Development of a Simple IFN-γ Release Whole Blood Assay for the Assessment of <i>Leishmania infantum</i> Specific Cellular Immunity in Dogs

Canine leishmaniosis (CanL) is a zoonotic disease caused by <i>Leishmania infantum</i>, where increased interferon-gamma (IFN-γ) levels are associated with controlling the infection and mild to moderate disease. Therefore, monitoring IFN-γ concentrations is essential for monitoring the i...

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Bibliographic Details
Main Authors: Anna Sára Molnár, Andrea Murillo-Picco, Clara Jiménez-Fortunato, Laia Solano-Gallego
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Animals
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Online Access:https://www.mdpi.com/2076-2615/14/23/3464
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Summary:Canine leishmaniosis (CanL) is a zoonotic disease caused by <i>Leishmania infantum</i>, where increased interferon-gamma (IFN-γ) levels are associated with controlling the infection and mild to moderate disease. Therefore, monitoring IFN-γ concentrations is essential for monitoring the immune responses in CanL. This study compared a faster, cost-effective IFN-γ release whole blood assay in tubes (WBA-T) with a standardized version (WBA-S) in 41 dogs at different states of <i>L. infantum</i> infection. WBA-T was performed at 24, 48, and 72 h of incubation with three conditions: blood, blood with <i>L. infantum</i>-soluble antigen (LSA), and blood with mitogen ConA. WBA-S was performed in plates, with blood diluted and incubated for five days using the same conditions. Supernatants (WBA-S) or plasma (WBA-T) were harvested for IFN-γ measurement by ELISA. No significant differences were observed in terms of IFN-γ concentration between WBA-T and WBA-S under LSA conditions. However, the 48 h incubation period during WBA-T showed the highest median of IFN-γ concentration compared to other incubation periods and WBA-S. The IFN-γ concentrations under ConA stimulation in WBA-S were significantly higher than in WBA-T at all incubation times studied. In conclusion, WBA-T stimulated with LSA at 48 h incubation time was shown to be the most appropriate for assessing IFN-γ production.
ISSN:2076-2615