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J Reprod Dev ; 66(6): 599-605, 2020 Dec 22.
Article in English | MEDLINE | ID: mdl-33012735

ABSTRACT

Autoimmune orchitis is a condition related to cellular immunity. A disease model involving transfer of T lymphocytes activated by known antigens would be useful for defining pathogenical molecules. Since no method for activating rat T cells using specific antigens is available, we started the study to develop the method. T cells were collected from draining lymph nodes of immunized rats, then co-cultured with syngeneic splenocytes as antigen-presenting cells (APC) in antigen-supplemented medium (= stimulation). The cells were then incubated in medium without antigens and APC (= resting). Repetitive stimulation and resting increased the number of the T cells more than 100-fold. The antigen-specific activation was demonstrated by cell proliferation assay and ELISA assay for interferon gamma. Flow cytometry revealed that > 95% of the cells expressed tumor necrosis factor alpha, a cytokine responsible for autoimmune orchitis. The present method will provide a new procedure to evaluate antigenicity of sperm molecules.


Subject(s)
Antigens/metabolism , Autoimmune Diseases/metabolism , Lymphocyte Activation , Orchitis/metabolism , Spermatozoa/physiology , T-Lymphocytes/cytology , Animals , Antigen-Presenting Cells/immunology , Cell Proliferation , Cell Survival , Coculture Techniques , Cytokines/metabolism , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Homozygote , Immunity, Cellular , In Vitro Techniques , Male , Rats , Rats, Wistar , Spermatozoa/immunology , Spleen/cytology , Spleen/metabolism , Tumor Necrosis Factor-alpha/metabolism
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