ABSTRACT
Monoclonal antibodies (mAbs) CC34 and CC41 recognize overlapping subsets of leukocyte immune-type receptors (LITRs). The mAb CC34 was raised against the clonal TS32.15 cytotoxic T cell line and the mAb CC41 was raised against the clonal NK cell line TS10.1. In this study, an in vitro model was developed to monitor CC34- and CC41-reactive cells in response to Edwardsiella ictaluri infection. Briefly, head kidney leukocytes and peripheral blood lymphocytes (PBL) were isolated from individual catfish and labeled with CellTrace Violet and CellTrace FarRed dye, respectively. Head kidney-derived macrophages were infected with E. ictaluri and then cocultured with autologous PBL. The combined cell cultures were then analyzed using flow cytometry. A significant increase in CC41 staining was observed in the PBL population at 2, 5 and 7 days after culture, which suggest that LITRs are involved in cell-mediated immunity to E. ictaluri.
Subject(s)
Catfishes/immunology , Edwardsiella ictaluri/physiology , Enterobacteriaceae Infections/immunology , Fish Proteins/genetics , Killer Cells, Natural/immunology , Macrophages/immunology , Receptors, Immunologic/genetics , T-Lymphocytes, Cytotoxic/immunology , Animals , Antibodies, Monoclonal/metabolism , Cell Line , Coculture Techniques , Fish Proteins/metabolism , Flow Cytometry , Head Kidney/pathology , Immunity, Cellular , Receptors, Immunologic/metabolismSubject(s)
Allergy and Immunology , Developmental Biology/methods , Societies, Scientific , Animals , Biological Evolution , Congresses as Topic , Developmental Biology/trends , Humans , Immune System Phenomena/physiology , Immunologic Techniques/methods , Immunologic Techniques/trends , International CooperationABSTRACT
The present study establishes the effectiveness of natural drug delivery mechanisms and investigates the interactions between drug and its natural carrier. The binding between the isoflavone diadzein (DZN) and the natural carrier hemoglobin (HbA) was studied using optical spectroscopy and molecular dynamics simulations. The inherent fluorescence emission characteristics of DZN along with that of tryptophan (Trp) residues of the protein HbA were exploited to elucidate the binding location and other relevant parameters of the drug inside its delivery vehicle HbA. Stern-Volmer studies at different temperatures indicate that static along with collisional quenching mechanisms are responsible for the quenching of protein fluorescence by the drug. Molecular dynamics and docking studies supported the hydrophobic interactions between ligand and protein, as was observed from spectroscopy. DZN binds between the subunits of HbA, â¼15 Å away from the closest heme group of chain α1, emphasizing the fact that the drug does not interfere with oxygen binding site of HbA.
