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Identification of CD3+ T lymphocytes in the green turtle Chelonia mydas
Authors:Fernando A Muoz  Sergio Estrada-Parra  Andres Romero-Rojas  Thierry M Work  Erik Gonzalez-Ballesteros  Iris Estrada-Garcia
Institution:aLaboratorio 8, Unidad de Posgrado, Campo 1, Facultad de Estudios Superiores Cuautitlan-UNAM, Av 1 de Mayo s/n, Cuautitlan Izcalli, Estado de Mexico, Mexico;bDepartamento de Inmunologia, Escuela Nacional de Ciencias Biologicas-IPN, Mexico City, Mexico;cUS Geological Survey, National Wildlife Health Center, Honolulu Field Station, Honolulu, HI, USA
Abstract:To understand the role of the immune system with respect to disease in reptiles, there is the need to develop tools to assess the host's immune response. An important tool is the development of molecular markers to identify immune cells, and these are limited for reptiles. We developed a technique for the cryopreservation of peripheral blood mononuclear cells and showed that a commercially available anti-CD3 epsilon chain antibody detects a subpopulation of CD3 positive peripheral blood lymphocytes in the marine turtle Chelonia mydas. In the thymus and in skin inoculated with phytohemagglutinin, the same antibody showed the classical staining pattern observed in mammals and birds. For Western blot, the anti-CD3 antibodies identified a 17.6 kDa band in membrane proteins of peripheral blood mononuclear cell compatible in weight to previously described CD3 molecules. This is the first demostration of CD3+ cells in reptiles using specific antibodies.
Keywords:Cryopreservation of mononuclear cells  Reptilian T lymphocyte  Chelonia mydas  TCR/CD3 complex  Delayed-type hypersensitivity  Flow cytometry
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