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Epidemiologic Trends in Malaria Incidence Among Travelers Returning to Metropolitan France, 1996-2016
research-article
Author(s):
Eric Kendjo , MSc
1
,
,
Sandrine Houzé , MD, PhD
2 ,
Oussama Mouri , PharmD
3 ,
Aida Taieb , PhD
4 ,
Frédérick Gay , MD, PhD
1 ,
Stéphane Jauréguiberry , MD, PhD
1 ,
Ilhame Tantaoui , PharmD, MSc
1 ,
Papa A. Ndour , PhD
5 ,
Pierre Buffet , MD, PhD
5 ,
Martine Piarroux , MD, PhD
6 ,
Marc Thellier , MD, MSc
1 ,
Renaud Piarroux , MD, PhD
1
Publication date (Electronic):
5 April 2019
Journal:
JAMA Network Open
Publisher:
American Medical Association
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Key Points
Question
Could changes in the population experiencing imported malaria in France over the past 2 decades explain the persistence of the high number of malaria cases despite national preventive measures?
Abstract
Importance
Despite annually adapted recommendations to prevent malaria in travelers to endemic areas, France is still the industrialized country reporting the highest number of imported cases of malaria. Better understanding of the epidemiologic context and evolution during the past 2 decades may help to define a better preventive strategy.
Objective
To study epidemiologic trends of imported cases of malaria in travelers in geographic territories of France on the European continent (metropolitan France) from 1996 through 2016 to potentially explain the persistence of high imported malaria incidence despite national preventive measures.
Design, Setting, and Participants
In a cross-sectional study, between January 1 and May 31, 2018, data were extracted from the French National Reference Center of Malaria Surveillance. Trends in patients with imported malaria in association with age, sex, ethnicity, purpose of travel, malaria species, severity of illness, case mortality rate, and endemic countries visited were analyzed in 43 333 malaria cases among civilian travelers living in metropolitan France.
Main Outcomes and Measures
Evolution of the main epidemiologic characteristics of patients with imported malaria.
Results
Among the 43 333 patients with imported malaria in civilian travelers included in the study, 24 949 were male (62.4%), and 8549 were younger than 18 years (19.9%). A total of 28 658 malaria cases (71.5%) were among African individuals, and 10 618 cases (26.5%) among European individuals. From 1996 through 2016, the number of confirmed malaria cases peaked at 3400 cases in 2000, then declined to 1824 cases in 2005 and stabilized thereafter to approximately 1720 malaria cases per year. A total of 37 065 cases (85.5%) were due to Plasmodium falciparum . The proportion of malaria cases among African individuals rose from 53.5% in 1996 to 83.4% in 2016, and the most frequent motivation for traveling was visiting friends and relatives (25 329 [77.1%]; P < .001). Despite an increase in the proportion of severe cases, which rose from 131 cases (8.9%) in 1996 to 279 cases (16.7%) in 2016 ( P < .001), mortality remained stable, being approximately 0.4% during the study period.
Conclusions and Relevance
Beyond the apparent stability of the number of imported malaria cases in France, significant changes appear to have occurred among the population who developed malaria infection following travel in endemic areas. These changes may imply that adaptation of the preventive strategy is needed to reduce the burden of the disease among travelers.
Abstract
This cross-sectional study of data from the French National Reference Center of Malaria Surveillance examines the epidemiologic characteristics associated with imported cases of malaria among travelers returning to metropolitan France.
Related collections
Most cited references31
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S. Cohen, I A McGregor, S Carrington (1961)
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The Plasmodium falciparum-Specific Human Memory B Cell Compartment Expands Gradually with Repeated Malaria Infections
Introduction To date, most successful vaccines have targeted pathogens that induce long-lived protective antibodies after a single infection, such as the viruses that cause smallpox, measles and yellow fever [1]. It has proved more difficult to develop highly effective vaccines against pathogens that do not induce sterile immunity such as the human immunodeficiency virus type-1 (HIV-1), Mycobacterium tuberculosis (Mtb), and Plasmodium falciparum malaria [2]. However, unlike HIV-1 and Mtb, clinical immunity to malaria can be acquired, but only after years of repeated Pf infections [3]. Passive transfer studies indicate that antibodies ultimately play a key role in protection from malaria [4], yet several studies show that antibodies to Pf antigens are inefficiently generated and rapidly lost in the absence of ongoing exposure to the parasite (reviewed in [5]). Elucidating the cellular basis of the inefficient acquisition of malaria immunity may ultimately prove critical to the design of an effective malaria vaccine. Despite the key role that antibodies play in protection from a variety of infectious diseases, remarkably little is known about the cellular basis of acquiring humoral immunity in response to natural infections in humans. This gap in our knowledge is due in large part to the difficulty in studying natural infections in humans when we cannot predict who within a population will be infected with a given pathogen at a given time. Thus, our current understanding of the acquisition of immunity is largely derived from animal models and studies of humans after vaccination. These studies have established that long-lived, antibody-based immunity requires the generation and maintenance of memory B cells (MBCs) and long-lived plasma cells (LLPCs) (reviewed in [6], [7]). This process begins when naïve B cells bind antigen near the interface of B and T cell areas of secondary lymphoid organs. Several studies suggest that high-affinity binding drives naïve B cells to differentiate into short-lived, isotyped switched plasma cells (PCs) within the extra-follicular region which contributes to the initial control of infection. In contrast, lower affinity binding selects for entry of naïve B cells into follicles where germinal centers are formed. After a period of 7–10 days, through the CD4+ T-cell dependent process of somatic hypermutation, the germinal center reaction yields MBCs and LLPCs of higher affinity than the initial wave of short-lived plasma cells (SLPCs). MBCs recirculate and mediate recall responses after re-exposure to their cognate antigen by rapidly expanding and differentiating into PCs, whereas LLPCs residing in the bone marrow constitutively secrete antibody and provide a critical first line of defense against re-infection. The mechanisms by which antibody responses are maintained over the human life-span remains an open question. In one model, LLPCs survive indefinitely in the bone marrow and independently maintain steady-state antibody levels [8]. Alternative models predict that PCs are replenished by MBCs that proliferate and differentiate in response to persistent [9] or intermittent exposure to antigen, and/or through non-specific by-stander activation (e.g. cytokines or TLR ligands) [10]. Unlike PCs, which are terminally-differentiated, MBCs may be maintained through homeostatic proliferation [11], possibly through exposure to polyclonal stimuli [10]. To address fundamental questions related to the generation and maintenance of MBCs and Abs specific for Pf malaria in children in malaria endemic areas, we conducted a year-long prospective study in a rural village of Mali that experiences an intense, sharply-demarcated six-month malaria season annually. We determined whether Pf infection generates MBCs specific for Pf blood stage antigens, and if so, whether they accumulate with age and cumulative Pf exposure, and also whether their frequency correlates with protection from malaria. In addition, we determined whether acute, symptomatic Pf infection resulted in an increase in the number of Pf-specific MBCs and the levels of specific antibodies, and if so, whether this increase remained stable over a six-month period of markedly reduced Pf transmission. By taking advantage of the tetanus immunization schedule in Mali in which infants and women of child-bearing age are vaccinated, we compared the relative efficiencies of the acquisition of tetanus toxoid (TT)- and Pf-specific MBCs and Ab, and also tested three hypotheses: 1) that growth of the MBC compartment depends on immunological experience rather than age, 2) that Pf infection induces non-specific activation of bystander B cells [12], [13], and 3) that polyclonal activation during heterologous immune responses is a general mechanism for maintaining MBCs and LLPCs [10]. Results Malaria immunity is acquired gradually despite intense exposure to the Pf parasite In May 2006 we initiated an observational cohort study in Mali to investigate the mechanisms underlying naturally-acquired malaria immunity. A detailed description of the study site and cohort has been reported elsewhere [14]. The study population was an age-stratified, random sample representing 15% of all individuals living in a small, rural, well-circumscribed, non-migratory community where antimalarial drugs were provided exclusively by the study investigators. During a two-week period one month prior to the abrupt onset of the six-month malaria season, we enrolled 225 individuals in four age groups: 2–4 years (n = 73), 5–7 years (n = 52), 8–10 years (n = 51), and 18–25 years (n = 49). Attendance at scheduled follow-up visits was >99% for children (2–10 years) and 82% for adults (18–25 years) during the one-year study period indicating a high degree of study awareness and participation. For the MBC analysis reported here, a subset of 185 individuals was randomly selected within each of the four age categories. All subsequent data and analysis refer to these 185 individuals. The baseline demographic and clinical characteristics of this subset are shown in Table 1, according to age group. As previously reported [14], only three of the characteristics shown in Table 1 were associated with decreased malaria risk in multivariate analysis—increasing age, sickle cell trait (HbAS), and asymptomatic Pf parasitemia at study enrollment. During the one-year study period there were 380 unscheduled clinic visits, during which 219 cases of malaria were diagnosed, five of which met the WHO criteria for severe malaria [15]. Malaria episodes were defined as an axillary temperature ≥37.5°C, Pf asexual parasitemia ≥5000 parasites/µL, and a non-focal physical exam by the study physician. As expected in this region of Mali, all malaria cases were confined to a six-month period that began in July, peaked in October, and ended by January (Fig. 1A). The incidence of malaria and the proportion of individuals experiencing at least one malaria episode decreased with age, whereas the time to the first malaria episode increased with age (Table 2 and Fig. 1B). Thus, despite intense annual Pf transmission at this study site, malaria immunity is acquired slowly. 10.1371/journal.ppat.1000912.g001 Figure 1 Malaria immunity is acquired gradually despite intense annual exposure to the Pf parasite. (A) Number of malaria episodes per day during the study period. Over a one-year surveillance period 185 individuals experienced 219 malaria episodes during a sharply-demarcated six-month malaria season. Malaria episodes were defined as fever ≥37.5°C and Pf asexual parasitemia ≥5000/µL blood. To track the B-cell response to acute malaria, and after a period of reduced Pf exposure, PBMCs and plasma were collected at points indicated by the arrows: before the malaria season, two weeks after the first malaria episode (arrow with asterisk indicates the mean time to first malaria episode, 132 days from enrollment), and six months after the end of the malaria season. (B) Kaplan-Meier estimates of the cumulative probability of malaria over the study period, according to age category. The number of individuals at risk over the study period is shown below the graph. The P value was obtained using the log rank test. 10.1371/journal.ppat.1000912.t001 Table 1 Baseline characteristics of the study cohort by age group. Age group, years All (n = 185) 2–4 (n = 59) 5–7 (n = 37) 8–10 (n = 42) 18–25 (n = 47) Gender, % female (no.) 66.1 (39) 48.7 (18) 33.3 (14) 61.7 (29) 54.1 (100) Ethnicity, % (no.) Bambara 62.7 (37) 51.4 (19) 54.8 (23) 66.0 (31) 59.5 (110) Sarakole 32.2 (19) 43.2 (16) 35.7 (15) 27.7 (13) 34.1 (63) Fulani 3.4 (2) 5.4 (2) 7.1 (3) 4.3 (2) 4.9 (9) Malinke 1.7 (1) 0.0 (0) 2.4 (1) 2.1 (1) 1.6 (3) Hemoglobin AS, % (no.)a– 13.6 (8) 8.1 (3) 7.1 (3) 10.6 (5) 10.3 (19) P. falciparum smear positive at enrollment, % (no.)b– 6.8 (4) 10.8 (4) 11.9 (5) 6.4 (3) 8.7 (16) Parasitemia if smear positive at enrollment, parasites/microliter [geometric mean (95% CI)] 1438 (159–12973) 3616 (1500–8715) 415 (134–1287) 953 (39–23381) 1137 (579–2232) GI helminth, % positive at enrollment (no.)c– 14.6 (8) 8.3 (3) 11.8 (4) 0 (0) 9.7 (15) Urine schistosomiasis, % positive at enrollment (no.)d– 0 (0) 0 (0) 5.3 (2) 29.0 (9) 7.4 (11) Distance lived from clinic, meters (mean ±SD) 395 (±116) 408 (±140) 365 (±83) 359 (±91) 382 (±110) Bed net use, % (no.)e– 27.3 (15) 41.2 (14) 17.1 (7) 39.5 (15) 30.4 (51) a–Data available for 177 subjects. b–All subjects were asymptomatic at enrollment. c–Data available for 154 subjects; GI = gastrointestinal. d–Data available for 148 subjects. e–Nightly bednet use self-reported at the end of the malaria season. 10.1371/journal.ppat.1000912.t002 Table 2 Malaria clinical outcomes by age group. Age group, years All (n = 185) 2–4 (n = 59) 5–7 (n = 37) 8–10 (n = 42) 18–25 (n = 47) Malaria incidence, mean (±SD)a– 1.86 (±1.28) 1.81 (±1.17) 0.95 (±1.08) 0.09 (±0.28) 1.19 (±1.27) Severe malaria incidence, no.b– 4 1 0 0 5 At least one malaria episode, % (no.) 83.1 (49) 81.1 (30) 57.1 (24) 8.5 (4) 57.8 (107) Time to first malaria episode, days (median)c– 101 121 124 153 118 Parasitemia at first malaria episode, parasites/microliter [geometric mean (95% CI)] 39084 (30579–49954) 26417 (19440–35896) 20561 (15683–26956) 8816 (4082–19037) 28678 (24334 –33799) a–Malaria episode defined as T≥37.5°C, asexual parasitemia ≥5000/microliter, and non-focal physical examination. b–WHO definition of severe malaria. c–Days since study enrollment. Analysis of Pf-specific and TT-specific MBCs and Abs in Pf-uninfected children and adults before the malaria season We first established baseline levels of IgG+ AMA1-, MSP1- and TT-specific MBCs and Abs in Pf-uninfected, healthy children and adults in May just before the malaria season, a point at which there had been little to no Pf transmission for five months. For this analysis we excluded individuals with asymptomatic Pf parasitemia (8.7% of total cohort; Table 1), because they showed a decreased risk of malaria and tended toward higher frequencies of AMA1- and MSP1-specific MBCs and levels of Ab (data not shown). We focused our analyses on MBCs and Abs specific for Pf blood-stage antigens because humoral responses are known to be critical to blood-stage immunity [4]. We examined the response to two blood stage proteins, Apical Membrane Antigen 1 (AMA1) and Merozoite Surface Protein 1 (MSP1), because we had previously studied the MBC and Ab responses to these antigens in vaccine trials of Pf-naïve individuals [16]. This afforded the opportunity to compare the acquisition of B cell memory to the same antigens after vaccination versus natural Pf infection. We express MBC data as ‘MBCs per 106 PBMCs’, where ‘MBCs’ refers to the number of antibody secreting cells derived from MBCs during the six-day culture, and ‘106 PBMCs’ refers to the number of PBMCs after culture. In the present study, the mean frequency of AMA1-specific MBCs per 106 PBMCs increased with age (Fig. 2A; 2–4 yr: 1.2 [95% CI: 0.45–1.9]; 5–7 yr: 5.0 [95% CI: −0.2–10.1]; 8–10 yr: 8.9 [95% CI: 4.9–12.9]; 18–25 yr: 37.8 [95% CI: 10.4–65.3]; P 50 years) MBCs in nearly all vaccinees [34], a remarkably high proportion of adults in the present study did not have detectable AMA1- or MSP1-specific MBCs despite annual exposure to 50–60 infective mosquito bites per person per month at the height of the malaria season [17], similar to what Dorfman et al. observed in a cross-sectional study in Kenya [35]. Importantly, most female adults in the present study had detectable TT-specific MBCs three to ten years after a single TT booster vaccine in adolescence. We previously reported that AMA1- and MSP1-specific MBCs were reliably generated in Pf-naïve U.S. adults following just two vaccinations [16]. Taken together, these observations indicate that the relatively inefficient generation and/or maintenance of Pf-specific MBCs in response to natural Pf infection cannot be ascribed entirely to inherent deficiencies in the antigens themselves. Collectively, these observations raise a central question: if AMA1 and MSP1-specific MBCs and Abs can be efficiently generated by vaccination of Pf-naïve adults, and TT-specific MBCs and Abs can be efficiently generated by vaccination of Pf-exposed individuals in this cohort, what underlies the inefficient acquisition and/or maintenance of AMA1 and MSP1-specific MBCs and Abs in response to natural Pf infection? One simple answer, in addition to parasite antigenic variation [36], [37], might be that the enormous number of antigens encoded by the over 5,400 Pf genes overwhelms the immune system's capacity to select for and commit a sufficient number of MBCs and LLPCs specific for any given Pf antigen to a long-lived pool [38]. If immunity to clinical malaria requires high levels of antibodies to a large number of Pf proteins, then the inability to commit large numbers of MBCs and LLPCs specific for any given Pf antigen during any given infection, as shown here, may explain, in part, why malaria immunity is acquired slowly. In this scenario the Pf-infected individual is capable of the normal generation and maintenance of MBCs and LLPCs, but acquiring a sufficient number of MBCs and LLPCs to a large number of antigens may simply take years. It is also possible that Pf infection disrupts the immune system's ability to generate or maintain MBCs or LLPCs. The differentiation of B cells into long-lived MBCs depends to a great extent on the affinity of their BCRs for antigen. Recently, evidence was presented that affinity maturation of B cells may fail to occur in the absence of adequate Toll-like receptor (TLR) stimulation [39]. We recently reported that Malian adults were relatively refractory to CpG, a TLR9 agonist incorporated into two subunit malaria vaccine candidates [40], raising the possibility that the slow acquisition of MBCs observed here may be due to a failure of B cells to undergo affinity maturation during Pf infection. Although our data do not directly address the role of apoptosis in the gradual acquisition of Pf-specific MBCs, it is worth noting that we found no evidence of Pf-induced ablation of Plasmodium-specific MBCs, as was observed in mice four days after Plasmodium yoelii infection [41]. The relatively inefficient response to natural Pf infection also does not appear to be due to a persistent, Pf-induced general immunosuppression as the frequency of TT-specific MBCs increased significantly in most adult females in response to a single TT booster vaccination, an increase that appeared to be maintained for years. In an experimental model of lymphocytic choriomeningitis virus (LCMV) infection, a high antigen-to-B cell ratio disrupted germinal center formation and the establishment of B cell memory [42]. It is plausible that a similar mechanism is at play during the blood stage of Pf infection when the immune system encounters high concentrations of parasite proteins. Indeed, germinal center disruption is observed in mice infected with P. berghei ANKA [43] and P. chabaudi [44]. It is also possible that specific parasite products selectively interfere with the regulation of B cell differentiation [45] or with the signals required for sustaining LLPCs in the bone marrow [46]. It is also conceivable that the disproportionately high level of class-switched SLPCs we observed in response to Pf infection arises from pre-diversified IgM+IgD+CD27+ (marginal zone) B cells—analogous to the rapid protective response against highly virulent encapsulated bacteria that do not elicit classical T-dependent responses [47]. These and other hypotheses could be tested by applying systems biology methods [48] and targeted ex vivo and in vitro assays to rigorously conducted prospective studies of Pf-exposed populations. We previously reported that Pf exposure is associated with a functionally and phenotypically distinct population of FCRL4+ hypo-responsive atypical MBCs [18], similar to the ‘exhausted’ MBCs described in HIV-infected individuals [19]. In this study, with a larger sample size, we confirmed that Pf exposure is associated with an expansion of FCRL4+ MBCs. The accumulation of atypical MBCs could be linked to the slow acquisition of Pf-specific MBCs, as naïve B cells in response to Pf infection could have a propensity to differentiate into atypical rather than classical MBCs. We also observed that the FCRL4+ MBC population decreased in the peripheral circulation two weeks after acute malaria suggesting that these MBCs are directly involved in the response to Pf infection, possibly trafficking to secondary lymphoid tissues. Although the function of FCRL4+ MBCs is not established, Moir et al. [19] suggested that FCRL4+ ‘exhausted MBCs’ contribute to the B cell deficiencies observed in HIV-infected individuals. In contrast, Ehrhardt et al.[49], who first described FCRL4+ ‘tissue-like MBCs’ in lymphoid tissues associated with epithelium, suggested that these cells may play a protective role during infections. At present, the factors that underlie the expansion of atypical MBCs in this study population are not known. Genetic or environmental factors that are associated with Pf transmission but not accounted for in this study could explain this observation. It will be of interest to understand the origin, antigen-specificity, and function of FCRL4+ MBCs in the context of Pf infection and the potential impact of these MBCs on the ability of children to respond to malaria vaccines. In multivariate analysis we found no correlation between the frequency of MBCs and levels of Abs specific for AMA1 or MSP1 and malaria risk. This is not necessarily unexpected in light of recent clinical trials that showed that vaccination with either AMA1 or MSP1 did not confer protection [20], [21]. Furthermore, we suspect that the frequency of MBCs per se may not reliably predict clinical immunity to malaria regardless of antigen specificity. Malaria symptoms only occur during the blood stages of Pf infection and can begin as early as three days after the blood stage infection begins [50].Because the differentiation of MBCs into PCs peaks ∼6–8 days after re-exposure to antigen [10], there may not be sufficient time for MBCs specific for Pf blood stage antigens to differentiate into the antibody-secreting cells that would prevent the onset of malaria symptoms. In contrast, the longer incubation period of other pathogens allows MBCs to differentiate into protective antibody-secreting cells before symptoms develop. For example, follow-up studies of hepatitis B vaccinees have shown that protection can persist despite the decline of hepatitis B-specific antibodies to undetectable levels [51], presumably due to the recall response of persistent MBCs. Thus, protection against the blood stages of malaria may depend on achieving and maintaining a critical level of circulating antibody that can rapidly neutralize the parasite. MBCs may contribute to the gradual acquisition of protective immunity by differentiating into LLPCs with each Pf infection. Here we also provide evidence concerning the mechanism by which MBCs and LLPCs are maintained. We observed a modest but statistically significant increase in TT-specific MBCs two weeks after acute malaria, in support of the hypothesis that MBCs are renewed by polyclonal or ‘bystander’ activation [10]. The stable frequency of TT-specific MBCs with age suggests that the small increases associated with Pf-induced polyclonal activation are matched by the rate of loss of senescent TT-specific MBCs. It has also been proposed that non-specific polyclonal stimulation maintains long-lived Ab responses by driving MBCs to differentiate into SLPCs or LLPCs [10]. Similarly, it has been hypothesized that Plasmodium infection generates large amounts of non-specific Ig [52] through polyclonal B cell activation [12], [13]. However, despite the presence of TT-specific MBCs and their expansion following Pf infection, we did not observe a concomitant increase in TT-specific IgG. This finding is consistent with recent human studies that demonstrate a lack of bystander IgG production after heterologous vaccination or viral infection [32], [53]; as well as studies in mice that demonstrate PC persistence after MBC depletion [54], and the failure of MBCs to differentiate into PCs in vivo upon TLR4 and 9 activation [55]. This finding does not represent an overt inability of TT-specific MBCs to differentiate into PCs, since adult females in this study had a sharp increase in tetanus IgG after a single tetanus booster. It is possible that bystander MBCs specific for antigens other than TT differentiate into PCs after Pf infection, but based on the results of this study we hypothesize that the preponderance of IgG produced in response to malaria is specific for the ∼2400 Pf proteins expressed during the blood-stage of infection [56], and that increases in ‘non-specific’ IgG reflect boosting of cross-reactive B cells [57], [58]. From a basic immunology perspective, these data support a model in which non-specific stimuli contribute to MBC self-renewal, but not to the maintenance of LLPCs. Studies of other Ab specificities and isotypes before and after malaria and other infections would test this hypothesis further. Although a recent mouse study showed that MBCs do not proliferate in vivo after immunization with an irrelevant antigen [59], this may reflect the difference in requirements for MBC maintenance in mammals with relatively short life spans. It is of general interest to determine which parasite products are responsible for the polyclonal activation of MBCs observed here. Studies in vitro suggest that Pf drives polyclonal MBC activation by the cysteine-rich interdomain regions 1α (CIDR1α) of the Pf erythrocyte membrane protein 1 (PfEMP1) [13], [60], but it is conceivable that Pf-derived TLR agonists [61], [62] or bystander T cell help [63], [64], [65] also contribute to MBC proliferation in the absence of BCR triggering [66]. Animal models have provided important insights into the immunobiology of Plasmodium infection [67], but ultimately, despite obvious experimental limitations, it is critical to investigate the human immune response to Pf in longitudinal studies since findings from animal models do not always mirror human biology or pertain to the clinical context [68], [69]. Key challenges for future studies will be to determine the molecular basis of the inefficient generation of MBCs and LLPCs in response to Pf infection and to determine the longevity of these cells in the absence of Pf transmission over longer periods of time. Greater insight into the molecular and cellular basis of naturally-acquired malaria immunity could open the door to strategies that ultimately prove useful to the development of a highly effective malaria vaccine. Materials and Methods Ethics statement The ethics committee of the Faculty of Medicine, Pharmacy, and Odonto-Stomatology, and the institutional review board at the National Institute of Allergy and Infectious Diseases, National Institutes of Health approved this study (NIAID protocol number 06-I-N147). Written, informed consent was obtained from adult participants and from the parents or guardians of participating children. Study site This study was carried out in Kambila, a small (<1 km2) rural village with a population of 1500, located 20 km north of Bamako, the capital of Mali. Pf transmission is seasonal and intense at this site from July through December. The entomological inoculation rate measured in a nearby village was approximately 50–60 infective bites per person per month in October 2000 and fell to near zero during the dry season [17]. A detailed description of this site and the design of the cohort study has been published elsewhere [14]. Sampling strategy, study participants, and malaria case definition In May 2006, during a two-week period just prior to the malaria season, 225 individuals aged 2–10 years and 18–25 years were enrolled after random selection from an age-stratified census of the entire village population. Enrollment exclusion criteria were hemoglobin <7 g/dL, fever ≥37.5°C, acute systemic illness, use of anti-malarial or immunosuppressive medications in the past 30 days, or pregnancy. All analysis in the present study pertains to an age-stratified subset of individuals (n = 185) randomly selected from those who had complete sets of PBMC samples over the entire study period. From May 2006 through May 2007, participants were instructed to report symptoms of malaria at the village health center, staffed 24 hours per day by a study physician. For individuals with signs or symptoms of malaria, blood smears were examined for the presence of Pf. Patients with positive smear results (i.e. any level of parasitemia) were treated with a standard 3-day course of artesunate plus amodiaquine, following the guidelines of the Mali National Malaria Control Program. Anti-malarial drugs were provided exclusively by the study investigators. Children with severe malaria were referred to Kati District Hospital after an initial parenteral dose of quinine. For research purposes, a malaria episode was defined as an axillary temperature ≥37.5°C, Pf asexual parasitemia ≥5000 parasites/µL, and a nonfocal physical examination by the study physician. Severe malaria, as defined by the WHO [15], was included in this definition. Three clinical endpoints were used to evaluate the relationship between Pf-specific immune responses and malaria risk: 1) whether or not malaria was experienced, 2) the incidence of malaria, and 3) the time to the first malaria episode. Blood smears were prepared and venous blood samples collected during the two-week enrollment period (month 0), 14 days after the first episode of malaria (convalescence), and during a two-week period at the end of the six-month dry season (month 12). Hemoglobin was typed from venous blood samples. Stool and urine were examined at enrollment for the presence of helminth infections. Venous blood samples from ten healthy U.S. adult blood bank donors were analyzed as controls. Travel histories for these U.S. adults were not available, but prior exposure to Pf is unlikely. PBMC and plasma collection Blood samples (8 ml for children and 16 ml for adults) were drawn by venipuncture into sodium citrate-containing cell preparation tubes (BD, Vacutainer CPT Tubes) and transported 20 km to the laboratory where they were processed within three hours of collection. Plasma and PBMCs were isolated according to the manufacturer's instructions. Plasma was stored at −80°C. PBMCs were frozen in fetal bovine serum (FBS) (Gibco, Grand Island, NY) containing 7.5% dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO), kept at −80°C for 24 hours, and then stored at −196°C in liquid nitrogen. For each individual, PBMC and plasma samples from all time points were thawed and assayed simultaneously. Measurement of peripheral blood Pf parasitemia Thick blood smears were stained with Giemsa and counted against 300 leukocytes. Pf densities were recorded as the number of asexual parasites/µl of whole blood, based on an average leukocyte count of 7500/µl. Each smear was evaluated separately by two expert microscopists blinded to the clinical status of study participants. Any discrepancies were resolved by a third expert microscopist. Hemoglobin typing Hemoglobin was typed by high performance liquid chromatography (HPLC; D-10 instrument; Bio-Rad, Hercules, CA) as previously described [14]. Stool and urine exam for helminth infection At enrollment, duplicate stool samples were examined for Schistosoma mansoni eggs and other intestinal helminths using the semi-quantitative Kato-Katz method. To detect Schistosoma haematobium eggs, 10 ml of urine were poured over Whatman filter paper. One or two drops of ninhydrine were placed on the filter and left to air dry. After drying, the filter was dampened with tap water and helminths were eggs detected by microscopy. Geographic information system data collection Latitude and longitude coordinates of study subjects' households were measured by a handheld global positioning system receiver (GeoXM; Trimble) and reported earlier [14]. Antibody detection by ELISA ELISAs were performed by a standardized method as described previously [70]. For both AMA1 and MSP1, a 1∶1 mixture of FVO and 3D7 AMA1 and MSP1 isotypes was used to coat the ELISA plates. The limit of detection for the AMA1 and MSP1 ELISA is based on the range of values that gives reproducible results at the Malaria Vaccine and Development Branch at NIAID where the assay is routinely performed. More specifically, the limit of detection is the ELISA unit value at the lowest point on the standard curve, multiplied by the dilution factor at which samples are tested. The minimal detection levels for the MSP1 and AMA1 ELISA assays were 11 and 33 ELISA units, respectively. For analysis, all data below the minimum detection level were assigned a value of one half the limit of detection (i.e. 6 units for MSP1, 17 units for AMA1). The limit of detection for the TT ELISA was not determined because we did not have access to TT-naïve serum. Memory B cell analysis Antigen-specific MBCs were quantified by a modified version of the method developed by Crotty et al [71]. We found that adding IL-10 to the cocktail of polyclonal activators resulted in a six-fold increase in the efficiency of the assay (Weiss et al., unpublished observation). Briefly, PBMCs were thawed and cultured in 24 well plates at 37°C in a 5% CO2 atmosphere for six days in media alone (RPMI 1640 with L-Glutamine, Penicillin/Streptomycin 100 IU/ml, 10% heat-inactivated FBS, 50 µM β-Mercaptoethanol) or media plus a cocktail of polyclonal activators: 2.5 µg/ml of CpG oligonucleotide ODN-2006 (Eurofins MWG/Operon, Huntsville, AL), Protein A from Staphylococcus aureus Cowan (SAC) at a 1/10,000 dilution (Sigma-Aldrich, St. Louis, MO), pokeweed mitogen at a 1/100,000 dilution (Sigma-Aldrich), and IL-10 at 25 ng/ml (BD Biosciences). Cells were washed and distributed on 96-well ELISPOT plates (Millipore Multiscreen HTS IP Sterile plate 0.45 um, hydrophobic, high-protein binding) to detect antibody-secreting cells (ASCs). ELISPOT plates were prepared by coating with either: a 10 µg/ml solution of polyclonal goat antibodies specific for human IgG (Caltag) to detect all IgG-secreting cells; a 1% solution of bovine serum albumin (BSA) as a non-specific protein control; or 5 µg/ml solutions of either tetanus toxoid (TT), AMA1, or MSP1 to detect antigen-specific ASCs. For AMA1 and MSP1, a 1∶1 mixture of FVO and 3D7 isotypes was used to coat the ELISPOT plates. Plates were blocked by incubation with a solution of 1% BSA. For the detection of antigen-specific ASCs, cells were plated in duplicate in eight serial dilutions beginning with 5×105 cells/well. For detection of total IgG ASCs cells were plated at six serial dilutions beginning at 4×104 cells/well. After a five hour incubation of the cells in the ELISPOT plates, plates were washed four times each in PBS and PBS-Tween 20 0.05% (PBST), and incubated overnight with a 1∶1000 dilution of alkaline phosphatase-conjugated goat antibodies specific for human IgG (Zymed) in PBST/1% FCS. Plates were washed four times each in PBST, PBS, and ddH2O; developed using 100 µl/well BCIP/NBT for 10 minutes; washed thoroughly with ddH2O and dried in the dark. ELISPOTS were quantified using Cellular Technologies LTD plate-reader and results analyzed using Cellspot software. Results are reported as frequencies of MBCs per 106 PBMCs after the six-day culture. The limit of detection of the MBC ELISPOT assay for this analysis was five ASCs per 106 PBMC based on the average number of ASCs on the BSA control. Assay failure was defined as fewer than 1000 IgG+ ASCs per 106 PBMCs after the six-day culture which resulted in the exclusion of 15% of individuals at month 0, 13.2% 14 days after the first malaria episode, and 7.3% at month 12. For individuals with a limited number of PBMCs, priority was given to performing the ELISPOT assay for MSP1, then TT, and then AMA1. Phenotypic analysis of B cell subsets All phenotypic analyses were performed using mouse mAbs specific for human B cell markers conjugated to fluorophores as previously reported [18]. Fluorophore-conjugated mAbs specific for the following markers were used: PECy7-CD19, PE-CD20, APC-CD10, APC-CD27, PE-IgG (BD Biosciences, San Jose, CA) and FITC-CD21 (Beckman Coulter, Fullerton, CA). A four-color, two-stain strategy was used to identify B cell subsets as follows: plasma cells/blasts (CD19+ CD21− CD20−), naive B cells (CD19+ CD27−CD10−), immature B cells (CD19+ CD10+), classical MBCs (CD19+ CD27+ CD21+), atypical MBCs (CD19+ CD21− CD27− CD10−) and activated MBCs (CD19+ CD21− CD27+CD20+). FACS analyses were performed on a FACSCalibur flow cytometer (BD Biosciences) using FlowJo software (Tree Star, Ashland, OR). Statistical analysis Data were analyzed using STATA (StataCorp LP, Release 10.0) and GraphPad Prism for Windows (GraphPad Software, version 5.01).The Kruskal-Wallis test was used to compare continuous variables between groups, and the Fisher's exact test was used to compare categorical variables. The Wilcoxon matched-pairs signed-rank test was used to compare measurements of the same parameter at two time points for the same individual. The correlation between different continuous measures was determined by using the Spearman correlation coefficient. The malaria-free probability over the twelve-month study period was estimated by the Kaplan-Meier curve, and the time to the first malaria episode was compared by the log rank test. Cox's proportional hazards model was used to assess the effect of the following factors on the hazard of malaria: age, gender, weight, ethnicity, distance lived from study clinic, self-reported bednet use, hemoglobin type, antigen-specific MBC frequencies and Ab levels. The same list of variables was included in logistic and Poisson regression models to determine their impact on the odds and incidence of malaria episodes, respectively. For all tests, two-tailed p values were considered significant if ≤0.05. Supporting Information Figure S1 Correlative analysis of antibody levels and memory B cell frequencies specific for AMA1, MSP1, and tetanus toxoid. Shown are scatterplots of antibody levels versus memory B cell frequencies specific for (A) AMA1 (n = 64), (B) MSP1 (n = 67) and (C) tetanus toxoid (n = 128). Data are derived from venous blood samples drawn before the malaria season. Only individuals with both antibody and memory B cell data are included. For AMA1 and MSP1 the plots include individuals with antibody levels at or above the limit of detection of the ELISA. Individuals with ‘failed' ELISPOT assays are not included. As described in ‘Materials and Methods', assay failure was defined as fewer than 1000 IgG+ ASCs per 106 PBMCs after the six-day culture. The Spearman's correlation coefficient is given for each plot. (3.84 MB TIF) Click here for additional data file.
- Record: found
- Abstract: found
- Article: found
Genetic polymorphisms linked to susceptibility to malaria
Adel Driss, Jacqueline Hibbert, Nana O. Wilson … (2011)
The influence of host genetics on susceptibility to Plasmodium falciparum malaria has been extensively studied over the past twenty years. It is now clear that malaria parasites have imposed strong selective forces on the human genome in endemic regions. Different genes have been identified that are associated with different malaria related phenotypes. Factors that promote severity of malaria include parasitaemia, parasite induced inflammation, anaemia and sequestration of parasitized erythrocytes in brain microvasculature. Recent advances in human genome research technologies such as genome-wide association studies (GWAS) and fine genotyping tools have enabled the discovery of several genetic polymorphisms and biomarkers that warrant further study in host-parasite interactions. This review describes and discusses human gene polymorphisms identified thus far that have been shown to be associated with susceptibility or resistance to P. falciparum malaria. Although some polymorphisms play significant roles in susceptibility to malaria, several findings are inconclusive and contradictory and must be considered with caution. The discovery of genetic markers associated with different malaria phenotypes will help elucidate the pathophysiology of malaria and enable development of interventions or cures. Diversity in human populations as well as environmental effects can influence the clinical heterogeneity of malaria, thus warranting further investigations with a goal of developing new interventions, therapies and better management against malaria.
Author and article information
Journal
Journal ID (nlm-ta): JAMA Netw Open
Journal ID (iso-abbrev): JAMA Netw Open
Journal ID (pmc): JAMA Netw Open
Title:
JAMA Network Open
Publisher:
American Medical Association
ISSN
(Electronic):
2574-3805
Publication date
(Electronic):
5
April
2019
Publication date Collection:
April
2019
Publication date PMC-release: 5
April
2019
Volume: 2
Issue: 4
Electronic Location Identifier: e191691
Affiliations
[1
]Sorbonne Université, Institut National de
la Santé et de la Recherche Médicale (INSERM), Institut Pierre–Louis
d’Epidémiologie et de Santé Publique, Assistance Publique
Hôpitaux de Paris (AP-HP), Centre National de Référence du Paludisme,
Hôpital Pitié–Salpêtrière, France
[2
]Sorbonne Université, Institut de Recherche
pour le Développement, AP-HP, Centre National de Référence du
Paludisme, Hôpital Bichât Claude–Bernard, Paris, France
Author notes
Article Information
Open Access: This is an open access
article distributed under the terms of the
CC-BY License. © 2019 Kendjo E et al.
JAMA Network
Open
.
Corresponding Author: Eric Kendjo, MSc,
Assistance Publique–Hôpitaux de Paris (AP-HP), Centre National de
Référence du Paludisme, Service de Parasitologie–Mycologie,
Hôpital Pitié-Salpêtrière; 47 boulevard de l'hôpital,
75013 Paris, Cedex 13, France (
eric.kendjo@
123456gmail.com
).
Author Contributions: Mr Kendjo
had full access to all of the data in the study and takes responsibility for the
integrity of the data and the accuracy of the data analysis.
Acquisition, analysis, or interpretation of data: Kendjo,
Houzé, Mouri, Taieb, Jauréguiberry, Tantaoui, Ndour, Buffet, M.
Piarroux, Thellier, R. Piarroux.
Critical revision of the manuscript for important intellectual
content:
Kendjo, Houzé, Gay, Jauréguiberry, Tantaoui,
Ndour, Buffet, M. Piarroux, Thellier, R. Piarroux.
Administrative, technical, or material support: Kendjo,
Jauréguiberry, Tantaoui, Ndour, Buffet, Thellier, R. Piarroux.
Conflict of Interest Disclosures: Drs
Houzé and Thellier reported grants from Santé Publique France during
the conduct of the study. Drs Buffet, Jauréguiberry, Ndour, and Thellier
reported grants from Guilin Pharmaceuticals during the conduct of the study. Dr
Jauréguiberry also reported grants from Walter Reed Army Institute of
Research during the conduct of the study. Dr Ndour also reported grants from
Institut National de la Santé et de la Recherche Médicale during the
conduct of the study. No other disclosures were reported.
Funding/Support: This project was
funded by the French Institute for Public Health Surveillance, Santé
Publique France.
Role of the Funder/Sponsor: Santé Publique France had no role
in the design or the conduct of the study; collection, management, analysis, or
interpretation of the data; preparation, review, or approval of the manuscript;
and decision to submit the manuscript for publication.
Group Information: Members of
the French Imported Malaria Study group are: Christophe Strady (CHU Reims),
Caroline Lohmann (CH du Moenchsberg, Mulhouse), Céline Arriuberge (CH
Trousseau, Paris), Emmanuel Grimprel (CH Trousseau, Paris), Jean-Marie
Délabre (CH du Moenchsberg, Mulhouse), Michel Thibault (CH René Dubos,
Pontoise), Mohamadou Niang (CHR Orléans), Alain Barrans (CH Sète),
Antoine Martin (CH Périgueux), André Spiegel (DESP Nord), Alexis
Valentin (CHU Toulouse), Anne-Sophie Le Guern (Institut Pasteur, Paris), Adela
Angoulvant (CHU Kremlin-Bicêtre, Paris), Adeline Dubois (CH Alès),
Adrien Genin (CH Pays d'Aix), Agathe Lebuisson (CHU Cochin), Agnès Riche
(CH Angoulême), Agnès Durand (Institut Pasteur, Paris), Agnès
Fromont (CH Auxerre), Ahmed Aboubacar (CHU de Strasbourg), Ahmed Fateh Ousser
(CH Louis Mourier), Aida Taieb (INTS, Paris), Alain Domergue (CH Alès -
Cévennes), Alain Gravet (CH du Moenchsberg, Mulhouse), Alain Lecoustumier
(CH de Cahors), Albert Faye (CHU Robert Debré, Paris), Alexander Pfaff (CHU
de Strasbourg), Alexandra Faussart (CHU Bichat-Claude Bernard, Paris), Alexandre
Chlilek (CHU Nîmes), Alice Borel (CHU Amiens), Alice Pérignon (CHU
Pitié-Salpêtrière, Paris), Ana Mendes-Mreira (CH La Rochelle),
André Gardrat (CH d’Evreux), Ange Kissila (CH Provins), Angèle
LI (CH Creil, Laënnec), Anne Cady (CH Bretagne Atlantique), Anne
Debourgogne (CHU de Nancy), Anne Delaval (CHI Robert Ballanger,
Aulnay-sous-Bois), Anne Goepp (CHI Villeneuve St Georges), Anne Marfaing-Koka
(Hôpital Antoine-Béclère), Anne Pauline Bellanger (CHU
Besançon, Jean Minjoz), Anne Vincenot-Blouin (CH Meaux), Anne-Marie
Teychene-Coutet (CH Bondy-Jean Verdier), Anne-Sophie Deleplancque (CH Lille),
Annick Verhaeghe (CH de Dunkerque), Annie Motard-Picheloup (CHI Fréjus St
Raphaël), Antoine Berry (CHU Toulouse), Antoine Huguenin (CHU Reims),
Arnaud Bouvet (CH Bretagne Atlantique), Audrey Merens (HIA Begin), Aurelie Roide
(CHU Lariboisière, Paris), Aurore Sanson (CH Jacques Cœur, Bourges),
Aurélie Fricot (CHU Necker), Aurélie Guigon (CHR Orléans),
Benfatallah Dhouha (CHU Necker, Paris), Benjamin Wyplosz (CHU
Kremlin-Bicêtre, Paris), Benoît Henri (INTS, Paris), Bernadette Buret
(CH Niort), Bernadette Cuisenier (CHU Dijon), Bernadette Worms (CHU Dijon),
Bernard Faugère (CH Timone, Marseille), Biligui Sylvestre (CHU
Pitié-Salpêtrière, Paris), Boualem Sendid (CH Lille), Bruno
Megarbane (CHU Lariboisière, Paris), Bruno Pradine (IMTSSA, Marseille),
Béatrice Quinet (CH Trousseau, Paris), Christine Braidy (CH du Sud Seine et
Marne, Fontainebleau), Cécile Farrugia (CH de Dourdan), C. Finot (CH de
Dreux), Camille Roussel (INTS, Paris), Camille Runel-Belliard (CHU de Bordeaux),
Caren Brump (CHU Lariboisière, Paris), Carine Dokoula (CH Jacques
Cœur, Bourges), Carmina Camal (CH Louis Mourier), Carole Mackosso (CHU
Bichat-Claude Bernard, Paris), Carole Poupon (CH de Gonesse), Caroline Garandeau
(CH Angoulême), Catherine Benoit (CH du Sud Seine et Marne), Catherine
Branger (CH Louis Mourier), Catherine Brehant (CH La Rochelle), Catherine
Desideri-Vaillant (HIA Clermont Tonnerre, Brest), Catherine Kauffmann Lacroix
(CH Poitiers), Catherine Lafaurie (CH d’Epernay), Cécile
Hombrouck-Alet (CH Blois), Cécile Ramade (Lyon-Croix-Rousse), Céline
Damiani (CHU Amiens), Céline Gourmel (CHU Lariboisière, Paris),
Chantal Duhamel (CHU Côte de Nacre), Chantal Garabedian (CH Pays d'Aix),
Chralotte Chambrion (INTS, Paris), Christelle Morelle (CHU Montpellier),
Christelle Pomares Estran (CH Universitaire de Nice), Christelle Prince (CH de
Cayenne Andrée Rosemon), Christian Durand (CH Provins), Christian Fulleda
(CHU Lariboisière, Paris), Christian Raccurt (CHU Amiens), Christine
Chaigneau (GHPSO, Creil), Christine Chemla (CHRU de Reims), Christine Van batten
(CH Laënnec, Creil), Christophe Martinaud (HIA Percy, Clamart), Christophe
Rapp (HIA Begin), Claire Augé (CHU Bichat-Claude Bernard, Paris), Claire
Malbrunot (CH Corbeil Essonne), Claudine Febvre (CH de Montbéliard),
Claudine Sarfati (Hôpital Saint-Louis, Paris), Coralie l'Ollivier (CH de la
Timone, Marseille), Corinne Huet (Hôpital Louis-Pasteur,
Cherbourg-Octeville), Cournac Jean-Marie (HIA Percy, Clamart), Cynthia Pianetti
(CH Gabriel Martin, La Réunion), Cécile Angebault (CHU Necker, Paris),
Cécile Ficko (HIA Begin), Cécile Garnaud (CHU de Grenoble),
Cécile Leprince (CHI Robert Ballanger, Aulnay-sous-Bois), Céliat Merat
(CHU Nantes), Céline Dard (CHU de Grenoble), Céline Nourrisson (CHRU
Clermont-Ferrand), Céline Tournus (Hôpital Delafontaine, Saint-Denis),
Daniel Azjenberg (CHU Dupuytren, Limoges), Daniel Camus (CH Lille), Daniel
Lusina (CHI Robert Ballanger, Aulnay-sous-Bois), Daniel Parzy (IMTSSA,
Marseille), Denis Pons (CHRU Clermont-Ferrand), Denis Filisetti (CHU
Strasbourg), Denis Malvy (CHU de Bordeaux), Didier Basset (CHU Montpellier),
Didier Jan (CH Laval), Didier Poisson (CHR Orléans), Didier Raffenot (CH
Chambéry), Dieudonné Bemba (CH Bondy-Jean Verdier), Dominique Maubon
(CHU de Grenoble), Dominique Mazier (CHU Pitié-Salpêtrière,
Paris), Dominique Popjora (CH Trousseau, Paris), Dominique Toubas (CHRU de
Reims), Dorothée Quino (CHRU Morvan, Brest), Alioune Ndour (INTS, Paris),
Ducout Louis (CH de la Côte Basque), Duong Thanh Hai (CHRU Bretonneau), E.
Boyer (CH Le Mans), Edgar Ombandza (CH Provins), Edith Mazars (CH de
Valenciennes), Elisabeth Buffet (CH de Epernay), Elodie Collin (CHI Robert
Ballanger, Aulnay-sous-Bois), Elodie Meynet (CH Annecy Genevois), Emeline
Scherer (CHU Besançon, Jean Minjoz), Emilie Fréalle (CH Lille), Emilie
Klein (CHU Lariboisière, Paris), Emilie Sitterle (CHU Necker, Paris), Emily
Ronez (CHU Lariboisière, Paris), Emmanuel Dutoit (CH Lille), Enrique
Casalino (CHU Bichat-Claude Bernard, Paris), Eric Caumes (CHU
Pitié-Salpêtrière, Paris), Eric Dannaoui (Hôpital
Européen Georges Pompidou, Paris), Eric Gardien (CH de Draguignan,
Bordeaux), Eric Kendjo (CHU Pitié-Salpêtrière, Paris), Eric
d'Ortenzio (CHU Bichat-Claude Bernard, Paris), Ermanno Candolfi (CHU de
Strasbourg), Estelle Perraud-Cateau (CH Poitiers), Eterne Twizeyimana (CH du
Cotentin), F. Roblot (CH Poitiers), Fabienne Pateyron (CH Provins), Fabrice
Bruneel (CH de Versailles, André Mignot), Fabrice Legros (CNR du
Paludisme), Fabrice Simon (HIA Laveran), Fakhri Jeddi (CHU Nantes), Farida M.
Benaoudia (CH Troyes), Faïzi Ajana (CH Tourcoing), Felix Djossou (CH de
Cayenne Andrée Rosemon), Firouze Banisadr (CHRU de Reims), Florent Morio
(CHU Nantes), Francis Derouin (Hôpital Saint-Louis, Paris), Francois
Moussel (CH François-Quesnay, Mantes-La-Jolie), Francoise Foulet (CHU Henri
Mondor), François Peyron (Lyon-Croix-Rousse), Françoise Benoit-Vical
(CHU Toulouse), Françoise Botterel (CHU Henri Mondor), Françoise
Gayandrieu (CHU Nantes), Françoise Schmitt (CH du Moenchsberg, Mulhouse),
Frederic Ariey (CHU Cochin, Paris), Frédéric Grenouillet (CHU Jean
Minjoz, Besançon), Frédéric Sorge (CHU Necker),
Frédérique Gay (CHU Pitié-Salpêtrière, Paris),
Frédérique Foudrinier (CHRU de Reims), G. Courrouble (CH Blois),
Gildas Gallou (CH de Falaise), Généviève Julienne (CH Belfort),
G. Philippon (Centre Médical CMETE, Paris), Gauthier Pean-de-Ponfilly (CHU
Lariboisière, Paris), Geneviève Grise (CH d’Elbeuf), Ghania
Belkacem Belkadi (CH Tenon), Gilbert Lorre (CHD La Roche-sur-Yon), Gilles
Gargala (CHU Rouen), Gilles Nevez (CHRU Morvan, Brest), Gisele Dewulf (CH de
Valenciennes), Guillaume Désoubeaux (CHRU Bretonneau, Tours), Guillaume
Escriou (CHU Bichat-Claude Bernard, Paris), Guillaume Le Loup (CH Tenon, Paris),
Guillaume Menard (HIA Saint-Anne, Toulon), Guy Carroger (CH Jacques Cœur,
Bourges), Guy Galeazzi (CH Louis Mourier), Gwénaël le Moal (CH
Poitiers), Hana Talabani (CHU Cochin, Paris), Hanene Abid (CHU Necker, Paris),
Helene Broutier (CHI Robert Ballanger, Aulnay-sous-Bois), Herve Pelloux (CHU de
Grenoble), Houria Ichou (CH Louis Mourier), Hugo Laurent (CHU Lariboisière,
Paris), Hélène Broutier (CH Meaux), Hélène Lapillonne (CH
Trousseau, Paris), Hélène Yera (CHU Cochin, Paris), Hélène
Savini (HIA Laveran), Isabelle Hermès (CH Saint-Malo), Ilhame Tantaoui (CHU
Pitié-Salpêtrière, Paris), Isabelle Poilane (CH Bondy-Jean
Verdier), Isabelle Amouroux (Hôpital Antoine-Béclère), Isabelle
Mazurier (Hôpitaux Civils de Colmar), Isabelle Salimbeni (CH de Cannes),
Isabelle Tawa (Centre Médical CMETE, Paris), Joseph Cuziat (CH
Saint-Nazaire), Jean-Bernard Poux (CH de Val d'Ariège - Foix), J. Heurtet
(CH Beauvais), J. Rome (CH de Fougères), Jennifer Truchot (CHU
Lariboisière, Paris), Jean-Marc Segalin (CHR Orleans), Jacques Gaillat (CH
Annecy Genevois), Jacques Le bras (CHU Bichat-Claude Bernard, Paris), Jacques
Thevenot (Centre Médical CMETE, Paris), Jacques Vaucel (CH Saint-Brieuc),
Jean Dunand (Hôpital Ambroise Paré), Jean-Benjamin Murat (CH de
Roanne), Jean-Marie Trapateau (CH Angoulême), Jean-Yves Peltier (CHI
Poissy-St-Germain), Jean-Etienne Pilo (HIA Begin), Jean-Francois Magnaval (CHU
Toulouse), Jean-François Faucher (CHU Jean Minjoz, Limoge), Jean-Paul
Boutin (DESP Sud), Jean-Paul Couaillac (CH de Cahors), Jean-Philippe Breux (CH
Cholet), Jean-Pierre Hurst (CH Jacques Monod, Le Havre), Jean-Yves Siriez (CHU
Robert Debré, Paris), Jean-philippe Bouchara (CHU Angers), Jérôme
Clain (CHU Bichat-Claude Bernard, Paris), Jérôme Naudin (CHU Robert
Debré, Paris), Jordan Leroy (CH Lille), Josette Jehan (CH du Cotentin),
Joudia Najid (CHU Pitié-Salpêtrière, Paris), Judith Gorlicki (CHU
Lariboisière, Paris), Julie Bonhomme (CHU Côte de Nacre), Julie Brunet
(CHU de Strasbourg), Jérôme Guinard (CHR Orleans), Karima Cheikh (CHU
Henri Mondor), Laurence Pougnet (HIA Clermont Tonnerre, Brest), Lauren Pull (CHU
Robert Debré, Paris), Laurence Millon (CHU Jean Minjoz, Besançon),
Laurence Campergue-Mayer (CH Avignon), Laurence Estepa (CH Blois), Laurence
Lachaud (CHU Nîmes), Laurent Aaron (CH Jacques Cœur, Bourges), Laurent
Bret (CHR Orléans), Laurent Guillaume (CH Blois), Liliane Cicéron (CHU
Pitié-Salpêtrière, Paris), Lionnel Bertaux (CNR du Paludisme),
Lise Musset (Institut Pasteur, Guyane), Louise Basmacyan (CHU Dijon), Loïc
Favennec (CHU Rouen), Luce Landraud (CH Louis Mourier), Lucile Cadot (CH
Alès - Cévennes), Ludovic de Gentile (CHU Angers), Luis Macias (CHU
Bichat-Claude Bernard, Paris), Luu-ly Pham (CHU Kremlin-Bicêtre, Paris), M.
Cambon (CHRU Clermont-Ferrand), Marie-France Biava (CHU de Nancy),
Marie-Hélène Kiefer (CH du Moenchsberg), M.P. Carlotti (CNR du
Paludisme), Madeleine Fontrouge (CH de Gonesse), Marc Pihet (CHU Angers), Marc
Thellier (CHU Pitié-Salpêtrière, Paris), Marie-Catherine Receveur
(CHU de Bordeaux), Marie-Claire Machouart (CHU de Nancy), Marie-Elisabeth
Bougnoux (CHU Necker, Paris), Marie-Laure Bigel (CH François-Quesnay,
Mantes-la-Jolie), Marie-Laure Darde (CHU Dupuyrien, Limoges), Marie-Nadège
Bachelier (CH Jacques Cœur, Bourges), Marion Almeras (CH Béziers),
Marion Leterrier (CHD La Roche-sur-Yon), Martin Danis (CHU
Pitié-Salpêtrière, Paris), Géraldine Martin (CH du
Cotentin), Martine Bloch (CH Louis Mourier), Martine Liance (CHU Henri Mondor,
Paris), Marylin Madamet (IMTSSA, Marseille), Matthieu Revest (CHU Pontchaillou,
Rennes), Matthieu Mechain (CHU de Bordeaux), Maxime Thouvenin (CH Troyes),
Mermond Sylvain (Institut Pasteur, Nouméa), Michel Develoux (CH Tenon,
Paris), Michel Miegeville (CHU Nantes), Milène Sasso (CHU Nîmes),
Mohamed Diaby (CH Vernon), Monique Marty (CH La Rochelle), Monique Greze (CH
Albi), Monique Lemoine (CHU Bichat-Claude Bernard, Paris), Mouri Oussama (CHU
Pitié-Salpêtrière, Paris), Muriel Cornet (Hôpital
Hôtel-Dieu, Paris), Muriel Mimoun Ayache (CH Trousseau), Muriel Nicolas
(CHU Pointe-à-Pitre / Abymes), Muriel Roumier (CH Arles), Muriel Silva (CH
Jacques Monod), Mylène Penot (CERBA), Myriam Gharbi (CHU Bichat-Claude
Bernard, Paris), Nadia Guennouni (CHU Bichat-Claude Bernard, Paris), Nadine
Godineau (Hôpital Delafontaine, Saint-Denis), Naima Dahane (CHU Cochin,
Paris), Nathalie Bourgeois (CHU Montpellier), Nathalie Désuremain (CH
Trousseau, Paris), Nathalie Fauchet (CHI de Créteil), Nathalie Parez (CH
Louis Mourier), Nathalie Wilhelm (CH de Cahors), Nawel Ait-Ammar (Hôpital
Ambroise Paré), Nayla Nassar (CH Auxerre), Nicolas Argy (CHU Bichat-Claude
Bernard, Paris), Nicolas Blondiaux (CH Tourcoing), Nicolas Taudon (CERBA),
Nicole Desbois-Nogard (CHU de la Martinique), Noura Hassouni (CHU Necker), Odile
Bouret-Dubouis (CH Bretagne Atlantique), Odile Eloy (CH de Versailles,
André Mignot), Odile Falguiere (CH Béziers), Odile Fenneteau (CHU
Robert Debré, Paris), Olivia Bandin (Hôpital
Saint-Camille/Bry-sur-Marne), Olivier Albert (CHU de Bordeaux), Olivier Bouchaud
(CH Bobigny-Avicenne), Olivier Patey (CHI Villeneuve St Georges), Olivier
Rogeaux (CH Chambéry), Philippe Clergeau (CH Sallanches), P. Daumain (CH de
Dourdan), Paul-Henry Consigny (Institut Pasteur, Paris), Paméla Chauvin
(CHU Toulouse), Pascal Delaunay (CH Universitaire de Nice), Pascal Hazera (CH
Saint-Lo), Pascal Houzé (Hôpital Saint Louis, Paris), Pascal Millet
(CHU de Bordeaux), Pascal Pouedras (CH Bretagne Atlantique), Pascale Penn (CH Le
Mans), Patrice Agnamey (CHU Amiens), Patrice Bourrée (CHU
Kremlin-Bicêtre, Paris), Patricia Barbut (CH Longjumeau), Patricia Brugel
(CH Antibes Juan-Les-Pins), Patricia Roux (CH Saint-Antoine, Paris), Patrick
Leguen (HIA Clermont Tonnerre, Brest), Patrick Valayer (CH Notre-Dame de la
Miséricorde), Pauline Caraux-Paz (CHI Villeneuve St Georges), Pauline
Touroultjupin (CH Cholet), Philippe Abboud (CHU Rouen), Philippe Cormier (CH
d’Evry), Philippe Minodier (CH Marseille Nord), Philippe Moskovtchenko
(Hôpitaux Civils de Colmar), Philippe Parola (CH Marseille Nord), Philippe
Poirier (CHRU Clermont-Ferrand), Philippe Stolidi (CH Aubagne), Pierre Patoz (CH
Tourcoing), Pierre Buffet (INTS, Paris), Pierre Buffet (CHU
Pitié-Salpêtrière, Paris), Pierre Flori (CH Saint-Etienne),
Pierre Marty (CH Universitaire de Nice), Pierre Mornand (CH Trousseau, Paris),
Pinel Claudine (CHU de Grenoble), Richard Dahan (CHU de Strasbourg), Rémi
Devallière (CH Saint-Nazaire), Richard Mazataud (CH Vitry le
François), Rahaf Haj Hamid (CH Louis Mourier), Régis Courtin (CHU
Pitié-Salpêtrière, Paris), Renaud Blonde (CHU Robert Debré,
Paris), René Nabias (CHI Poissy-St-Germain), Roland Fabre (HIA Begin),
Rose-Anne Lavergne (CHU Nantes), Roxane Courtois (CH Cholet), Rym Chouk Turki
(CHU Henri Mondor), Rémy Durand (CH Bobigny-Avicenne), Réné
Nabias (CHU Necker, Paris), Sabah Kubab (CH Corbeil Essonne), Sabine Lasserre
(CH Trousseau, Paris), Samia Hamane (Hôpital Saint-Louis, Paris), Sandrine
Cojean (CHU Bichat-Claude Bernard, Paris), Sandrine Houzé (CHU
Bichat-Claude Bernard, Paris), Sophie Matheron (CHU Bichat-Claude Bernard,
Paris), Sorya Belaz (CHU Pontchaillou, Rennes), Stephane Jauréguiberry (CHU
Pitié-Salpêtrière, Paris), Stephane Ranque (CH de la Timone,
Marseille), Stephanie Dulucq (CHU de Bordeaux), Stéphane Bretagne
(Hôpital Saint-Louis, Paris), Stéphane Pelleau (Institut Pasteur,
Guyane), Stéphane Picot (Hospices Civils de Lyon), Sylvain Clauser
(Hôpital Ambroise Paré), Sylviane Chevrier (CHU Pontchaillou, Rennes),
Sylviane Dydymski (CHRU Clermont-Ferrand), Sylvie Lariven (CHU Bichat-Claude
Bernard, Paris), Sylvie Lhopital (CH Vernon), Sylvie Maurellet Evrard (CHI
Villeneuve St Georges), Sylvie Roulaud (CH Angouleme), Sébastien
Larréché (HIA Begin), Thi-Hai-Chau Trinh (CHR Orléans), Thierry
Ancelle (CHU Cochin, Paris), Thierry Pistone (CHU de Bordeaux), Thomas Hanslik
(Hôpital Ambroise Paré), Thomas Guimard (CHD La Roche-sur-Yon),
Timothée Klopfenstein (CHU Besançon, Jean Minjoz), Valerie
Fuster-Dumas (CHU de Bordeaux), Veronique Blanc-Amrane (CH Antibes
Juan-Les-Pins), Veronique Delcey (CHU Lariboisière, Paris), Veronique
Sarrasin-Hubert (CHU Bichat-Claude Bernard, Paris), Vincent Foissaud (HIA Percy,
Clamart), Virginie Mouton-Rioux (CH Bretagne Atlantique), Virginie Vitrat (CH
Annecy Genevois), Véronique Jan-Lasserre (CH Lagny-sur-Marne), Xavier
Nicolas (HIA Clermont Tonnerre, Brest), Yannick Costa (CH Lagny-sur-Marne),
Yassamine Lazrek (Institut Pasteur, Guyane), Yaye Senghor (Hôpital Saint
Joseph, Paris), Yohann Le Govic (CHU Angers), Yves Guimard (CH Jacques
Cœur, Bourges), Yves Poinsignon (CH Bretagne Atlantique), Claude Flamand
(Institut Pasteur, Guyane), C.Nguyen (CH Trousseau, Paris), G. Noël (CH
Marseille Nord), Georges Soula (CH Marseille Nord), J.M.Didier (CH Vesoul),
Marie-Francois Raynaud (CH Antibes Juan-Les-Pins), M. Julien (CH Béziers),
Marc Morillon (HIA Laveran), Marie-Paule Carlotti (IMTSSA), Pascal Chantelat
(CHI Vesoul), Pascale Dussert (CH Belfort), Pascal Ralaimazava (CH
Bobigny-Avicenne), S. Zaouche (CHU Necker, Paris), Élodie Lesteven (CHU
Lariboisière, Paris).
Article
Publisher ID: zoi190081DOI: 10.1001/jamanetworkopen.2019.1691
PMC ID: 6523451
PubMed ID: 30951158
SO-VID: 0ce4b288-be9d-446b-b752-b74fc66ed0dd
Copyright © Copyright 2019 Kendjo E et al.
JAMA Network
Open
.
License:
This is an open access article distributed under the terms of the CC-BY License.
History
Date
received
: 20
September
2018
Date: 8
February
2019
Date
accepted
: 15
February
2019
Categories
Subject:
Research
Subject:
Original Investigation
Subject:
Online Only
Subject:
Infectious Diseases
Data availability: