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5. Clinical trial in healthy malaria-naïve adults to evaluate the safety, tolerability, immunogenicity and efficacy of MuStDO5, a five-gene, sporozoite/hepatic stage Plasmodium falciparum DNA vaccine combined with escalating dose human GM-CSF DNA

Author

Richie, Thomas L., Charoenvit, Yupin, Wang, Ruobing, Epstein, Judith E., Hedstrom, Richard C., Kumar, Sanjai, Luke, Thomas C., Freilich, Daniel A., Aguiar, Joao C., Sacci, John B., Jr., Sedegah, Martha, Nosek, Ronald A., Jr., De La Vega, Patricia, Berzins, Mara P., Majam, Victoria F., Abot, Esteban N., Ganeshan, Harini, Richie, Nancy O., Banania, Jo Glenna, Baraceros, Maria Fe B., Geter, Tanya G., Mere, Robin, Bebris, Lolita, Limbach, Keith, Hickey, Bradley W., Lanar, David E., Ng, Jennifer, Shi, Meng, Hobart, Peter M., Norman, Jon A., Soisson, Lorraine A., Hollingdale, Michael R., Rogers, William O., Doolan, Denise L., and Hoffman, Stephen L.

Published
2012

6. Novel malaria antigen Plasmodium yoelii E140 induces antibody-mediated sterile protection in mice against malaria challenge

Author

Smith, Emily C., primary, Limbach, Keith J., additional, Rangel, Nonenipha, additional, Oda, Kyosuke, additional, Bolton, Jessica S., additional, Du, Mengyan, additional, Gowda, Kalpana, additional, Wang, Jianyang, additional, Moch, J. Kathleen, additional, Sonawane, Sharvari, additional, Velasco, Rachel, additional, Belmonte, Arnel, additional, Danner, Rebecca, additional, Lumsden, Joanne M., additional, Patterson, Noelle B., additional, Sedegah, Martha, additional, Hollingdale, Michael R., additional, Richie, Thomas L., additional, Sacci, John B., additional, Villasante, Eileen D., additional, and Aguiar, Joao C., additional

Published
2020
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9. Cytomegalovirus vectors expressing Plasmodium knowlesi antigens induce immune responses that delay parasitemia upon sporozoite challenge

Author

Hansen, Scott G., primary, Womack, Jennie, additional, Scholz, Isabel, additional, Renner, Andrea, additional, Edgel, Kimberly A., additional, Xu, Guangwu, additional, Ford, Julia C., additional, Grey, Mikayla, additional, St. Laurent, Brandyce, additional, Turner, John M., additional, Planer, Shannon, additional, Legasse, Al W., additional, Richie, Thomas L., additional, Aguiar, Joao C., additional, Axthelm, Michael K., additional, Villasante, Eileen D., additional, Weiss, Walter, additional, Edlefsen, Paul T., additional, Picker, Louis J., additional, and Früh, Klaus, additional

Published
2019
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10. Discovery of Novel Plasmodium falciparum Pre-Erythrocytic Antigens for Vaccine Development

Author

Aguiar, Joao C., primary, Bolton, Jessica, additional, Wanga, Joyce, additional, Sacci, John B., additional, Iriko, Hideyuki, additional, Mazeika, Julie K., additional, Han, Eun-Taek, additional, Limbach, Keith, additional, Patterson, Noelle B., additional, Sedegah, Martha, additional, Cruz, Ann-Marie, additional, Tsuboi, Takafumi, additional, Hoffman, Stephen L., additional, Carucci, Daniel, additional, Hollingdale, Michael R., additional, Villasante, Eileen D., additional, and Richie, Thomas L., additional

Published
2015
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11. Transcriptional Analysis of In vivo Plasmodium yoelii Liver Stage Gene Expression

Author

MARYLAND UNIV BALTIMORE SCHOOL OF MEDICINE, Sacci, Jr., John B., Ribeiro, Jose M., Huang, Fengying, Alama, Uzma, Russell, Joshua A., Blair, Peter L., Witney, Adam, Carucci, Daniel J., Azada, Abdu F., Aguiar, Joao C., MARYLAND UNIV BALTIMORE SCHOOL OF MEDICINE, Sacci, Jr., John B., Ribeiro, Jose M., Huang, Fengying, Alama, Uzma, Russell, Joshua A., Blair, Peter L., Witney, Adam, Carucci, Daniel J., Azada, Abdu F., and Aguiar, Joao C.

Abstract

The transcriptional repertoire of the in vivo liver stage of Plasmodium has remained largely unidentified and seemingly not amenable to traditional molecular analysis because of the small number of parasites and large number of uninfected hepatocytes. We have overcome this obstruction by utilizing laser capture microdissection to provide a high quality source of parasite mRNA for the construction of a liver stage cDNA library. Sequencing and annotation of this library demonstrated expression of 623 different Plasmodium yoelii genes during development in the hepatocyte. Of these genes, 25% appear to be unique to the liver stage. This is the first comprehensive analysis of in vivo gene expression undertaken for the liver stage of P. yoelii, and provides insights into the differential expression of P. yoelii genes during this critical stage of development., The original document contains color images. Pub. in Molecular & Biochemical Parasitology, v142, p177-183, 2005.

Published
2005

12. Clinical trial in healthy malaria-naïve adults to evaluate the safety, tolerability, immunogenicity and efficacy of MuStDO5, a five-gene, sporozoite/hepatic stagePlasmodium falciparumDNA vaccine combined with escalating dose human GM-CSF DNA

Author

Richie, Thomas L., primary, Charoenvit, Yupin, additional, Wang, Ruobing, additional, Epstein, Judith E., additional, Hedstrom, Richard C., additional, Kumar, Sanjai, additional, Luke, Thomas C., additional, Freilich, Daniel A., additional, Aguiar, Joao C., additional, Sacci, Jr., John B., additional, Sedegah, Martha, additional, Nosek, Jr., Ronald A., additional, De La Vega, Patricia, additional, Berzins, Mara P., additional, Majam, Victoria F., additional, Abot, Esteban N., additional, Ganeshan, Harini, additional, Richie, Nancy O., additional, Banania, Jo Glenna, additional, Baraceros, Maria Fe B., additional, Geter, Tanya G., additional, Mere, Robin, additional, Bebris, Lolita, additional, Limbach, Keith, additional, Hickey, Bradley W., additional, Lanar, David E., additional, Ng, Jennifer, additional, Shi, Meng, additional, Hobart, Peter M., additional, Norman, Jon A., additional, Soisson, Lorraine A., additional, Hollingdale, Michael R., additional, Rogers, William O., additional, Doolan, Denise L., additional, and Hoffman, Stephen L., additional

Published
2012
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13. High-Throughput Generation of P. falciparum Functional Molecules by Recombinational Cloning

Author

NAVAL MEDICAL RESEARCH CENTER SILVER SPRING MD, Aguiar, Joao C., LaBaer, Joshua, Blair, Peter L., Shamailova, Victoria Y., Koundinya, Malvika, Russell, Joshua A., Huang, Fengying, Mar, Wenhong, Anthony, Robert M., Witney, Adam, Caruana, Sonia R., Brizuela, Leonardo, Sacci, Jr., John B., Hoffman, Stephen L., Carucci, Daniel J., NAVAL MEDICAL RESEARCH CENTER SILVER SPRING MD, Aguiar, Joao C., LaBaer, Joshua, Blair, Peter L., Shamailova, Victoria Y., Koundinya, Malvika, Russell, Joshua A., Huang, Fengying, Mar, Wenhong, Anthony, Robert M., Witney, Adam, Caruana, Sonia R., Brizuela, Leonardo, Sacci, Jr., John B., Hoffman, Stephen L., and Carucci, Daniel J.

Abstract

Large-scale functional genomics studies for malaria vaccine and drug development will depend on the generation of molecular tools to study protein expression. We examined the feasibility of a high-throughput cloning approach using the Gateway system to create a large set of expression clones encoding Plasmodium falciparum single-exon genes. Master clones and their ORFs were transferred en masse to multiple expression vectors. Target genes (n = 303) were selected using specific sets of criteria, including stage expression and secondary structure. Upon screening four colonies per capture reaction, we achieved 84% cloning efficiency. The genes were subcloned in parallel into three expression vectors: a DNA vaccine vector and two protein expression vectors. These transfers yielded a 100% success rate without any observed recombination based on single colony screening. The functional expression of 95 genes was evaluated in mice with DNA vaccine constructs to generate antibody against various stages of the parasite. From these, 19 induced antibody titers against the erythrocytic stages and three against sporozoite stages. We have overcome the potential limitation of producing large P. falciparum clone sets in multiple expression vectors. This approach represents a powerful technique for the production of molecular reagents for genome-wide functional analysis of the P. falciparum genome and will provide for a resource for the malaria resource community distributed through public repositories., The original document contains color images. Sponsored in part by the Office of Naval Research. Pub. in Genome Research, v14(10b) p2076-2082, Oct 2004. ISSN: 1088-9051/04.

Published
2004

14. Profiling humoral immune responses to P. falciparum infection with protein microarrays

Author

Doolan, Denise L., primary, Mu, Yunxiang, additional, Unal, Berkay, additional, Sundaresh, Suman, additional, Hirst, Siddiqua, additional, Valdez, Conrad, additional, Randall, Arlo, additional, Molina, Douglas, additional, Liang, Xiaowu, additional, Freilich, Daniel A., additional, Oloo, J. Aggrey, additional, Blair, Peter L., additional, Aguiar, Joao C., additional, Baldi, Pierre, additional, Davies, D. Huw, additional, and Felgner, Philip L., additional

Published
2008
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15. Novel Antigen Identification Method for Discovery of Protective Malaria Antigens by Rapid Testing of DNA Vaccines Encoding Exons from the Parasite Genome

Author

Haddad, Diana, primary, Bilcikova, Erika, additional, Witney, Adam A., additional, Carlton, Jane M., additional, White, Charles E., additional, Blair, Peter L., additional, Chattopadhyay, Rana, additional, Russell, Joshua, additional, Abot, Esteban, additional, Charoenvit, Yupin, additional, Aguiar, Joao C., additional, Carucci, Daniel J., additional, and Weiss, Walter R., additional

Published
2004
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16. A DNA vaccine encoding the 42 kDa C-terminus of merozoite surface protein 1 of Plasmodium falciparum induces antibody, interferon-γ and cytotoxic T cell responses in rhesus monkeys: immuno-stimulatory effects of granulocyte macrophage-colony stimulating factor

Author

Kumar, Sanjai, primary, Villinger, Francois, additional, Oakley, Miranda, additional, Aguiar, Joao C, additional, Jones, Trevor R, additional, Hedstrom, Richard C, additional, Gowda, Kalpana, additional, Chute, John, additional, Stowers, Anthony, additional, Kaslow, David C, additional, Thomas, Elaine K, additional, Tine, John, additional, Klinman, Dennis, additional, Hoffman, Stephen L, additional, and Weiss, Walter W, additional

Published
2002
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20. DNA vaccination against malaria

Author

Doolan, Denise L., primary, Sedegah, Martha, additional, Hedstrom, Richard C., additional, Aguiar, Joao C., additional, and Hoffman, Stephen L., additional

Published
1996
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21. Agglutination of Plasmodium falciparum-Infected Erythrocytes from East and West African Isolates by Human Sera from Distant Geographic Regions

Author

Aguiar, Joao C., primary, Howard, Russell J., additional, Lallinger, Gunter, additional, Jensen, James B., additional, McGregor, Ian A., additional, Sherwood, James A., additional, Martinez, Alberto, additional, Albrecht, Glenn R., additional, Minjas, J. N., additional, Greenwood, Brian M., additional, Patarroyo, M. Elkin, additional, Neequaye, Janet, additional, and Cegielski, Peter, additional

Published
1992
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22. Clinical trial in healthy malaria-naïve adults to evaluate the safety, tolerability, immunogenicity and efficacy of MuStDO5, a five-gene, sporozoite/ hepatic stage Plasmodium falciparum DNA vaccine combined with escalating dose human GM-CSF DNA.

Author

Richie, Thomas L., Charoenvit, Yupin, Wang, Ruobing, Epstein, Judith E., Hedstrom, Richard C., Kumar, Sanjai, Luke, Thomas C., Freilich, Daniel A., Aguiar, Joao C., Sacci Jr., John B., Sedegah, Martha, Nosek Jr., Ronald A., De La Vega, Patricia, Berzins, Mara P., Majam, Victoria F., Abot, Esteban N., Ganeshan, Harini, Richie, Nancy O., Banania, Jo Glenna, and Baraceros, Maria Fe B.

Published
2012
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24. Clinical trial in healthy malaria-naïve adults to evaluate the safety, tolerability, immunogenicity and efficacy of MuStDO5, a five-gene, sporozoite/hepatic stage Plasmodium falciparumDNA vaccine combined with escalating dose human GM-CSF DNA

Author

Richie, Thomas L., Charoenvit, Yupin, Wang, Ruobing, Epstein, Judith E., Hedstrom, Richard C., Kumar, Sanjai, Luke, Thomas C., Freilich, Daniel A., Aguiar, Joao C., Sacci, John B., Sedegah, Martha, Nosek, Ronald A., De La Vega, Patricia, Berzins, Mara P., Majam, Victoria F., Abot, Esteban N., Ganeshan, Harini, Richie, Nancy O., Banania, Jo Glenna, Baraceros, Maria Fe B., Geter, Tanya G., Mere, Robin, Bebris, Lolita, Limbach, Keith, Hickey, Bradley W., Lanar, David E., Ng, Jennifer, Shi, Meng, Hobart, Peter M., Norman, Jon A., Soisson, Lorraine A., Hollingdale, Michael R., Rogers, William O., Doolan, Denise L., and Hoffman, Stephen L.

Abstract

When introduced in the 1990s, immunization with DNA plasmids was considered potentially revolutionary for vaccine development, particularly for vaccines intended to induce protective CD8 T cell responses against multiple antigens. We conducted, in 1997−1998, the first clinical trial in healthy humans of a DNA vaccine, a single plasmid encoding Plasmodium falciparumcircumsporozoite protein (PfCSP), as an initial step toward developing a multi-antigen malaria vaccine targeting the liver stages of the parasite. As the next step, we conducted in 2000–2001 a clinical trial of a five-plasmid mixture called MuStDO5 encoding pre-erythrocytic antigens PfCSP, PfSSP2/TRAP, PfEXP1, PfLSA1 and PfLSA3. Thirty-two, malaria-naïve, adult volunteers were enrolled sequentially into four cohorts receiving a mixture of 500 μg of each plasmid plus escalating doses (0, 20, 100 or 500 μg) of a sixth plasmid encoding human granulocyte macrophage-colony stimulating factor (hGM-CSF). Three doses of each formulation were administered intramuscularly by needle-less jet injection at 0, 4 and 8 weeks, and each cohort had controlled human malaria infection administered by five mosquito bites 18 d later. The vaccine was safe and well-tolerated, inducing moderate antigen-specific, MHC-restricted T cell interferon-γ responses but no antibodies. Although no volunteers were protected, T cell responses were boosted post malaria challenge. This trial demonstrated the MuStDO5 DNA and hGM-CSF plasmids to be safe and modestly immunogenic for T cell responses. It also laid the foundation for priming with DNA plasmids and boosting with recombinant viruses, an approach known for nearly 15 y to enhance the immunogenicity and protective efficacy of DNA vaccines.

Published
2012
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25. In Vitro Rosetting, Cytoadherence, and Microagglutination Properties of Plasmodium falciparum-InfectedErythrocytes From Gambian and Tanzanian Patients

Author

Hasler, Thomas, Handunnetti, Shiroma M., Aguiar, Joao C., Schravendijk, Marie Rose van, Greenwood, Brian M., Lallinger, Gunther, Cegielski, Peter, and Howard, Russell J.

Abstract

To understand the molecular mechanisms that lead to sequestration of red blood cells infected with mature stages of Plasmodium falciparumand to examine the relevance of earlier studies on adherence properties of laboratory-derived P falciparumparasites to the natural parasite population, we analyzed Gambian and Tanzanian isolates for in vitro cytoadherence and antibody-mediated microagglutination. Eighteen cryopreserved isolates of ring-stage parasites were cultured for 20 to 30 hours in vitro, in the patients original erythrocytes, to the trophozoite and schizont stage. All parasites were positive in the microagglutination assay with at least one of four African hyperimmune sera. In a rosetting assay, only 2 of the 18 isolates were strongly positive (35% and 41 % of parasitized erythrocytes with more than two uninfected cells bound). Thirteen isolates showed either intermediate (5% to 18%) or low (<5%) rosetting while three isolates did not form rosettes. Infected cell-binding of the different isolates to immobilized CD36 or thrombospondin, or C32 melanoma cells correlated with the percentage of mature parasites in the blood samples (r= .932 for CD36, r = .946 for thrombospondin, and r= .881 for C32 melanoma cells). There was a high correlation between binding to CD36 and thrombospondin (r= .982). The extent of infected cell rosetting with uninfected cells in these blood samples was not correlated with these other receptor properties. We also observed coexpression of rosetting and cytoadherence receptors on the same parasitized erythrocytes.

Published
1990
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26. Protection of Mice against Plasmodium yoeliiSporozoite Challenge with P. yoeliiMerozoite Surface Protein 1 DNA Vaccines

Author

Becker, Sylvia I., Wang, Ruobing, Hedstrom, Richard C., Aguiar, Joao C., Jones, Trevor R., Hoffman, Stephen L., and Gardner, Malcolm J.

Abstract

ABSTRACTImmunization of mice with DNA vaccines encoding the full-length form and C and N termini of Plasmodium yoeliimerozoite surface protein 1 provided partial protection against sporozoite challenge and resulted in boosting of antibody titers after challenge. In C57BL/6 mice, two DNA vaccines provided protection comparable to that of recombinant protein consisting of the C terminus in Freund’s adjuvant.

Published
1998
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30. A DNA vaccine encoding the 42 kDa C-terminus of merozoite surface protein 1 of Plasmodium falciparum induces antibody, interferon-gamma and cytotoxic T cell responses in rhesus monkeys: immuno-stimulatory effects of granulocyte macrophage-colony stimulating factor.

Author

Kumar S, Villinger F, Oakley M, Aguiar JC, Jones TR, Hedstrom RC, Gowda K, Chute J, Stowers A, Kaslow DC, Thomas EK, Tine J, Klinman D, Hoffman SL, and Weiss WW

Subjects
Adjuvants, Immunologic pharmacology, Animals, Antibodies, Protozoan immunology, Cytotoxicity, Immunologic, Granulocyte-Macrophage Colony-Stimulating Factor pharmacology, Humans, Interferon-gamma immunology, Macaca mulatta, Merozoite Surface Protein 1 genetics, Merozoite Surface Protein 1 immunology, Plasmids, Plasmodium falciparum genetics, Recombinant Fusion Proteins immunology, T-Lymphocytes, Cytotoxic immunology, Granulocyte-Macrophage Colony-Stimulating Factor immunology, Malaria Vaccines genetics, Malaria Vaccines immunology, Malaria, Falciparum immunology, Malaria, Falciparum prevention & control, Plasmodium falciparum immunology, Vaccines, DNA genetics, Vaccines, DNA immunology
Abstract

We have constructed a DNA plasmid vaccine encoding the C-terminal 42-kDa region of the merozoite surface protein 1 (pMSP1(42)) from the 3D7 strain of Plasmodium falciparum (Pf3D7). This plasmid expressed recombinant MSP1(42) after in vitro transfection in mouse VM92 cells. Rhesus monkeys immunized with pMSP1(42) produced antibodies reactive with Pf3D7 infected erythrocytes by IFAT, and by ELISA against yeast produced MSP1(19) (yMSP1(19)). Immunization also induced antigen specific T cell responses as measured by interferon-gamma production, and by classical CTL chromium release assays. In addition, immunization with pMSP1(42) primed animals for an enhanced antibody response to a subsequent boost with the recombinant yMSP1(19). We also evaluated Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) as an adjuvant for pMSP1(42.) We tested both rhesus GM-CSF expressed from a DNA plasmid, and E. coli produced recombinant human GM-CSF. Plasmids encoding rhesus GM-CSF (prhGM-CSF) and human GM-CSF (phuGM-CSF) were constructed; these plasmids expressed bio-active recombinant GMCSF. Co-immunization with a mixture of prhGM-CSF and pMSP1(42) induced higher specific antibody responses after the first dose of plasmid, but after three doses of DNA monkeys immunized with or without prhGM-CSF had the same final antibody titers and T cell responses. In comparison, rhuGM-CSF protein did not lead to accelerated antibody production after the first DNA dose. However, antibody titers were maintained at a slightly higher level in monkeys receiving GM-CSF protein, and they had a higher response to boosting with recombinant MSP1(19). The GM-CSF plasmid or protein appears to be less potent as an adjuvant in rhesus monkeys than each is in mice, and more work is needed to determine if GM-CSF can be a useful adjuvant in DNA vaccination of primates.

Published
2002
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