes; Genes, MHC Class II; HLA Antigens; Humans; Isoantigens; Leukocytes; Lymphocytes; Major Histocompatibility Complex; Nitrobenzenes

Topics: Animals; Binding, Competitive; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Dose-Response Relationship, Immunologic; Epitopes; Immune Sera; Immunity, Cellular; Immunization, Passive; Immunoglobulin Idiotypes; Immunosorbent Techniques; Lymph Nodes; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Inbred CBA; Nitrobenzenes; Receptors, Antigen; Species Specificity

This report has examined the mechanisms by which major histocompatibility complex (MHC) non-restricted suppressor T cells (Ts), induced by the i.v. injection of 2,4-dinitropheny (DNP)-modified, syngeneic lymphoid cells (DNP-LC), suppress the passive transfer of contact sensitivity mediated by syngeneic and allogeneic immune delayed hypersensitivity T cells (TDH). In terms of suppression of syngeneic TDH, it was found that the suppressive action of the Ts was only blocked by pretreatment with soluble syngeneic DNP-LC membrane preparations. Monomeric DNP-lysine, polymeric DNP-protein conjugates, and syngeneic TNP-LC membranes did not inhibit Ts function. Further experiments showed that inhibition of syngeneic suppression could be achieved by DNP-modified-membrane preparations that were only H-2D-region compatible with the Ts donor. Thus, Ts antigen receptors in this system specifically recognize DNP-modified H-2D-region determinants. In contrast, it was found that pretreatment os syninduced Ts with syngeneic DNP-LC membranes did not inhibit the ability to suppress allogeneic TDH. However, pretreatment of Ts with DNP-allogeneic membranes which were H-2D-end compatible to the allogeneic target TDH eliminated their ability to suppress the specific allogeneic TDH, leaving intact suppression of syngeneic or third party TDH. It is proposed that perturbation of the immune system by i.v. injection of syngeneic NDP-LC leads to the induction of a polyclonal wave of DNP-specific Ts activity. Some members of this set of Ts recognize DNP-self MHC determinants with moderate affinity and are thus specifically inhibited after pretreatment with those DNP-self determinants. Other members of this set display receptors which cross-react with high affinity with DNP-allogeneic determinants and thus suppress allogeneic TDH cells. These allosuppressive clones can thus be specifically inhibited only by pretreatment with DNP-LC membranes, MHC-compatible with the target TDH. The data are discussed in terms of current models of T-cell cross-reactivity and T-cell-receptor recognition.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Female; H-2 Antigens; Histocompatibility; Immunization, Passive; Immunosuppression Therapy; Mice; Mice, Inbred Strains; Nitrobenzenes; T-Lymphocytes

Topics: Animals; Antigens, Surface; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Haptens; Immune Tolerance; Isoantigens; Lymphocytes; Macrophages; Major Histocompatibility Complex; Mice; Spleen; Time Factors

This study reports the induction of contact photodermatitis to musk ambrette, 2-methoxy-3,5-dinitro-4-methyl-t-butylbenzene, in guinea pigs. Photoallergic contact dermatitis was assayed using 2 alternative induction methods. Successful photosensitization was achieved only when the nuchal skin was stripped with scotch tape before application of musk ambrette and ultraviolet radiation. Induction methods utilizing nonstripped nuchal skin which induce photosensitivity to potent photoallergens were ineffective for musk ambrette. Phtotoxicity tests to musk ambrette at concentrations between 1 and 50% and a dose of 10.2 joules/cm2 from "Black Light" fluorescent tubes were all negative. Under identical irradiation conditions, anthracene at 0.9% and 8-methoxypsoralen at 1% were consistently positive. The mechanism of photosensitivity to musk ambrette appears to be photoallergic rather than phototoxic. The requirement for skin abrasion to induce photosensitization parallels the clinical reports of photosensitivity to musk ambrette in man.

Topics: Allergens; Animals; Dermatitis, Contact; Dinitrobenzenes; Female; Guinea Pigs; Nitrobenzenes; Perfume; Photosensitivity Disorders; Salicylanilides; Skin; Ultraviolet Rays

The contact sensitivity response to DNFB is decreased after adult thymectomy (ATX). This response decreases to 50% of the control response of normal age-matched mice as soon as 3 weeks after ATX and is not further depressed 9 to 16 weeks after ATX. These results suggest that two T cell subsets of different lifespan are involved in the anti-DNFB response. A circulating thymic factor (FTS) is able to restore the contact sensitivity response to DNFB when injected 3 to 9 weeks after ATX but not 16 weeks later. By contrast, FTS has a depressive effect on the contact sensitivity response to DNFB of normal mice through a cyclophosphamide-sensitive T cell subset. These results suggest that FTS regulates DNFB contact sensitivity by acting on a cyclophosphamide-sensitive T cell subset, still present 9 weeks after ATX but absent after 16 weeks. Thus although the T cell defect, causing a depression of the contact sensitivity reaction to DNFB is quantitatively similar 3 and 16 weeks after ATX, its nature is probably different.

Topics: Aging; Animals; Antibody Formation; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Hypersensitivity, Delayed; Immunoglobulin M; Immunosuppression Therapy; Male; Mice; Mice, Inbred CBA; Nitrobenzenes; Thymectomy; Thymic Factor, Circulating; Thymus Hormones

Contact sensitivity to dinitrochlorobenzene (DNCB) in guinea pigs could be rapidly suppressed by intravenous injection of dinitrobenzene sulfonic acid sodium salt (DNBSO3). This suppression is transient and antigen-specific. Macrophages from desensitized animals are not inactivated as shown by their ability to react, both in vivo and in vitro to lymphokines produced in a separate system. Therefore, effector lymphocytes are considered the target for the desensitizing antigen. Using an adoptive transfer system it was demonstrated that effector lymphocytes are inactivated by a direct effect of the hapten. Since this inactivation can be reversed by trypsin treatment, a receptor blockade of effector lymphocytes is proposed as the mechanism of desensitization of DNCB-contact sensitive guinea pigs. This does not exclude the possibility that additional mechanisms such as suppressor cells, compartmentalization or endogenous proliferation of lymph node lymphocytes may play an additional role.

Topics: Animals; Cell Migration Inhibition; Dermatitis, Contact; Desensitization, Immunologic; Dinitrobenzenes; Dinitrochlorobenzene; Epitopes; Female; Guinea Pigs; Haptens; Macrophages; Male; Nitrobenzenes; Sulfonic Acids; T-Lymphocytes

Genetic restrictions governing the induction and expression of suppressor T cells (Ts) in tolerance to 1-fluoro-2,4-dinitrogenzene (DNFB) contract sensitivity were studied. Tolerance was induced by using 2,4-dinitrophenyl (DNP)-modified lymphoid cells (DNP-LC) as tolerogen. Two kinds of Ts were found-those produced by DNP-LC syngeneic to the donor of the Ts (syninduced Ts), and those produced by DNP-LC allogeneic to the donor of Ts (alloinduced Ts). Studies employing congenic resistant mouse strains indicated that recognition of DNP-modified-major histocompatibility region determinants on the tolerogenic DNP-LC was essential for the induction of both types of Ts. Non-H-2 genetic background was irrelevant to Ts induction. Mapping studies indicated that induction of both syninduced and alloinduced Ts was associated with recognition of DNP-modified-MHC region determinants which map to the right of the H-2G region (i.e., H-2D gene products). Tolerization of donor mice with DNP-LC which were H-2D region compatible, but not with H-2K or I region compatible DNP-LC, was both sufficient and required for the induction of hapten-specific syninduced Ts. Tolerization of donor mice with DNP-LC which were incompatible only at the H-2D region was sufficient for the induction of alloinduced Ts. These Ts were capable of suppressing recipient mice only if the recipients shared the H-2D region with the strain providing the DNP-LC tolerogen, and were not capable of suppressing recipients sharing all but the H-2D region with the tolerogen.

Topics: Animals; Chromosome Mapping; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; H-2 Antigens; Haptens; Immune Tolerance; Immunity, Cellular; Immunosuppression Therapy; Mice; Nitrobenzenes; T-Lymphocytes

Pigs were skin painted with the contact sensitizing agent DNFB and afferent lymph was collected for the next 24 h. The lymph cells carried a small amount of DNP and were able to sensitize 40% of homologous recipients. Peripheral blood lymphocytes conjugated with DNFB in vitro to the same degree as afferent lymph cells sensitized 80% of autologous and 40% of homologous recipients: lymphocytes conjugated at higher levels sensitized 75% of animals in each group. Lightly conjugated cells were capable of survival and able to respond to stimulation by mitogens and their ability to sensitize autologous recipients was abolished by heat killing. Highly conjugated cells were not capable of survival, their sensitizing ability was not altered by heat killing and they were able to sensitize incompatible recipients. It is suggested that highly conjugated cells sensitize by a different mechanism which depends on the cooperation of non-lymphocytic cells, not easily mobilized from lymphoid tissue.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Histocompatibility; Immunization, Passive; Lymph Nodes; Lymphocyte Activation; Lymphocytes; Macrophages; Nitrobenzenes; Swine; T-Lymphocytes

Topics: Animals; Antigens; Cross Reactions; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Dose-Response Relationship, Immunologic; H-2 Antigens; Immune Tolerance; Immunization, Passive; Immunosuppression Therapy; Mice; Mice, Inbred BALB C; Mice, Inbred CBA; Species Specificity; T-Lymphocytes

We investigated the mechanism(s) by which MHC-restricted suppressor T cells (Ts) induced by i.v. injection of allogeneic DNP-modified lymphoid cells (alloinduced Ts) suppress the DNFB contact sensitivity response. It was shown that alloinduced Ts acted only during the early phases (afferent limb) of sensitization. They were incapable of suppressing previously sensitized recipients or of inhibiting the expression of DNFB-immune LN cells when co-transferred into normal recipients. The target of alloinduced Ts seems to be cell proliferation, i.e., inhibition of antigen-induced cell proliferation (DNA synthesis) in Ts recipient mice. The failure of recipients of alloinduced Ts to generate DNFB-immune LN cells capable of transferring contact sensitivity to normal recipients also suggests that these Ts act by preventing the development of an expanded clone of mature immune T cells. The suppressive effects of alloinduced Ts also were inhibited by prior in vitro treatment with anti-TNP serum. The data are discussed in terms of current models of suppression, and are compared to mechanisms of suppression in other contact sensitivity models.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; DNA; Dose-Response Relationship, Immunologic; Immunosuppression Therapy; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Mice, Inbred CBA; Species Specificity; T-Lymphocytes; Time Factors

DNP-conjugated lymph node cell plasma membranes, thymocyte plasma membranes and red cell ghosts were prepared and tested for their ability to induce contact sensitivity, using pigs as experimental animals. Lymph node cell membranes and red cell ghosts were able to sensitize, provided the dose of DNP was very large, but thymocyte membrane failed to sensitize most pigs. DNP-conjugated lymph proteins coming from the site of application of DNFB were also able to sensitize if large amounts were administered, but free DNFB itself, infused directly into an afferent lymphatic, was much more efficient. Since DNFB can often be detected in lymph folowing skin painting, it may conjugate cells within the node which later contact the recirculating population of lymphocytes and so sensitize the animal by a central mechanism. The best-equipped cells would be the macrophage-like lymph cells which are closely related to the epidermal Langerhans cells and are known to migrate to the paracortex of the node.

Topics: Animals; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Erythrocyte Membrane; Lymph; Lymph Nodes; Molecular Weight; Nitrobenzenes; Proteins; Serum Albumin; Swine; T-Lymphocytes

The distribution of DNP groups in various tissues of guinea-pigs following intravenous injection of DNBSO3Na was investigated by an immunofluorescent method using FITC-labelled anti-DNP antibody. DNP groups were detected either in the cytoplasms of epidermal cells or on/in lymphoid tissue cells. The epidermal distribution of DNP groups was shown in the skin obtained from 5 min to 3 days after injection. There was no fundamental difference in the epidermal localization of DNP groups between at the previously tested site and at virgin site in DNCB sensitized animals. Unreacted DNBSO3Na was demonstrated in the blood plasma of DNBSO3Na injected animals. The injection of either DNP-lysine or 2,4-dinitrophenol was found incapable of inducing flare up reaction and no epidermal localization of DNP groups in the sensitized animals was observed. A possibility that intravenously injected DNBSO3Na forms a complete antigen by conjugation with epidermal proteins and sensitized cells, which have been indicated by Polak, Turk & Frey (1973) to remain at the site of old contact reaction, react with the antigen resulting in flare up reaction, is suggested.

Topics: Animals; Antigens; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Guinea Pigs; Male; Skin; Skin Tests; Time Factors

Topics: Animals; Cell Membrane; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Histocompatibility Antigens; Immune Tolerance; Immunization, Passive; Immunosuppression Therapy; Kinetics; Lymphocyte Activation; Mice; Species Specificity; Structure-Activity Relationship; T-Lymphocytes

Immunoadsorbent columns charged with rabbit anti-dinitrophenyl (DNP) antibody were used to isolate and concentrate DNP-protein conjugates from the 105,000 g supernate of extracts prepared from guinea pig epidermis previously painted with dinitrofluorobenzene (DNFB). Conjugates isolated in this manner elicited positive delayed hypersensitivity skin tests when injected intradermally into DNFB-sensitized animals. Dinitrophenylated epidermal protein conjugates prepared in vitro were capable of specifically inhibiting the migration of peritoneal exudate cells obtained from DNFB-sensitized guinea pigs. These results demonstrate the application of immunoadsorbent chromatography and the migration inhibition assay to the further definition of the epitopes involved in contact sensitivity.

Topics: Animals; Cell Migration Inhibition; Dermatitis, Contact; Dinitrobenzenes; Female; Guinea Pigs; Hypersensitivity, Delayed; Immunosorbent Techniques; Macrophages; Nitrobenzenes; Proteins; Skin; Skin Tests

Active suppression of contact sensitivity to DNFB in mice is mediated by a soluble suppressor factor (SSF) which is released in vitro by cultures of LN cells that contain antigen-specific suppressor T cells. For SSF-mediated suppression to occur, identity is required among genes in the H-2 complex between the donor of SSF and the donar of DNFB-immune LN cells. Identity in either the left half or right half of the H-2 complex was found to be both sufficient and required. Additional experiments using congeneic strains with intra-H-2 recombinants showed that the genetic homology which was required mapped to genes contained in the H-2K and/or H-2D regions of the H-2 complex.

Topics: Animals; Antigens; Chromosome Mapping; Dermatitis, Contact; Dinitrobenzenes; Genes; Genetic Linkage; H-2 Antigens; Immune Tolerance; Mice; Mice, Inbred A; Mice, Inbred C3H; Mice, Inbred C57BL; Mice, Inbred CBA; Recombination, Genetic; Species Specificity

Topics: Animals; Antibody Formation; Cross Reactions; Dermatitis, Contact; Dinitrobenzenes; Dinitrofluorobenzene; Freund's Adjuvant; Haptens; Hemagglutination Tests; Hemocyanins; Immune Tolerance; Immunoglobulin G; Mice; Mice, Inbred BALB C; Nitrobenzenes; T-Lymphocytes

BALB/c mice were made tolerant to the T cell-dependent phenomenon of contact sensitivity to DNFB by i.v. injection of syngeneic lymphoid cells which had been previously modified with DNFB in vitro. The highly efficient unresponsiveness, as measured by ear challenge and in vitro antigen-induced cell proliferation, was shown to follow dose-response kinetics both in vivo and in vitro and to be exquisitely specific for the DNP moiety. The kinetics of tolerance induction were shown to be very rapid and had been previously shown to be long lasting. Unresponsiveness was more efficient when the hapten-modified cells were introduced by the i.v. route and tolerance could be increased by repeated injection of tolerogen. The tolerance could be transferred to normal syngeneic recipients by spleen and/or lymph node cells from tolerant donors. A wide variety of hapten-modified lymphoid cells, including mixed cell populations and enriched populations of T cells, B cells, and macrophages, were capable of inducing tolerance. The unresponsiveness was dependent merely on the association of DNP to the lymphoid membrane proteins and not upon the viability of the hapten-modified cells. These experiments support the hypothesis that in hapten-specific T cell sensitivity, as exemplified by contact sensitivity to DNFB, specific T cell tolerance in actively induced by hapten on self-membrane. In other hapten-specific antibody-forming systems, tolerance appears to be most readily induced by hapten on soluble self protein or on a nonimmunogeneic carrier.

Topics: Animals; Antigens; Dermatitis, Contact; Dinitrobenzenes; Dose-Response Relationship, Immunologic; Female; Haptens; Immune Tolerance; Mice; Mice, Inbred BALB C; Spleen; T-Lymphocytes; Transplantation, Homologous

Partial unresponsiveness to 2,4-dinitrofluorobenzene (DNFB) was produced by the epicutaneous application of 2 per cent 2,4-dinitrothiocyanatebenzene (DNTB) to the dorsum of the ear, 14 and 7 days before sensitization. This state of partial unresponsiveness could be broken by treatment with 300 mg/kg cyclophosphamide (CY) 3 days before the sensitization dose of DNFB. There was a significant increase in the uptake of 125I-labelled iododeoxyuridine in the draining lymph node, 6 days after the application of 2 per cent DNTB and in the contralateral nodes at 4 and 6 days. The state of partial unresponsiveness to DNFB was not associated with a decrease in T-cell proliferation in the draining lymph nodes. The generation of suppressor cells, capable of reacting in the periphery, was demonstrated by passive cell transfer into recipients sensitized to DNFB and depleted of normally produced suppressor cells by pretreatment with CY. These cells were found in spleen and peritoneal exudates of animals treated by the epicutaneous application of 2 per cent DNTB 14 and 7 days previously. It is suggested that the suppressor cells induced by contact with DNTB act by competition with effector cells at the periphery.

Topics: Animals; Cell Division; Cyclophosphamide; Dermatitis, Contact; Desensitization, Immunologic; Dinitrobenzenes; Dinitrofluorobenzene; Female; Guinea Pigs; Idoxuridine; Immune Tolerance; Immunity, Cellular; Immunization, Passive; Lymph Nodes; Male; Nitrobenzenes; Skin Tests; T-Lymphocytes; Thiocyanates; Time Factors

Topics: Animals; Dermatitis; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Erythema; Freund's Adjuvant; Guinea Pigs; Injections, Intravenous; Nitrobenzenes; Research; Skin Tests; Sulfonic Acids; Toxicology

Although induction of contact skin sensitivity by low molecular weight 2,4-dinitrobenzenes requires the formation in vivo of 2,4-dinitrophenyl-proteins, analogous protein conjugates prepared in vitro are unable to induce this hypersensitive state. Low molecular weight 2,4-dinitrobenzenes are concentrated by isolated lymph node cells, but a representative 2,4-dinitrophenyl-protein conjugate (2,4-dinitrophenyl-bovine serum albumin) was not taken up to a detectable extent by these cells. It is inferred that there exist large quantitative differences in the extent to which dinitrophenyl-proteins are localized within cells following the administration to an intact animal of (a) those simple dinitrobenzenes which are both concentrated by lymph node cells and have the capacity to form protein conjugates in vivo, and (b) 2,4-dinitrophenyl-protein conjugates prepared in vitro. It is suggested that this difference could account for the fact that a varietyof 2,4-dinitrophenyl-proteins prepared in vitro are unable to induce contact skin sensitivity.

Topics: Animals; Cell Separation; Dermatitis, Contact; Dinitrobenzenes; Dinitrophenols; Lymph Nodes; Nitrobenzenes; Pharmaceutical Preparations; Proteins; Serum Albumin, Bovine; Skin; Skin Physiological Phenomena

When one or two drops of a dilute, non-irritating solution of 2,4-dinitrochlorobenzene (DNCB) is applied to a small area of skin of the intact guinea pig, about 20 per cent of the applied material, or some derivative of it, is soon excreted in urine. In normal, as well as in specifically sensitized guinea pigs, DNCB at the site of local application becomes rapidly bound to skin protein through primary chemical bonds. Twenty-four hours after application roughly half of the material present at the local skin site is still extractable with organic solvents. Of the non-extractable dinitrophenyl groups, about 99 per cent are in epidermis, and about 85 per cent are substituted in epsilon-NH(2) groups of lysine residues. Only traces of bound dinitrophenyl groups were observed in the corium. It is uncertain whether these are formed in situ, or are experimental contaminants, or are migratory epidermally formed conjugates. Even when DNCB is injected intradermally it combines predominantly with overlying epidermis and with epidermal components of hair follicles, but only slightly with corium. The 2,4-dinitrophenyl conjugates which are localized in the deeper, viable half of the epidermis, close to the epidermal-dermal junction, are inferred to be the agents responsible for specifically evoking the allergic response in sensitized animals. Conjugates which are situated in the outer, cornified half of the epidermis are shown to be incapable of eliciting the allergic response. The results furnish a basis for interpreting a common pattern of lesions in allergic contact dermatitis as it occurs spontaneously in man.

Topics: Animals; Dermatitis; Dermatitis, Allergic Contact; Dermatitis, Contact; Dinitrobenzenes; Dinitrochlorobenzene; Epidermis; Guinea Pigs; Humans; Hypersensitivity; Male; Nitrobenzenes; Proteins; Skin; Skin Physiological Phenomena