Immunoreactivity to Cocoa and Nickel in Atopic and Allergic Contact Dermatitis

Comparison by leukocyte adherence inhibition of human immune response to cancer-associated immunogens, Thomsen-Friedenreich (T) and Tn, myelin basic protein, and organ-specific cancer neoantigens

Introduction. The combination of atopic dermatitis (AD) with allergic contact dermatitis (ACD) or the occurrence of ACD on the background of atopic dermatitis is called the overlap syndrome. Studies have demonstrated several reasons why patients with AD have a similar or even increased risk of developing ACD compared to those without AD. Allergens and haptens are trigger factors in a group of patients with AD and ACD overlap syndrome. The aim of the study. To confirm the diagnosis of ACD in a group of patients with AD – diagnose the overlap syndrome and analyze which allergens and haptens were the trigger factors in this group. Materials and methods. To confirm IgE-dependent sensitization in atopic dermatitis, skin prick tests or determination of specific IgE in blood serum were performed. Skin patch tests (European series S-1000) were performed to determine the mechanisms of delayed-type hypersensitivity. Results. It was found that the highest specific weight of positive allergic reactions has been recorded in response to the following allergens: ticks, ticks/ambrosia, birch and mold. The absolute majority of patients demonstrated positive specific IgE-dependent sensitization to Dermatophagoides pteronyssinus and Dermatophagoides farinae – 24 (50%), in turn, on Ambrósia – 14 (29.2%), and on Alternaria alternata – 8 (16.7%). Also, the reaction was most often recorded to haptens: cobalt, nickel, formaldehyde, PPD, textile dyes. Deterioration of the clinical course and shortening of AD remission periods were observed due to the formation of ACD against the background of impaired skin barrier function and the presence of chronic immune inflammation. Conclusions. Patients with AD are more often diagnosed with ACD, which predictably worsens the course of AD. Patients with confirmed overlap syndrome "AD + ACD" most often show reactions to haptens: Cobalt, Nikel, Formaldehyde, PPD, Textile dye mix – and in the vast majority to 2 haptens or more in one patient.

Mechanism of the LAI Test in Bladder Cancer Patients

The CD30 molecule has been proposed as a marker for a subset of CD4+CD45RO+ (memory) T cells with potent B cell helper activity producing IL-5 and IFN-gamma and as a specific marker for Th2 cells. Recently, an association has been demonstrated between elevated serum levels of soluble CD30, which is shed by CD30+ cells in vitro and in vivo, and atopic dermatitis but not respiratory atopic disorders or allergic contact dermatitis. We studied the expression of CD30 in the inflammatory infiltrate of atopic dermatitis compared with that of allergic contact dermatitis, with special regard to skin disease activity (acute vs subacute/ chronic). Biopsies were obtained from 16 patients suffering from atopic dermatitis (acute n = 6, subacute/ chronic n = 10), from 7 patients with acute allergic contact dermatitis and from 5 positive patch-test reactions. Paraffin-embedded as well as snap-frozen material was stained with anti-CD30 and anti-CD45RO mAbs according to standard procedures. Double-staining procedures for CD30CD3, CD30CD4, CD30CD45RO and CD30CD68 were also performed. Abundant CD45RO+ cells were detected both in atopic dermatitis and in allergic contact dermatitis lesions. We found scattered CD30+ cells in only one of six formalin-fixed paraffin-embedded acute atopic dermatitis biopsies, but in all of the respective snap-frozen specimens, possibly because CD30 expression on atopic dermatitis infiltrating cells is weak and sensitive to formalin fixation and paraffin embedding. CD30CD3 and CD30CD4 double staining identified CD30+ cells to be helper T lymphocytes. No significant CD30 expression (either in paraffin-embedded or in frozen material) could be found in subacute/chronic atopic dermatitis lesions or in any of the specimens of allergic contact dermatitis. The results suggest a specific regulatory function of CD30+ T cells in acute atopic dermatitis. With respect to the view that CD30 is a marker for Th2 cells, our observations confirm previous findings that Th2 cells predominate in the infiltrate particularly of acute atopic dermatitis. CD30 expression in acute atopic dermatitis but not in acute allergic contact dermatitis might be helpful in the histological differentiation of these disorders and in the further characterization of atopy patch testing.

Introduction. The combination of atopic dermatitis (AD) with allergic contact dermatitis (ACD) or the occurrence of ACD on the background of AD has been called overlap syndrome. Due to the debilitating symptom complex, which includes itching, discomfort, pain, sleep disturbances and psychological disorders, the overlap syndrome (OS) of ACD and AD has a significant negative impact on the quality of life. Aim. To determine the effectiveness of integrating a sedative component into the treatment regimen of ACD and AD overlap syndrome in the context of impact on patients’ quality of life. Materials and methods. Based at the Department of Dermatovenereology, Clinical and Laboratory Immunology of the Shupyk National Healthcare University of Ukraine, 68 patients with a confirmed diagnosis of AD and ACD overlap syndrome were treated. To assess the impact of the disease on the quality of life, the questionnaire "QLI-OS-AD/ACD-2024" was used. Results. Assessment of the patients' quality of life showed that both in the study group, whose patients received a sedative complex of magnesium and melatonin in the OS ACD and AD management scheme, and in the control group, after the treatment, the quality of life indicators predictably improved. Treatment significantly increases the proportion of patients who assess the impact of the disease on their quality of life as "insignificant" and reduces the proportion of those who recognize the impact as "significant" or as one that brings "significant deterioration." The proportion of patients with anxiety in the study group decreased by half, and the proportion of patients with insomnia – by 41.0%; on the other hand, in the control group, patients with anxiety decreased by 21.1%, and the number of patients suffering from insomnia decreased by only 14.4%. Conclusions. Integration of a sedative combination of magnesium, melatonin and vitamin B6 into the complex therapy regimen for OS ACD and AD leads to a more significant improvement in the quality of life of dermatological patients by reducing the manifestations of depression, anxiety and insomnia. The questionnaire "QLI-OS-AD/ACD-2024" allows the practicing physician to quickly and inexpensively assess the degree of impact of AD and ACD overlap syndrome on the patients’ quality of life.

Background: Burn-induced pain represents one of the most challenging clinical pain syndromes, characterized by persistent hyperalgesia and allodynia mediated in part through upregulation of Transient Receptor Potential Vanilloid 1 (TRPV1) channels. TRPV1, a non-selective cation channel activated by heat, acidic pH, and inflammatory mediators, plays a central role in nociceptive sensitization and the amplification of pain signals in both peripheral and central nervous system tissues. Current multimodal analgesia combining opioids and non-steroidal anti-inflammatory drugs (NSAIDs) remains the standard of care; however, concerns regarding adverse effects including gastric irritation, renal impairment, respiratory depression, and long-term dependence necessitate exploration of safer adjuvant alternatives. Cocoa (Theobroma cacao) contains bioactive flavanols, polyphenols, catechins, epicatechins, and methylxanthines with documented anti-inflammatory and analgesic properties, including reported suppression of TRPV1 expression in neuropathic pain models, making it a promising candidate as a natural analgesic adjuvant. This study aimed to compare the efficacy of cocoa extract versus ibuprofen as a tramadol adjuvant in a rat model of burn-induced pain, by assessing mechanical pain thresholds and TRPV1 protein levels in brain and spinal cord tissues. Methods: This experimental study utilized a randomized post-test only controlled design with 15 healthy male Wistar rats (Rattus norvegicus), weighing 140‒180 g and aged 4‒6 weeks, equally divided into three groups (n = 5). The control group received no analgesic therapy (placebo), the second group received tramadol 12.5 mg/kg intraperitoneally combined with ibuprofen 15 mg/kg orally, and the third group received tramadol 12.5 mg/kg intraperitoneally combined with cocoa extract 0.5 mg/kg orally. Following a 7-day acclimatization period, second-degree burn injuries were induced by immersing the right hind paw in a thermostatically controlled water bath maintained at 65°C for 3 seconds under ketamine-xylazineacepromazine anaesthesia. Treatments were administered immediately following burn induction. Mechanical pain thresholds were assessed using an electronic Von Frey anesthesiometer at 24 hours post-injury, while TRPV1 protein levels in brain and spinal cord tissues were quantified by enzyme-linked immunosorbent assay (ELISA). Statistical analyses included one- way ANOVA with Tukey HSD post-hoc for normally distributed data, Kruskal-Wallis with Mann-Whitney post-hoc for nonnormally distributed data, and Pearson correlation analysis, with significance set at p < 0.05. Results: Von Frey test results at 24 hours post-injury revealed significant differences in mechanical withdrawal thresholds among the three groups (one-way ANOVA, p = 0.003). The control group exhibited the lowest withdrawal threshold (mean 4.92 g), reflecting significant burn-induced hyperalgesia, whereas the tramadol-ibuprofen group (mean 13.92 g) and tramadol-cocoa group (mean 14.84 g) both showed markedly elevated thresholds compared to control (p = 0.003 for each). No significant difference was observed between the two treatment groups (p = 0.769), indicating equivalent analgesic efficacy. Kruskal-Wallis testing demonstrated significant intergroup differences in brain TRPV1 levels (p = 0.018), with the control group exhibiting the highest concentrations (mean 0.293 ng/mL). Mann-Whitney post-hoc analysis revealed a significant difference between the control and tramadol-ibuprofen groups (p = 0.012), but not between the control and tramadol-cocoa groups (p = 0.151), nor between the two treatment groups (p = 0.143). Spinal TRPV1 levels also differed significantly among groups (p = 0.034), with the tramadol-cocoa group demonstrating the greatest spinal TRPV1 suppression (mean 0.287 ng/mL) compared to control (mean 0.497 ng/mL; p = 0.024), while no significant difference was found between the control and tramadol-ibuprofen groups (p = 0.095) nor between the two treatment groups (p = 1.000). Pearson correlation analysis revealed a significant negative correlation between Von Frey values and brain TRPV1 levels (r = −0.632, p = 0.012) as well as a very strong negative correlation with spinal TRPV1 levels (r = −0.822, p = 0.0002), confirming TRPV1 as a key mediator of burn-induced hyperalgesia Conclusion: Cocoa extract, when used as an adjuvant to tramadol, provides equivalent analgesic efficacy and TRPV1 suppression to ibuprofen in a rat model of burn-induced pain, supporting its potential as a natural NSAID alternative in multimodal burn pain management strategies.

The aim of the review is to provide up-to-date information on the multiple roles of epidermal keratinocytes in the immune reactions associated with allergic contact dermatitis and atopic dermatitis skin diseases. In the last two decades, it has become clear that keratinocytes are highly active immunological cells, with major control over the acute and the chronic phases of skin inflammation by means of cytokine/chemokine production and surface molecule expression. Keratinocyte responses in skin allergic reactions are rather disease-specific and keratinocytes from genetically determined skin disorders, including atopic dermatitis, show intrinsic abnormalities in their capacity to respond to trigger factors. Lymphokines and cytokines released by T lymphocytes and other immune cells represent the most important stimuli that elicit the inflammatory activation of keratinocytes. Depending on the type and extent of T-cell infiltrate present in allergic contact dermatitis and atopic dermatitis skin lesions, keratinocytes are exposed to different cytokine micromilieu and, in turn, produce flogosis mediators qualitatively and quantitatively specific for each disease. Keratinocyte-derived inflammatory molecules amplificate skin immune responses associated with allergic contact dermatitis and atopic dermatitis, and contribute to the disease process and clinical phenotype development.

Background: Several publications report Dermatophagoides farinae allergens as responsible for several types of non-IgE-mediated allergic reactions.There is no standardized lab exam to endotype (quantify the participation of these mechanisms) in the context of allergic disease pathophysiology.Aims: To evaluate the potential of the Leukocyte Adherence Inhibition Test (LAIT) and the Tube Titration of Precipitins (TTP) on serum to discriminate non-IgE-mediated immunoreactivity against D. farinae in patients with non-IgE-mediated allergic phenotype with clinically suspected hypersensitivity to their allergens.Study Design: We retrospectively examined the medical charts of 134 allergic patients investigated with LAIT and 100 allergic patients investigated with TTP against an extract of D. farinae.Place and Duration of Study: Instituto Alergoimuno de Americana -So Paulo -Brazilbetween January 2018 and May 2024.Methodology: The registered results of the semi-quantitative serum TTP against D. farinae extract were distributed in ranges through a cascade distribution chart to outline the variability of the results.The registered results of the percentage of Leukocyte Adherence Inhibition (LAI) promoted by the ex vivo challenges with D. farinae extract were distributed in ranges through a cascade distribution chart to outline the variability of results.The statistical characteristics of these cohorts were calculated. Results:The TTP showed a wide distribution range, with four negative and most positive results concentrating on the higher dilutions.The mean was estimated at 1:242; the median was 1:128; the standard deviation was estimated at 1:210; the mode was 1:512 (appeared 36 times).The LAI ranged from 0% to 100%.The mean was 52%; the median was 55.5%; the standard deviation was 23.6%; the mode was 63% (appeared seven times).The cascade distribution demonstrates a wide range of LAI results.Conclusion: Our preliminary results support that the semi-quantitative TTP and the LAIT have the potential to endotype and clinically discriminate diverse degrees of cellular and humoral non-IgEmediated immunoreactivity against D. farinae in allergic patients.

Allergic Contact Dermatitis in Children With and Without Atopic Dermatitis

We have investigated 115 patients presenting with dermatoses, who worked with cutting oils. The study period was from 1986 to 1987. There were 111 men and 4 women, with ages ranging from 16 to 66 years (mean 42 ± 14.3 years). The final diagnoses were as follows: 28 patients had endogenous hand eczema and irritant contact dermatitis (ICD); 22 allergic contact dermatitis (ACD) and ICD; 21 ICD alone; 19 ACD alone; 12 endogenous hand eczema; 6 other (non-eczematous) conditions; 4 ACD, ICD and endogenous hand eczema; and 3 ACD and endogenous hand eczema. Formaldehyde-releasing biocides were the commonest cause of allergic reactions in the cutting oil series. Reactions were also seen to colophony, formaldehyde, nickel, rubber, dichromate and epoxy resin in the International Contact Dermatitis Research Group (ICDRG) standard series. As 42% of the patients had relevant positive patch tests, patch testing remains an essential investigation in the management of cutting oil dermatitis.

Atopic dermatitis (AD) and allergic contact dermatitis (ACD) are inflammatory skin conditions whose association is not clearly defined. To identify differences in ACD profile between patients with and without AD among those referred for patch testing. Additionally, to determine the prevalence of sensitisation to standard Spanish contact allergens in both groups. We analysed two groups (AD and non-AD) within the Spanish Registry of Research in Contact Dermatitis and Cutaneous Allergy (REIDAC). Contact allergy, clinical relevance and epidemiological data were compared between them. A total of 5055 patients were included. Among them, 23% (1168) had a history or final diagnosis of AD. At least one positive reaction was seen in 468 (40%) of AD patients and 1864 (48%) of non-AD patients. In both groups, the most common positive reactions were to nickel sulphate, methylchloroisothiazolinone/methylisothiazolinone and cobalt chloride. Age-adjusted OR for sensitisation to nickel sulphate was 0.72 (95% CI: 0.61-0.86), indicating a decreased likelihood of sensitisation in AD patients compared to non-AD individuals. We did not find an increased presence of ACD in patients with AD referred for patch testing, exhibiting similar profiles to non-AD population, except for a negative relationship between AD and sensitisation to nickel sulphate.

MECHANISM OF LEUKOCYTE ADHERENCE INHIBITION

After binding a sensitizing tumor antigen, human leukocytes undergo a series of changes that lead to a loss of their glass-adherent properties; a phenomenon called leukocyte adherence inhibition (LAI). After surgery or when patients have a large tumor burden, their test results become negative. This study shows that in vitro incubation of the leukocytes for 5 min with PGE2 converted to positive the negative test, in an immunologically specific manner. The effect was critically dose-dependent, too little or too much did not alter the result. The same effect was achieved with PGE2, PGI2, aminophylline or other drugs that raise intracellular nucleotides, including dibutyryl cyclic AMP and dibutyryl cyclic GMP. Dibutyryl cyclic AMP stimulated a stronger response and 100 times less was needed than of dibutyryl cyclic GMP. Prostaglandins did not mediate LAI since Indomethacin failed to inhibit a positive test. Nonetheless, arachidonate metabolites were critical for the LAI phenomenon since BPB and mepacrine, inhibitors of phospholipase A2, negated the LAI response. Moreover, ETYA, phenidone and NDGA, inhibitors of the lipoxygenase metabolic pathway, all negated the positive LAI response. The positive response was especially sensitive to nullification by ETYA. Although the last-named drugs inhibit other arachidonate metabolic pathways too, conclusive evidence that the metabolites of the lipoxygenase pathway, and leukotrienes in particular, mediate the LAI response was the fact that FPL 55712, a competitive antagonist of SRS, nullified a positive response at levels as low as 10(-13) M. The results imply that prostaglandins were able to modulate the expression of LAI by affecting intracellular nucleotides, but leukotrienes, it seems, were the metabolites that mediated leukocyte nonadherence after monocytes recognized and bound tumor antigen.

Reflectance confocal microscopy may differentiate acute allergic and irritant contact dermatitis in vivo

7 - Immunogens