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Purification and characterization of a mitogenic lectin from cephalosporium, a pathogenic fungus causing mycotic keratitis.

Nagre NN, Chachadi VB, Eligar SM, Shubhada C, Pujari R, Shastry P, Swamy BM, Inamdar SR - Biochem Res Int (2010)

Bottom Line: Ophthalmic mycoses caused by infectious fungi are being recognized as a serious concern since they lead to total blindness.CSL showed strong binding to normal human peripheral blood mononuclear cells (PBMCs) to elicit mitogenic activity.The sugar specificity of the lectin and its interaction with PBMCs to exhibit mitogenic effect indicate its possible role in adhesion and infection process of Cephalosporium.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Karnatak University, Dharwad 580 003, Karnataka, India.

ABSTRACT
Ophthalmic mycoses caused by infectious fungi are being recognized as a serious concern since they lead to total blindness. Cephalosporium is one amongst several opportunistic fungal species implicated in ophthalmic infections leading to mycotic keratitis. A mitogenic lectin has been purified from the mycelia of fungus Cephalosporium, isolated from the corneal smears of a keratitis patient. Cephalosporium lectin (CSL) is a tetramer with subunit mass of 14 kDa, agglutinates human A, B, and O erythrocytes, and exhibits high affinity for mucin compared to fetuin and asialofetuin but does not bind to simple sugars indicating its complex sugar specificity. CSL showed strong binding to normal human peripheral blood mononuclear cells (PBMCs) to elicit mitogenic activity. The sugar specificity of the lectin and its interaction with PBMCs to exhibit mitogenic effect indicate its possible role in adhesion and infection process of Cephalosporium.

No MeSH data available.


Related in: MedlinePlus

Mitogenic activity of CSL. PBMCs were isolated from blood of healthy donors and exposed to serial concentrations of CSL (0.625–10 μg/ml) and PHA-L (0.16–2.5 μg/ml) for 72 hours and proliferation was measured by tritiated thymidine incorporation assay as counts per minute (CPM). The data are presented as mean ± SE of four independent experiments done in triplicates.
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fig4: Mitogenic activity of CSL. PBMCs were isolated from blood of healthy donors and exposed to serial concentrations of CSL (0.625–10 μg/ml) and PHA-L (0.16–2.5 μg/ml) for 72 hours and proliferation was measured by tritiated thymidine incorporation assay as counts per minute (CPM). The data are presented as mean ± SE of four independent experiments done in triplicates.

Mentions: Mitogenic effect of CSL on human PBMCs was determined by tritiated thymidine incorporation assay. CSL stimulated the uptake of thymidine by human PBMCs in a dose-dependent manner, with maximum incorporation occurring at 10 μg/ml of the different doses tested (0.625–10 μg/ml), whereas PHA-L, a positive control exhibited maximum proliferation at 1.25 μg/ml (Figure 4).


Purification and characterization of a mitogenic lectin from cephalosporium, a pathogenic fungus causing mycotic keratitis.

Nagre NN, Chachadi VB, Eligar SM, Shubhada C, Pujari R, Shastry P, Swamy BM, Inamdar SR - Biochem Res Int (2010)

Mitogenic activity of CSL. PBMCs were isolated from blood of healthy donors and exposed to serial concentrations of CSL (0.625–10 μg/ml) and PHA-L (0.16–2.5 μg/ml) for 72 hours and proliferation was measured by tritiated thymidine incorporation assay as counts per minute (CPM). The data are presented as mean ± SE of four independent experiments done in triplicates.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3008968&req=5

fig4: Mitogenic activity of CSL. PBMCs were isolated from blood of healthy donors and exposed to serial concentrations of CSL (0.625–10 μg/ml) and PHA-L (0.16–2.5 μg/ml) for 72 hours and proliferation was measured by tritiated thymidine incorporation assay as counts per minute (CPM). The data are presented as mean ± SE of four independent experiments done in triplicates.
Mentions: Mitogenic effect of CSL on human PBMCs was determined by tritiated thymidine incorporation assay. CSL stimulated the uptake of thymidine by human PBMCs in a dose-dependent manner, with maximum incorporation occurring at 10 μg/ml of the different doses tested (0.625–10 μg/ml), whereas PHA-L, a positive control exhibited maximum proliferation at 1.25 μg/ml (Figure 4).

Bottom Line: Ophthalmic mycoses caused by infectious fungi are being recognized as a serious concern since they lead to total blindness.CSL showed strong binding to normal human peripheral blood mononuclear cells (PBMCs) to elicit mitogenic activity.The sugar specificity of the lectin and its interaction with PBMCs to exhibit mitogenic effect indicate its possible role in adhesion and infection process of Cephalosporium.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Karnatak University, Dharwad 580 003, Karnataka, India.

ABSTRACT
Ophthalmic mycoses caused by infectious fungi are being recognized as a serious concern since they lead to total blindness. Cephalosporium is one amongst several opportunistic fungal species implicated in ophthalmic infections leading to mycotic keratitis. A mitogenic lectin has been purified from the mycelia of fungus Cephalosporium, isolated from the corneal smears of a keratitis patient. Cephalosporium lectin (CSL) is a tetramer with subunit mass of 14 kDa, agglutinates human A, B, and O erythrocytes, and exhibits high affinity for mucin compared to fetuin and asialofetuin but does not bind to simple sugars indicating its complex sugar specificity. CSL showed strong binding to normal human peripheral blood mononuclear cells (PBMCs) to elicit mitogenic activity. The sugar specificity of the lectin and its interaction with PBMCs to exhibit mitogenic effect indicate its possible role in adhesion and infection process of Cephalosporium.

No MeSH data available.


Related in: MedlinePlus