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Enzyme-linked Immunospot Assay (ELISPOT): Quantification of Th-1 Cellular Immune Responses Against Microbial Antigens
Published on: November 24, 2010
Feline cytokine ELISPOT: issues in assay development
Sushila K Nordone1, Rosemary Stevens, Alora S LaVoy
1Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, USA.
Insights
The enzyme-linked immunospot (ELISPOT) assay is a powerful tool for measuring immune responses in cats. This technique is crucial for understanding feline diseases and developing effective vaccines.
Area of Science:
- Immunology
- Veterinary Medicine
- Cellular Biology
Background:
- The enzyme-linked immunospot (ELISPOT) assay is a widely used immunological tool for detecting cytokine production by cells.
- While extensively used in human and murine research, its application in feline immunology is recent.
- ELISPOT assays are valuable for studying feline disease immunopathogenesis and vaccine efficacy.
Purpose of the Study:
- To highlight the utility of ELISPOT assays in feline research.
- To discuss the application of ELISPOT for quantifying cytokine-producing cells in cats.
- To explore the potential of ELISPOT in advancing feline health through disease research and vaccine development.
Main Methods:
- The chapter focuses on the design parameters for developing feline-specific ELISPOT assays.
- It emphasizes the quantification of cytokine-producing cells, such as interferon (IFN)-gamma secreting T-cells.
- The methodology involves adapting ELISPOT techniques for feline samples.
Main Results:
- ELISPOT assays provide a sensitive method for assessing cellular immune responses in cats.
- The interferon (IFN)-gamma ELISPOT is particularly relevant for studying T-cell responses in feline immunodeficiency virus (FIV) infection and vaccination.
- The application of ELISPOT can enhance the utility of FIV as a model for human immunodeficiency virus.
Conclusions:
- ELISPOT assays are invaluable for investigating feline disease immunopathogenesis and vaccine efficacy.
- Broader application of ELISPOT in feline research will advance understanding of feline diseases.
- This technique supports the rational development of improved vaccines and therapeutics for feline health.
Abstract:
The enzyme-linked immunospot (ELISPOT) assay is a sensitive and relatively simple assay for detecting secreted cellular products such as cytokines and has become an invaluable immunological tool. The ELISPOT has been used extensively in human and murine research but has only recently been used to assess the feline immune system. For researchers studying feline disease or using the cat as a model of human disease, the quantification of cytokine-producing cells by ELISPOT is an invaluable technique for investigations of disease immunopathogenesis and vaccine efficacy. For example, use of the interferon (IFN)-gamma ELISPOT to measure the frequency of antigen-specific T-cells during feline immunodeficiency virus (FIV) infection or after immunization with candidate FIV vaccines is of particular interest. This application of the ELISPOT may serve to expand the utility of FIV as a model for human immunodeficiency virus. Broader applications of the ELISPOT should further our understanding of feline diseases and be useful in the rational development of more efficacious vaccines and therapeutic modalities for the enhancement of feline health. This chapter discusses important parameters of ELISPOT design that will enable researchers to develop and analyze the feline-specific assays within their own laboratory.
