BACKGROUND:Cannabis allergy is increasingly reported, with Can s 3 as a major allergen. Investigations into sensitisation to Can s 3 utilise sophisticated techniques, including the cytometric bead assay and the basophil activation test. This study aims to utilise a fluorescence enzyme immunoassay to quantify sIgE to Can s 3, employing a recombinant Can s 3 protein. METHODS:This study included 104 cannabis allergic patients, 20 healthy controls and 70 exposed atopic controls. Specific IgE by a fluorescence enzyme immunoassay or cytometric bead assay and the basophil activation test all used the same recombinant allergen. Two-graph ROC curves were used to determine the clinically validated allergen-specific cut-off for maximal sensitivity and specificity and to facilitate direct comparison of the test performances. RESULTS:Twenty-two individuals were non-responding in the basophil activation test and were excluded from all analyses involving basophil activation test results. The clinically validated cut-off points are > 0.16 kUA/L, ≥ 0.14 kUA/L and > 5% for a fluorescence enzyme immunoassay, cytometric bead assay and basophil activation test, respectively. Utilising these thresholds, the fluorescence enzyme immunoassay exhibited a sensitivity of 72% and specificity of 74%, the cytometric bead assay demonstrated a sensitivity of 49% and specificity of 89%. In responders, the basophil activation test exhibited a sensitivity of 51% and specificity of 82%. Remarkably, low positive fluorescence enzyme immunoassay results, particularly below 0.35 kUA/L, are negative for both cytometric bead assay and basophil activation test. Conversely, utilising the conventional threshold of > 0.35 kUA/L, the sIgE a fluorescence enzyme immunoassay results exhibited greater congruence with those of the cytometric bead assay and basophil activation test. CONCLUSION:This study underscores the complexity of establishing an optimal decision threshold for the sIgE rCan s 3 fluorescence enzyme immunoassay and indicates that the clinically validated decision cut-off may not always represent the most efficacious approach.

With increased access and decriminalization of cannabis use, cases of IgE-dependent cannabis allergy (CA) and cross-reactivity syndromes have been increasingly reported. However, the exact prevalence of cannabis allergy and associated cross-reactive food syndromes (CAFS) remains unknown and is likely to be underestimated due to a lack of awareness and insufficient knowledge of the subject among health care professionals. Therefore, this practical roadmap aims to familiarize the reader with the early recognition and correct management of IgE-dependent cannabis-related allergies. In order to understand the mechanisms underlying these cross-reactivity syndromes and to enable personalized diagnosis and management, special attention is given to the molecular diagnosis of cannabis-related allergies. The predominant signs and symptoms of CA are rhinoconjunctivitis and contact urticaria/angioedema. However, CA can also present as a life-threatening condition. In addition, many patients with CA also have distinct cross-reactivity syndromes, mainly involving fruits, vegetables, nuts and cereals. At present, five allergenic components of Cannabis sativa (Can s); Can s 2 (profilin), Can s 3 (a non-specific lipid protein), Can s 4 (oxygen-evolving enhancer protein 2 oxygen), Can s 5 (the Bet v 1 homologue) and Can s 7 (thaumatin-like protein) have been characterized and indexed in the WHO International Union of Immunological Sciences (IUIS) allergen database. However, neither of them is currently readily available for diagnosis, which generally starts by testing crude extracts of native allergens. The road to a clear understanding of CA and the associated cross-reactive food syndromes (CAFS) is still long and winding, but well worth further exploration.

PDF - 84KB, Joint effects of smoking and occupational exposure to aromatic amines and polycyclic aromatic hydrocarbons on the risk of bladder cancer (754 cases and 833 controls).

PDF - 84KB, Association of working in at-risk occupations with bladder cancer in men (609 cases and 665 controls) and women (145 cases and 168 controls) from EPIC.

PDF - 73KB, Joint effects of NAT2 acetylation status and occupational exposure to aromatic amines on the risk of bladder cancer (492 cases and 569 controls).

Mesothelioma is an aggressive cancer, strongly associated with prior exposure to asbestos. Commonly, tumors are detected at late stages of the disease. Detection at early stages might be meaningful, because therapies might be more effective when the tumor burden is relatively low and the tumor has not spread to distant sites. Circulating biomarkers in blood might be a promising tool to improve the early detection of mesothelioma, but for screening in asymptomatic subjects, candidate biomarkers need to be validated in appropriate studies. This study was conducted to assess the performance of biomarkers in liquid biopsies to detect mesothelioma at early stages. Over a period of 10 years, 2769 volunteers formerly exposed to asbestos were annually examined and liquid biopsies were collected. A follow-up was completed 17 months after the last blood collection. The article provides a detailed overview of our lessons learned and experiences of conducting a prospective, longitudinal, multicenter study. The existing cohort of individuals at risk is highly suitable for the validation of blood-based biomarkers for the early detection of mesothelioma as well as lung cancer.

PDF - 63KB, NAT2 variants and bladder cancer risk for 607 cases and 695 controls nested in EPIC.

PDF - 80KB, Association of smoking and occupational exposure with bladder cancer by NAT2 acetylation in cases and controls from EPIC.

Malignant mesothelioma (MM) is a severe disease mostly caused by asbestos exposure. Today, one of the best available biomarkers is the soluble mesothelin-related protein (SMRP), also known as mesothelin. Recent studies have shown that mesothelin levels are influenced by individual genetic variability. This study aimed to investigate the influence of three mesothelin (MSLN) gene variants (SNPs) in the 5′-untranslated promoter region (5′-UTR), MSLN rs2235503 C > A, rs3764246 A > G, rs3764247 A > C, and one (rs1057147 G > A) in the 3′-untranslated region (3′-UTR) of the MSLN gene on plasma concentrations of mesothelin in 410 asbestos-exposed males without cancer and 43 males with prediagnostic MM (i.e., with MM diagnosed later on) from the prospective MoMar study, as well as 59 males with manifest MM from Germany. The mesothelin concentration differed significantly between the different groups (p < 0.0001), but not between the prediagnostic and manifest MM groups (p = 0.502). Five to eight mutations of the four SNP variants studied were associated with increased mesothelin concentrations (p = 0.001). The highest mesothelin concentrations were observed for homozygous variants of the three promotor SNPs in the 5′-UTR (p < 0.001), and the highest odds ratio for an elevated mesothelin concentration was observed for MSLN rs2235503 C > A. The four studied SNPs had a clear influence on the mesothelin concentration in plasma. Hence, the analysis of these SNPs may help to elucidate the diagnostic background of patients displaying increased mesothelin levels and might help to reduce false-positive results when using mesothelin for MM screening in high-risk groups.

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Cannabis is a flowering plant in the family of Cannabaceae, and the most widespread species is Cannabis sativa. In the past decade, cannabis allergy (CA) has been recognized as an increasing health issue and its prevalence is likely underestimated because of cannabis's illegal status in many countries and the absence of reliable diagnostics. The clinical presentation of CA is very heterogeneous, varying from mild rhinoconjunctivitis, urticaria, and angioedema to life-threatening anaphylaxis. Not seldom, CA is associated with a broad cross-reactivity syndrome involving fruits, vegetables, nuts, cereals, and latex.

Introduction: IgE-mediated Hevea latex allergy and associated food-allergies constitute a significant health issue with serious consequences of diagnostic error. Hence, there is a need for more reliable confirmatory diagnostics.Areas covered: Here, we summarize the major limitations of conventional tests using native extracts and describe how piecing together the IgE reactivity profile can benefit correct diagnosis in difficult cases in whom conventional tests yield equivocal or negative results. A diagnostic algorithm integrating traditional sIgE and component-resolved diagnosis (CRD) is presented.Expert opinion: Moreover, it is clear that the discoveries in the field of the Hevea latex proteome will contribute to our understandings and accurate approach of sometimes complex cross-reactivity phenomena that extend beyond the 'latex-fruit syndrome.'

Evidence shows that cannabis can elicit IgE-mediated allergy with symptoms ranging from mild rhinoconjunctivitis to life-threatening anaphylaxis. Most studies point to Can s 3, the nonspecific lipid transfer protein (nsLTP) of Cannabis sativa as a major allergen, particularly in Europe.1 However, Can s 3 does not cover the entire IgE-reactivity profile as 42 of 120 of cannabis allergic (CA) patients in a recent study did not display a Can s 3 sensitization.2

Introduction: Although the use of cannabis dates back millennia, the first description of cannabis allergy is relatively recent (1971). Recent large-scale data show that cannabis allergy can manifest severe and generalized symptoms with extensive cross-reactions. Thus, it is essential to become familiarized with its clinical presentation, diagnostic aids, and adequate therapeutic guidance.Areas covered: Here we provide a hands-on overview on cannabis allergy focusing on symptomatology and the reliability of diagnostic options. Recent advances in proteomics are discussed in detail, elucidating the link with nsLTP-related allergies. The proteomics advancements have paved the way for more reliable diagnostics, especially component-based tools. Finally, the current experience in treatment options is highlighted.Expert opinion: Cannabis allergy is an allergy entity which can significantly impact the quality of life. For optimal diagnosis, we advise to start with a validated and standardized crude-extract based test such as sIgE hemp complemented by component-based diagnostics such as sIgE Can s 3 quantifications where available. Future research should lift the veil on the true prevalence of cannabis allergy and the importance of other cannabis allergens to further guide our practice.

PURPOSE:Malignant pleural mesothelioma (MPM) is a highly lethal cancer caused by exposure to asbestos. Currently, the diagnosis is a challenge, carried out by means of invasive methods of limited sensitivity. This is a case-control study to evaluate the individual and combined performance of minimally invasive biomarkers for the diagnosis of MPM.METHOD:A study of 166 incident cases of MPM and 378 population controls of Mestizo-Mexican ethnicity was conducted. Mesothelin, calretinin, and megakaryocyte potentiating factor (MPF) were quantified in plasma by ELISA. The samples were collected from 2011 to 2016.RESULTS:Based on ROC analysis and a preset specificity of 95%, the combination of the three biomarkers reached an AUC of 0.944 and a sensitivity of 82% in men. In women, an AUC of 0.937 and a sensitivity of 87% were reached. In nonconditional logistic regression models, the adjusted ORs in men were 7.92 (95% CI 3.02-20.78) for mesothelin, 20.44 (95% CI 8.90-46.94) for calretinin, and 4.37 (95% CI 1.60-11.94) for MPF. The ORs for women were 28.89 (95% CI 7.32-113.99), 17.89 (95% CI 3.93-81.49), and 2.77 (95% CI 0.47-16.21), respectively.CONCLUSIONS:To our knowledge, this is the first study evaluating a combination of mesothelin, calretinin, and MPF, and demonstrating a sex effect for calretinin. The biomarker panel showed a good performance in a Mestizo-Mexican population, with high sensitivity and specificity for the diagnosis of MPM.

The IgE-reactivity profile of cannabis remains incomplete and probably exhibits geographical variation. Both our own and other European data point to Can s 3, the non-specific lipid transfer protein (nsLTP) of cannabis as a major allergen. However, previous North-American research suggested other putative allergens such as Oxygen-Evolving-Enhancer Protein 2 (OEEP2). We aim at comparing the role of OEEP2 and Can s 3 in cannabis allergy in North-Western Europe. 109 patients were included; patients with a clinical history of cannabis allergy (CA = 42), healthy controls (HC = 30) and atopic controls with pollen and/or nsLTP-sensitizations (P+LTP-/+ = 37). An nCan s 3-rich skin prick test, specific (s)IgE rCan s 3 and rOEEP2 were performed. Positive results were followed by additional basophil activation tests (BATs) with rCan s 3 and/or rOEEP2. 26/42 (62%) CA-patients displayed a Can s 3-sensitization, two of them were also OEEP2-sensitized (both sIgE- and BAT-positive). 18/42 CA-patients were neither sensitized to OEEP2 nor to Can s 3 (of which 6/18 reported cannabis-mediated anaphylaxis). Three of the P+LTP+ controls displayed a Can s 3 sensitization (BAT or sIgE), all other controls had completely negative explorations. These preliminary experiments confirm that Can s 3 is a major allergen and also disclose that OEEP2 binds IgE-antibodies and triggers basophil degranulation. Therefore, aside from Can s 3, OEEP2 is likely to be another novel cannabis allergen. Nevertheless, it appears to be only a minor allergen (5% sensitized) in North-Western Europe. Future collaborative studies to further unveil and compare its clinical relevance in geographically different areas are needed.

Objectives Cannabis allergy has mainly been described following recreational use but some cases also point to cannabis sensitisation as a result of occupational exposure. As a consequence, little is known on the prevalence and clinical phenotype of occupational cannabis allergy. Therefore, this study aims to explore the allergy-associated health risks of occupational cannabis exposure in Belgian police force personnel. Methods 81 participants, active in the police force, reporting regular occupational cannabis exposure during the past 12 months, were included. History was combined with a standardised questionnaire on allergies and cannabis exposure. Basophil activation tests (BAT s) with a crude cannabis extract and rCan s 3 were performed. In addition, specific (s) IgE rCan s 3 as well as sIgE to house dust mite, six pollen and three mould allergens were quantified. Results Although 42% of the participants reported respiratory and/or cutaneous symptoms on occupational cannabis exposure, all cannabis diagnostics were entirely negative, except one symptomatic case demonstrating a borderline result. Furthermore, there is no significant difference between the groups with and without symptoms on cannabis exposure in terms of allergenic sensitisations. Conclusions The origins of the reported respiratory and cutaneous symptoms during cannabis exposure remain elusive but are probably due to non-immune reactions. It should be noted that the study was volunteer-based possibly reflecting an excessive number of symptomatic individuals. Nevertheless, as only one participant reported using fully protective gear, much improvement is needed for reducing the number of symptoms reported on duty, independent of their origin.

BACKGROUND:Cannabis allergy (CA) has mainly been attributed to Can s 3, the nonspecific lipid transfer protein (nsLTP) of Cannabis sativa. Nevertheless, standardized diagnostic tests are lacking and research on CA is scarce.OBJECTIVE:To explore the performance of 5 cannabis diagnostic tests and the phenotypic profile of CA.METHODS:A total of 120 patients with CA were included and stratified according to the nature of their cannabis-related symptoms; 62 healthy and 189 atopic controls were included. Specific IgE (sIgE) hemp, sIgE and basophil activation test (BAT) with a recombinant Can s 3 protein from Cannabis sativa (rCan s 3), BAT with a crude cannabis extract, and a skin prick test (SPT) with an nCan s 3-rich cannabis extract were performed. Clinical information was based on patient history and a standardized questionnaire.RESULTS:First, up to 72% of CA reporting likely-anaphylaxis (CA-A) are Can s 3 sensitized. Actually, the Can s 3-based diagnostic tests show the best combination of positive and negative predictive values, 80% and 60%, respectively. sIgE hemp displays 82% sensitivity but only 32% specificity. Secondly, Can s 3+CA reported significantly more cofactor-mediated reactions and displayed significantly more sensitizations to other nsLTPs than Can s 3-CA. Finally, the highest prevalence of systemic reactions to plant-derived foods was seen in CA-A, namely 72%.CONCLUSIONS:The most effective and practical tests to confirm CA are the SPT with an nCan s 3-rich extract and the sIgE rCan s 3. Can s 3 sensitization entails a risk of systemic reactions to plant-derived foods and cofactor-mediated reactions. However, as Can s 3 sensitization is not absolute, other cannabis allergens probably play a role.

BACKGROUND:Common wheat (Triticum aestivum) and durum wheat (T. turgidum) are both involved in Baker's asthma (BA) and food allergy (FA) including wheat-dependent exercise-induced asthma (WDEIA). However, allergens in durum wheat have not been described, and the over-expression of T. turgidum non-specific lipid-transfer protein (nsLTPs) is considered to increase resistance to phytopathogens.OBJECTIVE:To identify and assess the allergenicity of nsLTP from T. turgidum.METHODS:Recombinant T. turgidum nsLTP Tri tu 14 was generated and tested for structural integrity (circular dichroism-spectroscopy) and purity (SDS-PAGE). Thirty-two wheat allergic patients were enrolled: 20 Spanish patients (BA) with positive bronchial challenge to wheat flour, and 12 Italian patients (wheat FA/WDEIA) with positive double-blind placebo-controlled food challenge/open food challenge (OFC) to pasta. IgE values to wheat, Tri tu 14, Tri a 14 (T. aestivum) and Pru p 3 (P. persica) were determined by ImmunoCAP testing. Allergenic potency (in vitro mediator release) and IgE cross-reactivity were investigated.RESULTS:Tri tu 14 was found to share 49% and 52% amino acid identity with Tri a 14 and Pru p 3, respectively. Among 25 Tri a 14 CAP positive sera, 23 (92%) were reactive to wheat extract, 22 (88%) to Tri tu 14 and 20 (80%) to Pru p 3. The correlation between Tri a 14 and Tri tu 14 specific IgE levels was r = 0.97 (BA) and r = 0.93 (FA/WDEIA), respectively. FA/WDEIA patients showed higher specific IgE values to Tri tu 14 and Pru p 3 than BA patients. Tri tu 14 displayed allergenic activity by mediator release from effector cells and IgE cross-reactivity with Pru p 3. The degree of IgE cross-reactivity between the two wheat nsLTPs varied between individual patients.CONCLUSIONS AND CLINICAL RELEVANCE:Sensitization to Tri tu 14 likely appears to be more important in wheat FA/WDEIA than in BA. Over-expression of Tri tu 14 in wheat would represent a risk for patients with nsLTP-mediated FA.