Virulence Attenuation of a UDP-galactose/ N-acetylglucosamine β1,4 Galactosyltransferase Expressing Leishmania donovani Promastigote
EPrints@IICB
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Title |
Virulence Attenuation of a UDP-galactose/ N-acetylglucosamine β1,4 Galactosyltransferase Expressing Leishmania donovani Promastigote |
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Creator |
Bhaumik, S K
Singh, M Basu, R Bhaumik, S Roychoudhury, K Naskar, Khudiram Roy, Syamal De, Tripti |
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Subject |
Infectious Diseases and Immunology
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Description |
Protozoan parasites of the genus Leishmania are the causative agent of leishmaniasis, a disease whose manifestations in humans range from mild cutaneous lesions to fatal visceral infections. Human visceral leishmaniasis is caused by Leishmania donovani. Long-term culture in vitro leads to the attenuation of the parasite. This loss of parasite virulence is associated with the expression of a developmentally regulated UDP-Galactose/N-acetylglucosamine β 1–4 galactosyltransferase and galactose terminal glycoconjugates as determined by their agglutination with the pea nut agglutinin (PNA). Thus, all promastigotes passaged for more than 11 times were 100% agglutinated with PNA, and represent a homogeneous population of avirulent parasites. Identical concentrations of PNA failed to agglutinate promastigotes passaged for ≤5 times. These PNA− promastigotes were virulent. Promastigotes passaged from 5 to 10 times showed a mixed population. The identity of populations defined by virulence and PNA agglutination was confirmed by isolating PNA+ avirulent and PNA− virulent clones from the 7th passage promastigotes. Only the PNA+ clones triggered macrophage microbicidal activity. The PNA+ clones lacked lipophosphoglycan. Intravenous administration of [14C] galactose-labeled parasite in BALB/c mice resulted in rapid clearance of the parasite from blood with a concomitant accumulation in the liver. By enzymatic assay and RT-PCR we have shown the association of a UDP-Galactose/Nacetylglucosamine β1,4 galactosyltransferase with only the attenuated clones. By immunofluorescence we demonstrated that the enzyme is located in the Golgi apparatus. By western blot analysis and SDS-PAGE of the affinitypurified protein, we have been able to identify a 29 KDa galactose terminal protein from the avirulent clones. |
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Date |
2008
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Type |
Article
PeerReviewed |
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Format |
application/pdf
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Identifier |
http://www.eprints.iicb.res.in/295/1/GLYCOCONJUGATE_JOURNAL%2C_25(5)%2C459%2D472%2C2008[59].pdf
Bhaumik, S K and Singh, M and Basu, R and Bhaumik, S and Roychoudhury, K and Naskar, Khudiram and Roy, Syamal and De, Tripti (2008) Virulence Attenuation of a UDP-galactose/ N-acetylglucosamine β1,4 Galactosyltransferase Expressing Leishmania donovani Promastigote. Glycoconjugate Journal, 25 (5). pp. 459-472. |
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Relation |
http://dx.doi.org/10.1007/s10719-007-9098-0
http://www.eprints.iicb.res.in/295/ |
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