In Vitro Lymphocyte Adhesive Interactions With Cultured Rat Lacrimal Gland Acinar Epithelial Cells
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Authors
Bonner, Beth
Issue Date
1996
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
Immunity at the ocular surface is dependent upon the function of the lacrimal
glands and their association with the entire mucosal immune network, a relationship made
possible by the circulation of lymphocytes. The localization of lymphocytes within
lacrimal gland tissue contributes to the expression of immunoglobulin A (IgA) antibodies
in the tears and is regulated by cellular interactions between lacrimal gland acinar epithelial
cells and the lymphoid cells. This study sought to 1) characterize the lymphocyte
adhesive interactions with cultured lacrimal gland acinar epithelial cells, 2) compare these
interactions with those observed for lacrimal gland cryostat sections and primary
cultures, and 3) determine whether molecules mediating lymphocyte-lacrimal gland
binding could be isolated from cultured lacrimal acinar epithelial cells. Through use of
lymphocyte adherence assays it was found that thoracic duct lymphocytes (TDL) bound
to lacrimal gland acinar epithelial cells in greater numbers than thymocytes (294 cells/mm2
versus 8 cells/mm2). Adhesion was found to be not only dose dependent, but
temperature and time dependent as well. Use of the chemical inhibitors sodium azide,
cytochalasin B, and formaldehyde resulted in diminished lymphocyte adherence (12%,
4%, and 1% of control binding, respectively). Furthermore, binding was dependent on
the presence of divalent cations: Inhibition by ethylenediaminetetraacetic acid (32% of
untreated) was reversed by replacing the lymphocyte suspensions with calcium (106% of
control binding) and magnesium (76% of control binding). In addition, binding seemed to be enhanced when calcium and magnesium were replaced simultaneously (167% of contol
binding). TDL adherence was inhibited in the presence of fucoidin or phoshomannan
polysaccarides (47% and 65%, respectively). Treating the TDL with anti-selectin
monoclonal antibodies A.11.5, HRL-2 and HRL-3 gave very low inhibition (7%, 21%,
and 29%, respectively). Addition of anti-integrin monoclonal antibodies WT.l and HP2/1
to the TDL resulted in diminished adherence (38% and 52% of control binding,
respectively). 1A.29, an anti-ICAM1 antibody, reduced TDL adherence to only 38% of
control binding. Other anti-adhesion molecule antibodies, OX7, against Thy-I, and
OX49, against CD44, inhibited adherence by 55% and 43% respectively. Finally, greater
than 40% inhibition was observed when TDL were assayed in the presence of several
concentrated culture supernatants. Collectively, these results indicate that lymphocyte
binding to cultured lacrimal gland acinar epithelial cells requires viable lymphocytes with
unimpaired surface function and the presence of divalent cations. They further suggest
that the adhesion event involves the action ofintegrins, members of the immunoglobulin
superfamily, selectins, and other cellular adhesion molecules. Furthermore, these findings
suggest that the lacrimal gland epithelial cell cultures will be useful for isolating adhesion
molecules involved in lymphocyte localization. Finally, the results of the lymphocyte
adherence characterization studies indicate that thoracic duct lymphocyte adherence to
cultured lacrimal gland acinar epithelial cells showed good correlation with previously
reported adherence to lacrimal gland cryostat sections and primary cultures.
With honors.
With honors.
Description
vii, 55 p.
Citation
Publisher
Kalamazoo College
License
U.S. copyright laws protect this material. Commercial use or distribution of this material is not permitted without prior written permission of the copyright holder.