A reverse transcription-polymerase chain reaction method to analyze porcine cytokine gene expression

Veterinary Immunology and Immunopathology
C M DozoisIsabelle P Oswald

Abstract

A reverse transcription-polymerase chain reaction (RT-PCR) method was developed in order to provide a highly sensitive, rapid, and simple means of simultaneously measuring the expression of porcine cytokines in immune cell populations. Oligonucleotide primers were designed to amplify porcine cytokine cDNA from genes encoding IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IFN-gamma, TNF-alpha, TNF-beta and the housekeeping genes beta-actin and cyclophilin by PCR. Primers were chosen from different exons to detect for possible genomic DNA contamination of samples. To validate RT-PCR, unstimulated and concanavalin A (ConA) stimulated porcine peripheral blood mononuclear cells (PBMCs) were cultured from 2 h to 72 h, RNA was extracted and reverse transcribed, and cDNA was amplified using the different primer sets. Band intensities of PCR products were quantified by densitometric scanning and values were normalized against cyclophilin. For each of the cytokines, the kinetics of gene expression were similar among PBMCs isolated from different animals and could be grouped into two main patterns. Lymphocyte derived cytokines (IL-2, IL-4, IFN-gamma, and TNF-beta) exhibited low level expression in unstimulated cells and incr...Continue Reading

References

Oct 8, 1992·Nature·P D Siebert, J W Larrick
Jun 13, 1991·Biochimica Et Biophysica Acta·J C GoodallL Hall
Jul 25, 1990·Nucleic Acids Research·R DijkmansB Alfons
Jul 25, 1990·Nucleic Acids Research·C R MaliszewskiP E Baker
Feb 1, 1995·Veterinary Immunology and Immunopathology·H F SeowP R Wood
Mar 1, 1995·Veterinary Immunology and Immunopathology·J B RottmanM B Tompkins
Mar 28, 1995·Proceedings of the National Academy of Sciences of the United States of America·G BlanchoC LeGuern
Oct 1, 1994·Veterinary Immunology and Immunopathology·M P Murtaugh

❮ Previous
Next ❯

Citations

Dec 14, 2002·Journal of Biochemical and Molecular Toxicology·Carol L K SabourinJohn J Schlager
Apr 15, 2011·Tropical Animal Health and Production·Jean-Christophe BambouDavid Renaudeau
Dec 14, 1999·Journal of Immunological Methods·M Spagnuolo-WeaverG Allan
Jan 4, 2001·Veterinary Immunology and Immunopathology·S BastaK C McCullough
May 18, 2001·Veterinary Immunology and Immunopathology·R ThanawongnuwechE L Thacker
May 18, 2001·Veterinary Immunology and Immunopathology·H YancyM J Myers
Jun 8, 2001·Veterinary Immunology and Immunopathology·J HernándezE Zenteno
Aug 11, 2001·Veterinary Immunology and Immunopathology·T VerfaillieB M Goddeeris
Jun 8, 2002·Veterinary Immunology and Immunopathology·Ilja TrebichavskýYasuyuki Mori
Dec 3, 2002·Veterinary Immunology and Immunopathology·O MikamiY Nakajima
Feb 9, 1999·Veterinary Immunology and Immunopathology·S Giguère, J F Prescott
Aug 14, 1999·Immunology·S M KnoetigK C McCullough
Dec 1, 2010·Archives of Animal Nutrition·Ionelia TaranuIsabelle P Oswald
Nov 26, 1999·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Y MunetaY Mori
Jul 18, 2012·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·Xin ZhaoZhaoxin Zheng
Aug 23, 2005·Infection and Immunity·Francis GirardJohn M Fairbrother
Jan 5, 2002·Journal of Virology·Sang Heui Seo, Robert G Webster
Jul 14, 2005·Immunology·Johanna de GrootCornelis G van Reenen
May 26, 2004·Journal of Veterinary Medicine. A, Physiology, Pathology, Clinical Medicine·W S Cho, C Chae
Jul 21, 2004·Veterinary Immunology and Immunopathology·Wenjing TaoByram Bridle
Jul 18, 2006·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·Sandrine BouhetIsabelle P Oswald
Feb 7, 2006·Journal of Immunological Methods·Sangeeta BudhiaBarbara A Blacklaws
Oct 15, 2005·Journal of Immunological Methods·J C DuvigneauM Gemeiner
May 25, 2005·Veterinary Immunology and Immunopathology·Mhan-Pyo YangEui-Bae Jeung
Jun 21, 2011·Stress : the International Journal on the Biology of Stress·Y MunetaY Omata
Jan 5, 2002·The Journal of Veterinary Medical Science·I S ChoiH S Yoo

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.