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Biomedical subjects

E Oei

Publications and source records attributed to E Oei.

7 recordsLinked to original sources

Regulation of class II expression in monocytic cells after HIV-1 infection.

Human macrophage hybridoma cells were used to study HLA-DR expression after HIV-1 infection. HLA-DR surface expression was lost 2 wk after infection that was associated with decreased mRNA transcription. Transfecting HLA-DR-alpha and HLA-DR-beta cDNA driven by a nonphysiological CMV promoter restored expression, suggesting that regulatory DNA-binding proteins may be affected by HIV-1 infection. There was no protein binding to conserved class II DNA elements (W/Z/S box, X-1 and X-2 boxes, and Y box) in a HIV-1-infected human macrophage hybridoma cell line, 43(HIV), and in primary monocytes that lost HLA-DR expression after HIV-1(BaL) infection. PCR analysis of the HIV-1-infected cells that lost HLA-DR expression revealed mRNA for W/Z/S (RFX-5), X-1 (RFX-5), X-2 (hX-2BP), and one Y box DNA-binding protein (NF-YB), and CIITA, a non-DNA-binding protein necessary for class II transcription. There was no mRNA for the Y box-binding protein, NF-YA. However, HLA-DR expression could be restored by transfection with NF-YA driven by a CMV promoter, although HLA-DR failed to localize in either the late endosomes, lysosomes, or acidic compartments. This was associated with a loss of class II-associated invariant chain peptide and leupeptin-induced protein in the 43(HIV) cells. To address this further, non-HIV-1-infected 43 cells were infected with vaccinia virus containing HIV-1 gag, nef, pol, and env proteins. HLA-DR failed to localize in neither the late endosomes, lysosomes, or acidic compartments in the vaccinia-infected cells containing HIV-1 env protein. HIV-1 appears to have multiple effects on class II expression in monocytic cells that may contribute to the immune defects seen in HIV-1-infected patients.

Antigens, Differentiation, B-Lymphocyte↗

Antigen uptake and trafficking in human intestinal epithelial cells.

Primary intestinal epithelial cells, human colonic adenocarcinoma cell lines (DLD-1, Caco-2, and HT-29), and monocytes were used as model systems to study antigen uptake, antigen-presenting cell properties, as well as the kinetics of antigen uptake in intestinal epithelial cells (IEC). Intracellular staining of fluoresceinated tetanus toxoid was not evident in the IEC until after 30 min of incubation at 37 degrees C, whereas in monocytes intracellular punctate staining of fluoresceinated tetanus toxoid was evident after 5 mins. In polarized Caco-2 cells antigen could be internalized at both the apical and basolateral surfaces with polarized transport. When analyzed by electron microscopy, gold-labeled tetanus toxoid was internalized and found within endosomes and multivesicular bodies, but not within the lysosomal compartments by 60 min. By 2 hrs, gold-labeled tetanus toxoid was evident in the secondary lysosomes. These results demonstrate that tetanus toxoid follows an endocytic pathway in intestinal epithelial cells and that the kinetics of antigen uptake is slower than that of conventional antigen-presenting cells.

Antigens↗

Factors affecting antigen uptake by human intestinal epithelial cell lines.

We assessed the role of size, solubility, and prophagocytic cytokines interferon-gamma (IFN-gamma), and granulocyte-macrophage colony stimulatory factor (GM-CSF) in antigen uptake and kinetics by intestinal epithelial cells using keyhole limpet hemocyanin and ovalbumin. Both fluoresceinated keyhole limpet hemocyanin (3000-7500 kDa) and fluoresceinated ovalbumin (45 kDa) were internalized by human colonic epithelial cell lines, with kinetics similar to those of fluoresceinated tetanus toxoid, and there was decreased uptake of insoluble immune complexes and no enhancement in the uptake of soluble immune complexes. In addition, neither IFN-gamma nor GM-CSF altered the kinetics of uptake nor enhanced antigen internalization by the intestinal epithelial cell lines. These data suggest that regardless of the size of the soluble antigen, the presence of prophagocytic cytokines, or the formation of soluble immune complexes, fluid phase endocytosis of antigen by intestinal epithelial cells appears to be a relatively stable process.

Antigen-Antibody Complex↗

Antigen trafficking and accessory cell function in respiratory epithelial cells.

We investigated accessory cell function, antigen (Ag) trafficking, and uptake of immune complexes in isolated nasal epithelial cells (NEC) and airway epithelial cells (AEC), as well as in the two respiratory epithelial cell lines A549 and BEAS-2B. The NEC and AEC were capable of supporting Ag-specific as well as phytohemagglutinin-induced and anti-CD3 antibody-induced T-cell proliferation. We colocalized fluorescein isothiocyanate (FITC)-labeled Ags with human leukocyte antigen (HLA)-DR in A549 and BEAS-2B, utilizing laser confocal microscopy. Respiratory epithelial cells stimulated and unstimulated with interferon (IFN)-gamma were pulsed with FITC-labeled Ags for varying periods and evaluated for their ability to internalize Ag. In the unstimulated cells, intracellular punctate staining was evident at 60 min and persisted up to 120 min. In the IFN-gamma-stimulated cells (100 U/ml for 48 h), uptake occurred at 30 min, was maximal at 60 min, and diminished at 120 min. We conducted kinetic studies in the A549 and BEAS-2B cells, utilizing electron microscopy with colloidal gold-conjugated Ags (Au-OVA). At 15 min, Au-OVA was evident in the early compartments resembling the compartment of uncoupling of receptor and ligand. At 30 min, multivesicular bodies were labeled with Au-OVA, and by 60 min Au-OVA was present in the primary and secondary lysosomes. The FITC-labeled Ags colocalized with an early endosomal marker (anti-cathepsin D), a late endosomal marker (M6PR), a lysosomal marker (CD63), and with 3-(2, 4-dinitroanilino)-3'-aminomethyldipropylamine, a marker of acidic vesicles. The BEAS-2B and A549 cells, and NEC and AEC, expressed surface Fcgamma receptor and internalized IgG immune complexes. The NEC and AEC also expressed the costimulatory molecules CD80 and CD86 as determined with flow cytometry, the reverse transcription-polymerase chain reaction for RNA, and immunohistochemistry, and T-cell proliferation could be blocked by treating NEC and AEC with anti-CD80 and anti-CD86 antibodies. Our findings suggest that respiratory epithelial cells may have a role in local Ag presentation.

Antigen-Antibody Complex↗

In vivo antagonism of agonist actions at N-methyl-D-aspartate and N-methyl-D-aspartate-associated glycine receptors in mouse cerebellum: studies of 1-hydroxy-3-aminopyrrolidone-2.

Intracerebellar injections of either NMDA or D-serine dramatically elevated levels of cGMP in the cerebellum of the mouse, in vivo. These actions were both antagonized by simultaneous injection of the NMDA-associated glycine receptor antagonist, HA-966. Intracerebellar injections of D-serine were also antagonized by peripheral (s.c.) injections of HA-966, demonstrating the bioavailability of this glycine receptor antagonist. Parenteral administration of HA-966 was also effective in antagonizing the actions of intravenously injected harmaline, an activator of the cerebellar climbing fiber pathway, on cGMP in the cerebellum. An evaluation of the parenteral dose-response curve for HA-966, revealed no effect on basal activity within the cerebellum. This contrasts sharply with the abilities of both competitive and non-competitive NMDA antagonists to decrease basal levels of cGMP in the cerebellum. In summary, these studies demonstrate that HA-966 is a bioavailable antagonist of the NMDA-associated glycine receptor and that this compound can limit excessive stimulation of the NMDA receptor by exogenous application of agonist, with minimal effects on basal activity. These data suggest that antagonists of the NMDA-associated glycine receptor may be optimal therapies in the treatment of stroke and epilepsy.

Animals↗

Cholecystokinin attenuates basal and drug-induced increases of limbic and striatal dopamine release.

Subcutaneous administration to mice of the sulfated octapeptide of cholecystokinin (CCK; 0.2-1 mg/kg) lowered dopamine release and metabolism in the caudate-putamen and frontal cortex in a dose- and time-related manner. Twelve-fold higher doses of CCK were required to lower dopamine release and metabolism in the olfactory tubercle. Amphetamine-induced increases in dopamine release but not metabolism in the caudate-putamen and olfactory tubercle were attenuated in a dose-related manner by CCK. Increases in dopamine release and metabolism following haloperidol were also attenuated by CCK. These data are consistent with the potential antipsychotic action of CCK receptor agonists. CCK appears to be a suppressor of striatal, limbic and cortical dopamine release, especially when release is augmented.

3,4-Dihydroxyphenylacetic Acid↗

Dopamine autoreceptors modulate the in vivo release of dopamine in the frontal, cingulate and entorhinal cortices.

The regulation by autoreceptors of dopamine release in the rat neocortex was inferred from measurements of 3-methoxytyramine after the peripheral injection of a selective (CGS 15855A) or nonselective (apomorphine) dopamine autoreceptor agonist. Basal levels of dopamine release were greatly decreased in the frontal cortex and caudate putamen after the injection of CGS 15855A or apomorphine. The pargyline-induced accumulations of 3-methoxytyramine in the frontal cortex, cingulate cortex and caudate putamen were attenuated by 52 to 82% after the injection of either agonist. 3-Methoxytyramine accumulations in the entorhinal cortex were attenuated by 68% by apomorphine. Thus, as in the caudate putamen, dopamine autoreceptors modulate the in vivo release of dopamine in the frontal cortex and the turnover of the releasable dopamine pool in the frontal, cingulate and entorhinal cortices.

Animals↗