In vivo pretreatment with PGG-glucan fails to alter cytokine mRNA expression of equine peripheral blood mononuclear cells exposed to endotoxin ex vivo.

This study examines the effects of PGG-Glucan on immune response in healthy horses and finds that pretreatment with PGG-Glucan has no notable impact on the expression of certain cytokines when cells are later exposed to endotoxin.

Methodology

• The study involves 12 healthy horses, which were divided into a treatment group and a control group. The horses in the treatment group were given PGG-Glucan [ED-1] intravenously, 24 hours before extracting their peripheral blood mononuclear cells (PBMC).
• The PBMCs were then isolated and subjected to lipopolysaccharide (LPS), which triggers a strong immune response in animals.
• At varying time intervals – 0, 6, 12, 24, and 48 hours – the messenger RNA (mRNA) was extracted from these cells.

Data Collection

• After the extraction of mRNA, it was processed using reverse transcription polymerase chain reaction (PCR), a widely-used laboratory method for the amplification of specific DNA sequences.
• The cytokine mRNA expression for tumor necrosis factor alpha (TNFalpha), interleukin 1beta (IL-1beta), interleukin 10 (IL-10), and interferon gamma (IFN-gamma) were then determined by employing real-time PCR. The aforementioned cytokines generally play a crucial role in inducing inflammatory responses in the body.

Findings

• Significant effects were noted on TNFalpha, IL-1beta, IL-10, and IFN-gamma production due to LPS stimulation over time. This is likely due to the body’s natural immune response to the exposure of endotoxins.
• However, no significant difference was highlighted between the group pretreated with PGG-Glucan and the control group at each time point. This signifies that PGG-Glucan failed to make a discernible difference in modulating the cytokine expression in response to endotoxin exposure.

The results of the research thus indicate that the pretreatment with PGG-Glucan doesn’t affect the cytokine mRNA expression of equine PBMCs when later exposed to endotoxin in the observed conditions.