Daniel Lagasse
Katy Shaw
Eva Tse
Anidi, Ifeanyi
Spalding, Maroya
Overstreet, Michael
Jayabalasingham, Bamini
Kent, Rebekah
LeBlanc, Ralph
Noland, Greg
Ruben, Adam

Hughes, Grant
Brant, Gabriel
Bosch, Gundula
Souza-Neto, Jayme
Mlambo, Godfree
Cockburn, Ian
Cha, Sung-Jae
Abhai, Tripathi
Xi, Zhiyong
Kanzok, Stefan
Chowdhury, Debabani
Liu, Jeff Zhiqiang
Bano, Nazneen

Krairojananan, Panadda
Nyakundi, Ruth
Moreira, Luciano
Hawela, Moonga
Dhar, Ravi

Gregory Noland

Host Response to Malaria and Helminth Co-Infection: Implications for Natural and Vaccine-Induced Immunity

Aims

1. Evaluate host response to malaria during intestinal helminth co-infection

2. Compare relative malaria transmission potential between trematode-co-infected and malaria-only-infected mice

3. Characterize immunologic basis of interactions between intestinal trematodes and malaria parasites

4. Evaluate protection conferred by two candidate malaria vaccines in helminth-infected mice

Bamini Jayabalasingham

Elucidating the Molecular Function of Plasmodium falciparum Sexual Stage-Specific Protein Pfg27

Aims

1. Identify proteins with which Pfg27 interacts during the process of gametocytogenesis

2. Characterize the nature of the interaction between Pfg27 and interacting protein(s)

3. Investigate the possible role of Pfg27 in translational regulation in P. falciparum 

Rebekah J. Kent

Malaria Transmission in Macha, Zambia: Entomological Inoculation Rates of Anopheles arabiensis and Anopheles funestus S.S and Vectorial Capacity of An. arabiensis

Aims

1. Determine the relative importance of An. arabiensis and An. funestus to P. falciparum transmission in Macha by comparison of their entomological inoculation rates

2. Describe the blood feeding behavior of An. arabiensis in Macha

3. Estimate the efficiency of P. falciparum transmission by An. arabiensis

Ralph LeBlanc, MD, MPH

Development of a Malaria Transmission-Blocking Mono-and Multivalent Vaccine (TBV) Based on Pfs230, Pfs48/45 and Pfs25

Aims

1. To determine the immunogenicity of selected antigens by ELISA, IFA and immunoprecipitation (I.P.)

2. To assess functional transmission-blocking activity (TBA) by gamete lysis and membrane feeding assay (MFA)

3. To evaluate combinations of pre- and post-fertilization antigens in a multiple-antigen vaccine

4. To evaluate natural boosting potential of immune responses primed by DNA plasmid vaccine

Adam Ruben

Thermodynamic Characterization of Plasmepsin Family Inhibitors Adaptable against Multiple Targets

Aims

1.  Continue designing and screening potential inhibitors of Plm II

2.  Continue to determine the Ki of selected inhibitors against secondary targets Plm I and Plm IV

3.  Crystallize Plms in complex with selected inhibitors

4.  Based on the results of  Specific Aims 1-3, identify candidates for erythrocyte assays. Use the results of those assays to relate structural and thermodynamic properties with in vitro function

5.  Identify inhibitors to test in mouse and monkey models

March 2006

Michael Overstreet

Crosstalk between CD4+ T cells and CD8+ T cells in Immunity to Malaria

Aims

1.  Examine crosstalk between transgenic CD4+ T cells and CD8+ T cells in response to irradiated sporozoites using an adoptive transfer model

2.  Examine CD8+ T cells behavior in response to peptide immunization in presence or absence of CD4+ T cell activation

Maroya Spalding

The Role of Lipoate Metabolism in P. falciparum Survival

Aims

1.  Investigate whether lipoate synthesis is essential in the apicoplast through conditional disruption of lipoylation in this organelle and to use lipoate-disrupted parasites to detemrine the relationship between lipoate synthesis and fatty acide synthesis

2.  To invstigate whether lipoate scavenging is essential in the mitochondrion and the cytosol through conditional disruption of lipoylation, and if lipoate scavenging is essential in the cytosol, to identify the lipoylated protein(s) in this subcellular compartment

Ifeanyi Anidi

Malaria and Lung Macrophages

Aims

1.  To definte the phenotypic and functional consequences that malaria infection has on the lung-derived macrophages

2.  To determine the role that lung macrophages play in the pathogenesis of pulmonary malaria

January 2010

Daniel Lagasse

The role of lung macrophages in the control of malaria-induced pulmonary inflammation

Aims

1.  Dynamics of immune cell influx into thelungs upon malaria infection

2.  Selective depletion of lung machrophages

3.  Selective depletion of lung macrophages during P. berghei infection

4.  Adopotive transfer of macrophages into the lungs

Katy Shaw

Genetic Analysis of Plasmodium falciparum Gametocytogenesis

Aims

1.  Transposon-mediated mutagenesis and screening for genes involved in gametocytogenesis

2.  Interesting genes identified by the mutagenesis screen

3.  Hypothesis about the function of Class I genes

Eva Tse

Migration and Residency of Antigen Specific Memory CD8+ T cells in the Liver 

Aims

1.  To characterize the expression pattern of homing and adhesion molecules on antigen specific CD8+ T cells in the liver 

2. To investigate the functional role of selected homing and adhesion molecules of antigen specific CD8+ T cells in the liver

3. To investigate the proliferation, circulation and renewal properties of memory CD8+ T cells establishing residence in the liver Background and Significance: Our laboratory has demonstrated that the priming of antigen specific CD8+ T cells first occurs at skin-

Jeff Zhiqiang Lu, PhD

Structure and Inhibition Studies of Plasmodium falciparum KAS Enzymes

Aims

1. To investigate the mode of action of PfKASIII inhibitors using biochemical techniques

2. To study the activity of PkFASI/II and test thiolactomycin and the PfKASIII inhibitors for activity against this enzyme

3. To structurally characterize the atomic level interactions of novel KAS inhibitors with PfKASIII enzyme

Nazneen Bano

Identification of the Host Cellular Pathway/s and Parasite Transporter/s Involved in Host Cholesterol Trafficking to Intrahepatic Plasmodium

Aims

1.  Identification of the host cellular pathways mediating cholesterol delivery from host cell to intraheptic parasites

2.  Functional analysis of the role Plasmodium ABCG1 transporter homologues in parasite cholesterol homeostasis

Debabani Chowdhury

Evaluation of B cell Responses against Plasmodium falciparum Transmission-blocking Vaccine Antigen (Pfs25) in Naive and Helminth-infected Mice

Aims

1.  To evaluate the efficacy of Pfs25 DNA vaccine in production of memory B cells and long-lasting plasma cell (LLPC) responses

2.  To characterize the effectiveness of Pfs25 DNA vaccine in generation of memory B cells and LLPC responses, when the host (mice) is pre-infected with the helminth Heligmosomoides polygyrus (Hp)

3.  To determine the formatin of germinal center (GC) in response to Pfs25 DNA vaccine in mice, which are uninfected or pre-infected with Hp

Stefan Kanzok

Genetic Analysis of Plasmodium falciparum Gametocyte Formation

Aims

1.  Gametocyte reporter strain

2.  Transposon-mediated mutagenesis

Zhiyong Xi

The role of Anopheles gambiae MD2-like Genes in Innate Immune Defense Against the Human Malaria Parasite

Aims

1.  Determination of expression patterns upon infection

2.  Determination of defense specificity

3.  Assessing immune pathway regulatory functions of AGMLs

4.  Assessing the interaction of AGMLs with plasmodium

Abhai Tripathi

Antigen Presentation Following Infection with Malaria Sporozoites

Aims

1.  Identification of dendritic cell subsets involved in presentation of sporozoite antigens to CD8+ T cells

2.  Examine the role of innate immune system components in the development of CD8+ T cell responses

Sung-Jae Cha

Aims

1.  Identification and characterization of sporozoite ligands for Kupffer cell invasion

2.  Identification and characterization of putative Kupffer cell receptors

3.  Identification and characterization of human orthologs of Kupffer cell invasion

Ian Cockburn

Aims

1.  To identifiy the DCs that present sporozoite antigens to CD8 T cells and how the DC-T cell interactions are influenced by innate responses

Gabriel Brandt

Plastid DNA Replication in P. falciparum

Aims

1.  Production of apicoplast replication proteins: Express and purify the apicoplast-targeted polyprotein Prex in its full-length form, along with its helicase, primase, helicase-primase and polymerase domains. Express and purify the putative apicoplast single-stranded DNA binding protein

2.  Begin reconstitution of plastid genomic replication in vitro: Examine, using mini-circle models of the replication fork, the ability of the helicase-primase and polymerase domains to replicate DNA. Determine the effect of helicase activity and single-stranded DNA binding proteins on polymerase activity

3.  Undertake structural studies of the apicoplast replication proteins: Purify and concentrate Prex helicase, primase, helicase/primase and polymerase domains. Screen for crystallization conditions using sparse matrix methods and liquid-handling robot. Undertake co-crystallization of polymerase domain with primer and template DNA oligomers

Godfree Mlambo

Development of an Animal Model for Pre-clinical Evaluation of Plasmodium falciparum Transmission-blocking Vaccine (Pfs48/45) Using Transgenic P. berghei Parasites

Aims

1.  To make transgenic P. berghei parasites expressing the P. falciparum sexual antigen Pfs48/45

2.  To evaluate sera from non-human primates immunized with the Pfs48/45 vaccine

3.  To test whether natural infection with the transgenic parasite boosts antibodies to the antigen Pfs48/45

4.  To understand fertilization receptor functional conservation between P. berghei and P. falciparum

Jayme Souza-Neto

The Generation and Characterizatin of Immune Pathway Inducible Plasmodium Resistant Transgenic Mosquitoes

Aims

1.  Generate an inducible Rel2 construct for the generation of transgenic A. stephensi mosquitoes

2.  Characterize the dox-dependent rtTa-driven induction of the Rel2 factor

3.  Characterize the dox-induced Rel2 mediated resistance to Plasmodium infection at the ookinete stage

4.  Assess the impact of dox-mediated Rel2 activation on mosquito longevity and fecundity

Gundula Bosch

Structural and Functionl of Iron-Sulfur Proteins in Isoprenoid Biosynthesis and Utilization in Plasmodium falciparum

Aims

1.  To produce and characterize the IspG, IspH and MiaB enzymes

2.  To reveal the active site architecture and catalytic mechanism of IspG, IspH and MiaB

Grant Hughes

Introduction of Wolbachia into Anopheles gambiae and Modulation of Plasmodium Vector Competence

Aims

1. Artificially transinfect Anopheles gambiae with Wolbachia. Wolbachia will be microinjected into adults in conjunction with microbeads to suppress the mosquito immune response. Transinfections will be performed with two strains of Wolbachia (wMelPop and wAlbB), which have been pre-adapted to Anopheles in cell culture 1

2. Characterize the novel Wolbachia-Anopheles association. Transmission rates, life-shortening and reproductive manipulations on the host will be assessed in the mosquito. Additionally, the distribution of Wolbachia within the mosquito will be characterized. Anopheles gene expression in response to Wolbachia infection will be analyzed in whole mosquitoes by microarrays

3. Determine Wolbachia-Plasmodium interactions. Stably or transiently Wolbachia infected mosquitoes will be challenged with Plasmodium and development of the parasite analyzed to determine if Wolbachia interferes with parasite development and transmission

Ravi Dhar, PhD

Dr. Dhar is a scientist with the National Institute of Immunology, New Delhi, India. He spent five months in the laboratory of Nirbhay Kumar, learning advanced molecular techniques for genotype analysis of Plasmodium falciparum in India. With the knowledge he gained during his fellowship, he will study parasite diversity in severe and non-severe infections in Indian patients, apply this information to the study of responses to antimalarial drugs and build bridges for future collaboration with JHMRI investigators.

Moonga Hawela, Msc

Mr. Hawela is a Biomedical Scientist and Acting Chief Parasitologist with the National Malaria Control Centre in Zambia.  He spent three months in the laboratory of Dr. David Sullivan where he mastered the techniques of extraction of parasite DNA, ampliciation of MSP2 for P. falciparum, trained to use MSP2 primers and received extensive training in performance of electrophoresis and reading of gels.    

Luciano Moreira, PhD

Dr. Moreira is an Associate Researcher with the Malaria Laboratory of Centro de Pesquisas Rene Rachou/FIOCRUZ, Brazil.  He spent 1 month in the laboratory of Dr. Marcelo Jacobs-Lorena learning Plasmodium falciparum cutlure and mosquito transmission techniques. 

Ruth Nyakundi, Msc

Ms. Nyakundi is an Assistant Research Scientist with the Department of Tropical and Infectious Diseases, Institute of Primate Research in Nairobi, Kenya.  She spent 3 months in the laboratory of Dr. Nirbhay Kumar, to gain expertise in P. falciparum cutlure and mosquito transmission methods to introduce her and her institution to new technology and new techniques. 

Panadda Krairojananan, MS

Ms. Krairojanan is a PhD candidate and Assistant Chief with the Molecular Biology Section, Department of Entomology for the Armed Forces Research Institue of Medical  Sciences in Bangkok, Thailand.  She spent 3 months in the laboratory of Dr. Rhoel Dinglasan studying the molecular mechanisms of Plasmodium-mosquito interactions.  The experiences she gained not only benefited her but her Institution in molecular techniques as much of these techniques were not avaialble at the Mahidol University and AFRIMS.