Division of Parasitic Diseases and Malaria


1.    DPDM Genetics and Molecular Biology of Malaria Vector Mosquitoes  


R.A. Wirtz

We propose to reduce the incidence of malaria by understanding and interfering with host-parasite interactions and vector responses to control measures such as insecticide resistance. Our focus is on the African malaria vector Anopheles gambiae. In order to analyze and modify the insect vector to interfere with the transmission cycle, we are developing genetic tools including mosquito transgenesis. Utilizing proven morphological markers and transposable element vectors, development of various control strategies is feasible.  Wide-ranging awardee-defined opportunities are available to develop an innovative, related area of study utilizing the entomological, molecular, and parasite infection capabilities of the Division of Parasitic Disease.  

References

Benedict, M. Q., and A. S. Robinson. 2003. The first releases of transgenic mosquitoes: an argument for the sterile insect technique. Trends in Parasitology 19: 349-355.
Grossman, G. L., C. S. Rafferty, J. R. Clayton, O. Mukabayire, and M. Q. Benedict. 2001. Germline transformation of the malaria vector, Anopheles gambiae, with the piggyBac transposable element. Insect Molecular Biology 10: 597-604.
Jasinskiene, N., C. J. Coates, M. Q. Benedict, A. J. Cornel, C. S. Rafferty, A. A. James, and F. H. Collins. 1998. Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly. Proceedings of the National Academy of Sciences of the United States of America 95: 3743-3747. 

2.  DPDM Evaluation of malaria control under widely varying levels of transmission.

W. A. Hawley 

The vast archipelago of Indonesia has levels of malaria endemicity ranging from nil to highly endemic. CDC, working through UNICEF, the Indonesian MOH, and a network of university partners in Indonesia, aims to develop methods for rapid stratification of malaria endemicity in Indonesia. Evaluation of various combinations of malaria interventions at different transmission levels will also be undertaken.
 

3. DPDM Identification and Control of Insect Vectors of Parasitic Diseases 
 

R. A. Wirtz  

Research goals involve the development and field evaluation of malaria control strategies as well as studies of the physiology and behavior of insect vectors of parasitic diseases. Specific research opportunities include (i) identification of insect vectors by molecular, immunological, and electromagnetic methods; (ii) evaluation of insecticide control by chemical analysis; (iii) monitoring of antimalarial drugs and drug metabolites by chemical analysis; and (iv) studies of the physiology and behavior of malaria vectors and parasite-vector interactions.  

References

Bangs, M.J., S. Rusmiarto, R.A. Wirtz, R.L. Anthony, and B. Subianto. 1996. Malaria transmission by Anopheles punctulatus in the highlands of Irian Jaya, Indonesia. Ann. Trop. Med. Parasitol. 90:29-38.
Green, M.D., D.L. Mount, and G.D. Todd. 1995. Chemiluminescent detection of artemisinin. Novel endoperoxide analysis using luminol with out hydrogen peroxidase. J. Chrom. A. 695(2): 237-242.  

4. DPDM Schistosomiasis, Trichomoniasis, Opportunistic Protozoa


W. E. Secor

My laboratory studies various aspects of infection with the parasitic worm, Schistosoma mansoni. Schistosomiasis afflicts more than 200 million persons worldwide and presents with a range of morbidities, from moderate to life-threatening. Current projects focus mainly on immunologic aspects of human disease that range from development of better diagnostic tools to markers of morbidity to impact of helminth infection on susceptibility to other infections. This work is facilitated by the existence of the CDC field station in Kisumu, Kenya on the shores of Lake Victoria, where schistosomiasis is highly endemic. Ongoing projects/opportunities include:            
--identification of diagnostic tools that can distinguish active from previous schistosome infection            
--identification of biomarkers that can be used to detect morbidity during schistosomiasis            
--characterization of Treg and Th17 cells during schistosomiasis and association with morbidity            
--identification of what responses to which antigens are associated with resistance to reinfection            
--determination of treatment effects on morbidity following reinfection and resistance to reinfection             
--evaluation of schistosomiasis and its treatment on coinfection with HIV-1 or malaria           
--evaluation of schistosomiasis and its treatment on vaccine efficacy in children and adults           
--operational research on optimal frequency and distribution of mass treatment 

5.  DPDM Molecular and Immunologic Studies of Human Malaria
 

V. Udhayakumar (Kumar) 

The research program of the laboratory focuses on studies of immune responses to malaria, pathogenesis, host and parasite genetics and anti-malarial drug resistance. These programs are supported by ongoing cohort studies in Africa, Asia and South America. Our laboratory has unique biological samples available from several malaria cohort studies to investigate the role of antibodies, cytokines and innate immune factors in protection against both complicated and uncomplicated malaria.  We are also investigating the development of protective immune mechanisms against placental malaria and how HIV-1 infection impacts the development of this immunity. Recent advances in human genome research have opened up new opportunities for identifying host genetic factors associated with severe disease outcomes and our laboratory is making use of these developments to identify potential host genetic factors associated with the risk of severe malaria.  The emergence of anti-malarial drug resistance has become a major public health challenge.  Our laboratory is actively involved in developing molecular markers to track the emergence and spread of drug resistant parasites in different parts of the world.  All these studies involve collaborative work with many laboratories in U.S. and around the world.  We also support development and testing of novel tools for the diagnosis of malaria.

References

Ned RM, Moore JM, Chaisavaneeyakorn SJ, and Udhayakumar V.  Modulation of Immune Responses During HIV/Malaria Co-infection in Pregnancy. Trends in Parasitol. 2005, 21(6):284-91.

Hobbs MR, Udhayakumar V, Levesque MC, Booth J, Tkachuk AN, Pole A, Roberts JM, Kariuki S,  Nahlen BL, Mwaikambo ED, Lal AA, Granger DL, Anstey NM, and Weinberg JB.  A new NOS2 promoter polymorphism associated with protection from severe malaria in children from Tanzanian and Kenyan children. Lancet , 2002, 360(9344):1468-75.

Keen J, Serghides L, Ayi K, Patel SN,  Ayisi J, van Eijk A,  Steketee R,  Udhayakumar V, and Kain KC.  HIV Impairs Opsonic Phagocytic Clearance of Pregnancy-Associated Malaria Parasites. PLOS Medicine, 2007 in press.

McCollum AM, Mueller K, Villegas L, Udhayakumar V, Escalante A. Common origin and fixation of Plasmodium falciparum dhfr and dhps mutations associated with sulphadoxine-pyrimethamine resistance in a low transmission area in South America. Antimicrob Agents Chemother 2007,  51 (6): 2085-2091. 


6.  DPDM Molecular, Cellular, and Immune Biology of Host-Parasite Interactions in Malaria 

J. W. Barnwell  

Malaria is a disease of many complexities that arise through the influence of external ecological factors and from internal interactions between the parasite and elements of the host environment to affect transmission, pathogenesis, immunity, and parasite population structure. The research program of this laboratory seeks to investigate the biology of host-parasite relationships through the discovery and molecular, cellular, functional, and genetic characterization of parasite and host molecules that interact to effect tissue invasion, pathogenesis, or the modulation and evasion of immune responses. Other areas of investigation include drug resistance mechanisms and transmission blocking interventions.  The laboratory maintains or has access to all stages of the life cycle of the malaria parasite life cycle in the vertebrate and invertebrate hosts. The studies center on human and primate species of Plasmodium and make use of field studies as well as primate models to understand the complex biology and to assess the immunogenic potential of parasite antigens as components of malaria vaccines.
 

References

Barnwell, J.W. and Galinski, M.R. 1998. Invasion of Vertebrate Cells: Erythrocytes. In Malaria: Parasite Biology, Pathogenesis, Protection, Editor: Irwin W. Sherman, ASM Press, New York, Chapter 7, pp. 93-120.
Rayner, J.C., Vargas-Serrato, E., Huber, C.S., Galinski, M.R., Barnwell, J.W.  2001.  A Plasmodium falciparum homologue of Plasmodium vivax reticulocyte binding protein (PvRBP1) defines a trypsin-resistant erythrocyte invasion pathway.  J Exp Med. 194:1571-1581.
Darko, C.A., Angov, E., Collins, W.E., Bergmann-Leitner, E.S., Girouard, A.S., Hitt, S.L., McBride, J.S., Diggs, C.L., Holder, A.A., Long, C.A., Barnwell, J.W., Lyon, J.A. 2005. The clinical-grade 42-kilodalton fragment of merozoite surface protein 1 of Plasmodium falciparum strain FVO expressed in Escherichia coli protects Aotus nancymai against challenge with homologous erythrocytic-stage parasites. Infect Immun.  73:287-297.  
 

7. DPDM Immunology and Molecular Biology of Human Malaria

Y.P. Shi 

Research in this laboratory focuses on malaria immune protective mechanisms, host and parasite genetics, antimalaria drug resistance, impact of malaria interventions on host and parasite responses, and malaria/HIV interaction. Current projects include the following areas: (i) investigation of the impact of  transmission reduction by use of insecticide-treated bednets on parasite population genetic structure, distribution of genes conferring antimalarial drug resistance, complexity and virulence of parasite,  (ii) anitmalarial drug resistance molecular surveillance in pregnant women and adults, and investigation of the influence of host immunity factors  on selection of drug resistant parasites, (iii) determination of host immunogenetic risk factors for clinical outcomes of  malaria infection and malaria/HIV interaction in both children and pregnant women, (iv) standardization of in vitro immunological functional assays and identification of in vitro immune correlates of protection against malaria using the assays, and (v) development of low cost, field useable and sensitive  novel  tools for the diagnosis of malaria.

References

Gatei W, Kariuki S, Hawley W, ter Kuile F, Terlouw D, Phillips-Howard P, Nahlen B, Gimnig J, Lindblade K, Walker E, Hamel M, Crawford S, Williamson J, Slutsker L, Shi YP. Effects of transmission reduction by insecticide-treated bed nets (ITNs) on parasite genetics population structure: I. The genetic diversity of Plasmodium falciparum parasites by microsatellite markers in western Kenya. Malar J. 2010 Dec 6;9:353

OPTIMALVAC consortium, Cavanagh DR, Dubois PM, Kisser A, Leroy O, Locke E, Moorthy VS, Remarque  EJ, Shi YP. Towards validated assays for key immunological outcomes in malaria vaccine development.  Vaccine. 2011 Feb 3.

 

Othoro C, Julie Moore, Kathleen A. Wannemnehler, Sichangi Moses, Altaf Lal,  Juliana Otieno, Bernard Nahlen ,  Laurence Slutsker, and Shi,YP.  IFN-gamma producing NK cells, CD45RO memory-like T cells and CD4 T cells are associated with protection against malaria during pregnancy. Infect Immunity. 2008 76(4): 1678-85.

 

Brouwer KC, Lal AA, Mirel LB , Lal RB,  Van Eijk AM , Ayisia, Otieno J, Steketee R, Nahlen BL, Shi YP. Association between genetic polymorphism in  Fc receptor IIa  for IgG (Fc( RIIa) and placenta malaria in western Kenya. JID 2004, 196: 1192-8.

 

Kariuki SK, Lal AA, Terlouw DJ, ter Kuile FO, Ong'echa JMO, Phillips-Howard PA, Orago ASS, Kolczak MS,  Hawley BA, Nahlen BL, and Shi YP. Effects of permethrin-treated bed nets on immunity to malaria in  western Kenya II. antibody responses in young children in an area of intense malaria transmission. Am J  Trop Med & Hyg. 2003 68 (suppl): 108-114.


8. DPDM Molecular Biology, Epidemiology and Genetics of Parasitic Disease Agents
 

E.M. Dotson  

Research goals involve the development and application of molecular biological approaches for use in the control of parasitic disease agents and their insect vectors.  Specific research areas include the following: i) genetic modification of the symbiotic bacteria of insects as a means for controlling insect-transmitted parasitic diseases, ii) bacterial ecology in relation to acquisition and dispersal of symbiotic bacteria by insects and dispersal in artificial media, iii) phylogeny, population biology and genetics of insect vectors of disease agents. 

References

Dotson, E.M. & Beard, C.B. (2001) Sequence and organization of the mitochondrial genome of the Chagas disease vector, Triatoma dimidiata.  Insect Mol. Biol. 10(3): 205-215.

Beard, C.B., Dotson, E.M., Pennington, P.M., Eichler, S., Cordon-Rosales, C., & Durvasula, R.V. (2001) Bacterial symbiosis and paratransgenic control of vector-borne Chagas disease.  Internat. J. Parasitol.  31: 621-627.

Anderson, J.M, Lai, J., Dotson, E.M. and Beard, C.B. (2002) Identification and characterization of microsatellite markers in Triatoma dimidiata (Hemiptera, Reduviidae), a Chagas Disease vector. Infect. Genet. Evol. 1:243-248

Dotson, E.M. and C.B. Beard. (2002) Paratransgenic strategies for the control of Chagas disease, pp 147-155. In: Miles, M. & K.M. Tyler (eds), World Class Parasites, Vol IV.  Kluwer Academic Publishers, Norwell, MA.
 
Dotson, E.M., Plikaytis, B., Shinnick, T., Durvasula, R. and Beard, C. B. (2003) Transformation of Rhodoccocus rhodnii, a symbiont of the Chagas disease vector Rhodnius prolixus, with integrative elements of the L1 mycobacteriophage. Infect. Genet. Evol. 3: 103-109.

Costa, J., Dotson, E., Lins, A., Vinhaes, M., Siveira, A.C., and Beard, C. B. (2003) The emergence of Triatoma brasiliensis as the primary domestic vector of Chagas disease in northeastern Brazil.  Mem. Inst. Oswaldo Cruz, 98(4): 443-449

Marcet, PM, Lehmann, T, Groner, G, Gurtler, RE, Kitron, U & Dotson, EM. 2006. Identification and characterization of microsatellite markers in the Chagas disease vector Triatoma infestans. Infect. Genet. Evol. 6(1): 32-7.
 


9. DPDM Ecology and Control of African Vectors of Malaria
 

J.E. Gimnig 

My research goals focus on the ecology and behavior of Anopheles gambiae in both the larval and adult stages.  The ultimate aim is to find and exploit resources in the mosquito’s life cycle that are essential to its survival or to parasite transmission. Current research opportunities at our field station in western Kenya include the identification and characterization of productive Anopheles gambiae larval habitats, mapping of larval habitats and correlation with adult mosquito densities in houses, development of novel traps for monitoring adult mosquito populations, behavioral studies to assess the impact of insecticide based interventions, and monitoring of phenotypic, biochemical and molecular markers of insecticide resistance. 

References

Gimnig JE, Ombok M, Otieno S, Kaufman MG, Vulule JM & Walker ED. 2002. Density dependent development of Anopheles gambiae (Diptera: Culicidae) larvae in artificial habitats. J Med Entomol 39 (1): 162-172.

Mathenge EM, Gimnig JE, Kolczak M, Ombok M, Irungu LW & Hawley WA. 2001. The effects of permethrin-impregnated nets on exiting behavior, blood feeding success, and time of feeding of malaria mosquitoes (Diptera: Culicidae) in western Kenya. J Med Entomol 38 (4): 531-536.

Gimnig JE, Ombok M, Kamau L & Hawley WA. 2001. Characteristics of Larval Anopheline (Diptera: Culicidae) Habitats in Western Kenya. J Med Entomol 38 (2): 282-288.
 

 

10. DPDM Laboratory Tools for Elimination and Control of Onchocerciasis

V. A. Cama

 

Our laboratory works on the assessment, validation and improvement of laboratory tools used in the elimination and control of neglected tropical diseases (NTD) caused by parasites. The objective is to provide tools that would support the assessment of program activities. Our primary focus is the evaluation and validation of immunological methods for detecting exposure to Onchocerca volvulus, the etiology of onchocerciasis or River blindness. This parasite affects about 80 million people in Africa and Latin America, although the vast majority live in Africa. Onchocerciasis has several clinical manifestations, the most significant being blindness1 . Multidisciplinary efforts for controlling and eliminating onchocerciasis started in 19872. Several countries have made significant progress towards stopping transmission3, and robust methods to properly evaluate the cumulative efforts of programs are urgently needed. The foundation for our work is to be part of multidisciplinary teams, while collaboration with other NTD activities. Our research work on Onchocerca is funded by the Bill and Melinda Gates Foundation.

 

References

 

WHO Onchocerciasis, http://www.who.int/topics/onchocerciasis/en/.

Mectizan Donation Program, http://www.mectizan.org/history,

Carter Center, http://www.cartercenter.org/health/river_blindness/index.html

 

11. DPDM Development and Evaluation of Biological, Biochemical and Molecular Tools for the Detection, Assessment and Remediation of Insecticide Resistance in Arthropod-borne Disease Vectors

W.G. Brogdon

 

We have been developing bioassay, biochemical assay and molecular techniques for detecting and assessing the significance of insecticide resistance in arthropod vectors of human diseases, particularly those involved in transmission of malaria. This has been in response to the emerging crisis of resistance development in the large multinational programs that are using long-lasting insecticide impregnated nets (LLINs) and indoor residual spraying (IRS).

We are especially interested in the interaction of resistance genotypes through processes such as multiplicative synergistic epistasis since these interactions have led to particularly high resistance frequency and intensity phenotypes in many countries. Further complicating the situation is the focal nature of resistance and the independent selection of resistance mechanisms among the numerous sympatric vector species in many locations.

For successful management of insecticide resistance, we must fully understand resistance selection in terms of its biology, biochemistry, toxicology and molecular biology. Quite a challenge!

 

References

 

David, J. P., C. Strode et al. (2005). "The Anopheles gambiae detoxification chip: a highly

specific microarray to study metabolic-based insecticide resistance in malaria vectors." Proc

Natl Acad Sci U S A 102(11): 4080-4.

 

Fonseca-González I, Quiñones ML, McAllister J, Brogdon WG. (2009). Mixed-function oxidases and esterases associated with cross-resistance between DDT and lambda-cyhalothrin in Anopheles darlingi Root 1926 populations from Colombia. Mem Inst

Oswaldo Cruz. 2009 Feb;104(1):18-26.

 

Hardstone, M.C. et al (2008). Multiplicative interaction between the two major mechanisms

of permethrin resistance, kdr and cytochrome P450-monooxygenase detoxification, in

mosquitoes. J Evol Biol 22: 446-423

 

N'Guessan, R., Corbel, V., Akogbeto, M., & Rowland, M. (2007). Reduced efficacy of

insecticide-treated nets and indoor residual spraying for malaria control in pyrethroid

resistance area, Benin. Emerg Infect Dis, 13(2), 199-206.

 

Zamora Perea E, Balta León R, Palomino Salcedo M, Brogdon WG, Devine GJ. (2009).

Adaptation and evaluation of the bottle assay for monitoring insecticide resistance in disease vector mosquitoes vector mosquitoes in the Peruvian Amazon. Malar J. 2009 Sep 3; 8:208.

 

12. DPDM Maintaining the Efficacy of Insecticides for the Control of Mosquito-borne Diseases: Understanding and Managing the Development of Insecticide Resistance

A. Lenhart

 

We are interested in measuring genotypic and phenotypic changes in naturally isolated Anopheles darlingi malaria vector populations and Aedes aegypti dengue vector populations in the Peruvian Amazon currently under selection pressure by different insecticide-based strategies. The investigation of incipient insecticide resistance can provide insight into mechanisms and rates of insecticide resistance development in the field, and their potential operational implications. It can also provide a scientific evidence base for selecting strategies for the management of insecticide resistance and/or the amelioration of its effects. Fieldwork would be done in collaboration with the Peruvian Ministry of Health and the US Naval Medical Research Unit-6 field station in Iquitos, Peru, and the research project would be linked with CDC’s role as a technical partner in the Amazon Malaria Initiative.  

 

References

David, J. P., C. Strode et al. (2005). "The Anopheles gambiae detoxification chip: a highly specific - microarray to study metabolic based insecticide resistance in malaria vectors." Proc Natl Acad Sci U SA 102(11): 4080-4.

Fonseca-Gonzalez, I., Quinones, M. L., McAllister, J., & Brogdon, W. G. (2009). Mixed-function oxidases and esterases associated with cross-resistance between DDT and lambda-cyhalothrin in Anopheles darlingi Root 1926 populations from Colombia. Mem Inst Oswaldo Cruz, 104(1), 18-26.

Fonseca-González I, Quiñones ML, Lenhart A and Brogdon WG. (2011) Insecticide resistance status of Aedes aegypti (L.) from Colombia. Pest Management Science 67(4):430-7.

Grisales, N., Poupardin, R., Fonseca-Gonzalez, I., Ranson, H., & Lenhart A.E. Mechanisms and operational implications of temephos resistance in Aedes aegypti from Colombia. In preparation.

Lenhart, A.E., Vasquez La Torre, G., Goodson, D., Zamora, E., Astete, H. Scott, T.W., Morrison, A.C. & McCall P.J. Rapid development of insecticide resistance in Aedes aegypti in Iquitos Peru in a population under deltamethrin pressure. In preparation.

Marcombe, S., A. Carron, et al. (2009). "Reduced efficacy of pyrethroid space sprays for dengue control in an area of Martinique with pyrethroid resistance." Am J Trop Med Hyg 80(5): 745-51.

N'Guessan, R., Corbel, V., Akogbeto, M., & Rowland, M. (2007). Reduced efficacy of insecticide-treated nets and indoor residual spraying for malaria control in pyrethroid resistance area, Benin. Emerg Infect Dis, 13(2), 199-206.

 

 

TPL_asm2013_SEARCH