Quantitative cytokine level of TNF-α, IFN-γ, IL-10, TGF-β and circulating Epstein-Barr virus DNA load in individuals with acute Malaria due to P. falciparum or P. vivax or double infection in a Malaria endemic region in Indonesia

Abstract

Plasmodium falciparum Malaria and Epstein-Barr Virus (EBV) infection are risk factors in the development of Burkitt’s lymphoma. In Indonesia, 100% of the population is persistently infected with EBV early in life and at risk of developing EBV-linked cancers. Currently, 10.7 million people in Indonesia are living in Malaria-endemic areas. This cross-sectional study was initiated to investigate how acute Malaria dysregulates immune control over latent EBV infection. Using blood and plasma samples of 68 patients with acute Malaria and 27 healthy controls, we measured the level of parasitemia for each plasmodium type (P. falciparum, P. vivax, and mixed) by microscopy and rapid test. The level of 4 regulatory cytokines was determined by quantitative ELISA and the level of circulating EBV genome by real-time PCR targeting the single copy EBNA-1 sequence. All Plasmodium-infected cases had high-level parasitemia (>1000 parasites/ul blood) except for one case. EBV-DNA levels were significantly more elevated in P. falciparum and P. vivax infections (P<0.05) compared to controls. EBV-DNA levels were not related to age, gender, Malaria symptoms, or plasmodium type. TNF-α and IL-10 levels were increased in Malaria cases versus controls, but IFN-γ and TGF- β levels were comparable between the groups. Only TNF-α levels in P. falciparum cases showed a clear correlation with elevated EBV DNA levels (R2 = 0.8915). This is the first study addressing the relation between EBV (re)activation and cytokine responses during acute Malaria, revealing a clear correlation between pro-inflammatory cytokine TNF-α and EBV-DNA levels, specifically in P. falciparum cases, suggesting this cytokine to be key in dysregulating EBV homeostasis during acute P. falciparum Malaria.

Transforming Growth Factor-β Concerning Malarial Infection and Severity: A Systematic Review and Meta-Analysis

“…For specific reasons, after the screening of full texts, the following articles were excluded: 42 were TGF-β in vivo studies, 33 were TGF-β in vitro studies, 10 were TGF-β gene/protein expression studies, 7 were mosquito experiments, 7 were reviews, 6 were TGF-β in pregnancy/cord blood, 6 were TGF-β in uncomplicated malaria only, 6 were TGF-β gene polymorphism studies, 3 were conference abstracts, 2 were TGF-β in co-infection, 1 was about TGF- β in severe malaria only, 1 had no full-text available, 1 was a systematic review, and 1 was a duplicate study. Finally, 13 studies were included [ 26 , 27 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 ]: 7 studies [ 33 , 34 , 35 , 36 , 37 , 38 , 39 ] comparing TGF-β levels between uncomplicated malaria and healthy controls and 6 studies [ 26 , 27 , 40 , 41 , 42 , 43 ] comparing TGF-β levels between severe and uncomplicated malaria ( Figure 1 ).…”

“…These studies were prospective observational studies (six studies; 46.2%) [ 33 , 39 , 40 , 41 , 42 , 43 ], case–control studies (three studies; 23.1%) [ 27 , 35 , 37 ], cross-sectional studies (three studies; 23.1%) [ 34 , 36 , 38 ] and a cohort study (one study; 7.69%) [ 26 ]. The included studies were conducted in Africa (53.8%; Uganda [ 26 , 35 , 37 ], Gabon [ 33 ], Burkina Faso [ 40 ], Kenya and Uganda [ 41 ] and Gabon [ 42 ]), Asia (30.8%; Thailand [ 27 , 39 ], Indonesia [ 34 ], India [ 43 ]) and South America (15.4%; Brazil [ 36 , 38 ]). Most included studies (10 studies; 76.9%) enrolled patients with P. falciparum [ 26 , 27 , 33 , 35 , 37 , 39 , 40 , 41 , 42 , 43 ].…”

Impact of Epstein-Barr virus co-infection on natural acquired Plasmodium vivax antibody response

“…vivax individuals, EBV-DNA levels were not related to age, gender, or malaria symptoms [66]. To properly address the influence of the persistence of EBV-DNA detection in the course of vivax malaria, a non-immune symptomatic P. vivax population should be investigated that was out of the scope of the current MS.…”

“…As expected, few malaria cases were detected in our malaria semi-immune study population, which precluded our ability to evaluate an association between acute P. vivax infection and persistent detection of EBV-DNA. Although the current study was not designed to investigate whether EBV can change the course of an acute P. vivax malaria infection, scant data from Indonesia suggest that whereas EBV-DNA levels were significantly elevated in high parasitemic P. vivax individuals, EBV-DNA levels were not related to age, gender, or malaria symptoms [66]. To properly address the influence of the persistence of EBV-DNA detection in the course of vivax malaria, a non-immune symptomatic P. vivax population should be investigated that was out of the scope of the current MS.…”

Impact of Epstein-Barr virus co-infection on natural acquired Plasmodium vivax antibody response

“…Although the current study was not designed to investigate whether EBV can change the course of an acute P . vivax malaria infection, scant data from Indonesia suggest that whereas EBV-DNA levels were significantly elevated in high parasitemic P. vivax individuals, EBV-DNA levels were not related to age, gender, or malaria symptoms [ 75 ]. To properly address the influence of the persistence of EBV-DNA detection in the course of vivax malaria, a non-immune symptomatic P. vivax population should be investigated that was out of the scope of the current study.…”