Molecular allergology: a clinical laboratory tool for precision diagnosis, stratification and follow-up of allergic patients.

Identification of the molecular culprits of allergic reactions leveraged molecular allergology applications in clinical laboratory medicine. Molecular allergology shifted the focus from complex, heterogeneous allergenic extracts, e.g. pollen, food, or insect venom, towards genetically and immunologically defined proteins available for in vitro diagnosis. Molecular allergology is a precision medicine approach for the diagnosis, stratification, therapeutic management, follow-up and prognostic evaluation of patients within a large range of allergic diseases. Exclusively available for in vitro diagnosis, molecular allergology is nonredundant with any of the current clinical tools for allergy investigation. As an example of a major application, discrimination of genuine sensitization from allergen cross-reactivity at the molecular level allows the proper targeting of the culprit allergen and thus dramatically improves patient management. This review aims at introducing clinical laboratory specialists to molecular allergology, from the biochemical and genetic bases, through immunological concepts, to daily use in the diagnosis and management of allergic diseases.

Reinforcement during the COVID-19 pandemic: Perception of nursing students and impact on intention to drop-out of nursing education.

Background: Although the COVID-19 pandemic's impact on students has already been studied, its impact on nursing students' perception of their training and their conception of their future profession is unknown. Aims: To describe nursing students' perception of their involvement in reinforcement during the COVID-19 pandemic and the impact of working as reinforcement staff during the COVID-19 pandemic on nursing students. Design: Cross-sectional, comparative case/non-case study. Setting: nurse training institutions in France. Participants: "Cases" defined as nursing students who worked as reinforcement staff during the COVID-19 pandemic; "non-cases" defined as people who were in final year of nursing studies in 2018-2019 or 2019-2020 and so did not work as reinforcement staff during their nursing studies. Methods: questionnaire about representations of the nursing profession, role of the nurse in society, previous thinking of dropping out of nursing education. Results: 534 subjects included (310 cases; 214 non-cases). Cases reported feeling useful (38.6%) or very useful (25.7%) as reinforcement workers, while 91.5% concurred that nurses had an important role in the management of COVID-19 patients. Cases more frequently reported that the nursing profession is one where you save lives (61.5% vs 52.5%, p = 0.05). The desire to work as a nurse for a whole life had been more frequently expressed by cases (45.3% vs 34.8%, p = 0.05). Nursing education drop-out has been considered by 63.4% of subjects, without difference between "cases" and "non-cases" (p = 0.63). Subjects who considered dropping out of nursing education were younger (p = 0.01) and less often prone to think that the nursing profession was a profession personally rewarding (p = 0.01) and a life-saving profession (p = 0.03). Conclusion: The majority of nursing students reported feeling useful during the pandemic, and underlined the importance of the nurse's role in management of COVID-19 patients. Participation in reinforcement staff during the pandemic had no influence on dropping out of nursing education.

Practices of French General Practitioners Regarding Vaccination of Boys Against Human Papillomavirus (HPV), One Year After the Application of Its Official Recommendation.

In 2019, French health authorities extended the recommendation for human papillomavirus (HPV) vaccination to include boys aged 11 to 19 years. We describe HPV vaccination practices among French general practitioners (GPs) since this recommendation wasapplied. We also identified factors associated with the propensity to propose HPV vaccination to boys. Cross-sectional study, between May and August 2022, among French GPs using a questionnaire asking about the GPs, their practices, and opinions regarding HPV vaccination, including whether they systematically proposed HPV vaccination to eligible boys or not. We investigated factors associated with systematic proposal of HPV vaccination, using univariate and multivariate logistic regression. In total, 360 GPs participated (76.6% females; mean age 34.7 ± 7.8 years; 22.9% had additional training in gynecology or pediatrics); 5.5% reported that they systematically offered HPV vaccination to boys prior to the recommendation, whereas 61.2% do so systematically since the recommendation. Factors associated with systematic proposal to boys (post recommendation) were female GP sex (78.6% versus 66.2%; OR = 2.0 [95% confidence interval (CI) 1.2-3.3]; p = 0.007) and systematic proposal prior to the recommendation (8.5% versus 0.7%; OR = 13.3 [1.7-101.7]; p = 0.01). Protection against HPV-induced cancer was cited as an argument to vaccinate girls (98.3% versus 89.2%; p < 0.0001); while reducing the risk of transmission was more commonly an argument to vaccinate boys (78.1% versus 51.8%; p < 0.0001). This study underlines the positive impact of the official recommendation for HPV vaccination of boys on the attitude of GPs, with an increase in the systematic proposal of HPV vaccination to boys.

Impact of a dTcaP booster vaccine awareness campaign initiated by the French national health insurance for adults aged 25 years in 2021.

BACKGROUND: Vaccination schedules differ from country to country. In France, the diphtheria, tetanus, pertussis, poliomyelitis (dTcaP) booster vaccine coverage for adults aged 25 has been lower than those recommended. We evaluated the impact of an awareness campaign undertaken by the French national health insurance system in 2021. METHODS: A randomized, controlled study with adults residing in the Ardennes region was conducted to evaluate the effect on vaccine coverage of the booster vaccine reminder campaign carried out via letter and/or email and/or SMS. The randomization unit was the municipal administrative area (canton). Ten cantons were grouped into the intervention group (INT) and nine were the control group (CON). Outcomes were the booster vaccine delivery and the consultation of a general practitioner (GP) within 12 months (since the French national health insurance running the campaign suggested patients to consult their GP). RESULTS: A total of 1,975 adults were included (INT: 67.3% vs. CON: 32.7%). Of them, 331 received a booster vaccine (INT: 17.4% vs. CON: 15.5%; p = 0.29), and 1,442 consulted a GP (INT: 73.7% vs. CON: 76.8%; p = 0.14). Those who consulted a GP had more frequent vaccine delivery (INT: 19.1% vs. CON: 10.5%; p < 0.0001). CONCLUSIONS: This study found that the awareness campaign run by the French national health insurance did not improve the uptake of the dTcaP booster and that there was a low rate of vaccinated adults aged 25 years. A GP consultation was associated with dTcaP booster vaccine delivery which may show that there is a need of involving GPs in vaccination follow-ups. Patients recognize GPs as providers of credible information and they may play a key role in individualized preventive healthcare actions. Systematic consultations with GPs for follow-up could be proposed to insured adults aged 25 years in the future.

Characterization of the novel HLA-A*33:220 allele by next-generation sequencing.

HLA-A*33:220 differs from HLA-A*33:03:01:01 by one nucleotide substitution in codon 245 in exon 4.

Does the use of color coding facilitate parents' understanding of body mass index curves?

BACKGROUND: In France, national guidelines recommend early detection and management of overweight and obesity in children, with multi-year systematic generation of children's body mass index (BMI) curves in primary care. It is important for the parents to understand the BMI curves displayed in the child's health notebook and to become involved in the care with health professionals. OBJECTIVE: The aim of the current study was to compare parents' understanding of a BMI curve displayed using color coding versus their understanding of the same curve displayed without color coding. METHODS: An observational, cross-sectional, comparative study was performed between February 1, 2021 and November 15, 2021. Adult parents with at least one child attending primary school were included. Two questionnaires testing parents' understanding were completed: one showing BMI curves without color coding (five questions) and one showing BMI curve with color coding (five questions). The primary endpoint was the proportion of parents achieving fully correct answers. Comparisons of endpoints between the color-coded and non-color-coded curve were performed using the McNemar test. Factors associated with the primary endpoint were investigated by mixed logistic regression models with the subject as a random effect. RESULTS: The 109 participants (45.4% response rate) had an average age of 39.4 ± 6.6 years; 81.7% were women. A total of 214 complete questionnaires were compared: The proportion of participants with fully correct responses was significantly higher using the BMI curve with color coding compared to the curve without color coding (86.0% vs. 54.2%, p<0.0001). In multivariate analysis, the use of color coding was significantly associated with a higher likelihood of achieving fully correct responses (odds ratio: 5.9, 95% CI: 3.0-11.2, p<0.0001). CONCLUSION: The use of color coding improved parents' understanding of BMI curves. Further research should explore equally the benefits and risks associated with weight loss and mental health when using a colored BMI curve for the detection and management of overweight and obese children.

Characterization of the novel HLA-A*01:383 allele by next-generation sequencing.

HLA-A*01:383 differs from HLA-A*01:01:01:01 by two nucleotide substitutions at positions 28 and 48 in exon 1.

Early plasma interferon-ß levels as a predictive marker of COVID-19 severe clinical events in adult patients.

We assessed relationships between early peripheral blood type I interferons (IFN) levels, clinical new early warning scores (NEWS), and clinical outcomes in hospitalized coronavirus disease-19 (COVID-19) adult patients. Early IFN-ß levels were lower among patients who further required intensive care unit (ICU) admission than those measured in patients who did not require an ICU admission during severe acute respiratory syndrome coronavirus type 2 infection. IFN-ß levels were inversely correlated with NEWS only in the subgroup of patients who further required ICU admission. To assess whether peripheral blood IFN-ß levels could be a potential relevant biomarker to predict further need for ICU admission, we performed receiver operating characteristic (ROC) curve analyses that showed for all study patients an area under ROC curve of 0.77 growing to 0.86 (p = 0.003) when the analysis was restricted to a subset of patients with NEWS ≥5 at the time of hospital admission. Overall, our findings indicated that early peripheral blood IFN-ß levels might be a relevant predictive marker of further need for an ICU admission in hospitalized COVID-19 adult patients, specifically when clinical score (NEWS) was graded as upper than 5 at the time of hospital admission.

Exploring the impact of prior spontaneous miscarriage on stress among pregnant women during the first trimester: an observational study.

BACKGROUND: Spontaneous miscarriage (SM) is the most common complication of pregnancy. Its psychological repercussions are widely documented but few studies have investigated its effect on women's experience of a subsequent pregnancy. AIM: To evaluate the impact of prior SM on the level of stress experienced by pregnant women during the first trimester of pregnancy. DESIGN AND SETTING: Cross-sectional, observational study, which was conducted between June and October 2021 in France. METHOD: A self-report questionnaire was distributed to women in the first trimester of pregnancy. Stress was assessed using the Antenatal Perceived Stress Inventory to yield an overall score and a score for three dimensions ('medical and obstetric risks or fetal health'; 'psychosocial changes during pregnancy'; and the 'prospect of childbirth'). Women with a history of prior SM and those without were compared. RESULTS: In total, 93 women were included; 63 without and 30 with a history of prior SM. Prior SM was not associated with the overall score. The score for the dimension 'medical and obstetric risks or fetal health' was significantly higher in women with prior SM (3.00±0.86 versus 2.34±0.80; ß = 0.61 [95% confidence interval {CI} = 0.25 to 0.96]; P = 0.001). Prior SM was significantly associated with the items 'the baby's health' (P = 0.048) and 'the echography' (P = 0.002). CONCLUSION: This study shows a significant impact of prior SM on the level of stress of pregnant women during the first trimester, particularly relating to the medical and obstetric risks or fetal health, underlining the need for appropriate psychological support to be provided to women who experience SM.

Characterization of the novel HLA-B*53:64 allele by next-generation sequencing.

HLA-B*53:64 differs from HLA-B*53:01:01:01 by one nucleotide substitution at position 1617 in exon 4.

Characterization of the novel HLA-C*07:1001N allele by next-generation sequencing.

HLA-C*07:1001N differs from HLA-C*07:01:01:01 by a deletion of 17 nucleotides in exon 1.

Characterization of the novel HLA-C*03:04:94 allele by next-generation sequencing.

HLA-C*03:04:94 differs from HLA-C*03:04:01:01 by one nucleotide substitution at position 737 in exon 4.

Characterization of the novel HLA-C*16:184 allele by next-generation sequencing.

HLA-C*16:184 differs from HLA-C*16:02:01:01 by one nucleotide substitution at position 737 in exon 3.

Characterization of the novel HLA-C*05:255 allele by next-generation sequencing.

HLA-C*05:255 differs from HLA-C*05:01:01:02 by one nucleotide substitution at position 2013 in exon 5.

Characterization of the novel HLA-C*06:317 allele by next-generation sequencing.

HLA-C*06:317 differs from HLA-C*06:02:01:01 by one nucleotide substitution at position 921 in exon 3.

Characterization of the novel HLA-B*35:29:03 allele by next-generation sequencing.

HLA-B*35:29:03 differs from HLA-B*35:29:01 by one nucleotide substitution at position 374 in exon 2.

Absence of Anti-Glomerular Basement Membrane Antibodies in 200 Patients With Systemic Lupus Erythematosus With or Without Lupus Nephritis: Results of the GOODLUPUS Study.

Introduction: Anti-glomerular basement membrane (GBM) antibodies are pathogenic antibodies first detected in renal-limited anti-GBM disease and in Goodpasture disease, the latter characterized by rapidly progressive crescentic glomerulonephritis combined with intra-alveolar hemorrhage. Studies have suggested that anti-GBM antibody positivity may be of interest in lupus nephritis (LN). Moreover, severe anti-GBM vasculitis cases in patients with systemic lupus erythematosus (SLE) have been described in the literature, but few studies have assessed the incidence of anti-GBM antibodies in SLE patients. Objective: The main study objective was to determine if positive anti-GBM antibodies were present in the serum of SLE patients with or without proliferative renal damage and compared to a healthy control group. Methodology: This retrospective study was performed on SLE patients' sera from a Franco-German European biobank, developed between 2011 and 2014, from 17 hospital centers in the Haut-Rhin region. Patients were selected according to their renal involvement, and matched by age and gender. The serum from healthy voluntary blood donors was also tested. Anti-GBM were screened by fluorescence enzyme immunoassay (FEIA), and then by indirect immunofluorescence (IIF) in case of low reactivity detection (titer >6 U/ml). Results: The cohort was composed of 100 SLE patients with proliferative LN (27% with class III, 67% with class IV, and 6% with class V), compared to 100 SLE patients without LN and 100 controls. Patients were mostly Caucasian and met the ACR 1997 criteria and/or the SLICC 2012 criteria. Among the 300 tested sera, no significant levels of anti-GBM antibodies were detected (>10 U/ml) by the automated technique, three sera were found "ambivalent" (>7 U/ml): one in the SLE with LN group and two in the SLE without LN group. Subsequent IIF assays did not detect anti-GBM antibodies. Conclusion: Anti-GBM antibodies were not detected in the serum of Caucasian patients with SLE, even in case of renal involvement, a situation favoring the antigenic exposure of glomerular basement membranes. Our results reaffirm the central role of anti-GBM antibodies as a specific diagnostic biomarker for Goodpasture vasculitis and therefore confirm that anti-GBM antibody must not be carried out in patients with SLE (with or without LN) in the absence of disease-suggestive symptoms.

Neutrophil Extracellular Traps Generation Relates with Early Stage and Vascular Complications in Systemic Sclerosis.

Systemic sclerosis (SSc) is a systemic disease characterized by a great clinical and immunological heterogeneity whose pathophysiology is still being unraveled. Recently, innate immunity has been proposed to participate to the pathogenesis of SSc. In this study, we investigated the release of neutrophil extracellular traps (NETs) according to patient phenotype. Polymorphonuclear neutrophils (PMN) from 34 SSc patients and 26 healthy controls were stimulated by serum from SSc or healthy subject. NETs were visualized using epifluorescence microscope after DNA, myeloperoxidase, and Histone H3 tagging. Area of NETs were quantified using an original macro running in ImageJ® software. PMN from SSc patients were significantly more prone to releasing NETs than control PMN after autologous stimulation. PMN from patients with severe vascular complications (pulmonary arterial hypertension, digital ulcers) produced more NETs than PMN from other SSc patients and their aberrant NET production appeared to be sustained over time. In patients with pulmonary interstitial disease or extensive cutaneous fibrosis, NET production was high at an early stage of the disease before progressively decreasing. Both serum factors and PMN activation status were involved in the enhanced production of NETs in SSc. Consequently, neutrophils and especially NETosis represent new physiopathological and therapeutic fields in SSc.

NET Formation in Bullous Pemphigoid Patients With Relapse Is Modulated by IL-17 and IL-23 Interplay.

Background: DNA extracellular traps (ETs), released by neutrophils (NETs), or eosinophils (EETs), play a pathogenic role in several autoimmune disorders. However, to date, NETs have never been investigated in bullous pemphigoid (BP) with respect to clinical and immunological activities, both at baseline and at time of relapse which have been characterized with specific IL-17 and IL-23 patterns. Objective: We sought to assess whether ETs were associated with BP as well as the relative contribution of IL-17 axis cytokines to NET induction. Methods: Skin biopsy specimens were obtained from 11 patients with BP. Immuno-detection of neutrophils and eosinophils combined to DNA staining allowed us to investigate the in-situ presence of NETs and EETs using confocal scanning microscopy. NETs release was evaluated ex vivo by stimulating polymorphonuclear cells from BP patients with BP biological fluids in presence of IL-17A and IL-23 or of glucocorticoids. Results: At baseline, ETs were observed in BP lesions at the site of dermal-epidermal cleavage. Despite an important infiltrate of eosinophils, ETs were essentially associated with neutrophils in situ and were not related to BP clinical activity at diagnosis. In situ observation of NETs was associated in 6 among 8 patients with serum capacity of NET induction. Notably both blister fluid and sera from BP patients at diagnosis and at time of relapse could induce NET formation ex vivo. In contrast, a longitudinal investigation showed a decrease of NET formation with time of treatment in patients undergoing remission. Mimicking relapse, complementation of sera from BP patients with ongoing remission with either IL-17A or IL-23 increased NET formation. Conversely, IL-17A inhibited NET formation induced by serum from BP patients with relapse supplemented or not with IL-23. Finally, glucocorticoids also inhibited NET formation ex vivo in BP. Conclusion: NET formation is an associated phenomenon with BP. Furthermore, we showed that IL-23 favored NET formation, whereas the effects of IL-17A are environment dependent. Indeed, IL-17A displayed a protective effect on NET formation when associated with IL-23, showing for the first-time differential effects of these two cytokines in BP.

Diagnostic Utility of Separate Anti-Ro60 and Anti-Ro52/TRIM21 Antibody Detection in Autoimmune Diseases.

Anti-SS-A antibodies are often sought for in autoimmune diseases diagnosis. Two different target proteins have actually been identified: Ro52 and Ro60. Clinical and immunological associations seem different depending on anti-Ro52 or anti-Ro60 antibodies presence. However, due to a heterogeneous presentation in the literature, some immunology laboratories in France have stopped providing anti-Ro52 antibody findings. We report here a new hospital study designed to determine the diagnostic utility of the separate detection of anti-Ro52 and anti-Ro60 antibodies. We conducted a retrospective, observational study, including every adult patient with positive antinuclear antibodies (ANA) tested in our immunology laboratory, and associated with anti-Ro52 and/or anti-Ro60 antibodies, between 2011 and 2014. Out of 13032 sera tested for ANA, 399 adults had antibodies to Ro52 and/or Ro60; 81.7% were female, with a mean age of 54.5 ± 17.0 years. Anti-Ro52 antibodies were found in 75.7% of the patients and anti-Ro60 antibodies in 56.9%. Among them, 43.1% were classified in the Ro52 + Ro60- group, 32.6% in the Ro52 + Ro60 + group and 24.3% in the Ro52-Ro60+ group. In the Ro52-Ro60+ group, systemic lupus was the most frequent diagnosis (48.5%), with a possible association with antiphospholipid antibodies (anti-cardiolipin antibodies: OR 2.5 (CI95 [1.0-5.0], p = 0.05) and lupus anticoagulant {OR 3.6 (CI95 [1.10-10.0] p = 0.02)}. In the Ro52+Ro60+, primary Sjögren Syndrome was the most likely (OR 4.2 95% CI [2.1-8.3] p < 10-4), especially in patients Ro52+Ro60+La+. Patients with isolated anti-Ro52 had a wider variety of diseases associated, but among auto-immune diseases they were more prone to inflammatory myositis (OR 10.5 [1.4-81.7], p = 0.02) and inflammatory rheumatism (OR 4.6 [1.6-13.8], p = 0.006) in contrast to systemic lupus (OR 0.2 [0.1-0.3], p < 10-4) or primary Sjögren's syndrome (OR 0.1 [0.06-0.2], p < 10-4). We therefore suggest that, when anti-ENA antibodies are prescribed, it should include separate anti-Ro52 and anti-Ro60 antibodies determination. To go even further, we would like to suggest a change in ENA nomenclature to avoid confusion, abandoning the anti-SS-A label in favor of the anti-Ro52/TRIM21 or anti-Ro60 antibody for a clearer designation.