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9-O-acetylated sialic acids enhance entry of virulent Leishmania donovani promastigotes into macrophages

Published online by Cambridge University Press:  15 December 2008

A. GHOSHAL
Affiliation:
Infectious disease and Immunology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Jadavpur, Kolkata700 032, India
S. MUKHOPADHYAY
Affiliation:
Infectious disease and Immunology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Jadavpur, Kolkata700 032, India
A. K. CHAVA
Affiliation:
Infectious disease and Immunology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Jadavpur, Kolkata700 032, India
G. J. GERWIG
Affiliation:
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, Padualaan 8, NL-3584CHUtrecht, The Netherlands
J. P. KAMERLING
Affiliation:
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, Padualaan 8, NL-3584CHUtrecht, The Netherlands
M. CHATTERJEE
Affiliation:
Department of Pharmacology, Institute of Postgraduate Medical Education & Research, 244B A.J.C. Bose Road, Kolkata, 700 020, India
C. MANDAL*
Affiliation:
Infectious disease and Immunology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Jadavpur, Kolkata700 032, India
*
*Corresponding author: Infectious disease and Immunology Division, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Jadavpur, Kolkata -700 032, India. Tel: 91 33 2429 8861. Fax: 91 33 2473 5197 or 91 33 2473 0284. E-mail: [email protected] or [email protected]

Summary

Distribution of 9-O-acetylated sialic acids (9-O-AcSA) on Leishmania donovani has been previously reported. Considering their role in recognition, the differential distribution of sialic acids especially 9-O-acetylated sialic acids in avirulent (UR6) versus virulent (AG83 and GE1) promastigotes of Leishmania donovani and its role in entry into macrophages was explored. Fluorimetric-HPLC, fluorimetric determination and ELISA revealed 14-, 8- and 5-fold lower sialic acids in UR6 as compared to AG83. Interestingly, on UR6, flow cytometry indicated lower (α2→6)-linked sialoglycoproteins along with minimal 9-O-acetylated sialoglycoproteins by Scatchard analysis. Further, UR6 demonstrated a 9- and 14·5-fold lower infectivity and phagocytic index than AG83. Additionally, de-O-acetylation and de-sialylation of AG83 demonstrated a 3- and 1·5-fold reduced phagocytic index. The role of 9-O-AcSA in entry was further confirmed by pre-blocking the macrophage surface with a cocktail of sugars followed by microscopic quantification. The phagocytic index of AG83 exclusively through 9-O-AcSA was significantly high. Interestingly, AG83 produced higher metacyclic promastigotes containing increased 9-O-AcSA as compared to avirulent UR6 supporting its virulent nature. Taken together; our results conclusively demonstrate the increased presence of 9-O-acetylated sialic acid on promastigotes of virulent Leishmania donovani as compared to avirulent UR6 and their subsequent role in entry within macrophages.

Type
Research Article
Copyright
Copyright © 2008 Cambridge University Press

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