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Antiproliferative effect of T/Tn specific Artocarpus lakoocha agglutinin (ALA) on human leukemic cells (Jurkat, U937, K562) and their imaging by QD-ALA nanoconjugate

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Abstract

T/Tn specificity of Artocarpus lakoocha agglutinin (ALA), isolated from the seeds of A. lakoocha (Moraceae) fruit and a heterodimer (16 kD and 12 kD) of molecular mass 28 kD, was further confirmed by SPR analysis using T/Tn glycan containing mammalian glycoproteins. N-terminal amino acid sequence analysis of ALA showed homology at 15, 19–21, 24–27, and 29 residues with other lectin members of Moraceae family viz., Artocarpus integrifolia (jacalin) lectin, Artocarpus hirsuta lectin, and Maclura pomifera agglutinin. It is mitogenic to human PBMC and the maximum proliferation was observed at 1 ng/ml. It showed an antiproliferative effect on leukemic cells, with the highest effect toward Jurkat cells (IC50 13.15 ng/ml). Synthesized CdS quantum dot-ALA nanoconjugate was employed to detect the expression of T/Tn glycans on Jurkat, U937, and K562 leukemic cells surfaces as well as normal lymphocytes by fluorescence microscopy. No green fluorescence was observed with normal lymphocytes indicating that T/Tn determinants, which are recognized as human tumor associated structures were cryptic on normal lymphocyte surfaces, whereas intense green fluorescent dots appeared during imaging of leukemic cells, where such determinants were present in unmasked form. The above results indicated that QD-ALA nanoconjugate is an efficient fluorescent marker for identification of leukemic cell lines that gives rise to high quality images.

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Abbreviations

ALA:

Artocarpus lakoocha agglutinin

BSM:

Bovine submandibular gland mucin

CdS:

Cadmium sulphide

ELLSA:

Enzyme - linked lectinsorbent assay

ESI-MS:

Electron spray ionization mass spectrometry

FPLC:

Fast protein liquid chromatography

HBS:

Hepes buffered saline

HEPES:

N-(2-hydoxyethyl) piperizine-N′-(2-hydroxypropane sulfonic acid)

HSM:

Hamster submaxillary mucin

PBMC:

Peripheral blood mononuclear cells

PVDF:

Polyvinylidene difluoride

Q-ToF:

Quadrupole-time of flight

QD:

Quantum dot

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

2 ME:

2-Mercaptoethanol

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Acknowledgements

We sincerely thank Dr. Sujata Sharma of Department of Biophysics, All India Institutes of Medical Sciences, New Delhi for performing amino acid sequence analysis. The assistance of Dr. Syamal Roy of Indian Institute of Chemical Biology, Kolkata and Mr. Gautam Mondal of West Bengal University of Technology, Kolkata is gratefully acknowledged. This study was financially supported by a research grant (BT/PR4462/BRB/10/1350/2003) of the Department of Biotechnology, Government of India, New Delhi to B.P.C.

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Author notes

  1. Bishnu P. Chatterjee

    Present address: Department of Biotechnology, West Bengal University of Technology, BF 142, Sector 1, Salt Lake, Kolkata, 700 064, India

  2. Urmimala Chatterjee

    Present address: Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Science, Uppsala, Sweden

  3. Partha Pratim Bose

    Present address: Department of Biochemistry and Organic Chemistry, Uppsala University, Uppsala, Sweden

Authors and Affiliations

  1. Department of Biological Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata, 700 032, India

    Urmimala Chatterjee, Partha Pratim Bose & Bishnu P. Chatterjee

  2. Department of Biophysics, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029, India

    Sharmistha Dey & Tej P. Singh

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Correspondence to Bishnu P. Chatterjee.

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Chatterjee, U., Bose, P.P., Dey, S. et al. Antiproliferative effect of T/Tn specific Artocarpus lakoocha agglutinin (ALA) on human leukemic cells (Jurkat, U937, K562) and their imaging by QD-ALA nanoconjugate. Glycoconj J 25, 741–752 (2008). https://doi.org/10.1007/s10719-008-9134-8

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