碩士 / 東吳大學 / 微生物學系 / 85 / In shrimp, hemocytes constitute mainly the first line of defence
against the pathgenic infections. In order to realize the
defence functions of hemocytes, they must be separated into the
different subpopulations based on the cell surface markers. in
order to get the monoclonal antibodies (Mabs)against the
granular cells (GC), hemocyte from tiger shrimp (Penaeus
monodon) were separated by a negative selection with concanalin
A (Con A)-linked bead. It could rise up the Gc content from 31%
to 78% in the total hemocytes. FourMabs, which were disignated
as Z5E10 (IgG2a), Z6A5 (IgG1), Z6A6 (IgG1), andZ6H8 (IgG2b),
were produced via the Balb/c mouse immunized with the hemocytes
with a rich GC content. Their reactivities to hemocytes were
assayed with the enzymeimmuno staining. The molecular mass of
the corresponding epitopes weredetermined with either the
Western-blotting or the immunoprecipitation. A 14kDa protein,
present in both GC and semigranular cells (SGC), could be
recognized by Mab Z5E10, no matter was in a reducing or a non-
reducing forms. The molecular mass of the non-reducing epitopes,
present in SGC, were largerthan 205 kDa which was determined by
the recognition of the other there Mabs. Epitope recognized by
Mab Z6A6 was show to be a glycoprotein proven with themethod of
periodate oxidation, however, those epitopes recognized by the
other three Mabs were protein in nature. These results imply
that four Mabs could recognize different epitopes of the
hemocytes. Besides, observing the indirect immunofluorescence
with the confocal microscope, antigens thatdistributed on the
hemocytes could cross-link to the capping or ring forms once
they were recognized by the four Mab. Following the negative
selection with each of the four Mab-Link beads, the
concentration of hyaline cells increased in hemocyte suspension.
These results suggested that antigens recognized by the four
Mabs were surface location.
| Identifer | oai:union.ndltd.org:TW/085SCU00381014 |
| Date | January 1996 |
| Creators | Wu, Pin-Yi, 吳品逸 |
| Contributors | Hin-Chung Wong, 宋宏紅, 宋延齡 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 2 |
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