Is humoral and cellular response to SARS-CoV-2 vaccine modified by DMT in patients with multiple sclerosis and other autoimmune diseases?

BACKGROUND: The effect of disease-modifying therapies on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine response is unclear. OBJECTIVES: We aim to determine the immunological responses to SARS-CoV-2 in multiple sclerosis (MS) and anti-CD20-treated patients with other autoimmune diseases (AID). METHODS: Humoral and cellular responses we determined before and 30-90 days after vaccination in patients with MS and anti-CD20-treated patients with other AID in two Catalan centers. RESULTS: 457 patients were enrolled. Findings showed that humoral response decreased under anti-CD20s or sphingosine 1-phosphate receptor modulators (S1PRM) and with longer treatment duration and increased after 4.5 months from the last anti-CD20 infusion. Cellular response decreased in S1PRM-treated. Patients on anti-CD20 can present cellular responses even in the absence of antibodies. CONCLUSION: Anti-CD20s and S1PRM modify the immunological responses to SARS-CoV-2 vaccines.

Endogenous antisense RNA curbs CD39 expression in Crohn's disease.

CD39 is an ectonucleotidase that initiates conversion of extracellular nucleotides into immunosuppressive adenosine. CD39 is expressed by regulatory T (Treg)-cells, where it mediates immunosuppression, and by a subset of T-helper (Th) 17-cells, where it limits pathogenicity. CD39 is regulated via single-nucleotide-polymorphisms and upon activation of aryl-hydrocarbon-receptor and oxygen-mediated pathways. Here we report a mechanism of CD39 regulation that relies on the presence of an endogenous antisense RNA, transcribed from the 3'-end of the human CD39/ENTPD1 gene. CD39-specific antisense is increased in Treg and Th17-cells of Crohn's disease patients over controls. It largely localizes in the cell nucleus and regulates CD39 by interacting with nucleolin and heterogeneous-nuclear-ribonucleoprotein-A1. Antisense silencing results in CD39 upregulation in vitro and amelioration of disease activity in a trinitro-benzene-sulfonic-acid model of colitis in humanized NOD/scid/gamma mice. Inhibition/blockade of antisense might represent a therapeutic strategy to restore CD39 along with immunohomeostasis in Crohn's disease.

Análisis reproductivo de vacas Suizo Pardo x Cebú y Simmental x Cebú en condiciones tropicales / Reproductive analysis of Brown Swiss x Zebu and Simmental x Zebu cows in tropical conditions

RESUMEN Objetivo. Comparar la fertilidad de vacas cruzadas Suizo Pardo x Cebú y Simmental x Cebú criadas en un ambiente tropical. Materiales y métodos. Se evaluaron características reproductivas de 185 vacas cruzadas Suizo Pardo x Cebú y Simmental x Cebú con diversos porcentajes de raza europea. El pastoreo de las vacas fue rotacional. El ordeño fue dos veces al día con la ayuda (amamantamiento) del becerro, el cual se mantuvo atado cerca de la vaca mientras ella se ordeñaba. Las características se evaluaron ajustando un modelo de mediciones repetidas (excepto para edad a primer parto). Periodo interparto, edad a primer parto, días abiertos, periodo parto-primer servicio y peso al parto fueron analizados con PROC MIXED de SAS. Tasa de gestación a primer servicio y servicios por concepción, se analizaron con PROC GENMOD del mismo programa. Resultados. Las vacas Simmental x Cebú se sirvieron después del parto 39 días antes (p<0.05) y tuvieron 47 días abiertos menos (p<0.05) que las Suizo Pardo x Cebú. El periodo interparto de las vacas Simmental x Cebú fue 45 días más corto (p<0.05) que el de las Suizo Pardo x Cebú. Las vacas Simmental x Cebú pesaron 34 kg más al parto (p<0.05) que las Suizo Pardo x Cebú. Conclusiones. Las vacas Simmental x Cebú tuvieron mejor fertilidad que las Suizo Pardo x Cebú.

ABSTRACT Objective. Compare the fertility of Brown Swiss x Zebu and Simmental x Zebu crossbred cows reared in a tropical environment. Materials and methods. Reproductive traits of 185 Brown Swiss x Zebu and Simmental x Zebu crossbred cows with diverse percentages of European breed were evaluated. Grazing of cows was rotational. The milking was twice daily with the help (suckling) of the calf, which was kept tied next to the dam while she was milked. Traits were evaluated fitting a repeated measures model (except for age at first calving). Calving interval, age at first calving, days open, interval from calving to first service, and weight at calving were analyzed with PROC MIXED of SAS. Pregnancy rate at first service and services per conception were analyzed with PROC GENMOD of the same software. Results. Simmental x Zebu cows started to re-bred 39 days earlier after calving (p<0.05) and had 47 fewer days open (p<0.05) than Brown Swiss x Zebu cows. The calving interval of the Simmental x Zebu cows was 45 days shorter (p<0.05) than that of the Brown Swiss x Zebu cows. Simmental x Zebu cows were 34 kg heavier at calving (p<0.05) than Brown Swiss x Zebu cows. Conclusions. Simmental x Zebu cows had better fertility than Brown Swiss x Zebu cows.

Modulation of CD39 and Exogenous APT102 Correct Immune Dysfunction in Experimental Colitis and Crohn's Disease.

BACKGROUND AND AIMS: CD39/ENTPD1 scavenges pro-inflammatory nucleotides, to ultimately generate immunosuppressive adenosine, which has a central role in immune homeostasis. Global deletion of Cd39 increases susceptibility to experimental colitis while single nucleotide polymorphisms within the human CD39 promoter, and aberrant patterns of expression during experimental hypoxia, predispose to Crohn's disease. We aimed to define the impact of transgenic human CD39 [hTG] overexpression in experimental colitis and to model therapeutic effects using the recombinant apyrase APT102 in vivo. We also determined the in vitro effects of APT102 on phenotypic and functional properties of regulatory T-lymphocytes derived from patients with Crohn's disease. METHODS: Colitis was induced by administration of dextran sulfate sodium in wild-type [WT] or hTG mice, and, in another model, by adoptive transfer of CD45RBhigh cells with or without WT or hTG regulatory T cells [Treg]. In additional experiments, mice were treated with APT102. The effects of APT102 on phenotype and function of Treg and type-1 regulatory T [Tr1] cells were also evaluated, after purification from peripheral blood and lamina propria of Crohn's disease patients [n = 38]. RESULTS: Overexpression of human CD39 attenuated experimental colitis and protected from the deleterious effects of systemic hypoxia, pharmacologically induced by deferoxamine. Administration of APT102 in vivo enhanced the beneficial effects of endogenous Cd39 boosted by the administration of the aryl hydrocarbon receptor [AhR] ligand unconjugated bilirubin [UCB]. Importantly, supplemental APT102 restored responsiveness to AhR stimulation by UCB in Treg and Tr1 cells, obtained from Crohn's disease patients. CONCLUSIONS: hCD39 overexpression ameliorated experimental colitis and prevented hypoxia-related damage in vivo. Exogenous administration of APT102 boosted AhR-mediated regulatory effects in vivo while enhancing Treg functions in Crohn's disease in vitro.

Exome sequencing study in patients with multiple sclerosis reveals variants associated with disease course.

BACKGROUND: It remains unclear whether disease course in multiple sclerosis (MS) is influenced by genetic polymorphisms. Here, we aimed to identify genetic variants associated with benign and aggressive disease courses in MS patients. METHODS: MS patients were classified into benign and aggressive phenotypes according to clinical criteria. We performed exome sequencing in a discovery cohort, which included 20 MS patients, 10 with benign and 10 with aggressive disease course, and genotyping in 2 independent validation cohorts. The first validation cohort encompassed 194 MS patients, 107 with benign and 87 with aggressive phenotypes. The second validation cohort comprised 257 patients, of whom 224 patients had benign phenotypes and 33 aggressive disease courses. Brain immunohistochemistries were performed using disease course associated genes antibodies. RESULTS: By means of single-nucleotide polymorphism (SNP) detection and comparison of allele frequencies between patients with benign and aggressive phenotypes, a total of 16 SNPs were selected for validation from the exome sequencing data in the discovery cohort. Meta-analysis of genotyping results in two validation cohorts revealed two polymorphisms, rs28469012 and rs10894768, significantly associated with disease course. SNP rs28469012 is located in CPXM2 (carboxypeptidase X, M14 family, member 2) and was associated with aggressive disease course (uncorrected p value < 0.05). SNP rs10894768, which is positioned in IGSF9B (immunoglobulin superfamily member 9B) was associated with benign phenotype (uncorrected p value < 0.05). In addition, a trend for association with benign phenotype was observed for a third SNP, rs10423927, in NLRP9 (NLR family pyrin domain containing 9). Brain immunohistochemistries in chronic active lesions from MS patients revealed expression of IGSF9B in astrocytes and macrophages/microglial cells, and expression of CPXM2 and NLRP9 restricted to brain macrophages/microglia. CONCLUSIONS: Genetic variants located in CPXM2, IGSF9B, and NLRP9 have the potential to modulate disease course in MS patients and may be used as disease activity biomarkers to identify patients with divergent disease courses. Altogether, the reported results from this study support the influence of genetic factors in MS disease course and may help to better understand the complex molecular mechanisms underlying disease pathogenesis.

HIF-1α-induced xenobiotic transporters promote Th17 responses in Crohn's disease.

In Crohn's disease, pathogenic Th17-cells express low levels of CD39 ectonucleotidase and are refractory to the immunosuppressive effects of unconjugated bilirubin (UCB), an endogenous ligand for aryl-hydrocarbon-receptor (AhR). This resistance to AhR ligation might be associated with alterations in responses to hypoxia. Limited exposure to hypoxia appears beneficial in acute tissue injury. However, in protracted inflammation, hypoxemia may paradoxically result in Th17-cell activation. We report here that in vitro exposure of Th17-cells from Crohn's disease patients to hypoxia limits responsiveness to AhR stimulation by UCB, as reflected by lower CD39 levels. Blockade of hypoxia-inducible-factor-1alpha (HIF-1α) upregulates CD39 and favors Th17-cell regulatory responses. Resistance of Th17-cells to AhR signaling results, in part, from HIF-1α-dependent induction of ATP-binding cassette (ABC) transporters: multidrug-resistance-protein-1 (MDR1) and multidrug-resistance-associated-protein-4 (MRP4). Increased ABC transporters promote efflux of suppressive AhR ligands, such as UCB, from Th17-cells. Inhibition of MDR1, MRP4 and/or HIF-1α with ritonavir (RTV) reconstitutes AhR function in Th17-cells, enhancing therapeutic effects of UCB in dextran-sulfate-sodium-induced experimental colitis. Deleterious effects of hypoxia on Th17-cells in Crohn's disease can be ameliorated either by inhibiting HIF-1α or by suppressing ABC transporters to increase UCB availability as an AhR substrate. Targeting HIF-1α-ABC transporters could provide innovative therapeutic pathways for IBD.

Neurofilament light chain and oligoclonal bands are prognostic biomarkers in radiologically isolated syndrome.

The prognostic role of cerebrospinal fluid molecular biomarkers determined in early pathogenic stages of multiple sclerosis has yet to be defined. In the present study, we aimed to investigate the prognostic value of chitinase 3 like 1 (CHI3L1), neurofilament light chain, and oligoclonal bands for conversion to clinically isolated syndrome and to multiple sclerosis in 75 patients with radiologically isolated syndrome. Cerebrospinal fluid levels of CHI3L1 and neurofilament light chain were measured by enzyme-linked immunosorbent assay. Uni- and multivariable Cox regression models including as covariates age at diagnosis of radiologically isolated syndrome, number of brain lesions, sex and treatment were used to investigate associations between cerebrospinal fluid CHI3L1 and neurofilament light chain levels and time to conversion to clinically isolated syndrome and multiple sclerosis. Neurofilament light chain levels and oligoclonal bands were independent risk factors for the development of clinically isolated syndrome (hazard ratio = 1.02, P = 0.019, and hazard ratio = 14.7, P = 0.012, respectively) and multiple sclerosis (hazard ratio = 1.03, P = 0.003, and hazard ratio = 8.9, P = 0.046, respectively). The best cut-off to classify cerebrospinal fluid neurofilament light chain levels into high and low was 619 ng/l, and high neurofilament light chain levels were associated with a trend to shorter time to clinically isolated syndrome (P = 0.079) and significant shorter time to multiple sclerosis (P = 0.017). Similarly, patients with radiologically isolated syndrome presenting positive oligoclonal bands converted faster to clinically isolated syndrome and multiple sclerosis (P = 0.005 and P = 0.008, respectively). The effects of high neurofilament light chain levels shortening time to clinically isolated syndrome and multiple sclerosis were more pronounced in radiologically isolated syndrome patients with ≥37 years compared to younger patients. Cerebrospinal fluid CHI3L1 levels did not influence conversion to clinically isolated syndrome and multiple sclerosis in radiologically isolated syndrome patients. Overall, these findings suggest that cerebrospinal neurofilament light chain levels and oligoclonal bands are independent predictors of clinical conversion in patients with radiologically isolated syndrome. The association with a faster development of multiple sclerosis reinforces the importance of cerebrospinal fluid analysis in patients with radiologically isolated syndrome.

Respuesta al tratamiento con interferón beta en pacientes con esclerosis múltiple. Validación del Rio Score / Response to treatment with interferon beta in patients with multiple sclerosis. Validation of the Rio Score

Introducción. Se han propuesto diferentes criterios de respuesta al tratamiento con interferón beta, y el Rio Score es uno de los más utilizados. El objetivo de este estudio fue validar la utilidad del Rio Score en una cohorte independiente. Pacientes y métodos. Estudio multicéntrico, prospectivo y longitudinal de pacientes con esclerosis múltiple remitente recurrente tratados con interferón beta. Los pacientes fueron clasificados basándose en la presencia de brotes, lesiones activas (nuevas en T2 o lesiones que captaban gadolinio) en la resonancia magnética, incremento confirmado de la discapacidad o combinaciones de estas variables (brotes, incremento en la Expanded Disability Status Scale y lesiones activas) tras un año de tratamiento. Se utilizó un análisis de regresión con el fi n de identificar las variables de predicción de respuesta después de un seguimiento de tres años. Resultados. Se incluyó a 249 pacientes con esclerosis múltiple remitente recurrente. El modelo logístico confirmó que la presencia de dos (odds ratio = 6,6; IC 95% = 2,7-16,1; p < 0,0001) o tres (odds ratio = 8,5; IC 95% = 1,6-46; p < 0,01) variables positivas durante el primer año de tratamiento confería un riesgo significativo de actividad (brotes o progresión) en los siguientes dos años. Conclusiones. Se confirma, en una cohorte independiente, la utilidad del Rio Score para identificar a pacientes con un mayor riesgo de desarrollar actividad clínica o progresión de la discapacidad durante el tratamiento con interferón beta (AU)

Introduction. Different criteria have been proposed for the response to treatment with interferon beta, and the Rio Score is one of the most widely used. The aim of this study was to validate the usefulness of the Rio Score in an independent cohort. Patients and methods. A multi-centre, prospective, longitudinal study was conducted on patients with relapsing-remitting multiple sclerosis treated with interferon beta. The patients were classified according to the presence of attacks, active lesions (new in T2 or gadolinium enhancing lesions) in magnetic resonance imaging, a confirmed increase in disability or combinations of these variables (attacks, increase on the Expanded Disability Status Scale and active lesions) after one year’s treatment. Regression analysis was used in order to identify the response-predicting variables after a three-year follow-up. Results. The sample consisted of 249 patients with relapsing-remitting multiple sclerosis. The logistic model confirmed that the presence of two (odds ratio = 6.6; CI 95% = 2.7-16.1; p < 0.0001) or three (odds ratio = 8.5; CI 95% = 1.6-46; p < 0.01) positive variables during the first year of treatment were indicative of a significant risk of activity (attacks or progression) in the next two years. Conclusions. The usefulness of the Rio Score is confirmed, in an independent cohort, as a means of identifying patients with a higher risk of developing clinical activity or progression of disability during treatment with interferon beta (AU)

Estimación de parámetros genéticos para características de fertilidad en ganado Suizo Pardo bajo condiciones subtropicales en México / Estimation of genetic parameters for fertility traits in Brown Swiss cattle under subtropical conditions of Mexico

Pedigree and reproductive records of 151 Brown Swiss cows were analyzed with the objective of estimating the heritability (h²) and the repeatability (r) for different measures of fertility. Cows were daughters of 49 sires and 102 dams. The study was carried out at Las Margaritas research station located in Puebla, Mexico. The pedigree was the same for all traits; it consisted of 287 individuals, and included dams without records and sires. Genetic parameters were estimated for conception rate at first service (CR), days to first registered heat after calving (DH), days to first registered service after calving (DS), days open (DO), and calving interval (CI). Estimations were made independently for each trait with a repeatability animal model, which included the direct additive genetic effect, as well as the permanent environmental effect of the cow. Genetic parameters were estimated with restricted maximum likelihood. In general, estimates of permanent environmental variance were greater than estimates of additive genetic variance, so that r was mainly determined by permanent environmental effects. Estimates of h² and r were: 0.02 ± 0.06 and 0.16, 0.00 ± 0.05 and 0.15, 0.03 ± 0.03 and 0.03, 0.00 ± 0.03 and 0.14, and 0.03 ± 0.07 and 0.12 for DH, DS, CR, DO and CI. Artificial selection as a tool to induce genetic change in measures of fertility studied would be slightly effective because additive genetic variance is scarce.

Se analizaron los registros genealógicos y reproductivos de 151 vacas de la raza Suizo Pardo para estimar la heredabilidad (h²) y el índice de constancia (r) de diferentes indicadores de fertilidad. Las vacas fueron hijas de 49 sementales y 102 madres. El estudio se realizó en el sitio experimental Las Margaritas, en Puebla, México. El pedigrí fue el mismo para todas las características, consistió de 287 animales e incluyó madres sin registros y padres. Los parámetros genéticos fueron estimados para tasa de gestación a primer servicio (TG), días al primer calor registrado después del parto (DC), días al primer servicio registrado después del parto (DS), días abiertos (DA) e intervalo entre partos (IEP). Las estimaciones se hicieron en forma independiente para cada característica, con un modelo de repetibilidad que incluyó el efecto genético aditivo individual, así como el efecto ambiental permanente de la vaca. Los parámetros genéticos fueron estimados por medio de máxima verosimilitud restringida. Los estimadores de la varianza del ambiente permanente, en general, fueron mayores que los de la varianza genética aditiva, por lo que r estuvo determinado principalmente por el ambiente permanente. Los estimadores de h² y r fueron: 0.02 ± 0.06 y 0.16, 0.00 ± 0.05 y 0.15, 0.03 ± 0.03 y 0.03, 0.00 ± 0.03 y 0.14, y 0.03 ± 0.07 y 0.12 para DC, DS, TG, DA e IEP, respectivamente. La selección artificial como una herramienta para inducir cambio genético en las diferentes mediciones de fertilidad sería poco efectiva, ya que la variación genética es escasa.