Aris, Farizan
(2016)
Identification Of Antigens Recognized By The Monoclonal Antibody, Ibmr3 On Lymphocytes And Established Cell Lines.
PhD thesis, Universiti Sains Malaysia.
Abstract
Antibodi monoklon menunjukkan kegunaan yang tinggi dalam penyelidikan dan klinikal. Antibodi monoklon IBMR3 (IBMR3 MAb) telah menunjukkan keupayaan mengcam molekul yang mungkin berhubungkait dengan kompleks reseptor interleukin-4 (IL-4R). Selama ini, IL-4R sering dikaitkan dengan kanser. Tujuan penyelidikan ini adalah untuk mengesan pengekspresan antigen IBMR3 (IBMR3 Ag) pada sel mononuklear darah periferal (PBMC) dan panel titisan sel. Kaedah pewarnaan imuno menunjukkan IBMR3 MAb boleh mengcam antigen pada beberapa jenis sel termasuk sel epithelial, fibroblas, haematopoitik, kanser dan normal. Yang menariknya, IBMR3 Ag diekpres dengan signifikan dalam ruang sitoplasma tetapi tidak diekspres pada permukaan sel kecuali pada sel limfosit normal. Analisis pengekspresan IBMR3 Ag menunjukkan pengekspresannya adalah tinggi pada limfosit teraktif berbanding limfosit tidak teraktif tanpa mengambilkira jenis ejen mitogenik yang digunakan. Limfosit yang diaktifkan oleh CD3 MAb menunjukkan pengekspresan tertinggi (60.00 ± 1.94%) diikuti oleh limfosit yang diaktifkan oleh rangsangan PHA (41.23 ± 0.63%).
Monoclonal antibodies have demonstrated significant utilities in both research and clinical applications. IBMR3 monoclonal antibody (IBMR3 MAb) has been shown to recognize a molecule that may be associated with the human interleukin-4 receptor (IL-4R) complex. IL-4 has been shown to be extensively associated with cancer. The aim of this study is to identify the IBMR3 antigen (IBMR3 Ag) expressed in normal peripheral blood mononuclear cells (PBMC) and in a panel of established cell lines. Immunostaining revealed that IBMR3 MAb can recognize antigen in a variety of cell types, including epithelial, fibroblast, haemopoietic, cancer, and normal cells. Interestingly, IBMR3 Ag was found to be expressed in the cytoplasm but was absent on the cell surface except for normal lymphocytes. The analysis of IBMR3 Ag expression on lymphocytes showed that IBMR3 Ag expression was higher on activated lymphocytes compared to resting lymphocytes regardless of the type of mitogenic agents used. The lymphocytes activated by CD3 MAb demonstrated the highest expression (60.00 ± 1.94%) followed by those activated only by phytohaematoglutinin (PHA) stimulation (41.23 ± 0.63%).
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