scholarly journals Analysis of mRNA and circRNA Expression Profiles of Bovine Monocyte-Derived Macrophages Infected With Mycobacterium avium subsp. paratuberculosis

2022 ◽  
Vol 12 ◽  
Author(s):  
Yanhong Bao ◽  
Yu Yao ◽  
Zi Wang ◽  
Shuiyin Wu ◽  
Xiuyun Jiang ◽  
...  
Keyword(s):  
Immune Response ◽  
Mycobacterium Avium ◽  
Rna Binding ◽  
Expression Profiles ◽  
Johne's Disease ◽  
The Body ◽  
Johne’S Disease ◽  
Differentially Expressed ◽  
Economic Losses ◽  
Signal Pathways

Mycobacterium avium subsp. paratuberculosis (MAP) is the pathogen of Johne’s disease (paratuberculosis), which mainly causes chronic infectious granulomatous enteritis in ruminants and has brought huge economic losses to animal husbandry. As a specific intracellular pathogen, when MAP invades the body, it is internalized by macrophages where it is able to replicate by inhibition of the phagosome maturation, escaping the host immune system and surviving, which leads to the spread of the disease. More recent studies have shown that circRNA is involved in many pathological and physiological processes of the body as the molecular sponge of miRNA, the scaffold of RNA binding protein and having the characteristic of being able to translate into protein. In this study, the mRNA and circRNA expression profiles of MAP-infected bovine monocyte-macrophages and uninfected bovine cells were analyzed by high-throughput sequencing. A total of 618 differentially expressed mRNA were screened out, including 322 upregulated mRNA and 296 downregulated mRNA. In addition, the analysis of circRNA differential expression profile showed 39 differentially expressed genes including 12 upregulated and 27 downregulated genes. Moreover, differential genes belonging to cytokine activity, chemokine activity, inflammatory reaction, apoptosis, and other functional groups related to macrophage immune response were significantly enriched in Gene Ontology (GO). Multiple signal pathways including NF-κB, MAPK, Toll-like receptor, IL-17, JAK-STAT, and other signaling pathways related to activating macrophage immune response were significantly enriched in Kyoto Encyclopedia of Genes and Genomes (KEGG). In addition, RT-qPCR technology verified the accuracy of the mRNA sequencing results. In this study, we have obtained the transcriptome information of mRNA and circRNA of bovine monocyte-macrophage infected with MAP. These results will provide data support for the further study of mRNA–miRNA–circRNA network and immune escape mechanism of MAP and will enrich the knowledge of the molecular immune mechanisms of Johne’s disease as well.

scholarly journals Mycobacterium avium subspecies paratuberculosis: An Emerging Bacterial Disease of Global Public Health Significance

2016 ◽  
Vol 4 (1) ◽  
pp. 4-13
Author(s):  
Mahendra Pal ◽  
Md Tanvir Rahman
Keyword(s):  
Public Health ◽  
Crohn’S Disease ◽  
Crohn's Disease ◽  
Mycobacterium Avium ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Economic Losses ◽  
Public Health Significance ◽  
Health Significance

Mycobacterium avium subspecies paratuberculosis (MAP) is the etiological agent of chronic enteric disease of ruminant known as paratuberculosis (Johne's disease). The disease causes considerable economic losses worldwide due to reduced milk production and eventually, diarrhoea, weight loss and death. Johne's disease (JD) has some pathological similarities with Crohn's disease (CD) in humans, and the role of MAP in the causation of CD has been under investigation for last 100 years. Animals infected with JD shed viable MAP in the blood, and tissues. Consequently, transmission to humans may occur via consumption of animal derived foods. In developing countries, limited information is available on the occurrence of MAP infection in animals and humans. MAP infection has been established in animals and humans may get the MAP exposure through food chain or contaminated environment. Presently, MAP is of great public health significance because it is speculated to be involved in Crohn's disease in humans. The present review summarizes the information primarily on the nature of MAP in animals and humans, economic losses and morbidity and mortality due to JD and CD at global level. Current concept on the possible relationship between MAP and Crohn's disease has also been reviewed.Microbes and Health, January 2015. 4(1): 4-13

scholarly journals Isolation of High-Affinity Single-Chain Antibodies against Mycobacterium avium subsp. paratuberculosis Surface Proteins from Sheep with Johne's Disease

2006 ◽  
Vol 13 (9) ◽  
pp. 1022-1029 ◽  
Author(s):  
Sven Berger ◽  
Dominik Hinz ◽  
John P. Bannantine ◽  
J. Frank T. Griffin
Keyword(s):  
Mycobacterium Avium ◽  
Magnetic Bead ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Surface Proteins ◽  
Broth Culture ◽  
Economic Losses ◽  
Single Chain ◽  
Single Chain Antibodies ◽  
Magnetic Bead Separation

ABSTRACT Johne's disease, caused by infection with Mycobacterium avium subsp. paratuberculosis, causes significant economic losses to the livestock farming industry. Improved investigative and diagnostic tools—necessary to understand disease processes and to identify subclinical infection—are much sought after. Here, we describe the production of single-chain antibodies with defined specificity for M. avium subsp. paratuberculosis surface proteins. Single-chain antibodies (scFv) were generated from sheep with Johne's disease by cloning heavy-chain and lambda light-chain variable regions and expressing these in fusion with gene III of filamentous phages. Two scFv clones (designated SurfS1.2 and SurfS2.2) were shown to be immunoreactive against M. avium subsp. paratuberculosis surface targets by flow cytometry, and immunoblotting identified specificity for a 34-kDa proteinase-susceptible determinant. Both antibodies were cross-reactive against Mycobacterium avium subsp. avium but nonreactive against Mycobacterium bovis or Mycobacterium phlei cells and were shown to be capable of enriching M. avium subsp. paratuberculosis cells by a factor of approximately 106-fold when employed in magnetic bead separation of mixed Mycobacterium sp. cultures. Further, magnetic bead separation using SurfS1.2 and SurfS2.2 was capable of isolating as few as 103 M. avium subsp. paratuberculosis cells from ovine fecal samples, indicating the diagnostic potential of these reagents. Finally, inclusion of SurfS1.2 or SurfS2.2 in in vitro broth culture with M. avium subsp. paratuberculosis indicated that surface binding activity did not impede bacterial growth, although colony clumping was prevented. These results are discussed in terms of the potential use of single-chain phage display monoclonal antibodies as novel diagnostic reagents.

scholarly journals Comprehensive Analyses of circRNA Expression Profiles and Function Prediction in Chicken Cecums After Eimeria tenella Infection

2021 ◽  
Vol 11 ◽  
Author(s):  
Hailiang Yu ◽  
Changhao Mi ◽  
Qi Wang ◽  
Wenbin Zou ◽  
Guojun Dai ◽  
...  
Keyword(s):  
Immune Response ◽  
High Throughput Sequencing ◽  
Inflammatory Responses ◽  
Expression Profiles ◽  
Killer Cell ◽  
Eimeria Tenella ◽  
Differentially Expressed ◽  
Economic Losses ◽  
Circular Rnas ◽  
Poultry Production

Coccidiosis is an important intestinal parasitic disease that causes great economic losses to the global poultry production industry. Circular RNAs (circRNAs) are long non-coding RNAs that play important roles in various infectious diseases and inflammatory responses. However, the expression profiles and functions of circRNAs during Eimeria tenella (E. tenella) infection remain unclear. In this study, high-throughput sequencing was carried out to detect circRNAs in chicken cecal tissues from the control (JC), resistant (JR), and susceptible (JS) groups on day 4.5 postinfection (pi), respectively. A total of 104 circRNAs were differentially expressed, including 47 circRNAs between the JS and JC groups, 38 between the JR and JS groups, and 19 between the JR and JC groups. Functional analyses indicated that these differentially expressed circRNAs were involved in pathways related to E. tenella infection; the adaptive immune response was enriched in the JS vs JC group, the NF-kappa B signaling and natural killer cell-mediated cytotoxicity pathways were enriched in the JS vs JC and JR vs JC groups, while the B cell receptor signaling pathway was enriched in only the JR vs JC group. Moreover, the coexpression network of differentially expressed circRNAs and mRNAs suggested that circRNA2202 and circRNA0759 associated with DTX1 in the JS vs JC group, circRNA4338 associated with VPREB3 and CXCL13L3 in the JR vs JC group, and circRNA2612 associated with IL8L1 and F2RL2 in the JR vs JS group were involved in the immune response upon E. tenella infection. In conclusion, our results provide valuable information on the circRNAs involved in the progression of chicken E. tenella infection and advance our understanding of the circRNA regulatory mechanisms of host resistance and susceptibility to E. tenella infection in chickens.

scholarly journals Early Induction of Humoral and Cellular Immune Responses during Experimental Mycobacterium avium subsp. paratuberculosis Infection of Calves

2003 ◽  
Vol 71 (9) ◽  
pp. 5130-5138 ◽  
Author(s):  
W. R. Waters ◽  
J. M. Miller ◽  
M. V. Palmer ◽  
J. R. Stabel ◽  
D. E. Jones ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Mononuclear Cells ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Cell ◽  
Ifn Γ ◽  
Inoculation Route

ABSTRACT Johne's disease (paratuberculosis) of cattle is widespread and causes significant economic losses for producers due to decreased production and poor health of affected animals. The chronic nature of the disease and the lack of a reproducible model of infection hinder research efforts. In the present study, instillation of Mycobacterium avium subsp. paratuberculosis into the tonsillar crypts of neonatal calves resulted in peripheral colonization as detected by antemortem culture of feces and postmortem (320 days postchallenge) culture of intestinal tissues. Antigen-specific blastogenic, gamma interferon (IFN-γ), and nitric oxide responses by blood mononuclear cells from infected calves exceeded prechallenge responses beginning 194 days postchallenge. Upon in vitro stimulation with paratuberculosis antigens, CD4+ cells from infected calves proliferated, produced IFN-γ, and increased expression of CD26 and CD45RO (indicative of an activated memory phenotype). Utilizing a lipoarabinomannan-based enzyme-linked immunosorbent assay, specific serum immunoglobulin was detected as early as 134 days postchallenge and generally increased after this time point. Two antigens of ∼50 and ∼60 kDa were particularly immunodominant early in infection, as shown by immunoblot with serum collected within 2 weeks postchallenge. Findings indicate that the intratonsillar inoculation route will prove useful as an experimental model for paratuberculosis infection. Additionally, this study confirms that mycobacteria-specific antibody is detectable early in the course of experimental Johne's disease, even preceding the development of specific cell-mediated responses.

scholarly journals Differential proteome analysis of Mycobacterium avium subsp. paratuberculosis grown in vitro and isolated from cases of clinical Johne's disease

Microbiology ◽  
2011 ◽  
Vol 157 (2) ◽  
pp. 557-565 ◽  
Author(s):  
Mathias Weigoldt ◽  
Jochen Meens ◽  
Klaus Doll ◽  
Isabel Fritsch ◽  
Petra Möbius ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Mycobacterium Avium ◽  
Proteome Analysis ◽  
Economic Problem ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Differentially Expressed ◽  
Associated Proteins ◽  
Mycobacterium Avium Subsp Paratuberculosis

Bovine Johne's disease (paratuberculosis), caused by Mycobacterium avium subspecies paratuberculosis, poses a significant economic problem to the beef and dairy industry worldwide. Despite its relevance, however, pathogenesis of Johne's disease is still only partially resolved. Since mycobacterial membrane proteins expressed during infection are likely to play an important role in pathogenesis, membrane-enriched fractions, namely mucosa-derived membranes (MDM) and culture-derived membranes (CDM), of M. avium subsp. paratuberculosis from three cows with clinical paratuberculosis were investigated. An initial analysis by 2D difference gel electrophoresis (2D DIGE) and MALDI-TOF-MS analysis revealed four differentially expressed proteins with only one predicted membrane protein. Due to this limited outcome, membrane preparations were subjected to a tube–gel trypsin digestion and investigated by using nanoflow-liquid-chromatography-coupled tandem MS. Based on this approach a total of 212 proteins were detected in MDM including 32 proteins of bovine origin; 275 proteins were detected in CDM; 59 % of MDM and CDM proteins were predicted to be membrane-associated. A total of 130 of the proteins were detected in both MDM and CDM and 48 predicted membrane proteins were detected in MDM from at least two cows. Four of these proteins were not detected in CDM, implying differential expression in the host. All membrane-associated proteins, especially the four identified as being differentially expressed, might be relevant targets for further analyses into the pathogenesis of bovine paratuberculosis.

Evaluation of methods for Mycobacterium avium ssp. paratuberculosis detection in milk samples from the cattle herd showing low seroprevalence of Johne’s disease

2014 ◽  
Vol 17 (3) ◽  
pp. 459-463 ◽  
Author(s):  
J. Szteyn ◽  
A. Wiszniewska-Łaszczych ◽  
A. Smolińska
Keyword(s):  
Mycobacterium Avium ◽  
Genetic Material ◽  
Johne's Disease ◽  
Reliability Estimation ◽  
Johne’S Disease ◽  
Economic Losses ◽  
Specific Method ◽  
Diagnostic Reliability ◽  
Milk Samples ◽  
Sensitivity Specificity

Abstract Mycobacterium avium ssp. paratuberculosis (MAP) is the cause of chronic gastroenteritis in cattle called the Johne’s disease (JD). The disease causes significant economic losses in cattle production. MAP is also supposed to be involved in the Crohn’s disease and inflammatory bowel disease (IBD) in people. The detection of the cattle infection based on investigations of milk samples and evaluation of the capacity of the methods used to detect the disease was the objective of the present study. Following methods were applied for milk samples testing: detection of MAP in bacterial culture, detection of the specific IS-900 fragment of MAP in the genetic material isolated directly and detection of MAP antibodies. The results obtained were compared with the “golden standard” results, i.e. the isolation of MAP from the faeces. PQStat-the program for diagnostic reliability estimation, was used for evaluation of the sensitivity, specificity and predictive value. The method based on detection of the specific IS-900 fragment of MAP in the genetic material isolated directly from milk samples was found to possess the highest sensitivity. Detection of anti-MAP antibodies on the other hand showed the lowest sensitivity. The method of detecting anti-MAP antibodies in milk was the most specific while detection of the IS-900 fragment in the genetic material was the least specific method. These results obtained may serve as a guide to choose the most appropriate method for diagnosis of MAP infections by milk sample testing.

scholarly journals Membrane and Cytoplasmic Proteins of Mycobacterium avium subspecies paratuberculosis that Bind to Novel Monoclonal Antibodies

2018 ◽  
Vol 6 (4) ◽  
pp. 127 ◽  
Author(s):  
John Bannantine ◽  
Judith Stabel ◽  
John Lippolis ◽  
Timothy Reinhardt
Keyword(s):  
Monoclonal Antibodies ◽  
Mycobacterium Avium ◽  
Mycobacterium Avium Subspecies Paratuberculosis ◽  
Cell Extract ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mass Spectrometry Analysis ◽  
Lambda Phage ◽  
Expression Library ◽  
Spectrometry Analysis

Monoclonal antibodies against Mycobacterium avium subspecies paratuberculosis (Map) proteins are important tools in Johne’s disease research and diagnostics. Johne’s disease is a chronic inflammatory intestinal disease of cattle, sheep, and other ruminant animals. We have previously generated multiple sets of monoclonal antibodies (mAbs) in different studies; however, because many were generated and screened against a whole-cell extract of Map, the antigens that bind to these antibodies remained unknown. In this study, we used three different approaches to identify the corresponding Map antigens for 14 mAbs that could not be identified previously. In the first approach, a new Map-lambda phage expression library was screened to identify corresponding antigens for 11 mAbs. This approach revealed that mAbs 7C8, 9H3, 12E4, 3G5, and 11B8 all detect MAP_3404 encoding the biotin carboxylase subunit of acetyl-CoA carboxylase, while mAbs 7A6, 11F8, and 10C12 detect the GroEL2 chaperonin (MAP_3936), 6C9 detects electron transfer flavoprotein (MAP_3060c), and 14G11 detects MAP_3976, a lipoprotein anchoring transpeptidase. The epitopes to a selection of these mAbs were also defined. In a second approach, MAP_2698c bound monoclonal antibody (mAb) 14D4 as determined using protein arrays. When both of these approaches failed to identify the antigen for mAb 12C9, immunoprecipitation, mass spectrometry analysis, and codon optimization was used to identify the membrane protein, MAP_4145, as the reacting antigen. Characterized antibodies were used to quickly interrogate mycobacterial proteomic preps. We conclude by providing a complete catalog of available mAbs to Map proteins, along with their cognate antigens and epitopes, if known. These antibodies are now thoroughly characterized and more useful for research and diagnostic purposes.

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