| Background and objective: Human secretory leukocyte protease inhibitor (hSLPI), is a 12 kD nonglycosylated proteinase inhibitor with a high capacity for inhibiting neutrophil elastase (NE) present in mucous secretions. SLPI is locally produced by epithelial cells, mast cells, neutrophils, monocytes, macrophages and is widely distributed at skin, respiratory tract, gastrointestinal tract, salivary gland and genital gland. SLPI possesses potent inhibitory activity against neutrophil elastase, cathepsin G, chymase, trypsin and chymotrypsin and play an important role in allergic and inflammatory diseases. Measuring the levels of SLPI help us understand the protease-antiprotease balance in allergic and inflammatory diseases well. Therefore, it is meaningful to prepare and identify monoclonal antibodies (mAbs) against human secretory leukocyte protease inhibitor.Methods: BALB/c mice were immunized with hSLPI, and hybridoma cell lines were obtained by fusing mouse spleen cells with myeloma NS-1 cells. The specificity of mAbs were characterized by enzyme-linked immunosorbent assay (ELISA), Western blot, immuno-histochemical staining, flow cytometry (FCM) and confocal laser scanning microscopy (CLSM).Results: 10 hybridoma cells which secreted the mAbs to hSLPI were obtained. They were identified as the IgM isotype. Western blot analysis showed that the mAbs could recognize a target molecule with relative molecular mass (Mr) of 12 000. Immunohistochemical staining revealed that the reactivities of 10 mAbs to the epithelial cells in lung, colon and foreskin tissues, mast cell-like cells in lung, colon, tonsil and foreskin tissues were positive. The result of FCM showed that the 10 mAbs recognized SLPI expressed in A549 cells. CLSM examination confirmed that the fluorescent markers were mainly localized in the cytoplasm of A549 cells.Conclusion: The mAb preparations in the present study will be raised valuable tools for studies on allergic and inflammatory diseases.
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