Stable Transfection of the BovineNRAMP1Gene into Murine RAW264.7 Cells: Effect onBrucella abortusSurvival

Barthel, Robert; Feng, Junjie; Piedrahita, Jorge A.; McMurray, David N.; Templeton, Joe W.; Adams, L. Garry

doi:10.1128/iai.69.5.3110-3119.2001

Cited by 88 publications

(67 citation statements)

References 58 publications

(54 reference statements)

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“…5,10 NRAMP1 is primarily expressed in MØs and tissues of the recticuloendothelial system, but its role in the resistance of or Vet Pathol 43: 709-717 (2006) susceptibility to mycobacterial infection remains undetermined. 3,4,35 In a recent study we demonstrated high expression of NRAMP1 in granulomas of M. bovis-infected cattle; 15 since granulomas are protective inflammatory reactions, 17,20,21 it was thought that NRAMP1 plays a defensive role against mycobacteria locally. This finding, in addition to recent studies showing a regulatory role for NRAMP1 in iNOS production, 2,18 prompted this study, which aims to analyze the expression of NRAMP1 and iNOS in M. bovis-infected cattle.…”

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Coexpression of NRAMP1, iNOS, and Nitrotyrosine in Bovine Tuberculosis

et al. 2006

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Abstract. In murine models the inducible nitric oxide synthase (iNOS) and the natural resistance associated macrophage protein (NRAMP1) play major roles in host defense against mycobacteria. iNOS regulates nitric oxide (NO) production, which is noxious for ingested mycobacteria, and NRAMP1 displays pleiotropic antimicrobial effects, including upregulation of iNOS expression. Little is known about the role of these molecules in bovine tuberculosis (TB). In this work we demonstrate by Western blot a high expression of NRAMP1 in peripheral blood mononuclear cells (PBMCs), alveolar macrophages (obtained by bronchioalveolar lavage), and lymph node granulomas from 8 HolsteinFreisian cattle with autopsy-proven bovine TB. Immunohistochemistry revealed the abundant expression of NRAMP1 and iNOS in lymph node and lung granulomas. Immunoreactivity was abundant in the cytoplasm of many epithelioid macrophages and multinucleated giant cells of the Langhans type. A striking accumulation of nitrotyrosine (NT), an indicator of iNOS activity and local NO production, was observed in granuloma cells, particularly in multinucleated Langhans cells. This study shows that the expression of NRAMP1 and iNOS is costimulated in granulomas, which are protective T-cell reactions against mycobacteria.

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Coexpression of NRAMP1, iNOS, and Nitrotyrosine in Bovine Tuberculosis

et al. 2006

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“…En estudios funcionales se ha demostrado que el gen Nramp1 regula la actividad antimicrobial en macrófagos de ratón, a través de la expresión de citoquinas como el factor de necrosis tumoral (TNFα) e interferón gama (IFN-γ) (Ables et al, 2001). También se ha reportado que los macrófagos de bovinos genéticamente resistentes a un desafío infeccioso in vivo con Brucella abortus (cepa 2308) son superiores en su capacidad para controlar su multiplicación bacterial intracelular, al igual que de Salmonella dublin y Mycobacterium bovis; este mecanismo es mediado también por el gen Nramp1 (Barthel et al, 2001).…”

Section: N T R O D U C C I ó Nunclassified

“…El gen presenta un polimorfismo en la región terminal 3' no traducida (UTR), el cual fue reportado por Horin y colaboradores (1999) como una variación en el número de repeticiones de la secuencia GT (guanina-timina), y se ha demostrado una asociación del alelo GT/13 con resistencia a la enfermedad en bovinos (Barthel et al, 2001) y en búfalos (Boriello et al, 2006;Caparelli et al, 2007).…”

Section: N T R O D U C C I ó Nunclassified

“…El gen Nramp1 es uno de los principales candidatos para resistencia natural a brucelosis en bovinos (Templeton et al, 1990;Feng et al, 1996;Horin et al, 1999;Barthel et al, 2001) y búfalos (Borriello et al, 2006;Capparelli et al, 2007). En este gen se han encontrado variaciones tipo microsatélite (Horin et al, 1999) y polimorfismos tipo SNP en regiones codificantes y no codificantes (Ables et al, 2002;Coussens et al, 2004;Martínez et al, 2008a), por esto es interesante encontrar posibles asociaciones entre las variaciones presentes en el gen y la resistencia a enfermedades.…”

Section: I S C U S I ó Nunclassified

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Frecuencias alélicas para variantes SNP en el gen Nramp1 en bovinos infectados con Brucella abortus o clasificados por resistencia al patógeno

Cerquera¹,

Martínez

Toro³

et al. 2009

CTA

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La resistencia natural a la brucelosis en bovinos ha sido asociada a factores genéticos, principalmente a algunos polimorfismos de nucleótido simple ubicados dentro del gen Nramp1. La presente investigación evalúa el efecto de variantes tipo polimorfismos de nucleótido simple presentes en regiones codificantes y en la región 3’UTR del gen Nramp1, en la clasificación de los animales como resistentes o susceptibles; además se determinan los genotipos predominantes en animales naturalmente infectados y comprobados como positivos por la presencia de anticuerpos anti Brucella abortus. Se establecieron las frecuencias genotípicas y alélicas para cinco polimorfismos de nucleótido simple identificados dentro del gen Nramp1 en animales de las razas blanco orejinegro (Bos taurus taurus) y cebú (Bos taurus indicus) y en muestras serológicamente positivas provenientes de animales cruzados (Bos taurus x Bos indicus). La determinación de genotipos se realizó mediante la metodología polimorfismo conformacional de cadena sencilla. Se realizó un ensayo de desafío infeccioso in vitro, para estimar la capacidad de los macrófagos bovinos para controlar la sobrevivencia bacterial, lo que permitió definir los individuos como resistentes o susceptibles. Los resultados sugieren una asociación significativa del SNP4 (p = 0,0506) con la variación para el fenotipo de susceptibilidad, pues se encontró el genotipo homocigoto (BB) en alta frecuencia en animales catalogados como resistentes y el genotipo heterocigoto (AB) en alta frecuencia en animales catalogados como susceptibles y en animales con títulos de anticuerpos anti Brucella abortus. Allelic frequencies for SNP variants in the gene Nramp1 in bovine infected with Brucella abortus or classified by resistance to the pathogen The natural resistance to brucellosis in cattle has been associated to genetic factors mainly to some single nucleotide polymorphism (SNP), located within Nramp1 gen. The current research has studied the effect of nucleotide variants to be found in coding regions and other one located in 3 non translated region of Nramp1 gene, on the animal classification as resistant or susceptible, moreover was identified the main genotypes to be found on the infected animals, confirmed as positives by antibody antibrucella titles. Was established the genotypic and allelic frequencies for five single nucleotide polymorphism in animals from blanco orejinegro (Bos taurus taurus) and zebu breeds (Bos taurus indicus) and serum samples belonging to positive crossbred animals (Bos taurus x Bos indicus). The genotype was defined by the methodology known as “single strand conformational polymorphism”. To estimate the macrophage capacity to control the bacterial survival, an in vitro assay was performed, which allowed define the phenotype as resistant or susceptible. The results suggest a significant association for SNP4 (p = 0.0506) with the phenotypic variation for resistant or susceptibility, because was found the genotype (BB) at higher frequency in susceptible animals and naturally infected animals, than those resistant animals.

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“…Likewise, our Bovine Slc11a1 diversity 505 Barthel et al (2001) have shown that transfected murine macrophages can be used to measure the functionality of two bovine Slc11a1 alleles, one of which correlates with enhanced expression and the in vitro control of Brucella abortus replication but not with Salmonella enterica. Searle et al (1999) have reported that polymorphism of the human Slc11a1 promoter (which encodes a Z-DNA forming di-nucleotide repeat) affects the level of gene expression and correlates with autoimmune versus infectious disease susceptibility in different human populations.…”

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A microsatellite study of bovine solute carrier family 11 a1 (Slc11a1) gene diversity in Mexico in relation to bovine tuberculosis

Vázquez-Flores¹,

Alonso

Villegas‐Sepúlveda³

et al. 2006

Genet. Mol. Biol.

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Bovine tuberculosis, caused by Mycobacterium bovis, is a disease of socio-economic and public health importance and of significance to international trade regulation. Allelic variants of several genes have been implicated in the genetic susceptibility to tuberculosis in some human populations, but little is known in cattle. We surveyed 34 European, 18 Asian, 20 Creole and 23 hybrid bovines for polymorphisms of the bovine solute carrier family 11 a1(Slc11a1) gene, formerly known as natural resistance associated macrophage protein (Nramp1), gene by typing the cattle using two microsatellite loci closely linked to this gene. The microsatellites used were 311-22, located at the 3' untranslated region (3' UTR) of the Slc11a1 gene, and ARO28 situated about 0.6 cM upstream of the same gene Based on allele size in base pairs (bp) we determined five 311-223 locus variants (221, 223, 225, 227 and 229 bp) and 12 ARO28 loci. There was marked diversity and a very high level of heterozygosity in most of the cattle surveyed except the Europeans bovines and especially Holsteins in relation to the 3' UTR microsatellite locus.

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Stable Transfection of the BovineNRAMP1Gene into Murine RAW264.7 Cells: Effect onBrucella abortusSurvival

Cited by 88 publications

References 58 publications

Coexpression of NRAMP1, iNOS, and Nitrotyrosine in Bovine Tuberculosis

Coexpression of NRAMP1, iNOS, and Nitrotyrosine in Bovine Tuberculosis

Frecuencias alélicas para variantes SNP en el gen Nramp1 en bovinos infectados con Brucella abortus o clasificados por resistencia al patógeno

A microsatellite study of bovine solute carrier family 11 a1 (Slc11a1) gene diversity in Mexico in relation to bovine tuberculosis

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