The colorimetric assay for swine lymphocyte blastogenesis

Area of Science:

• Veterinary Immunology
• Cellular Immunology
• Biotechnology

Background:

• Lymphocyte blastogenesis is a key indicator of cellular immune response in swine.
• Accurate measurement of lymphocyte proliferation is crucial for immunological studies.
• The 3H-thymidine incorporation assay is a standard method, but alternative colorimetric assays are being explored.

Purpose of the Study:

• To compare the efficacy of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay with the 3H-thymidine incorporation assay.
• To evaluate these assays in the context of swine lymphocyte blastogenesis induced by phytohemagglutinin-P (PHA) and concanavalin A (ConA).

Main Methods:

• Colorimetric MTT assay and 3H-thymidine incorporation assay were performed on swine lymphocytes.
• Lymphocyte proliferation was induced using PHA and ConA.
• Correlation coefficients, optimal cell concentrations, mitogen concentrations, and incubation times were determined.

Main Results:

• High correlation coefficients were observed between the MTT assay and 3H-thymidine assay (0.70 for PHA, 0.82 for ConA).
• Maximal lymphocyte responses were achieved with 2.0 x 10^6 cells/ml and specific mitogen concentrations for 72 hours.
• The stimulation index for peripheral blood lymphocytes from healthy 6-month-old pigs was comparable between the two assays.

Conclusions:

• The MTT assay is a valid and reliable alternative to the 3H-thymidine incorporation assay for assessing swine lymphocyte blastogenesis.
• This finding supports the use of the MTT assay for cost-effective and efficient immunological evaluations in swine.
• Optimal culture conditions were established for reliable lymphocyte proliferation assays in pigs.