Background
Adiponectin and leptin are pivotal in the regulation of metabolism. Pediatric lupus nephritis (pLN), a manifestation of childhood systemic lupus erythematosus (SLE) affecting the kidneys, ...is associated with impaired adipokine levels, suggesting a role in pLN pathogenesis. The aim of this study was to explore the potential relationship between specific single-nucleotide polymorphisms (SNPs)—methylenetetrahydrofolate reductase (MTHFR) rs1801131 and fibrinogen gamma chain (FGG) rs2066865—and the serum levels of leptin and adiponectin in patients with pLN.
Methods
Ninety-eight pLN patients and one hundred controls were enrolled in the study. Serum leptin and adiponectin levels were measured using ELISA. DNA extraction and real-time PCR genotyping were performed for
MTHFR
rs1801131 and
FGG
rs2066865 SNPs.
Results
Compared to healthy controls, pLN patients exhibited significantly greater serum leptin (11.3 vs. 18.2 ng/mL,
p
< 0.001) and adiponectin (18.2 vs. 2.7 ug/mL,
p
< 0.001). Adiponectin levels were positively correlated with proteinuria (
p
< 0.05), while leptin levels positively correlated with proteinuria, SLE disease activity index-2000 (SLEDAI-2K), and cyclophosphamide usage (all
p
< 0.05). There was no significant association between
MTHFR
rs1801131 or
FGG
rs2066865 SNPs and pLN in either codominant or allelic models (all
p
> 0.05). However, the AG genotype of
FGG
gene rs2066865 SNP was significantly associated with high leptin levels (> 15 ng/mL) (
p
= 0.01).
Conclusion
Serum adiponectin and leptin levels are associated with pathological manifestations of pLN. High leptin levels are associated with the AG genotype of
FGG
rs2066865 SNP in pLN patients, suggesting direct involvement in disease progression and potential utility as a disease biomarker.
Introduction: Multi-epitope vaccines in recent years have shown promising immunomodulatory effects on different alterations of the immune system; being allergic diseases a field to be explored, ...particularly with house dust mite allergens (HDM) and nematodes such as A. lumbricoides which are common in allergic patients in tropical regions. Objective: the objective of the present study was to design a multi-epitope protein from A. lumbricoides and APD allergens and to evaluate its IgE reactivity preliminarily. Methos: using computational tools, a molecule containing multiple "T" epitopes of allergens derived from A. lumbricoides and APD was designed "in silico" This multi-epitope protein (MP1) was expressed using an E. coli system and purified by affinity chromatography using Ni-NTA agarose. Anti-MP1 and anti-HDM extract IgE reactivity was evaluated by Dot-Blot and indirect ELISA from sera of HDM-allergic patients and non-allergic individuals from Barranquilla-Colombia. Allergic individuals had a positive skin test to a standardized battery of inhaled allergens (EUROLINE - Ref: DP 3704-1601-1 E) and mite-specific IgE. Results: multi-epitope (MP1) protein was expressed and purified with high purity. Dot-Blot result showed that all sera from allergic patients showed lower IgE reactivity to MP1 compared to HDM extract. By ELISA, significantly lower concentrations of anti-MP1 IgE (Median: 270.86 ng/ml; IQR: 90.3) were observed in contrast to anti-HDM IgE levels (Median: 988.5 ng/ml; IQR: 1117.6) in sera of patients allergic to HDM. Conclusions: A protein composed of multiple epitopes of A. lumbricoides and HDM allergens was designed, expressed, and purified. Preliminary Dot-Blot results suggest that this molecule shows hypoallergenic properties with very low IgE reactivity compared to mite extract. Further functional studies are needed to understand better the immune response induced by this molecule.
Objetivo: Diseñar una proteína multiepítope a partir de alérgenos de A. lumbricoides y APD; y evaluar preliminarmente su reactividad IgE. Métodos: Mediante herramientas computacionales se diseñó In Silico, una molécula que contiene múltiples epítopos T, de alérgenos derivados de A. lumbricoides y APD. Esta proteína multiepítope (MP1) se expresó utilizando un sistema de E. coli, y se purificó mediante cromatografía de afinidad, empleando agarosa Ni-NTA. La reactividad IgE anti-MP1 y anti-extracto de APD, se evaluó mediante Dot-Blot y ELISA indirecta, a partir de suero de pacientes alérgicos a APD, e individuos no alérgicos procedentes de Barranquilla, Colombia. Los individuos alérgicos contaron con prueba cutánea positiva a una batería estandarizada de alérgenos inhalados (EUROLINE - Ref: DP 3704-1601-1 E) e IgE específica para ácaros. Resultados: La proteína multiepítope MP1 se expresó y purificó con alta pureza. El resultado del Dot-Blot, mostró que todos los sueros de pacientes alérgicos tuvieron una reactividad IgE menor a MP1 en comparación al extracto de APD. Por ELISA, se observaron concentraciones significativamente menores de IgE anti-MP1 (Mediana: 270,86 ng/ml; RIQ: 90,3), en contraste a los niveles de IgE anti-APD (Mediana: 988,5 ng/ml; RIQ: 1117,6), en suero de pacientes alérgicos a APD. Conclusiones: Se diseñó, expresó y purificó una proteína compuesta por múltiples epítopes de alérgenos de A. lumbricoides y APD. Los resultados preliminares de Dot-Blot sugieren que esta molécula muestra propiedad hipoalergénica con una reactividad IgE muy baja, en comparación con el extracto de ácaros. Se necesita continuar con estudios funcionales para comprender mejor la respuesta inmune inducida por esta molécula.
Background: Helminth infections for Ascaris lumbricoides and exposure to mite allergens stimulate a Th2 immune response present in allergic diseases. Helminth proteins are well known can modulate the ...host immune response and lessen the inflammation response. We evaluate the Ig E reactivity to a molecule constructed from A. lumbricoides T epitopes (MP1) in sera from patients suffering from T1D, LN, and JIA Methods: We designed and expressed a synthetic multi-epítope protein named MP1 from A. lumbricoides and house dust mites allergens. By indirect ELISA, we evaluated IgE-reactivity to MP1 and to the whole-body extract of Ascaris lumbricoidesin 45 sera from Colombian Caribbean patients with lupus nephritis (LN; n=25), type 1 diabetes (T1D; n=10) and Juvenil idiopathic arthritis (JIA; n=10). Individuals with poly autoimmunity were excluded. All patients were referred to the study by their specialist doctor. Results: IgE to whole-body extract of A. lumbricoides showed the following median concentrations: 484.2 ng/ml (IQR: 203.4) in JIA patients, 325.6 ng/ml (IQR: 179.3) in individuals with LN, and 424.7 ng/ml (IQR: 80.1) in T1D group. On the other hand, IgE-reactivity to MP1 was 126.4 ng/ml (IQR: 90.9) in JIA patients, 130.7 ng/ml (IQR: 94.8) in an individual with LN, and 148.8 ng/ml (IQR: 102.1) in T1D group. Although no statistical differences were observed between patient groups, the IgE to MP1 in all patients (n: 45) (IgE median: 134.2 ng/ml; IQR: 100) were significantly less compared to Ascaris extract (IgE median: 380.7 ng/ml; IQR: 175.8); (W:0.732; p-value: 1.034x10-7). Conclusions: These preliminary results suggest that MP1 showed antigenic properties with low IgE-reactivity compared to Ascaris lumbricoides extract in individuals with autoimmune diseases. Further studies are needed to understand better the immune response induced by this molecule.
Objetivo: Evaluar la reactividad IgE de sueros en pacientes que padecen diabetes tipo 1 (DT1), nefritis lúpica (NL) y artritis idiopática juvenil (AIJ) frente a una molécula construida a partir de epítopes T de alérgenos de A. lumbricoides. Métodos: Se diseñó y expresó una proteína multi-epítopes sintética (MP1), a partir de alérgenos de A. lumbricoides y ácaros del polvo doméstico. Mediante ELISA indirecto, se evaluaron las reactividades IgE anti-MP1 y al extracto de cuerpo entero de Ascaris lumbricoides, en sueros de pacientes con nefritis lúpica (NL; n=25), diabetes tipo 1 (T1D; n=10) y artritis idiopática juvenil (AIJ; n=10), procedentes del Caribe colombiano. Se excluyeron los individuos con poliautoinmunidad. Todos los pacientes fueron remitidos al estudio por su médico especialista. Resultados: La IgE frente al extracto de cuerpo completo de A. lumbricoides mostró concentraciones de 484,2 ng/ml (RIQ: 203,4) en pacientes con AIJ; 325,6 ng/ml (RIQ: 179,3) en individuos con NL; y 424,7 ng/ml (RIQ: 80,1) en el grupo con DT1. Por otra parte, la reactividad de IgE anti-MP1 fue de 126,4 ng/ml (RIQ: 90,9) en los pacientes con AIJ; 130,7 ng/ml (RIQ: 94,8) en los individuos con NL; y 148,8 ng/ml (RIQ: 102,1) en el grupo con DT1. Aunque no se observaron diferencias estadísticas entre los grupos de pacientes, la reactividad IgE anti- MP1 en todos los pacientes (n: 45) (mediana de IgE: 134,2 ng/ml; RIQ: 100), fue significativamente inferior en comparación con el extracto de Ascaris (mediana de IgE: 380,7 ng/ml; RIQ: 175,8); (W: 0,732; p-valor: 1,034x10-7). Conclusiones: Estos resultados preliminares sugieren que MP1 mostró propiedades antigénicas con baja reactividad IgE, en comparación con el extracto de Ascaris lumbricoides en individuos con enfermedades autoinmunes. Se necesitan más estudios para comprender mejor la respuesta inmunitaria inducida por esta molécula.
Introduction: tropomyosin is a pan-allergen that shows cross-reactivity between different species. In tropical countries, allergy to arthropods (mites, shrimp, cockroaches, mosquitoes) or nematodes ...(Ascaris spp) is common. IgE epitopes of tropomyosin may be the source of sensitization and the development of allergic symptoms. However, T epitopes (MHC-II) polarize the Th2 response. Objective: This study aimed to identify by in silico methods tropomyosin consensus B and T epitopes of shrimp species, house dust mites, insects, and nematodes associated with allergic diseases in tropical countries. Methods: in silico analysis included tropomyosin from mites (Der p 10, Der f 10, Blo t 10), insects (Aed a 10, Per a 7, Bla g 7), shrimp (Lit v 1, Pen m 1, Pen a 1), and nematode (Asc l 3); all sequences were taken from the UniProt database. Linear IgE epitopes were predicted with AlgPred 2.0 and validated with BepiPred 3.0. MHC-II binding T cell epitopes were predicted using the IEDB server, which implements nine predictive methods (consensus method, combinatorial library, NN-align-2.3, NN-align-2.2, SMM-align, Sturniolo, NetMHCIIpan -3.1, and NetMHCIIpan -3.2) these predictions focused on 10 HLA-DR and 2 HLA-DQ alleles associated with allergic diseases. Subsequently, consensus B and T epitopes present in all species were identified. Results: we identified 12 sequences that behaved as IgE-epitopes and B-cell epitopes, three of them: 160RKYDEVARKLAMVEA174, 192ELEEELRVVGNNLKSLEVSEEKAN215, 251KEVDRLEDELV261 were consensus in all species. Eleven peptides (T-epitopes) showed strong binding (percentile rank ≤ 2.0) to HLA-DRB1*0301, *0402, *0411, *0701, *1101, *1401, HLA- DQA1*03:01/DQB1*03:02, and HLA- DQA1*05:01/DQB1*02:01. Only two T-epitopes were consensus in all species: 167RKLAMVEADLERAEERAEt GEsKIVELEEELRV199, and 218EEeY KQQIKT LTaKLKEAEARAEFAERSV246. Subsequently, we identified 2 B and T epitope sequences and reached a consensus between species 167RKLAMVEA174 and 192ELEEELRV199. Conclusions: These data describe three sequences that may explain the IgE cross-reactivity between the analyzed species. In addition, the consensus B and T epitopes can be used for further in vitro investigations and may help to design multiple-epitope protein-based immunotherapy for tropomyosin-related allergic diseases.
Objetivo: Este estudio tuvo como objetivo identificar mediante métodos in silico epítopes B y T consenso de tropomiosina de especies de camarón, ácaros del polvo doméstico, insectos y nematodos asociados a enfermedades alérgicas en países tropicales. Métodos: El análisis in silico incluyó tropomiosina de ácaros (Der p 10, Der f 10, Blo t 10), insectos (Aed a 10, Per a 7, Bla g 7), camarones (Lit v 1, Pen m 1, Pen a 1), y nematodo (Asc l 3). Todas las secuencias se tomaron de la base de datos UniProt. Los epítopes IgE lineales se predijeron con AlgPred 2.0 y se validaron con BepiPred 3.0. Los epítopes de células T de unión a MHC-II se predijeron utilizando el servidor IEDB, que implementa nueve métodos predictivos (método de consenso, biblioteca combinatoria, NN-align-2.3, NN-align-2.2, SMM-align, Sturniolo, NetMHCIIpan -3.1 y NetMHCIIpan -3.2). Estas predicciones se centraron en diez alelos HLA-DR y 2 HLA-DQ asociados con enfermedades alérgicas. Posteriormente, se identificaron epítopes consenso B y T presentes en todas las especies. Resultados: Se identificaron 12 secuencias que se comportaron como epítopes de IgE y, también, como epítopes de células B. Tres de ellas: 160RKYDEVARKLAMVEA174, 192ELEEELRVVGNNLKSLEVSEEKAN213 y 251KEVDRLEDELV261, fueron consenso en todas las especies. Once péptidos mostraron una fuerte unión (rango percentil ≤ 2,0) a HLA-DRB1*0301, *0402, *0411, *0701, *1101, *1401 y a HLA HLA-QA1*03:01/DQB1*03:02, o HLA-DQA1*05:01/DQB1*02:01. Solo se encontraron dos secuencias: 167RKLAMVEADLERAEERAEtGEsKIVELEEELRV199 con fuerte afinidad por HLADQA1* 03:01/DQB1*03:02, y HLA-DQA1*05:01/DQB1*02:01. Se identificaron dos secuencias que son epítopos B y T, y son consenso entre especies: 167RKLAMVEA174 y 192ELEEELRV199. Conclusiones: Estos datos describen tres secuencias que pueden explicar la reactividad cruzada de IgE entre las especies analizadas. Además, los epítopos B y T consenso se pueden usar para investigaciones in vitro adicionales, y pueden ayudar a diseñar inmunoterapia basada en proteínas de múltiepítopes para enfermedades alérgicas relacionadas con la tropomiosina.
Introduction: Chronic spontaneous urticaria (CSU) is an inflammatory skin disease related to poor quality of life. Previous studies have found that vitamin D deficiency and vitamin D recep-tor (VDR) ...TaqI, BsmI, FokI, and ApaI gene single-nucleotide polymorphisms (SNPs) influence immune response and susceptibility to skin disorders.
Aim: To explore the role of VDR SNPs, and the association of vitamin D serum levels in a sample of Colombian Caribbean CSU patients.
Methods: It is a case-control study. A group of CSU patients (n = 100) was compared with healthy individuals as a control group (n = 100). VDR polymorphisms were genotyped by quantitative polymerase chain reaction and Taqman® probes. Allelic, genotypic, and haplotype associations were estimated. Serum vitamin D levels were measured using enzyme-linked-immunosorbent serologic assay.
Results: Compared to the control group, the presence of G allele in TaqI and A allele in FokI SNPs of VDR gene was found to be a risk factor for CSU (odds ratio (OR) estimated using logistic regression adjusted by gender: 2.08 and 1.61, respectively, all P values < 0.05). The individuals who carry GCCA haplotype showed decrease in vitamin D levels (11.34 ng/mL; P = 0.002) with the G allele of TaqI and A allele of FokI gene SNPs.
Conclusion: We reported for the first time the association of TaqI rs731236 and FokI rs2228570 VDR gene SNPs showing as a risk factor for CSU in a sample of multiethnic patients from the Colombian Caribbean population.
Background
Although chronic urticaria (CU) is a common, cause of medical consulting both in general practitioners and allergist specialists worldwide, there is little information about its behavior ...and management in Latin America. Currently, national and international guidelines recommend using Omalizumab for cases refractory to management with antihistamines. Despite advances in the knowledge of Omalizumab for the management of CU, although there are few studies in underdeveloped countries, there are many studies evaluating the impact of Omalizumab treatment. There is not clinical information related with CSU-Omalizumab in patient settled in the Caribbean area. This research aims to evaluate the management of CU with Omalizumab in a real-life scenario in Colombia.
Methodology
We conducted an observational, descriptive, and retrospective study with patient recruitment between 2014 and 2017 of individuals diagnosed with Chronic Urticaria (CU) treating allergology specialists in five Colombian cities. We included patients with CU who failed to achieve disease control after treatment for 4 weeks with fourfold doses of second-generation H1-antihistamines, as recommended by the EAACI/GA
2
LEN/EDF/WAO guidelines and who received treatment with Omalizumab.
Results
We included 123 patients, 73.1% (
n
= 90) were women. The mean age was 47.1 years (Standard Deviation, SD: 16.2). The median of the total months of disease evolution was 30 (IQR = 13–58). 81.3 % (
n
= 100) of patients were diagnosed with chronic spontaneous urticarial (CSU). 4.8% (
n
= 6) had inducible CU (CIndU), and 13.8% (
n
= 17) reported mixed urticaria (spontaneous CU with at least one inducible component). Regarding emotional factors, 34.9% (
n
= 43) of subjects indicated anxiety symptoms, 34.1% (
n
= 42) had exacerbations associated with stress, and 14.6% (
n
= 18) manifested episodes of sadness. The percentage of patients with CSU controlled according to medical criteria at 3 months with Omalizumab were 80% (
n
= 80/100) and at 6 months 87% (
n
= 87/100). The frequency of adverse events was 29.2% (
n
= 36), with headache being the most frequent adverse event.
Conclusions
This real-life study with Omalizumab at CU describes percentages of effectiveness and safety similar to those observed in pivotal and real-life studies conducted in other regions around the world.
Los alérgenos de Blomia tropicalis son un factor de riesgo para el desarrollo de alergias en países tropicales. Los extractos alergénicos son reactantes valiosos para el diagnóstico de la ...hipersensibilidad alérgica a los ácaros y su control biológico. El objetivo fue evaluar las propiedades inmunoquímicas de extractos de B. tropicalis producidos desde cultivos in vitro. Este trabajo no tuvo como propósito producir extractos de aplicación clínica. Los ejemplares de B. tropicalis se establecieron a partir de ácaros aislados del polvo doméstico en la ciudad de Barranquilla, Colombia. Los ácaros libres de medio de cultivo se usaron para la obtención de los extractos. El rendimiento de la extracción se determinó mediante ensayo Bradford y el perfil electroforético por SDS-PAGE. La presencia de enzimas hidrolasas se determinó por Api® Zym y la inmunogenicidad se evaluó mediante Western blot donde se identificaron proteínas de unión a IgE sérica. La producción máxima de ácaros en los cultivos fue de 0,9 g/300 g medio. El rendimiento de extracción de proteínas fue de 3,61 % (± 0,42). Todos los extractos presentaron actividad enzimática tripsina, alfa-amilasa y quimotripsina. Western blot confirmó la presencia de proteínas de unión a IgE sérica. Presentamos un método sencillo para obtener extractos del ácaro B. tropicalis con actividad enzimática y alergénica.
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