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In Vitro Invasion Inhibition Assay Using Antibodies Against Plasmodium Knowlesi Duffy Binding Protein Alpha and Apical Membrane Antigen Protein 1 in Human Erythrocyte-Adapted P. Knowlesi A1-H.1 Strain

In Vitro Invasion Inhibition Assay Using Antibodies Against Plasmodium Knowlesi Duffy Binding Protein Alpha and Apical Membrane Antigen Protein 1 in Human Erythrocyte-Adapted P. Knowlesi A1-H.1 Strain

Fauzi Muh, Seong-Kyun Lee, Mohammad Rafiul Hoque, Jin-Hee Han, Ji-Hoon Park, Egy Rahman Firdaus, Robert W Moon, Yee Ling Lau, Eun-Taek Han

Malar J. 2018 Jul 27;17(1):272.

PMID: 30049277

Abstract:

Background:
The rapid process of malaria erythrocyte invasion involves ligand-receptor interactions. Inducing antibodies against specific ligands or receptors that abrogate the invasion process is a key challenge for blood stage vaccine development. However, few candidates were reported and remain to be validated for the discovery of new vaccine candidates in Plasmodium knowlesi.
Methods:
In order to investigate the efficacy of pre-clinical vaccine candidates in P. knowlesi-infected human cases, this study describes an in vitro invasion inhibition assay, using a P. knowlesi strain adapted to in vitro growth in human erythrocytes, PkA1-H.1. Recombinant proteins of P. knowlesi Duffy binding protein alpha (PkDBPα) and apical membrane antigen 1 (PkAMA1) were produced in Escherichia coli system and rabbit antibodies were generated from immune animals.
Results:
PkDBPα and PkAMA1 recombinant proteins were expressed as insoluble and produced as a functional refolded form for this study. Antibodies against PkDBPα and PkAMA1 specifically recognized recombinant proteins and native parasite proteins in schizont-stage parasites on the merozoite organelles. Single and combination of anti-PkDBPα and anti-PkAMA1 antibodies elicited strong growth inhibitory effects on the parasite in concentration-dependent manner. Meanwhile, IgG prevalence of PkDBPα and PkAMA1 were observed in 13.0 and 46.7% in human clinical patients, respectively.
Conclusion:
These data provide support for the validation of in vitro growth inhibition assay using antibodies of DBPα and AMA1 in human-adapted P. knowlesi parasite PkA1-H.1 strain.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR4247837	Anti-Munc-18 Interacting Protein 1 antibody produced in rabbit			Price

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