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Comparative Study
. 2009 Jun 2:8:115.
doi: 10.1186/1475-2875-8-115.

Effective and cheap removal of leukocytes and platelets from Plasmodium vivax infected blood

Kanlaya Sriprawat  1 , Supaporn Kaewpongsri, Rossarin Suwanarusk, Mara L Leimanis, Usa Lek-Uthai, Aung Pyae Phyo, Georges Snounou, Bruce Russell, Laurent Renia, François Nosten
Affiliations
Comparative Study

Effective and cheap removal of leukocytes and platelets from Plasmodium vivax infected blood

Kanlaya Sriprawat et al. Malar J. .

Abstract

Background: Investigations of Plasmodium vivax are restricted to samples collected from infected persons or primates, because this parasite cannot be maintained in in vitro cultures. Contamination of P. vivax isolates with host leukocytes and platelets is detrimental to a range of ex vivo and molecular investigations. Easy-to-produce CF11 cellulose filters have recently provided us with an inexpensive method for the removal of leukocytes and platelets. This contrasted with previous reports of unacceptably high levels of infected red blood cell (IRBC) retention by CF11. The aims of this study were to compare the ability of CF11 cellulose filters and the commercial filter Plasmodipur at removing leukocyte and platelet, and to investigate the retention of P. vivax IRBCs by CF11 cellulose filtration.

Methods and results: Side-by-side comparison of six leukocyte removal methods using blood samples from five healthy donor showed that CF11 filtration reduced the mean initial leukocyte counts from 9.4 x 103 per microl [95%CI 5.2-13.5] to 0.01 x 103 [95%CI 0.01-0.03]. The CF11 was particularly effective at removing neutrophils. CF11 treatment also reduced initial platelet counts from 211.6 x 103 per microl [95%CI 107.5-315.7] to 0.8 x 103 per microl [95%CI -0.7-2.2]. Analysis of 30 P. vivax blood samples before and after CF11 filtration showed only a minor loss in parasitaemia (<or= 7.1% of initial counts). Stage specific retention of P. vivax IRBCs was not observed.

Conclusion: CF11 filtration is the most cost and time efficient method for the production of leukocyte- and platelet-free P. vivax-infected erythrocytes from field isolates.

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Figures

Figure 1
Figure 1
The comparative efficacy of six methods for removing leukocytes (A) and platelets (B) from five healthy donor blood samples (repeated measures) prior to buffy coat removal. Leukocyte counts and platelet mean counts × 103μl of whole blood [+/-95% CI]; analysis by Automated Hematology Analyzer (Model pocH-100i, Sysmex Company).
Figure 2
Figure 2
Comparison of leukocyte composition of five healthy donor blood samples before and after treatment with Lymphoprep and Plasmodipur and CF11 methods. Leukocyte counts are expressed as Mean Count per 250 Thick Films (100× oil immersion).
Figure 3
Figure 3
The effect of CF11 filtration on the parasitaemia (A) and developmental stage composition (B) of 30 Thai Plasmodium vivax isolates. The effect of filtration is expressed as a change in the percentage parasitaemia (IRBC per 10 thin film fields at 100× oil immersion) or percentage mature P. vivax erythrocytic stages [(mature stages/(early stages + mature stages)*100)] relative to the blood stages prior to CF11 filtration (no change denoted by the red broken line).
Figure 4
Figure 4
The effect of CF11 filtration on thick and thin Giemsa stained films. Photomicrographs of thick (A&C) and thin (B&D) films before and after CF11 filtration (100× oil immersion, Scale bar = 20 μm).
See this image and copyright information in PMC

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