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Production and Monoclonal Antibodies (Mabs) Against Surface Antigens on Israeli Carp Lymphocytes and Their Applications |
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Jong-Kyu woo, Han-Na Jang, Young-Hye Cho, Yong-Suk Jang, Sang-Hoon Choi |
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| Abstract |
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In fish both humoral and cell mediated immune responses have been reported whereas antibodies recognizing specific cellular populations have not yet been developed except for ones recognizing surface Ig molecules on B lymphocytes. Our aim was to develop and characterize monoclonal antibodies (Mabs) specific for the immune-related cells. Mabs were produced by fusion of myeloma cells (SP2/0) with Bslb/c mouse spleen cells previously sensitized against Israeli carp (I. carp) kidney mononuclear cells. We obtained 44 Mabs positively reacting with I. carp kidney monocuclear cells and partially characterized 7 Mabs in the morphological and mitogen -based proliferative aspects. Fluorescence-activated cell sorter (FACS) analysis against I. carp kidney cells by using 7 different Mabs showed 80.3% for ICK 17-4, 65.1% for ICK 2-3, 64.1% for ICK 25-1, 67.5% for ICK 22-1, 70.8% for ICK 16-2, 76.8% for ICK 13-2, 79.7% for ICK 11-1. Panning method was used for the isolation of Mabs specific mononuclear carp spleen cells followed by Wright''''s stain. The stained cell populations were identified as monocytes (ICK 17-4, ICK 2-3, ICK 25-1, ICK 22-1 and ICK 16-2), lymphocytes (ICK 11-1), and a mixed cell population of monocytes and lymphocytes (ICK 13-2). In cell proliferation assay, monocytes purified by ICK 17-4, 2-3 and 22-1 efficiently responded to Con A and PHA, while ones separated by ICK 25-1 did not react with any mitogens. Lymphocytes isolated by ICK 11-1, though it is not known whether they are T or B cells, were more responsive to Corn A than PHA or LPS, suggesting that fish immune cells are somewhat different from mammalian cells in responding to mammalian T or B cell mitogens. |
| Keywords:
Israeli Carp; Mabs; Cell Production; Wright''s Stain; Lymphocytes |
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