Identification of CD4 isoforms by two anti-CD4 monoclonal antibodies
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Abstract
Background: CD4 isoforms expressed on leukocyte surface have been reported. The function of the CD4 isoforms is, however, unknown. Several studies are conducted aiming to uncover the functions of the CD4 isoforms which will lead to a better understanding of the immune responses.
Objectives: To identify CD4 isoforms expressed on leukocyte surface using two anti-CD4 monoclonal antibodies (mAbs) clones MT4 and MT4/3.
Materials and methods: Anti-CD4 mAbs MT4 and MT4/3 were purified by affinity chromatography. Specificity of the obtained purified mAbs were verified by 293T transfection and cell depletion experiment. Cellular distribution profiles of mAbs MT4 and MT4/3 were determined by immunofluorescence and flow cytometry. Identification of CD4 isoforms was performed by confocal microscopic analysis.
Results: Anti-CD4 mAbs MT4 and MT4/3, generated in our research center, were purified and confirmed their specificity by CD4-DNA transfection and cell depletion experiment. Cellular distribution profiles obtained from mAbs MT4 and MT4/3 were similar to those obtained using standard anti-CD4 mAb. By confocal microscopic analysis, mAbs MT4 and MT4/3 were demonstrated to recognize different CD4 molecule expressed on cell surface.
Conclusion: Anti-CD4 mAbs MT4 and MT4/3 were demonstrated to react with different CD4 isoforms. To the best of our knowledge, this is the first report showing that CD4 isoforms could be determined by specific mAbs. These mAbs will be an important tool for employing in characterization of structure and function of the CD4 isoforms.
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