Interaction of mycobacteria with Plasmin(ogen) affects phagocytosis and granuloma development.

Plasminogen and plasmin are fundamental components of the fibrinolytic system that interact with microorganisms generating different immunopathological effects. The molecules of Mycobacterium tuberculosis interplaying with plasminogen have already been identified and characterized. In this work, we studied the effects of plasmin(ogen) bound toMycobacterium bovisCalmette-Guérin (BCG) on phagocytosis in THP1 macrophages as well as in granuloma formation and development on in vitrohuman granuloma model. For this purpose, BCG was coated with plasminogen and plasmin, obtained after activation of zymogen by tissue plasminogen activator. The results showed a significant reduction in the number of bacteria phagocytosed by macrophages in presence of plasminogen or plasmin on BCG surface. On the other hand, at 3 days BCG/plasminogen/plasmin induced an increase granuloma numbers with respect to those induced by uncoated bacteria. BCG/plasminogen/environments also showed a significant increase of IL-6 secretion. At 7 days, a reduced number of granulomas and an increased number of bacteria was observed with respect to uncoated BCG environment. Altogether, these results showed that plasmin(ogen) on the mycobacterial surface affects phagocytosis, granuloma development and the cytokine context, thus resulting in an increased number of bacteria in granulomas.

High-content screening technology combined with a human granuloma model as a new approach to evaluate the activities of drugs against Mycobacterium tuberculosis.

Tuberculosis remains a major health problem due to the emergence of drug-resistant strains of Mycobacterium tuberculosis. Some models have provided valuable information about drug resistance and efficacy; however, the translation of these results into effective human treatments has mostly proven unsuccessful. In this study, we adapted high-content screening (HCS) technology to investigate the activities of antitubercular compounds in the context of an in vitro granuloma model. We observed significant shifts in the MIC50s between the activities of the compounds under extracellular and granuloma conditions.

Yellow fever and dengue fever serotype 3 viruses cocirculation in Côte d'Ivoire in 2008

Abstract Aim of study. To describe the emergence of dengue 3 virus in Côte d'Ivoire during a yellow fever outbreak which occurred in 2008. Materials and methods. Sera from suspected cases of yellow fever as well as contacts of yellow fever confirmed cases and imported dengue fever cases were tested for immunoglobulin M (IgM) antiyellow fever virus and anti-dengue virus (for IgM antibodies to yellow fever and dengue viruses) and by a specific real time RT-PCR (Bio-Rad) for yellow fever virus and dengue virus viral RNA detection. Results. Of the 511 sera from suspected cases of yellow fever tested, 21 (4.1%) were confirmed positive for yellow fever virus antibody, while 33 (7.6%) of the 432 sera tested were positive for dengue virus antibody. Thirteen viremic subjects, one for yellow fever virus and 12 for dengue 3 virus, were detected by RT-PCR. The majority of the confirmed cases of yellow fever (85%) and dengue 3 fever (93%)were adults, and resided in the city of Abidjan and its regions. Conclusion. These results indicate the existence of transmission foci of these arboviruses diseases in Cˆote d'Ivoire and the essential contribution of molecular tests for their diagnosis

Investigation of an outbreak of acute respiratory disease in côte d'ivoire in april 2007.

BACKGROUND: This study aim was to investigate an outbreak of human cases of unexplained influenza-like illness and fatal acute respiratory infection (ARI), with simultaneous poultry illness and high mortality raising concerns of possible influenza A (H5N1), virus in Cote d'Ivoire in February and March 2007. MATERIALS AND METHODS: To investigate the outbreak, we conducted active surveillance in the community and reviewed health registries. Persons meeting the case definition were asked to provide nasopharyngeal specimens. On the basis of clinical and epidemiological information, specimens were tested using conventional RT-PCR for the M gene of the influenza viruses and hemagglutinin H5 of avian influenza A (H5N1), virus; negative samples were tested for other respiratory viruses. Specimens from healthy animals were also collected. RESULTS: Between October 2006, and February 2007, 104 suspected cases of Acute Respiratory Disease that included; 31 deaths recorded. We collected and tested 73 nasopharyngeal specimens; of which, 2, were positive for human Coronavirus OC43 and 1 for influenza C virus. No pathogens were identified in animal specimens. CONCLUSIONS: The investigation quickly ruled out influenza A (H5N1), virus as the cause and found laboratory-confirmed cases of influenza C virus and human Coronavirus OC 43 for the first time in both Côte d'Ivoire and in a Sub-Saharan African country. However we were not able to show that these viruses caused the outbreak. Monitoring of influenza viruses must be a priority but other respiratory viruses and non-viral causes may be of interest too.

Comparing influenza positivity rates by real-time rt-pcr, elisa and viral culture methods in Cote d'Ivoire, West Africa, in 2009

Detection of circulating influenza strains is a key public health concern especially in limited-resource settings where diagnosis capabilities remain a challenge. As part of multi-site surveillance in Cote d'Ivoire during the 2009 influenza A(H1N1) pandemic; we had the opportunity to test respiratory specimens collected from patients with acute respiratory illness (ARI). We analyzed and compared the percentage of specimens testing positive using three laboratory methods (rtRT-PCR; ELISA; viral culture). From January to October 2009; 1;356 respiratory specimens were collected from patients with acute respiratory illness and shipped at the WHO NIC (Institut Pasteur) Cote d'Ivoire; and 453 (33) tested positive for influenza by one or more laboratory methods. The proportion of positive influenza tests did not differ by the sex or age of the patient or presenting symptoms; but did differ depending on the timing and site of specimen collection. Of the 453 positive specimens; 424 (93.6) were detected by PCR; 199 (43.9) by ELISA and 40 (8.8) by viral culture. While seasonal influenza A(H1N1) virus strains were prominent; only four 2009 pandemic influenza A(H1N1) cases were detected. Use of molecular biology method (rtRT-PCR) increased sensitivity and diagnosis capabilities. Among all three methods used; rRT-PCR was the most sensitive and rapid method. More capacity building is still required for viral culture. Need to collect denominator data in order to have an accurate estimate of the burden of influenza. There was delayed introduction of pandemic influenza A(H1N1)2009 in Cote d'Ivoire

Investigation of an outbreak of acute respiratory disease in Côte d'Ivoire in april 2007

Background: This study aim was to investigate an outbreak of human cases of unexplained influenza-like illness and fatal acute respiratory infection (ARI); with simultaneous poultry illness and high mortality raising concerns of possible influenza A (H5N1); virus in Cote d'Ivoire in February and March 2007. Materials and Methods: To investigate the outbreak; we conducted active surveillance in the community and reviewed health registries. Persons meeting the case definition were asked to provide nasopharyngeal specimens. On the basis of clinical and epidemiological information; specimens were tested using conventional RT-PCR for the M gene of the influenza viruses and hemagglutinin H5 of avian influenza A (H5N1); virus; negative samples were tested for other respiratory viruses. Specimens from healthy animals were also collected. Results: Between October 2006; and February 2007; 104 suspected cases of Acute Respiratory Disease that included; 31 deaths recorded. We collected and tested 73 nasopharyngeal specimens; of which; 2; were positive for human Coronavirus OC43 and 1 for influenza C virus. No pathogens were identified in animal specimens. Conclusions: The investigation quickly ruled out influenza A (H5N1); virus as the cause and found laboratory-confirmed cases of influenza C virus and human Coronavirus OC 43 for the first time in both Cote d'Ivoire and in a Sub-Saharan African country. However we were not able to show that these viruses caused the outbreak. Monitoring of influenza viruses must be a priority but other respiratory viruses and non-viral causes may be of interest too

Emergence in Western African countries of MDR-TB, focus on Côte d'Ivoire.

Tuberculosis (TB) is responsible for a high mortality rate (2.5%) worldwide, mainly in developing countries with a high prevalence of human immunodeficiency virus (HIV). The emergence of multiresistant strains of TB poses an extreme risk for TB outbreaks and highlights the need for global TB control strategies. Among Western African countries, Côte d'Ivoire (CI) represents a specific example of a country with great potential to prevent TB. Specifically, CI has a promising healthcare system for monitoring diseases, including vaccination programs. However, military and political conflict in CI favors the spread of infectious diseases, TB being among the most devastating. Compilation of the studies identifying common causes of TB would be extremely beneficial for the development of treatment and prevention strategies. Therefore, the purpose of this comprehensive review is to evaluate the epidemiology of TB in CI, describe the factors involved in pathogenesis, and suggest simple and applicable prevention strategies.

Comparing Influenza Positivity Rates by Real-Time RT-PCR, Elisa and Viral Culture Methods in Côte D'Ivoire, West Africa, in 2009.

Detection of circulating influenza strains is a key public health concern especially in limited-resource settings where diagnosis capabilities remain a challenge. As part of multi-site surveillance in Côte d'Ivoire during the 2009 influenza A(H1N1) pandemic, we had the opportunity to test respiratory specimens collected from patients with acute respiratory illness (ARI). We analyzed and compared the percentage of specimens testing positive using three laboratory methods (rtRT-PCR, ELISA, viral culture). From January to October 2009, 1,356 respiratory specimens were collected from patients with acute respiratory illness and shipped at the WHO NIC (Institut Pasteur) Cote d'Ivoire, and 453 (33%) tested positive for influenza by one or more laboratory methods. The proportion of positive influenza tests did not differ by the sex or age of the patient or presenting symptoms, but did differ depending on the timing and site of specimen collection. Of the 453 positive specimens, 424 (93.6%) were detected by PCR, 199 (43.9%) by ELISA and 40 (8.8%) by viral culture. While seasonal influenza A(H1N1) virus strains were prominent, only four 2009 pandemic influenza A(H1N1) cases were detected. Use of molecular biology method (rtRT-PCR) increased sensitivity and diagnosis capabilities. Among all three methods used, rRT-PCR was the most sensitive and rapid method. More capacity building is still required for viral culture. Need to collect denominator data in order to have an accurate estimate of the burden of influenza. There was delayed introduction of pandemic influenza A(H1N1)2009 in Cote d'Ivoire.

Sentinel surveillance for influenza and other respiratory viruses in Côte d'Ivoire, 2003-2010.

BACKGROUND: Many countries in Africa have lacked sentinel surveillance systems for influenza and are under-represented in data used for global vaccine strain selection. OBJECTIVES: We describe 8 years of sentinel surveillance data and the contribution of influenza and other viruses to medically attended influenza-like illness (ILI) in Côte d'Ivoire. METHODS: Sentinel surveillance was established in 2003. Nasopharyngeal (NP) specimens and epidemiologic data are collected from persons of all ages presenting with ILI at sentinel sites. Respiratory specimens have been tested for influenza using various viral and molecular diagnostic methods. A subset of 470 specimens collected from children aged 0-5 years were tested for multiple respiratory viruses using RT-PCR. RESULTS: From 2003 to 2010, 5074 NP specimens were collected from patients with ILI. Overall, 969/5074 (19%) of these specimens tested positive for influenza. Seasonal influenza A(H1N1) viruses predominated during 5 years and influenza A(H3N2) viruses predominated during 3 years. Influenza B viruses cocirculated with influenza A viruses during each year from 2004 to 2010. Seasonal peaks in influenza circulation were observed during the months of May, June, and October, with the largest peak corresponding with the primary rainfall season. Of 470 specimens collected from children under aged 5 who were tested for multiple respiratory viruses, a viral respiratory pathogen was detected in 401/470 (85%) of specimens. Commonly detected viruses were RSV (113 of 470 specimens, 24%), rhinoviruses (85/470, 18%), influenza (77/470, 16%), and parainfluenza (75/470, 16%). CONCLUSION: In Côte d'Ivoire, there is a significant annual contribution of influenza and other respiratory viruses to medically attended ILI.

Spatiotemporal circulation of influenza viruses in 5 African countries during 2008-2009: a collaborative study of the Institut Pasteur International Network.

BACKGROUND: Although recent work has described the spatiotemporal diffusion of influenza viruses worldwide, comprehensive data on spatiotemporal patterns of influenza from the African continent and Madagascar are still lacking. METHODS: National Influenza Centers from 5 countries-Cameroon, Côte d'Ivoire, Madagascar, Niger, and Senegal--collected specimens from patients presenting with influenza-like illness who visited sentinel surveillance clinics during a 2-year period (2008-2009). Isolates were genetically and antigenically characterized. RESULTS: Overall, 8312 specimens were tested. Seasonal influenza A virus subtypes H1N1 and H3N2 and influenza B viruses were detected in 329, 689, and 148 specimens, respectively. In 2009, pandemic influenza A virus subtype H1N1 was detected in Madagascar most commonly (98.5% of cases). Influenza activity was either significant year-round or occurred during a specific period of the year in the African countries we evaluated. CONCLUSIONS: Our results demonstrate that, from Madagascar to Senegal, the epidemiologic and virologic characteristics of influenza viruses are diverse in terms of spatiotemporal circulation of the different virus types, subtypes, and strains. Our data highlight the importance of country-specific surveillance and of data and virus sharing, and they provide a rational basis to aid policy makers to develop strategies, such as vaccination at the right moment and with the right formulation, aimed at reducing the disease burden in Africa and Madagascar.

Multilocus VNTR analysis of Mycobacterium ulcerans strains isolated in Côte d'Ivoire.

INTRODUCTION: Buruli ulcer, caused by Mycobacterium ulcerans, is endemic in more than 30 countries worldwide, with Côte d'Ivoire being among the most affected countries. METHODOLOGY: We used seven variable number of tandem repeats (VNTR) markers and analyzed 114 samples from 11 Ivorian localities consisting of 33 bacterial strains and 81 clinical samples. Complete data sets at loci 1, 6, 9 and 33 were obtained for 18 of these strains (n = 15) and samples (n = 3) collected in each of the localities. RESULTS: All the strains had allelic profile [3113], corresponding to the previously described Atlantic Africa genotype. CONCLUSION: Sequencing of PCR products at all loci showed no variation in sequence or repeat number, underlining the genetic monomorphism of M. ulcerans in Côte d'Ivoire.

Induction of dnaK through its native heat shock promoter is necessary for intramacrophagic replication of Brucella suis.

The heat shock protein DnaK is essential for intramacrophagic replication of Brucella suis. The replacement of the stress-inducible, native dnaK promoter of B. suis by the promoter of the constitutively expressed bla gene resulted in temperature-independent synthesis of DnaK. In contrast to a dnaK null mutant, this strain grew at 37 degrees C, with a thermal cutoff at 39 degrees C. However, the constitutive dnaK mutant, which showed high sensitivity to H(2)O(2)-mediated stress, failed to multiply in murine macrophage-like cells and was rapidly eliminated in a mouse model of infection, adding strong arguments to our hypothesis that stress-mediated and heat shock promoter-dependent induction of dnaK is a crucial event in the intracellular replication of B. suis.

Functional analysis of the ClpATPase ClpA of Brucella suis, and persistence of a knockout mutant in BALB/c mice.

The protein ClpA belongs to a diverse group of polypeptides named ClpATPases, which are highly conserved, and which include several molecular chaperones. In this study the gene encoding the 91 kDa protein b-ClpA of the facultative intracellular pathogen Brucella suis, which showed 70% identity to ClpA of Rhodobacter blasticus, was identified and sequenced. Following heterologous expression in Escherichia coli strains SG1126 (DeltaclpA) and SG1127 (Deltalon DeltaclpA), b-ClpA replaced the function of E. coli ClpA, participating in the degradation of abnormal proteins. A b-clpA null mutant of B. suis was constructed, and growth experiments at 37 and 42 degrees C showed reduced growth rates for the null mutant, especially at the elevated temperature. The mutant complemented by b-clpA and overexpressing the gene was even more impaired at 37 and 42 degrees C. In intracellular infection of human THP-1 or murine J774 macrophage-like cells, the clpA null mutant and, to a lesser extent, the strain of B. suis overexpressing b-clpA behaved similarly to the wild-type strain. In a murine model of infection, however, the absence of ClpA significantly increased persistence of B. suis. These results showed that in B. suis the highly conserved protein ClpA by itself was dispensable for intramacrophagic growth, but was involved in temperature-dependent growth regulation, and in bacterial clearance from infected BALB/c mice.