Investigation of Immnune Function in Naval Marine Mammals.
Abstract
Tremendous progress has been made over the past year in the development of cetacean-specific reagents to investigate immune function in Navy marine mammals. We have focused most of our efforts in studying and developing molecular reagents and antibodies to cD4, a cell surface recognition molecule present on T helper lymphocyte that is critical for the immune response. Previously, we cloned cetacean cD4 from a beluga cDNA library using the polymerase chain reaction and cDNA library screening techniques. To generate antibodies against cetacean CD4, we synthesized peptides to different regions of the molecule and expressed the CD4 protein in a bacteria expression vector, and subsequently, injected coupled peptides or the purified cD4 protein into rabbits. Both sets of antibodies were affinity purified, and characterized by Western blot analysis and flow cytometry. These antibodies recognized beluga and bottlenose dolphin lymphocyte lysates by Western blot analysis, but did not recognize the native form of the cD4 protein on the cell surface by flow cytometry. Immunofluorescence on tissue sections is currently being tested with these antibodies. In addition we have generated and purified antibodies to a variant form of cD4 we found in beluga when cloning the full-length cD4. The significance of this potential splice variant of CD4 in the beluga is currently being investigated with the antibodies we have generated.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 1998
- Accession Number
- ADA349490
Entities
People
- Tracy A. Romano
Organizations
- Texas A&M University