scholarly journals Cytokine RT-qPCR and ddPCR for immunological investigations of the endangered Australian sea lion (Neophoca cinerea) and other mammals

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e10306
Author(s):  
María-Ignacia Meza Cerda ◽  
Rachael Gray ◽  
Damien P. Higgins
Keyword(s):  
Gene Expression ◽  
Dynamic Range ◽  
Quantitative Polymerase Chain Reaction ◽  
Ovis Aries ◽  
Cytokine Gene Expression ◽  
Cytokine Gene ◽  
Sea Lion ◽  
Wildlife Species ◽  
Neophoca Cinerea ◽  
Australian Sea Lion

Measurement of cytokine gene expression by reverse transcription quantitative polymerase chain reaction (RT-qPCR) is used widely to assess the immune system of animals and to identify biomarkers of disease, but its application is limited in wildlife species due to a lack of species-specific reagents. The free-ranging endangered Australian sea lion (Neophoca cinerea) experiences significant clinical disease and high pup mortality due to intestinal hookworm infection. Developing immunological tools specific to the species will aid in the assessment of drivers of disease and its impact in population demographics. This study describes the development and validation of cross-reactive RT-qPCR assays to measure five important cytokines involved in innate and Th1/Th2 responses (IL-6, TNFα, IFNγ, IL-4 and IL-10) in unstimulated blood samples from a range of different mammalian species including the Australian sea lion. All RT-qPCR assays efficiencies ranged between 87% (Ovis aries TNFα) and 111% (Bos taurus IL-10) and had strong linearity (R2). IL-4 and IFNγ gene expression for N. cinerea fell below the dynamic range (and therefore quantifiable limits) of RT-qPCR assays but were able to be quantified using the novel droplet digital PCR (ddPCR). This study delivers new immunological tools for eco-immunologists studying cytokine gene expression in wildlife species and is to our knowledge, the first cytokine ddPCR approach to be reported in a pinniped species.

scholarly journals Cytokine gene expression assay as a diagnostic tool for detection of Mycobacterium bovis infection in warthogs (Phacochoerus africanus)

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Eduard O. Roos ◽  
Leeré A. Scott ◽  
Sedzani Ndou ◽  
Francisco Olea-Popelka ◽  
Peter E. Buss ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mycobacterium Bovis ◽  
Diagnostic Tool ◽  
Target Genes ◽  
Cytokine Gene Expression ◽  
Cytokine Gene ◽  
Wildlife Species ◽  
Cell Mediated Immune Response ◽  
Phacochoerus Africanus ◽  
Relative Gene

Abstract Mycobacterium bovis infection has been described in many wildlife species across Africa. However, diagnostic tests are lacking for many of these, including warthogs (Phacochoerus africanus). Most literature on suids has focused on using serological tools, with few studies investigating the use of cell-mediated immune response (CMI) assays. A recent study showed that warthogs develop measurable CMI responses, which suggests that cytokine gene expression assays (GEAs) may be valuable for detecting M. bovis-infection, as shown in numerous African wildlife species. Therefore, the aim of the study was to develop GEAs capable of distinguishing between M. bovis-infected and uninfected warthogs. Whole blood was stimulated using the QuantiFERON-TB Gold (In-Tube) system, using ESAT-6 and CFP-10 peptides, before determining the relative gene expression of five reference (B2M, H3F3A, LDHA, PPIA and YWHAZ) and five target (CXCL9, CXCL10, CXCL11, IFNG and TNFA) genes through qPCR. The reference gene H3F3A was the most stably expressed, while all target genes were significantly upregulated in M. bovis-infected warthogs with the greatest upregulation observed for CXCL10. Consequently, the CXCL10 GEA shows promise as an ante-mortem diagnostic tool for the detection of M. bovis-infected warthogs.

scholarly journals Cytokine gene expression in skin of susceptible guinea‐pig infected withTreponema pallidum

Immunology ◽  
1998 ◽  
Vol 95 (2) ◽  
pp. 242-247 ◽  
Author(s):  
WICHER ◽  
SCAROZZA ◽  
RAMSINGH ◽  
WICHER
Keyword(s):  
Gene Expression ◽  
Guinea Pig ◽  
Cytokine Gene Expression ◽  
Cytokine Gene

scholarly journals Epigenetic Regulation of Cytokine Gene Expression in T Lymphocytes

2009 ◽  
Vol 50 (3) ◽  
pp. 322 ◽  
Author(s):  
Choong-Gu Lee ◽  
Anupama Sahoo ◽  
Sin-Hyeog Im
Keyword(s):  
Gene Expression ◽  
T Lymphocytes ◽  
Epigenetic Regulation ◽  
Cytokine Gene Expression ◽  
Cytokine Gene

scholarly journals Immune modulators in disease: integrating knowledge from the biomedical literature and gene expression

2015 ◽  
Vol 23 (3) ◽  
pp. 617-626 ◽  
Author(s):  
Nophar Geifman ◽  
Sanchita Bhattacharya ◽  
Atul J Butte
Keyword(s):  
Gene Expression ◽  
Large Scale ◽  
Biomedical Literature ◽  
Cytokine Gene Expression ◽  
Future Research ◽  
Cytokine Gene ◽  
Medical Subject Headings ◽  
Expression Arrays ◽  
Gene Expression Arrays ◽  
Subject Headings

Abstract Objective Cytokines play a central role in both health and disease, modulating immune responses and acting as diagnostic markers and therapeutic targets. This work takes a systems-level approach for integration and examination of immune patterns, such as cytokine gene expression with information from biomedical literature, and applies it in the context of disease, with the objective of identifying potentially useful relationships and areas for future research. Results We present herein the integration and analysis of immune-related knowledge, namely, information derived from biomedical literature and gene expression arrays. Cytokine-disease associations were captured from over 2.4 million PubMed records, in the form of Medical Subject Headings descriptor co-occurrences, as well as from gene expression arrays. Clustering of cytokine-disease co-occurrences from biomedical literature is shown to reflect current medical knowledge as well as potentially novel relationships between diseases. A correlation analysis of cytokine gene expression in a variety of diseases revealed compelling relationships. Finally, a novel analysis comparing cytokine gene expression in different diseases to parallel associations captured from the biomedical literature was used to examine which associations are interesting for further investigation. Discussion We demonstrate the usefulness of capturing Medical Subject Headings descriptor co-occurrences from biomedical publications in the generation of valid and potentially useful hypotheses. Furthermore, integrating and comparing descriptor co-occurrences with gene expression data was shown to be useful in detecting new, potentially fruitful, and unaddressed areas of research. Conclusion Using integrated large-scale data captured from the scientific literature and experimental data, a better understanding of the immune mechanisms underlying disease can be achieved and applied to research.

IL-4 down-regulates IL-2-, IL-3-, and GM-CSF-induced cytokine gene expression in peripheral blood monocytes

1994 ◽  
Vol 68 (6) ◽  
pp. 293-298 ◽  
Author(s):  
F. H. M. Cluitmans ◽  
B. H. J. Esendam ◽  
J. E. Landegent ◽  
R. Willemze ◽  
J. H. F. Falkenburg
Keyword(s):  
Gene Expression ◽  
Peripheral Blood ◽  
Cytokine Gene Expression ◽  
Cytokine Gene ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Gm Csf

Cytokine gene expression and synthesis by human megakaryocytic cells

1992 ◽  
Vol 10 (2) ◽  
pp. 70-79 ◽  
Author(s):  
Hava Avraham ◽  
Edouard Vannier ◽  
Sherry Y. Chi ◽  
Charles A. Dinarello ◽  
Jerome E. Groopman
Keyword(s):  
Gene Expression ◽  
Cytokine Gene Expression ◽  
Cytokine Gene
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