Differential Surface Deposition of Complement Proteins on Logarithmic and Stationary Phase Leishmania Chagasi Promastigotes

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2012-12-01
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Ramer-Tait, Amanda
Lei, Soi Meng
Bellaire, Bryan
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Beetham, Jeffrey
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Entomology

The Department of Entomology seeks to teach the study of insects, their life-cycles, and the practicalities in dealing with them, for use in the fields of business, industry, education, and public health. The study of entomology can be applied towards evolution and ecological sciences, and insects’ relationships with other organisms & humans, or towards an agricultural or horticultural focus, focusing more on pest-control and management.

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The Department of Entomology was founded in 1975 as a result of the division of the Department of Zoology and Entomology.

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Previous works demonstrated that various species of Leishmania promastigotes exhibit differential sensitivity to complement-mediated lysis (CML) during development. Upon exposure to normal human serum (NHS), cultures of Leishmania chagasi promastigotes recently isolated from infected hamsters (fewer than 5 in vitro passages) are CML-sensitive when in the logarithmic growth phase but become CML-resistant upon transition to the stationary culture phase. Visualization by light and electron microscopy revealed dramatic morphological differences between promastigotes from the 2 culture phases following exposure to NHS. Flow cytometric analysis demonstrated that surface deposition of the complement components C3, C5, and C9 correlated inversely with promastigote CML-resistance. The highest levels of complement protein surface accumulation were observed for logarithmic phase promastigotes, while stationary phase promastigotes adsorbed the least amount of complement proteins. Additionally, fluorescence microscopy revealed that C3 and C5 localized in a fairly uniform pattern to the plasma membrane of promastigotes from logarithmic phase cultures, while the staining of promastigotes from stationary phase cultures was indistinguishable from background. By Western blot analysis, high levels of the complement proteins C3, C5, and C9 were detected in the total lysates of NHS-exposed logarithmic phase L. chagasi promastigotes, relative to NHS-exposed stationary phase promastigotes; this finding indicates that the low levels of C3 and C5 seen on the surface of stationary phase promastigotes were not due to protein uptake/internalization. Together, these data demonstrate the differential deposition of complement proteins on the surfaces of logarithmic and stationary phase L. chagasi promastigotes. The data support a model wherein stationary phase L. chagasi promastigotes resist CML by limiting the deposition of C3 and its derivatives, which, in turn, limit surface levels of complement proteins (including C5 and C9) that form the lytic membrane attack complex.

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This is a manuscript of an article from Journal of Parasitology 98 (2012): 1109, doi:10.1645/GE-3130.1. Posted with permission.

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Sun Jan 01 00:00:00 UTC 2012
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