Oral myeloid cells uptake allergoids coupled to mannan driving Th1/Treg responses upon sublingual delivery in mice.

BACKGROUND: Polymerized allergoids coupled to nonoxidized mannan (PM-allergoids) may represent novel vaccines targeting dendritic cells (DCs). PM-allergoids are better captured by DCs than native allergens and favor Th1/Treg cell responses upon subcutaneous injection. Herein we have studied in mice the in vivo immunogenicity of PM-allergoids administered sublingually in comparison with native allergens. METHODS: Three immunization protocols (4-8 weeks long) were used in Balb/c mice. Serum antibody levels were tested by ELISA. Cell responses (proliferation, cytokines, and Tregs) were assayed by flow cytometry in spleen and lymph nodes (LNs). Allergen uptake was measured by flow cytometry in myeloid sublingual cells. RESULTS: A quick antibody response and higher IgG2a/IgE ratio were observed with PM-allergoids. Moreover, stronger specific proliferative responses were seen in both submandibular LNs and spleen cells assayed in vitro. This was accompanied by a higher IFNγ/IL-4 ratio with a quick IL-10 production by submandibular LN cells. An increase in CD4+ CD25high FOXP3+ Treg cells was detected in LNs and spleen of mice treated with PM-allergoids. These allergoids were better captured than native allergens by antigen-presenting (CD45+ MHC-II+ ) cells obtained from the sublingual mucosa, including DCs (CD11b+ ) and macrophages (CD64+ ). Importantly, all the differential effects induced by PM-allergoids were abolished when using oxidized instead of nonoxidized PM-allergoids. CONCLUSION: Our results demonstrate for the first time that PM-allergoids administered through the sublingual route promote the generation of Th1 and FOXP3+ Treg cells in a greater extent than native allergens by mechanisms that might well involve their better uptake by oral antigen-presenting cells.

Safety of immunotherapy with glutaraldehyde modified allergen extracts in children and adults.

BACKGROUND: Glutaraldehyde-modified natural allergen extracts show significant reduction in the IgE-binding capacity and proteolytic activity. This allows the administration of higher doses in a shorter period of time, and to mix different allergen extracts. OBJECTIVE: Evaluate the safety of different concentrations and mixtures of glutaraldehyde-modified allergen extracts in a large group of paediatric and adult patients undergoing specific immunotherapy treatment. MATERIALS AND METHODS: 1855 patients (1156 adults and 699 children), suffering from rhinoconjunctivitis and/or asthma, participated in an observational multicentre cohort study, to evaluate the safety of immunotherapy using vaccines containing modified allergen extracts. Patients were monosensitised, or polysensitised, and received a therapeutic vaccine containing polymerised allergen extracts adsorbed onto aluminium hydroxide. Safety was assessed by recording all side reactions related to immunotherapy. RESULTS: The clinically relevant local reactions totalled 120, (90 immediate and 30 delayed) (1.02% of injections). Of them, 31 (0.26% of injections) occurred in children (26 immediate and 5 delayed) and 89 (0.76% of injections) in adults (64 immediate and 25 delayed). There were 38 systemic reactions. Eleven reactions were immediate (9 of grade 1 and 2 of grade 2) and 27 delayed (22 of grade 1 and 5 of grade 2). There were seven grade 2 systemic reactions (0.06% of the injections). No differences (P>0.05) in the number of reactions were observed between adults and children and between treatments were found in systemic reactions. All systemic reactions were mild and resolved spontaneously without the need of medication. CONCLUSION: Specific immunotherapy using natural modified allergen vaccines is safe to treat allergic patients, even at higher doses and in mixtures of unrelated allergen extracts. The percentage of adverse reactions detected is lower than those reported in the literature with native unmodified allergen extracts.

PET/CT in nuclear endocrinology: Results of the survey diffused through different social networks of the SEMNIM in Spain.

AIM: To know the current use of the PET/CT technology applied in the field of endocrinology, the Endocrinology Working Group of SEMNIM proposed conducting a survey. The objective was to obtain a snapshot of the use of PET/CT in nuclear endocrinology, to know if it is being used properly and detect possible needs. MATERIAL AND METHODS: During the first quarter of 2022, we analyzed the data obtained from a survey that was distributed through different social networks throughout the second half of 2021. The survey asked for the use of the different PET/CT techniques available in Spain in different endocrinological pathologies like differentiated thyroid carcinoma, medullary thyroid carcinoma, neuroendocrine tumors and hyperparathyroidism. RESULTS: A total of 15 centers responded to the survey. A percentage of 79 of hospitals used 18F-FDG PET/CT in the diagnosis and/or follow-up of differentiated thyroid carcinoma (mean annual studies: 36.9; range 10-100); 85% used 18F-DOPA PET/CT for the study of biochemical recurrence of medullary thyroid carcinoma (mean annual studies: 7.8; range 2-20); 77% used 18F-DOPA PET/CT for the study of neuroendocrine tumors: 77% used 18F-DOPA PET/CT (mean of 10 scans per year; range 2-30) and 69% used 68Ga-DOTA-SA (mean of 24.7 studies per year; range 2-127); 79% used 18F-choline PET/CT for the study of hyperparathyroidism (mean of 30.1 annual studies; range 10-120). CONCLUSIONS: We detected that the use of the PET/CT technique in endocrinology is not yet widespread, however, we saw that the indications in which it is used are, in general, those reported in the different consensus.

Our experience in the short-term diagnostic-therapeutic management of the patient with anal dysplasia.

INTRODUCTION: Anal intraepithelial neoplasia (AIN) is a premalignant lesion of anal squamous cell carcinoma. HIV-positive males who have sex with males, are the most affected at-risk population. Cytology and anuscopy are the best accepted methods for its diagnosis, although it is controversial which patients should complete it with a biopsy. Neither which patients should undergo treatment nor which is the best treatment is not well established. With this study, we would like to present our experience in the diagnostic-therapeutic management of AIN in the short term. METHODS: Retrospective observational study of patients at risk of AIN with altered anal cytology who underwent high-resolution anuscopy with biopsy. After histological confirmation of dysplasia, they started treatment with trichloroacetic acid. Its effectiveness was verified by subsequent cytology. The demographic variables of the sample and the results of both diagnostic and treatment tests were analyzed. RESULTS: The majority were HIV-positive males (104/115) and 50% had sexual relations with other men. We included 115 patients with altered anal cytology, of whom 92% had dysplasia on biopsy. 97% with atypia of uncertain significance on cytology had histological dysplasia. Cytology normalized after treatment in 60% of patients. CONCLUSION: Early detection of AIN should be routinely considered in known at-risk populations. Any cytological abnormality should be biopsied. Tricholoroacetic acid can be an effective treatment achieving a high percentage of regression, although currently, the information we have is of low level of evidence.

Evaluation of the prognostic value of the metabolic volumetric parameters calculated with 18F-FDG PET/CT and its value added to the molecular characteristics in patients with diffuse large B-cell lymphoma.

INTRODUCTION AND OBJECTIVES: Metabolic tumor volume (MTV) and total lesion glycolysis (TLG) are prognostic predictors in patients with diffuse large B-cell lymphoma (DLBCL). The objective of this study is to evaluate the prognostic impact of the baseline volumetric parameters calculated with positron emission tomography/computed tomography with 18F-fluorodeoxyglucose (18F-FDG PET/CT) and its added value to the molecular characteristics in patients with DLBCL not otherwise specified (NOS). METHODOLOGY: This is a retrospective observational study, which included 35 patients who underwent a baseline 18F-FDG PET/CT prior to treatment. A univariate analysis of the volumetric parameters (MTV and TLG), immunohistochemical study and chromosomal translocations were performed. The method for calculating the volumetric parameters was the SUV 2.5 threshold. The comparison between the predictive models was selected based on the information criterion value of Akaike (AIC), bayesian (BIC) and Harrell's C, after performing a Cox proportional hazards regression model. In addition, a univariate analysis of the volumetric parameters was performed according to the data of the immunohistochemical study using the Wilcoxon-Mann-Whitney test. RESULTS: A univariate analysis revealed that VMT and TLG are predictors of progression-free survival (PFS) and overall survival (OS), with a high discrimination capacity. Adding VMT and TLG to the immunohistochemical study and chromosomal translocation provided a better prognostic value for PFS and OS in patients diagnosed with DLBCL-NOS. Likewise, it was evidenced that the values of the volumetric parameters were lower in patients who presented a germinal center B cell phenotype (GCB) compared to patients with an activated B cell phenotype (ABC) who presented higher values. CONCLUSION: MTV and TLG added to the immunohistochemical study and chromosomal translocation provided a better prognostic value for PFS and OS in patients diagnosed with DLBCL-NOS.

The utility HBA1c test as a screening biomarker for detecting gestational diabetes mellitus.

BACKGROUND: Glycated Hb (HbA1c) has not been used for the diagnosis of gestational diabetes mellitus (GDM). Measurement of HbA1c levels is less complicated and more comfortable than glucose challenge test (GCT) for pregnant women. We studied HbA1c as a biomarker of GDM and as a screening test to avoid the use of GCT. METHODS: A prospective case-control study involves 745 pregnant women between 24th and 28th gestation week. HbA1c levels were measured and GDM was diagnosed according to Carpenter-Coustan criteria. Mean and SD were calculated for GCT value, HbA1c, age, and body mass index (BMI). A receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic performance of HbA1c test in diagnosing GDM. Cut-off points were calculated to rule out GDM and sensitivity (Se) and specificity (Sp) were also determined. A study of the implementation of HbA1c cut-offs was performed to avoid the GCT or to perform the confirmatory oral glucose tolerance test (OGTT). RESULTS: The area under the curve (AUC) was 0.67 (0.58-0.76). Using 4.6% HbA1c as a cut-off prevented false negatives but only decreased the number of GCTs performed by 7.2%. However, using 4.7% HbA1c resulted in one false negative (reduction of 15.0%). Finally, by selecting 4.8% HbA1c, we found two false negatives, but there were 25.9% who do not require a GCT. CONCLUSIONS: Adoption of HbA1c as a screening test for GDM may eliminate the need of GCT. Although the HbA1c test does not have sufficient Se and Sp to be used as the only diagnostic test, the use of a rule-out strategy in combination with the OGTT could be useful.

Multidisciplinary practice guidelines for the diagnosis, genetic counseling and treatment of pheochromocytomas and paragangliomas.

Pheochromocytomas and paragangliomas (PPGLs) are rare neuroendocrine tumors that arise from chromaffin cells of the adrenal medulla and the sympathetic/parasympathetic neural ganglia, respectively. The heterogeneity in its etiology makes PPGL diagnosis and treatment very complex. The aim of this article was to provide practical clinical guidelines for the diagnosis and treatment of PPGLs from a multidisciplinary perspective, with the involvement of the Spanish Societies of Endocrinology and Nutrition (SEEN), Medical Oncology (SEOM), Medical Radiology (SERAM), Nuclear Medicine and Molecular Imaging (SEMNIM), Otorhinolaryngology (SEORL), Pathology (SEAP), Radiation Oncology (SEOR), Surgery (AEC) and the Spanish National Cancer Research Center (CNIO). We will review the following topics: epidemiology; anatomy, pathology and molecular pathways; clinical presentation; hereditary predisposition syndromes and genetic counseling and testing; diagnostic procedures, including biochemical testing and imaging studies; treatment including catecholamine blockade, surgery, radiotherapy and radiometabolic therapy, systemic therapy, local ablative therapy and supportive care. Finally, we will provide follow-up recommendations.

Evaluation of the prognostic value of different methods of calculating the tumour metabolic volume with 18F-FDG PET/CT, in patients with diffuse large cell B-cell lymphoma. / Evaluación del valor pronóstico de los diferentes métodos de cálculo del volumen metabólico tumoral con 18F-FDG PET/TC, en pacientes con linfoma B difuso de células grandes.

INTRODUCTION AND OBJECTIVES: Metabolic tumor volume (MTV) is a promising indicator of prognosis in diffuse large B-cell lymphoma (DLBCL). The aim of the present study is to evaluate the different methods for the calculation of the basal metabolic tumor volume with 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) in the patients with DLBCL, relating each one of the volumes measured with progression-free survival (PFS) and overall survival (OS). METHODOLOGY: This is a retrospective analytical cohort study, in which 34 patients underwent to 18F-FDG PET/CT baseline prior to treatment. We compared three SUV thresholds 2.5, SUV 40% of the maximum SUV and SUV mean hepatic uptake (PERCIST) for the calculation of MTV and total lesion glycolysis (TLG) biomarkers, relating them to the PFS and OS. The best predictive model was selected based on the Akaike's information criterion (AIC) after performing a Cox proportional hazards regression. RESULTS: In relation to the PFS, they show statistically significant differences: MTV 2.5, TLG 2.5, MTV 40, TLG 40, MTV and TLG calculated with the PERCIST threshold. Among these, the one that has a lower AIC is MTV 2.5, so it is considered the best parameter to predict the PFS. With respect to OS, it shows statistically significant differences: MTV 2.5, VMT and TLG calculated with the PERCIST threshold. Among these three, the one with the lowest AIC is MTV 2.5, which is why it is considered the best parameter to predict OS. In addition, a higher value of MTV and total tumor glycolysis (TLG), is associated with worse PFS and OS CONCLUSION: The MTV calculated with the threshold SUV 2.5 seems to be the best parameter to predict PFS and OS in patients diagnosed with DLBCL with 18F-FDG PET/CT.

[Prenatal diagnosis of hydronephrosis: utility of diuretic renography]. / Diagnóstico prenatal de hidronefrosis: utilidad del renograma isotópico diurético.

OBJECTIVE: To investigate the value of diuretic renography in patients with prenatally diagnosed unilateral hydronephrosis. MATERIAL AND METHOD: We reviewed 44 patients who underwent ultrasonography in the first week of life and 1 month after birth, and cystography and diuretic renography at 1 month to evaluate differential renal function and the diuretic washout pattern. Patients with vesicoureteral reflux were followed-up according to the protocol for this disorder. In the remaining patients, ultrasonography and/or diuretic renography was performed every 3-6 months. RESULTS: In 4/44 patients vesicoureteral reflux was detected as cause a of hydronephrosis. In the remaining 40 patients, diuretic renography showed a washout pattern not suggestive of obstruction in 32 (only one patient needed surgery due to pyohydronephrosis). An indeterminate washout pattern was detected in one patient (who required surgery due to worsening of the washout pattern). A pattern suggestive of obstruction was detected in seven patients, four of whom required surgery (three due to a decrease in differential renal function and one due to worsening of the degree of pelvic dilatation). CONCLUSIONS: Diuretic renography is highly useful in risk stratification and in the management of newborn infants with hydronephrosis since infants with washout patterns not suggestive of obstruction will rarely develop obstructive hydronephrosis and can initially be followed-up with ultrasonography alone. In indeterminate and obstructive patterns, however, close monitoring that includes diuretic renography is mandatory.

MV140, a sublingual polyvalent bacterial preparation to treat recurrent urinary tract infections, licenses human dendritic cells for generating Th1, Th17, and IL-10 responses via Syk and MyD88.

Recurrent urinary tract infections (RUTIs) are one of the most common bacterial infectious diseases, especially in women. Antibiotics remain the mainstay of treatment, but their overuse is associated with antibiotic-resistant infections and deleterious effects in the microbiota. Therefore, alternative approaches are fully demanded. Sublingual immunization with MV140 (Uromune), a polyvalent bacterial preparation (PBP) of whole heat-inactivated bacteria, demonstrated clinical efficacy for the treatment of RUTIs, but the involved immunological mechanisms remain unknown. Herein, we demonstrated that MV140 endorses human dendritic cells (DCs) with the capacity to generate Th1/Th17 and IL-10-producing T cells by mechanisms depending on spleen tyrosine kinase (Syk)- and myeloid differentiation primary response gene 88 (MyD88)-mediated pathways. MV140-induced activation of nuclear factor κB (NF-κB) and p38 in human DCs is essential for the generated Th1/Th17 and IL-10 immune responses whereas c-Jun N-terminal Kinase (JNK) and extracellular-signal regulated kinase (ERK) contribute to Th1 and IL-10 responses, respectively. Sublingual immunization of BALB/c mice with MV140 also induces potent systemic Th1/Th17 and IL-10 responses in vivo. We uncover immunological mechanisms underlying the way of action of MV140, which might well also contribute to understand the rational use of specific PBPs in other clinical conditions with potential high risk of recurrent infections.

18F-Choline PET/CT scan in staging and biochemical recurrence in prostate cancer patients: Changes in classification and radiotherapy planning. / La PET/TC con 18F-Colina en la estadificación y recidiva bioquímica de pacientes con cáncer de próstata: cambios en la clasificación y planificación de radioterapia.

OBJECTIVE: To evaluate the role of the 18F-Choline PET/CT in prostate cancer management when detecting distant disease in planning radiotherapy and staging and to evaluate the therapy changes guided by PET/TC results. MATERIAL AND METHODS: A retrospective evaluation was performed on 18F-Choline PET/CT scans of patients with prostate cancer. Staging and planning radiotherapy scans were selected in patients with at least 9 months follow up. There was a total of 56 studies, 33 (58.93%) for staging, and 23 (41.07%) for planning radiotherapy. All scans were obtained using a hybrid PET/CT scanner. The PET/CT acquisition protocol consisted of a dual-phase procedure after the administration of an intravenous injection of 296-370MBq of 18F-Choline. RESULTS: There were 43 out of 56 (76.8%) scans considered as positive, and 13 (23.2%) were negative. The TNM staging was changed in 13 (23.2%) scans. The PET/CT findings ruled out distant disease in 4 out of 13 scans, and unknown distant disease was detected in 9 (69.3%) scans. CONCLUSIONS: 18F-Choline PET/CT is a useful technique for detecting unknown distant disease in prostate cancer when staging and planning radiotherapy. The inclusion of 18F-choline PET/CT should be considered in prostate cancer management protocols.

Skin tests with native, depigmented and glutaraldehyde polymerized allergen extracts.

INTRODUCTION: Dose-response skin prick tests are an important tool to standardise allergen extracts and to evaluate changes in skin test response as a consequence of allergen modifications. OBJECTIVES: To evaluate in vivo and in vitro characteristics of 3 different types of extracts of Phleumpratense, Olea europaea, Parietaria judaica and Dermatophagoides pteronyssinus. MATERIAL AND METHODS: Three types of extracts were used: native unmodified extracts (N), depigmented extracts (DP) (extracts subjected to a mild acid treatment under controlled conditions and dialysis), and a depigmented glutaraldehyde polymerised extract (DPP). Adult patients were skin tested in duplicate with the 3 types of extracts. The dose-response relationship between the geometric mean of the wheal areas and the allergen concentrations was calculated for each patient using regression line analysis. The amount of freeze-dried allergen preparation needed to produce the same wheal size as histamine was calculated in each patient (individual 10 HEP) and for each of the 3 types of extracts. In vitro analysis consisted of major allergen determinations and specific IgE and IgG inhibitions. RESULTS: The respective 10 HEP values for N, DP and DPP preparations were 0.20 mg, 0.15 and 2.11 for D. pteronyssinus. For P. pratense, these values were 0.02 mg, 0.02 and 0.99; for O. europaea 0.15, 0.44 and 4.9; and for P. judaica 0.01, 0.008 and 1.78 mg. CONCLUSIONS: The polymerised depigmented extracts are significantly less allergenic than the corresponding native and depigmented extracts. This could provide a greater safety margin for the administration of higher doses of immunotherapy in a shorter period of time.

Determination of Ole e 1 by enzyme immunoassay and scanning densitometry. Validation by skin-prick testing.

Ole e 1 is an important allergen in Olea europaea pollen extracts. This study describes the development of two new methods that can be used to estimate the Ole e 1 content in olive tree pollen extracts. They are based on (1) an enzyme immunoassay that uses rabbit polyclonal, monospecific antibodies and purified Ole e 1, and (2) scanning densitometry of SDS-PAGE gels. Twelve extracts were evaluated by in vivo and in vitro methods. The in vivo biological potency was estimated by prick skin testing 17 allergic individuals; the in vitro allergenic potency by direct IgE and IgE inhibition assays. The enzyme immunoassay showed an operative range of 0.03-100 microg/ml and demonstrated to be specific for Ole e 1. The Ole e 1 content ranged from 1% to 5% of the total protein in the 12 extracts. The amount of Ole e 1, assessed by gel scanning densitometry significantly correlated with the Ole e 1 content obtained by the immunoassay (r = 0.92; p < 0.001). The Ole e 1 content showed a significant correlation with the total allergenic potency of the extracts, evaluated by direct IgE, specific IgE inhibition and skin-prick testing. These two methods can be used to determine the Ole e 1 content in olive pollen extracts. The content of Ole e 1 can vary from 1% to 5% of the total protein in the extracts.

Estimation of IgE antibodies to the common allergens by reverse enzyme immunoassay. Comparison with the radioallergosorbent test.

A reverse enzyme immunoassay (REINA) is described, in which polystyrene microtiter wells are sensitized with murine monoclonal anti-human IgE, and then sequentially allowed to react with patient's serum, peroxidase-labeled allergens and substrate. The results obtained with the sera of patients allergic to Lolium perenne grass pollen, the tree pollens of Betula alba and Olea europea, the epithelia of cat and dog, the mite Dermatophagoides pteronyssinus, or to the foodstuffs cow's milk, chicken eggwhite or peanut were compared with the analytical data from the ratio allergosorbent test (RAST). The results show a good correlation between these two laboratory techniques.

Determinations of Par j 1 by a competitive enzyme immunoassay using human specific IgE and IgG. Validation by skin prick testing.

Par j 1 is the major allergen of Parietaria judaica. The objectives of this study were the following: 1) to purify Par j 1; 2) to develop an enzyme immunoassay based on the bivalent properties of specific IgE and IgG to determine the Par j 1 content in several batches of P. judaica extracts; and, 3) to study the contribution of Par j 1 to the total allergenicity and antigenicity of P. judaica extracts. P. judaica pollen was extracted and subjected to hydrophobic interaction and gel filtration chromatography for the purification of Par j 1. Inhibition enzyme immunoassays, SDS-PAGE and immunoblotting were used to characterize the allergen content. The in vivo biological potency of the extracts was estimated by skin prick testing 26 P. judaica clinically sensitive patients. The new enzyme immunoassay showed a high degree of specificity and sensitivity, detecting from 2 to 100 ng Par j 1/ml. The range of Par j 1 content in nine batches ranged from 23% to 78% of the total protein in the extracts. The Par j 1 content showed a significant correlation with the allergenic potency of these extracts evaluated by specific IgE inhibition and skin prick testing; the correlation with the specific IgG inhibition capacity was not significant. Purified Par j 1 shows great specific IgE and IgG binding capacity; its content can be determined using this newly developed enzyme immunoassay. Par j 1 levels exhibit a significant correlation with the biological potency of the extracts. This method allows the detection of Par j 1 isoforms.