Malaria vaccines: Focus on adenovirus based vectors

Vaccine

Volumn 30, Issue 35, 2012, Pages 5191-5198

  Schuldt, Nathaniel J ^a   Amalfitano, Andrea ^a,b,c  

^a MICHIGAN STATE UNIVERSITY   (United States)

^b MICHIGAN STATE UNIVERSITY   (United States)

^c NONE   (United States)

Author keywords

Adenovirus; Circumsporozoite protein; Malaria; Review; Vaccine

Indexed keywords

ADENOVIRUS VECTOR; APICAL MEMBRANE ANTIGEN 1; CHLOROQUINE; CIRCUMSPOROZOITE PROTEIN; HEPATITIS B SURFACE ANTIGEN; MALARIA VACCINE; PHARMACEUTICAL VEHICLES AND ADDITIVES; PHOSPHORYL LIPID A; RTS,S VACCINE; THROMBOSPONDIN; UNCLASSIFIED DRUG;

DRUG FORMULATION; DRUG RESEARCH; HUMAN; IMMUNE RESPONSE; MALARIA; NONHUMAN; PLASMODIUM FALCIPARUM; PLASMODIUM KNOWLESI; PLASMODIUM MALARIAE; PLASMODIUM OVALE; PLASMODIUM VIVAX; PRIORITY JOURNAL; REVIEW;

ADENOVIRIDAE; ANTIBODIES, PROTOZOAN; ANTIGENS, PROTOZOAN; GENETIC VECTORS; HUMANS; MALARIA VACCINES; MALARIA, FALCIPARUM; PROTOZOAN PROTEINS; VACCINATION;

EID: 84863828594     PISSN: 0264410X     EISSN: 18732518     Source Type: Journal    
DOI: 10.1016/j.vaccine.2012.05.048     Document Type: Review

References (95)

1. World malaria report: 2010 2010, WHO, World Health Organization, Geneva.
2. World malaria report 2009 2008, WHO, World Health Organization, Geneva.
3. Lars H. Development of vaccines against Plasmodium falciparum malaria: taking lessons from naturally acquired protective immunity. Microbes Infect 2007, 9:772-776.
4. Cohen S., McGregor I.A., Carrington S. Gamma-globulin and acquired immunity to human malaria. Nature 1961, 192:733-737.
5. Kinyanjui S., Bejon P., Osier F., Bull P., Marsh K. What you see is not what you get: implications of the brevity of antibody responses to malaria antigens and transmission heterogeneity in longitudinal studies of malaria immunity. Malar J 2009, 8:242.
6. Anders R.F. Multiple cross-reactivities amongst antigens of Plasmodium falciparum impair the development of protective immunity against malaria. Parasite Immunol 1986, 8:529-539.
7. Dorfman J.R., Bejon P., Ndungu F.M., Langhorne J., Kortok M.M., Lowe B.S., et al. B cell memory to 3 Plasmodium falciparum blood-stage antigens in a malaria-endemic area. J Infect Dis 2005, 191:1623-1630.
8. Butler N.S., Moebius J., Pewe L.L., Traore B., Doumbo O.K., Tygrett L.T., et al. Therapeutic blockade of PD-L1 and LAG-3 rapidly clears established blood-stage Plasmodium infection. Nat Immunol 2012, 13:188-195.
9. Doolan D.L. Plasmodium immunomics. Int J Parasitol 2011, 41:3-20.
10. Clyde D.F., McCarthy V.C., Miller R.M., Woodward W.E. Immunization of man against Falciparum and Vivax malaria by use of attenuated sporozoites. Am J Trop Med Hyg 1975, 24:397-401.
11. Rieckmann K.H., Beaudoin R.L., Cassells J.S., Sell K.W. Current considerations in vaccination of humans against malaria: use of attenuated sporozoites in the immunization of human volunteers against falciparum malaria. Bull World Health Organ 1979, 57:261-265.
12. Hoffman Stephen L., Goh Lucy M.L., Luke Thomas C., Schneider I., Le Thong P., Doolan Denise L., et al. Protection of humans against malaria by immunization with radiation-attenuated Plasmodium falciparum sporozoites. J Infect Dis 2002, 185:1155-1164.
13. Roestenberg M., Teirlinck A.C., McCall M.B.B., Teelen K., Makamdop K.N., Wiersma J., et al. Long-term protection against malaria after experimental sporozoite inoculation: an open-label follow-up study. Lancet 2011, 377:1770-1776.
14. Roestenberg M., McCall M., Hopman J., Wiersma J., Luty A.J.F., van Gemert G.J., et al. Protection against a malaria challenge by sporozoite inoculation. N Engl J Med 2009, 361:468-477.
15. Phillips-Howard P.A. Efficacy of drug prophylaxis. J R Soc Med 1989, 82(Suppl. 17):23-29.
16. Overstreet M.G., Cockburn I.A., Chen Y.C., Zavala F. Protective CD8T cells against Plasmodium liver stages: immunobiology of an 'unnatural' immune response. Immunol Rev 2008, 225:272-283.
17. Epstein J.E., Tewari K., Lyke K.E., Sim B.K.L., Billingsley P.F., Laurens M.B., et al. Live attenuated malaria vaccine designed to protect through hepatic CD8+ T cell immunity. Science 2011, 334:475-480.
18. Schmidt N.W., Butler N.S., Badovinac V.P., Harty J.T. Extreme CD8 T cell requirements for anti-malarial liver-stage immunity following immunization with radiation attenuated sporozoites. PLoS Pathog 2010, 6:e1000998.
19. Reyes-Sandoval A., Wyllie D.H., Bauza K., Milicic A., Forbes E.K., Rollier C.S., et al. CD8+ T effector memory cells protect against liver-stage malaria. J Immunol 2011.
20. Romero P., Maryanski J.L., Corradin G., Nussenzweig R.S., Nussenzweig V., Zavala F. Cloned cytotoxic T cells recognize an epitope in the circumsporozoite protein and protect against malaria. Nature 1989, 341:323-326.
21. Daubenberger C.A. First clinical trial of purified, irradiated malaria sporozoites in humans. Expert Rev Vaccines 2011, 11:31-33.
22. Butler Noah S., Schmidt Nathan W., Vaughan Ashley M., Aly Ahmed S., Kappe Stefan H.I., Harty John T. Superior antimalarial immunity after vaccination with late liver stage-arresting genetically attenuated parasites. Cell Host Microbe 2011, 9:451-462.
23. Hamilton A.J., Suhrbier A., Nicholas J., Sinden R.E. Immunoelectron microscopic localization of circumsporozoite antigen in the differentiating exoerythrocytic trophozoite of Plasmodium berghei. Cell Biol Int Rep 1988, 12:123-129.
24. Arun Kumar K., Sano G.-i., Boscardin S., Nussenzweig R.S., Nussenzweig M.C., Zavala F., et al. The circumsporozoite protein is an immunodominant protective antigen in irradiated sporozoites. Nature 2006, 444:937-940.
25. Grüner A.C., Mauduit M., Tewari R., Romero J.F., Depinay N., Kayibanda M., et al. Sterile protection against malaria is independent of immune responses to the circumsporozoite protein. PLoS ONE 2007, 2:e1371.
26. Mauduit M., Gruner A.C., Tewari R., Depinay N., Kayibanda M., Chavatte J.-M., et al. A role for immune responses against non-CS components in the cross-species protection induced by immunization with irradiated malaria sporozoites. PLoS ONE 2009, 4:e7717.
27. Kester Kent E., Cummings James F., Ofori-Anyinam O., Ockenhouse Christian F., Krzych U., Moris P., et al. Randomized, double-blind, phase 2a trial of Falciparum malaria vaccines RTS,S/AS01B and RTS,S/AS02A in malaria-naive adults: safety, efficacy, and immunologic associates of protection. J Infect Dis 2009, 200:337-346.
28. Bejon P., Lusingu J., Olotu A., Leach A., Lievens M., Vekemans J., et al. Efficacy of RTS,S/AS01E vaccine against malaria in children 5 to 17 months of age. N Engl J Med 2008, 359:2521-2532.
29. Kester K.E., Cummings J.F., Ockenhouse C.F., Nielsen R., Hall B.T., Gordon D.M., et al. Phase 2a trial of 0, 1, and 3 month and 0, 7, and 28 day immunization schedules of malaria vaccine RTS,S/AS02 in malaria-naïve adults at the Walter Reed Army Institute of Research. Vaccine 2008, 26:2191-2202.
30. Coler R.N., Carter D., Friede M., Reed S.G. Adjuvants for malaria vaccines. Parasite Immunol 2009, 31:520-528.
31. Gregson A.L., Oliveira G., Othoro C., Calvo-Calle J.M., Thorton G.B., Nardin E., et al. Phase I trial of an alhydrogel adjuvanted hepatitis B core virus-like particle containing epitopes of Plasmodium falciparum circumsporozoite protein. PLoS ONE 2008, 3:e1556.
32. Casares S., Brumeanu T.D., Richie T.L. The RTS,S malaria vaccine. Vaccine 2010, 28:4880-4894.
33. Vekemans J., Marsh K., Greenwood B., Leach A., Kabore W., Soulanoudjingar S., et al. Assessment of severe malaria in a multicenter, phase III, RTS,S/AS01 malaria candidate vaccine trial: case definition, standardization of data collection and patient care. Malar J 2011, 10:221.
34. Olotu A., Moris P., Mwacharo J., Vekemans J., Kimani D., Janssens M., et al. Circumsporozoite-specific T cell responses in children vaccinated with RTS, S/AS01E and protection against P. falciparum clinical malaria. PLoS ONE 2011, 6:e25786.
35. Patterson L.J., Robert-Guroff M. Replicating adenovirus vector prime/protein boost strategies for HIV vaccine development. Expert Opin Biol Ther 2008, 8:1347-1363.
36. First results of phase 3 trial of RTS,S/AS01 malaria vaccine in African children. N Engl J Med 2011, 365:1863-1875. The RTS SCTP.
37. Schwenk R., Asher L.V., Chalom I., Lanar D., Sun P., White K., et al. Opsonization by antigen-specific antibodies as a mechanism of protective immunity induced by Plasmodium falciparum circumsporozoite protein-based vaccine. Parasite Immunol 2003, 25:17-25.
38. Sun P., Schwenk R., White K., Stoute J.A., Cohen J., Ballou W.R., et al. Protective immunity induced with malaria vaccine, RTS,S, is linked to Plasmodium falciparum circumsporozoite protein-specific CD4+ and CD8+ T cells producing IFN-{gamma}. J Immunol 2003, 171:6961-6967.
39. Stewart V.A., McGrath S.M., Dubois P.M., Pau M.G., Mettens P., Shott J., et al. Priming with an adenovirus 35-circumsporozoite protein (CS) vaccine followed by RTS,S/AS01B boosting significantly improves immunogenicity to Plasmodium falciparum CS compared to that with either malaria vaccine alone. Infect Immun 2007, 75:2283-2290.
40. Limbach K.J., Richie T.L. Viral vectors in malaria vaccine development. Parasite Immunol 2009, 31:501-519.
41. Carey J.B., Pearson F.E., Vrdoljak A., McGrath M.G., Crean A.M., Walsh P.T., et al. Microneedle array design determines the induction of protective memory CD8+ T cell responses induced by a recombinant live malaria vaccine in mice. PLoS ONE 2011, 6:e22442.
42. Jiang G., Shi M., Conteh S., Richie N., Banania G., Geneshan H., et al. Sterile protection against Plasmodium knowlesi in rhesus monkeys from a malaria vaccine: comparison of heterologous prime boost strategies. PLoS ONE 2009, 4:e6559.
43. Amalfitano A.P.R. Separating fact from fiction: assessing the potential of modified adenovirus vectors for use in human gene therapy. Curr Gene Ther 2002, 2:111-133. (123).
44. Anderson R.D., Haskell R.E., Xia H., Roessler B.J., Davidson B.L. A simple method for the rapid generation of recombinant adenovirus vectors. Nat Gene Ther 2000, 7:1034-1038.
45. He T-C., Zhou S., da Costa L.T., Yu J., Kinzler K.W., Vogelstein B. A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci USA 1998, 95:2509-2514.
46. Croyle M.A., Patel A., Tran K.N., Gray M., Zhang Y., Strong J.E., et al. Nasal delivery of an adenovirus-based vaccine bypasses pre-existing immunity to the vaccine carrier and improves the immune response in mice. PLoS ONE 2008, 3:e3548.
47. Wang L., Cheng C., Ko S-Y., Kong W-P., Kanekiyo M., Einfeld D., et al. Delivery of human immunodeficiency virus vaccine vectors to the intestine induces enhanced mucosal cellular immunity. J Virol 2009, 83:7166-7175.
48. Gonin P., Oualikene W., Fournier A., Eloit M. Comparison of the efficacy of replication-defective adenovirus and Nyvac poxvirus as vaccine vectors in mice. Vaccine 1996, 14:1083-1087.
49. Hangalapura B.N., Oosterhoff D., Gupta T., de Groot J., Wijnands P.G.J.T.B., van Beusechem V.W., et al. Delivery route, MyD88 signaling and cross-priming events determine the anti-tumor efficacy of an adenovirus based melanoma vaccine. Vaccine 2011, 29:2313-2321.
50. Rodrigues E.G., Zavala F., Eichinger D., Wilson J.M., Tsuji M. Single immunizing dose of recombinant adenovirus efficiently induces CD8+ T cell-mediated protective immunity against malaria. J Immunol 1997, 158:1268-1274.
51. Gabitzsch E.S., Xu Y., Yoshida L.H., Balint J., Amalfitano A., Jones F.R. Novel Adenovirus type 5 vaccine platform induces cellular immunity against HIV-1 Gag, Pol, Nef despite the presence of Ad5 immunity. Vaccine 2009, 27:6394-6398.
52. Gabitzsch E.S., Xu Y., Yoshida L.H., Balint J., Gayle R.B., Amalfitano A., et al. A preliminary and comparative evaluation of a novel Ad5 [E1-, E2b-] recombinant-based vaccine used to induce cell mediated immune responses. Immunol Lett 2009, 122:44-51.
53. Weaver E.A., Nehete P.N., Buchl S.S., Senac J.S., Palmer D., Ng P., et al. Comparison of replication-competent, first generation, and helper-dependent adenoviral vaccines. PLoS ONE 2009, 4:e5059.
54. Appledorn D.M., Patial S., McBride A., Godbehere S., Van Rooijen N., Parameswaran N., et al. Adenovirus vector-induced innate inflammatory mediators, MAPK, signaling, as well as adaptive immune responses are dependent upon both TLR2 and TLR9 in vivo. J Immunol 2008, 181:2134-2144.
55. Appledorn D.M., McBride A., Seregin S., Scott J.M., Schuldt N., Kiang A., et al. Complex interactions with several arms of the complement system dictate innate and humoral immunity to adenoviral vectors. Gene Ther 2008, 15:1606-1617.
56. Hartman Z.C., Appledorn D.M., Amalfitano A. Adenovirus vector induced innate immune responses: impact upon efficacy and toxicity in gene therapy and vaccine applications. Virus Res 2008, 132:1-14.
57. Barlan A.U., Danthi P., Wiethoff C.M. Lysosomal localization and mechanism of membrane penetration influence nonenveloped virus activation of the NLRP3 inflammasome. Virology 2011, 412:306-314.
58. Iwasaki A., Medzhitov R. Regulation of adaptive immunity by the innate immune system. Science 2010, 327:291-295.
59. Xiang Z.Q., Yang Y., Wilson J.M., Ertl H.C.J. A replication-defective human adenovirus recombinant serves as a highly efficacious vaccine carrier. Virology 1996, 219:220-227.
60. Shott J.P., McGrath S.M., Pau M.G., Custers J.H.V., Ophorst O., Demoitié M-A., et al. Adenovirus 5 and 35 vectors expressing Plasmodium falciparum circumsporozoite surface protein elicit potent antigen-specific cellular IFN-[gamma] and antibody responses in mice. Vaccine 2008, 26:2818-2823.
61. Naslund T.I., Uyttenhove C., Nordstrom E.K.L., Colau D., Warnier G., Jondal M., et al. Comparative prime-boost vaccinations using Semliki forest virus, adenovirus, and ALVAC vectors demonstrate differences in the generation of a protective central memory CTL response against the P815 tumor. J Immunol 2007, 178:6761-6769.
62. Bruna-Romero O., Rocha C.D., Tsuji M., Gazzinelli R.T. Enhanced protective immunity against malaria by vaccination with a recombinant adenovirus encoding the circumsporozoite protein of Plasmodium lacking the GPI-anchoring motif. Vaccine 2004, 22:3575-3584.
63. Rodrigues E.G., Zavala F., Nussenzweig R.S., Wilson J.M., Tsuji M. Efficient induction of protective anti-malaria immunity by recombinant adenovirus. Vaccine 1998, 16:1812-1817.
64. Bruder J.T., Stefaniak M.E., Patterson N.B., Chen P., Konovalova S., Limbach K., et al. Adenovectors induce functional antibodies capable of potent inhibition of blood stage malaria parasite growth. Vaccine 2010, 28:3201-3210.
65. Tamminga C., Sedegah M., Regis D., Chuang I., Epstein J.E., Spring M., et al. Adenovirus-5-vectored P. falciparum vaccine expressing CSP and AMA1. Part B: safety, immunogenicity and protective efficacy of the CSP component. PLoS ONE 2011, 6:e25868.
66. Tamminga C. Lack of protective efficacy of an adenovirus-vectored P. falciparum malaria vaccine in the absence of DNA priming. 60th Annual American Society of Tropical Medicine and Hygiene Meeting 2011.
67. Sedegah M., Tamminga C., McGrath S., House B., Ganeshan H., Lejano J., et al. Adenovirus 5-vectored P. falciparum vaccine expressing CSP and AMA1. Part A: safety and immunogenicity in seronegative adults. PLoS ONE 2011, 6:e24586.
68. Barouch D.H., Kik S.V., Weverling G.J., Dilan R., King S.L., Maxfield L.F., et al. International seroepidemiology of adenovirus serotypes 5, 26, 35, and 48 in pediatric and adult populations. Vaccine 2011, 29:5203-5209.
69. Abbink P., Lemckert A.A.C., Ewald B.A., Lynch D.M., Denholtz M., Smits S., et al. Comparative seroprevalence and immunogenicity of six rare serotype recombinant adenovirus vaccine vectors from subgroups B and D. J Virol 2007, 81:4654-4663.
70. McCoy K., Tatsis N., Korioth-Schmitz B., Lasaro M.O., Hensley S.E., Lin S-W., et al. Effect of preexisting immunity to adenovirus human serotype 5 antigens on the immune responses of nonhuman primates to vaccine regimens based on human- or chimpanzee-derived adenovirus vectors. J Virol 2007, 81:6594-6604.
71. Haveman L.M., Bierings M., Legger E., Klein M.R., de Jager W., Otten H.G., et al. Novel pan-DR-binding T cell epitopes of adenovirus induce pro-inflammatory cytokines and chemokines in healthy donors. Int Immunol 2006, 18:1521-1529.
72. Joshi A., Tang J., Kuzma M., Wagner J., Mookerjee B., Filicko J., et al. Adenovirus DNA polymerase is recognized by human CD8+ T cells. J Gen Virol 2009, 90:84-94.
73. Zaiss A.K., Machado H.B., Herschman H.R. The influence of innate and pre-existing immunity on adenovirus therapy. J Cell Biochem 2009, 108:778-790.
74. Osada T., Yang X.Y., Hartman Z.C., Glass O., Hodges B.L., Niedzwiecki D., et al. Optimization of vaccine responses with an E1, E2b and E3-deleted Ad5 vector circumvents pre-existing anti-vector immunity. Cancer Gene Ther 2009, 16:673-682.
75. Amalfitano A., Hauser M.A., Hu H., Serra D., Begy C.R., Chamberlain J.S. Production and characterization of improved adenovirus vectors with the E1, E2b, and E3 genes deleted. J Virol 1998, 72:926-933.
76. Ophorst O.J., Radosevic K., Havenga M.J., Pau M.G., Holterman L., Berkhout B., et al. Immunogenicity and protection of a recombinant human adenovirus serotype 35-based malaria vaccine against Plasmodium yoelii in mice. Infect Immun 2006, 74:313-320.
77. Gaggar A., Shayakhmetov D.M., Lieber A. CD46 is a cellular receptor for group B adenoviruses. Nat Med 2003, 9:1408-1412.
78. Rodríguez A., Mintardjo R., Tax D., Gillissen G., Custers J., Pau M.G., et al. Evaluation of a prime-boost vaccine schedule with distinct adenovirus vectors against malaria in rhesus monkeys. Vaccine 2009, 27:6226-6233.
79. Radosevic K., Rodriguez A., Lemckert A.A., van der Meer M., Gillissen G., Warnar C., et al. The Th1 immune response to Plasmodium falciparum circumsporozoite protein is boosted by adenovirus vectors 35 and 26 with a homologous insert. Clin Vaccine Immunol 2010, 17:1687-1694.
80. Reyes-Sandoval A., Sridhar S., Berthoud T., Moore A.C., Harty J.T., Gilbert S.C., et al. Single-dose immunogenicity and protective efficacy of simian adenoviral vectors against Plasmodium berghei. Eur J Immunol 2008, 38:732-741.
81. Sheehy S.H., Duncan C.J.A., Elias S.C., Collins K.A., Ewer K.J., Spencer A.J., et al. Phase Ia clinical evaluation of the Plasmodium falciparum blood-stage antigen MSP1 in ChAd63 and MVA vaccine vectors. Mol Ther 2011, 19:2269-2276.
82. Biswas S., Dicks M.D.J., Long C.A., Remarque E.J., Siani L., Colloca S., et al. Transgene optimization, immunogenicity and in vitro efficacy of viral vectored vaccines expressing two alleles of Plasmodium falciparum AMA1. PLoS ONE 2011, 6:e20977.
83. Draper S.J., Biswas S., Spencer A.J., Remarque E.J., Capone S., Naddeo M., et al. Enhancing blood-stage malaria subunit vaccine immunogenicity in rhesus macaques by combining adenovirus, poxvirus, and protein-in-adjuvant vaccines. J Immunol 2010, 185:7583-7595.
84. Reyes-Sandoval A., Berthoud T., Alder N., Siani L., Gilbert S.C., Nicosia A., et al. Prime-boost immunization with adenoviral and modified vaccinia virus Ankara vectors enhances the durability and polyfunctionality of protective malaria CD8+ T-Cell responses. Infect Immun 2010, 78:145-153.
85. Goodman A.L., Blagborough A.M., Biswas S., Wu Y., Hill A.V., Sinden R.E., et al. A viral vectored prime-boost immunization regime targeting the malaria Pfs25 antigen induces transmission-blocking activity. PLoS ONE 2011, 6:e29428.
86. Hartman Z.C., Appledorn D.M., Serra D., Glass O., Mendelson T.B., Clay T.M., et al. Replication-attenuated human adenoviral type 4 vectors elicit capsid dependent enhanced innate immune responses that are partially dependent upon interactions with the complement system. Virology 2008, 374:453-467.
87. Appledorn D.M., Kiang A., McBride A., Jiang H., Seregin S., Scott J.M., et al. Wild-type adenoviruses from groups A-F evoke unique innate immune responses, of which HAd3 and SAd23 are partially complement dependent. Gene Ther 2008, 15:885-901.
88. Appledorn D.M., Aldhamen Y.A., DePas W., Seregin S.S., Liu C.-J.J., Schuldt N., et al. A new adenovirus based vaccine vector expressing an Eimeria tenella derived TLR agonist improves cellular immune responses to an antigenic target. PLoS ONE 2010, 5:e9579.
89. Schuldt N.J., Aldhamen Y.A., Appledorn D.M., Seregin S.S., Kousa Y., Godbehere S., et al. Vaccine platforms combining circumsporozoite protein and potent immune modulators, rEA or EAT-2, paradoxically result in opposing immune responses. PLoS ONE 2011, 6:e24147.
90. Bayer W., Tenbusch M., Lietz R., Johrden L., Schimmer S., Uberla K., et al. Vaccination with an adenoviral vector that encodes and displays a retroviral antigen induces improved neutralizing antibody and CD4+ T-cell responses and confers enhanced protection. J Virol 2010, 84:1967-1976.
91. Lu S. Heterologous prime-boost vaccination. Curr Opin Immunol 2009, 21:346-351.
92. Rerks-Ngarm S., Pitisuttithum P., Nitayaphan S., Kaewkungwal J., Chiu J., Paris R., et al. Vaccination with ALVAC and AIDSVAX to prevent HIV-1 infection in Thailand. N Engl J Med 2009, 361:2209-2220.
93. Withers M.R., Mckinney D., Ogutu B.R., Waitumbi J.N., Milman J.B., Apollo O.J., et al. Safety and reactogenicity of an MSP-1 malaria vaccine candidate: a randomized phase Ib does-escalation trial in jenyan children. PLoS Clin Trials 2006, 1(7):e32.
94. Cummings J.F., Spring M.D., Schwenk R.J., Ockenhouse C.F., Kester K.E., Polhemus M.E., et al. Recombinant liver stage antigen-1 (LSA-1) formulated with AS01 or AS02 is safe, elicits high titer antibody and induces IFN-gamma/IL-2 CD4+ T cells but does not protect against experimental Plasmodium falciparum infection. Vaccine 2010, 28(31):135-5144.
95. Pattnaik P., Shakri A.R., Singh S., Goel S., Mukherjee P., Chitnis C.E. Immunogenicity of a recombinant malaria vaccine based on receptor binding domain of Plasmodium falciparum EBA-175. Vaccine 2007, 25(5):6-813.