1. The Characterization of Monoclonal Antibodies to Mouse TLT-1 Suggests That TLT-1 Plays a Role in Wound Healing.
- Author
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Manfredi B, Morales-Ortíz J, Díaz-Díaz LM, Hernandez-Matias L, Barreto-Vázquez D, Menéndez-Pérez J, Rodríguez-Cordero JA, Villalobos-Santos JC, Santiago-Rivera E, Rivera-Dompenciel A, Lozada-Delgado EL, Kuchibhotla M, Carrasquillo-Carrión K, Roche-Lima A, and Washington AV
- Subjects
- Administration, Cutaneous, Amino Acid Sequence, Animals, Antibodies, Monoclonal biosynthesis, Antibodies, Monoclonal isolation & purification, Blood Platelets chemistry, Blood Platelets metabolism, Blotting, Western, Burns pathology, Clone Cells, Cytoplasmic Granules chemistry, Cytoplasmic Granules metabolism, Gene Expression, Immunization, Immunoprecipitation, Male, Mice, Peptides administration & dosage, Peptides chemistry, Peptides immunology, Rabbits, Receptors, Immunologic chemistry, Skin immunology, Skin pathology, Wound Healing immunology, Antibodies, Monoclonal pharmacology, Burns immunology, Platelet Aggregation drug effects, Receptors, Immunologic immunology, Skin drug effects, Wound Healing drug effects
- Abstract
Platelets play a vital role in hemostasis and inflammation. The membrane receptor TREM-like transcript-1 (TLT-1) is involved in platelet aggregation, bleeding, and inflammation, and it is localized in the α-granules of platelets. Upon platelet activation, TLT-1 is released from α-granules both in its transmembrane form and as a soluble fragment (sTLT-1). Higher levels of sTLT-1 have been detected in the plasma of patients with acute inflammation or sepsis, suggesting an important role for TLT-1 during inflammation. However, the roles of TLT-1 in hemostasis and inflammation are not well understood. We are developing the mouse model of TLT-1 to mechanistically test clinical associations of TLT-1 in health and disease. To facilitate our studies, monoclonal murine TLT-1 (mTLT-1) antibodies were produced by the immunization of a rabbit using the negatively charged region of the mTLT-1 extracellular domain
122 PPVPGPREGEEAEDEK139 . In the present study, we demonstrate that two selected clones, 4.6 and 4.8, are suitable for the detection of mTLT-1 by western blot, immunoprecipitation, immunofluorescent staining, flow cytometry and inhibit platelet aggregation in aggregometry assays. In addition, we found that the topical administration of clone 4.8 delayed the wound healing process in an experimental burn model. These results suggest that TLT-1 plays an important role in wound healing and because both clones specifically detect mTLT-1, they are suitable to further develop TLT-1 based models of inflammation and hemostasis in vivo.- Published
- 2018
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