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Application of Long-term cultured Interferon-γ Enzyme-linked Immunospot Assay for Assessing Effector and Memory T Cell Responses in Cattle
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1Infectious Bacterial Diseases Research Unit, National Animal Disease Center, Agricultural Research Service,United States Department of Agriculture, 2Department of Veterinary Pathology, College of Veterinary Medicine,Iowa State University, 3UK Veterinary Laboratories Agency, 4Visual Services, National Centers for Animal Health, Animal and Plant Health Inspection Service,United States Department of Agriculture
Capítulos
- 00:00Título
- 01:40Methods
- 02:05Long-term Culture
- 03:38Elispot Assay Day 1
- 04:30Elispot Assay Day 2
- 05:15Adherent Cell Isolation
- 05:37Flow Cytometry Plate Preparation
- 06:15Harvest the 13 Day Long-term Cells
- 09:05Elispot Assay Day 3
- 10:04Alkaline Phosphatase Preparation
- 11:08Substrate Solution Preparation
- 11:55Reading the Plate
- 12:12Performing Flow Cytometry
- 12:25Resultados
- 14:37Conclusions
Long-term cultured interferon-γ enzyme-linked immunospot assay is used as a measure of central memory responses and correlates with protective anti-mycobacterial vaccine responses. With this assay, peripheral blood mononuclear cells are stimulated with mycobacterial antigens and interleukin-2 for 14 days, enabling differentiation and expansion of central memory T cells.
