IL-8 release from human neutrophils cultured with pro-haptenic chemical sensitizers.

Cytokine release from dendritic cells in vitro is a useful marker to discriminate between sensitizing and irritant haptenic chemicals. Unfortunately, pro-haptens, which gain reactivity following metabolic/auto activation, yield negative results. To overcome this, we exposed human neutrophils and THP-1 cells to haptens/pro-haptens and measured IL-8 release. Haptenic compounds stimulated IL-8 release in neutrophils and THP-1 cells. In contrast, the pro-haptens eugenol, isoeugenol, and 2-aminophenol stimulated high levels of IL-8 release from neutrophils alone. Neutrophil cytokine release was reduced when glutathione was added. Cyp1A1/1B1/3A4 were not detectable in THP-1 cells or neutrophils; however, neutrophils expressed high levels of myeloperoxidase.

Cross-sensitization to haptens: formation of common haptenic metabolites, T cell recognition of cryptic peptides, and true T cell cross-reactivity.

To analyze T cell cross-reactivity to para-compounds, we established CD4(+) T cell hybridomas from mice immunized with adducts of self-globin and one of three different para-compounds: p-aminophenol, p-phenylenediamine, or Bandrowski's base. Some of the hybridomas obtained reacted not only to the immunizing antigen, but also to metabolically related para-compounds, bound to the same protein, thus suggesting formation of common metabolites. Other hybridomas cross-reacted to globin adducts of metabolically unrelated para-compounds, which denotes them as truly cross-reactive cells whose TCR failed to distinguish among the different haptens. One of these hybridomas also reacted against a non-haptenated, cryptic peptide of hemoglobin but not to the full-length native protein. As this hybridoma reacted even more strongly to the respective peptide after it was haptenated, recognition of the native, cryptic peptide was apparently due to true cross-reactivity. To conclude, true T cell cross-reactivity to haptens does occur, as well as the formation of a common reactive metabolite, and T cell recognition of cryptic self-peptides may underlie cross-sensitization to chemicals.

The spectrum of allergic (cross-)sensitivity in clinical patch testing with 'para amino' compounds.

BACKGROUND: Allergic contact sensitization to 'para amino' compounds is frequent and the spectrum of cross-reactivity between members of this chemical group is variable. METHODS: A retrospective analysis of clinical patch test data obtained with a special test series in the centres of the Information Network of Departments of Dermatology (IVDK) between 1995 and 1999. RESULTS: In the 638 patients tested with the above test panel positive reactions were observed most often to p-aminoazobenzene (16.2%), p-phenylenediamine (14.1%), p-toluylenediamine (10.0%), followed by 4,4'-diaminodiphenylmethane (8.5%), Disperse Orange 3 (8.4%) and p-aminophenol (3.1%). Among the 544 patients tested with p-phenylenediamine and all seven additional 'para amino' compounds, concordance between reactions varied greatly. The stronger the positive test reaction to p-phenylenediamine, p-toluylenediamine or p-aminoazobenzene, the more frequently additional positive reactions to the other compounds were observed. CONCLUSIONS: A screening employing several 'para amino' compounds is necessary to describe the individual spectrum of allergic contact sensitization, as there is no reliable marker substance. Further research should aim at (i) establishing the mechanism of cross-reactivity to 'para amino' compounds and (ii) identifying exposures in the environment.

Experimental study on skin sensitization potencies and cross-reactivities of hair-dye-related chemicals in guinea pigs.

In screening patch testing of hairdressers with occupational contact dermatitis, multiple positive reactions to hair dye-related chemicals, such as p-phenylenediamine (PPD), p-toluenediamine x 2HCl (PTD) and p-aminophenol (PAP), a fabric dye p-aminoazobenzene (PAB), and a tar dye Sudan III, were frequently encountered. To investigate individual skin sensitization potency and the cross-reactivities among above chemicals, a guinea pig maximization test with the above 5 chemicals was performed. In each group, 6 animals were induced with one of the chemicals at 0.1% concentration by intradermal injection and at 1.0% by topical application. The animals were challenged with all 5 chemicals in concentrations of dilution by 10 from 0.1% to 0.001%. Under the conditions of 0.1% challenges, similar sensitization potencies were observed in PPD (6/6), PTD (6/6), PAP (5/6) and PAB (6/6) groups, but no positive reactions were elicited in the Sudan III group. The cross-reactivities to PPD were confirmed in the animals challenged with PTD (6/6), PAP (6/6), PAB (6/6) and Sudan III (3/6). In the PTD-induced group, positive responses to cross-challenges were elicited by PPD (5/6), PAP (3/6), PAB (5/6) and Sudan III (1/6). The cross-reactivities to PAP were observed only with PPD (2/5) and PAB (5/5). PAB-induced animals responded only to PPD (1/6). The results indicate that all these chemicals except Sudan III are strong sensitizers. Their cross-reactivities are different in sensitized conditions, respectively. The cross-reactivities to PPD were higher than those to PTD, PAP and PAB.

Study on cross-reactivity to the para group.

In 80 patients, positive to at least one hapten of the para group (para-phenylenediamine, diaminodiphenylmethane, benzocaine, PPD mix), patch tests were carried out with freshly prepared solutions of para-phenylenediamine (PPD) and of 3 selected aromatic compounds related structurally to PPD (para-aminophenol, ortho-aminophenol, hydroquinone). The number of positive reactions correlated with the rate of decomposition of the substances as evaluated by high-pressure liquid chromatography. PPD, which was almost decomposed after 24 h, gave the highest number of positive reactions, followed by ortho-aminophenol and by para-aminophenol, while hydroquinone, which was oxidized to the extent of 35%, did not give any reactions. To evaluate if a different rate of oxidation can modify the patch test response, in the same patients and in 10 normal volunteers, tests were carried out with PPD solutions containing the oxidizing agent silver oxide (0.1%). By this procedure a significant increase in the number of positive responses was observed. The results suggest that the rate of decomposition and therefore the amount of quinone(s) generated, might be the key to eliciting patch test responses to oxidizable aromatic haptens.

Design of the immunogen and label for use in a fluoroimmunoassay for paracetamol.

The design of immunogens for raising antisera containing antibodies specifically directed against paracetamol is discussed. A derivative of the drug with minimal structural alteration was synthesised, coupled to the carrier protein key-hole limpet haemocyanin, and used to immunise sheep. The evaluation of the resulting antisera is described. Preparation of a fluorescent label with reduced bridge-binding then allowed the development of a polarisation fluoroimmunoassay for paracetamol in buffer, suitable for conversion to a direct serum assay.

Cross immunological reactions between three haptens of the "para" group and 4-aminoantipyrine.

Cross passive hemagglutination reactions between sulfanilic acid (SA), para-aminobenzoic acid (PABA), p-phenethidine (PT) and 4-aminoantipyrine (4-AA) haptens have been investigated using conjugates of diazo derivaties of these compounds with homologous serum proteins. Intense cross passive hemagglutination reactions were found between PABA, SA and 4-AA haptens. Although anti-PT sera cross reacted intensely with PABA, SA as well as with 4-AA haptens, antibodies cross reacting with PT were not detected in any of the anti-PABA, anti-SA and anti-4-AA antisera. The conjugates containing a heterologous hapten de not substantially alter the titer of the hemagglutination reactions performed with erythrocytes coated with the homologous hapten, but completely suppress the hemagglutination reactions performed with erythrocytes coated with this hapten as well as with other, but not all, heterologous, haptens. The results support the conclusion that the apparent polysensitization to drugs may actually rely upon cross reactions with haptens chemically related to the immunizing one.