Changes in total and differential leukocyte counts during the clinically silent liver phase in a controlled human malaria infection in malaria-naïve Dutch volunteers

Abstract

Background: Both in endemic countries and in imported malaria, changes in total and differential leukocyte count during Plasmodium falciparum infection have been described. To study the exact dynamics of differential leukocyte counts and their ratios, they were monitored in a group of healthy non-immune volunteers in two separate Controlled Human Malaria Infection (CHMI) studies.

Methods: In two CHMI trials, CHMI-a and CHMI-b, 15 and 24 healthy malaria-naïve volunteers, respectively, were exposed to bites of infected mosquitoes, using the P. falciparum research strain NF54 and the novel clones NF135.C10 and NF166.C8. After mosquito bite exposure, twice-daily blood draws were taken to detect parasitaemia and to monitor the total and differential leukocyte counts. All subjects received a course of atovaquone-proguanil when meeting the treatment criteria.

Results: A total of 39 volunteers participated in the two trials. Thirty-five participants, all 15 participants in CHMI-a and 20 of the 24 volunteers in CHMI-b, developed parasitaemia. During liver stage development of the parasite, the median total leukocyte count increased from 5.5 to 6.1 × 10⁹ leukocytes/L (p = 0.005), the median lymphocyte count from 1.9 to 2.2 (p = 0.001) and the monocyte count from 0.50 to 0.54 (p = 0.038). During the subsequent blood stage infection, significant changes in total and differential leukocyte counts lead to a leukocytopenia (nadir median 3.3 × 10⁹ leukocytes/L, p = 0.0001), lymphocytopenia (nadir median 0.7 × 10⁹ lymphocytes/L, p = 0.0001) and a borderline neutropenia (nadir median 1.5 × 10⁹ neutrophils/L, p = 0.0001). The neutrophil to lymphocyte count ratio (NLCR) reached a maximum of 4.0. Significant correlations were found between parasite load and absolute lymphocyte count (p < 0.001, correlation coefficient - 0.46) and between parasite load and NLCR (p < 0.001, correlation coefficient 0.50). All parameters normalized after parasite clearance.

Conclusions: During the clinically silent liver phase of malaria, an increase of peripheral total leukocyte count and differential lymphocytes and monocytes occurs. This finding has not been described previously. This increase is followed by the appearance of parasites in the peripheral blood after 2-3 days, accompanied by a marked decrease in total leukocyte count, lymphocyte count and the neutrophil count and a rise of the NLCR.

Keywords: Controlled Human Malaria Infection; Leukocyte count; Liver phase; Lymphocyte count; Lymphocytopenia; Monocytes; Neutropenia; Neutrophil to lymphocyte count ratio; Plasmodium falciparum.

Fig. 1

Changes in total and differential leukocyte counts in the 20 subjects who developed malaria in CHMI-b. The data are shown as medians (dots) and interquartile ranges (whiskers). The data from DT − 2 until DT + 3 were synchronized on DT

Fig. 2

Changes in differential cell count ratios in the 20 subjects who developed malaria in CHMI-b. The data are shown as medians (dots) and interquartile ranges (whiskers). The data from DT − 2 until DT + 3 were synchronized on DT