Measurement of multiple cytokines in mouse serum using the flowcytometry
- VernacularTitle:Урсгал эс тоолуур ашиглан хулганы ийлдсэнд олон төрлийн цитокиныг зэрэг хэмжсэн дүн
- Author:
Gansukh Choijilsuren
1
;
Enkhtushig Gavaabaljir
1
;
Anand Altankhuyag
1
;
Munkhjargal Davaajargal
1
;
Nyambayar Dashtsoodol
1
;
Tsogtsaikhan Sandag
1
;
Khongorzul Togoo
1
,
2
Author Information
1. Department of Immunology, School of Biomedicine, Mongolian National University of Medical Sciences
2. Laboratory of Immunobiology, Institute of Biomedical Sciences, Mongolian National University of Medical Sciences
- Publication Type:Journal Article
- Keywords:
capture beads;
concentration;
immunoassay;
LEGENDplex;
Sandwich ELISA
- From:Mongolian Medical Sciences
2024;207(1):3-7
- CountryMongolia
- Language:Mongolian
-
Abstract:
Introduction:Accurate measurement of multiple cytokines concurrently is essential for comprehensively understanding
immune responses in various diseases.
Goal:To develop a technological methodology for the simultaneous quantification of multiple cytokines in
mouse serum using a flowcytometry
Materials and Methods:Utilizing a C57BL/6 mouse model, we induced cytokine production by intraperitoneal injection of
alpha-galactosylceramide (α-GalCer) and analyzed cytokine levels 24 hours later using a MACSQuant
analyzer 10 equipped with LEGENDplex™ Mouse Th Panel (13-plex) assay. Means between groups
were compared using the independent sample t test, and statistically significant differences were
considered at p ≤ 0.05.
Results:Our results demonstrate that the emitted intensity increased proportionally with standard concentration,
with slightly elevated cytokine staining observed for MCP-1 and IL-10.
The concentration of each cytokine was determined using a regression equation based on the mean
fluorescence intensity value of the cytokine in each standard. Importantly, we observed significantly
highest levels of IFN-γ (p<0.05) in α-GalCer-injected mice, indicating successful induction of cytokine
production, particularly from NKT cells. Notably, our investigation revealed that levels of IFN-b, IL-17A
and IL-27 in α-GalCer-injected mice increased (p<0.05) those of other cytokines, compared to negative
controls. The determination of multiple cytokines simultaneously via flowcytometry, facilitated by the
LEGENDplex assay, holds significant importance in enabling comprehensive comparison of cytokine
concentrations.
Conclusion:An experimental methodology for the simultaneous determination of multiple cytokines by flowcytometry
was successfully developed.
- Full text:2025060517423458496MMS-2024-207(1)-3-7.pdf
