Molecular dissection of the human antibody response to the structural repeat epitope of Plasmodium falciparum sporozoite from a protected donor

Jonathan A Chappel, William O Rogers, Stephen L Hoffman, Angray S Kang, Jonathan A Chappel, William O Rogers, Stephen L Hoffman, Angray S Kang

Abstract

Background: The circumsporozoite surface protein is the primary target of human antibodies against Plasmodium falciparum sporozoites, these antibodies are predominantly directed to the major repetitive epitope (Asn-Pro-Asn-Ala)n, (NPNA)n. In individuals immunized by the bites of irradiated Anopheles mosquitoes carrying P. falciparum sporozoites in their salivary glands, the anti-repeat response dominates and is thought by many to play a role in protective immunity.

Methods: The antibody repertoire from a protected individual immunized by the bites of irradiated P. falciparum infected Anopheles stephensi was recapitulated in a phage display library. Following affinity based selection against (NPNA)3 antibody fragments that recognized the PfCSP repeat epitope were rescued.

Results: Analysis of selected antibody fragments implied the response was restricted to a single antibody fragment consisting of VH3 and VkappaI families for heavy and light chain respectively with moderate affinity for the ligand.

Conclusion: The dissection of the protective antibody response against the repeat epitope revealed that the response was apparently restricted to a single VH/VL pairing (PfNPNA-1). The affinity for the ligand was in the microM range. If anti-repeat antibodies are involved in the protective immunity elicited by exposure to radiation attenuated P. falciparum sporozoites, then high circulating levels of antibodies against the repeat region may be more important than intrinsic high affinity for protection. The ability to attain and sustain high levels of anti-(NPNA)n will be one of the key determinants of efficacy for a vaccine that relies upon anti-PfCSP repeat antibodies as the primary mechanism of protective immunity against P. falciparum.

Figures

Figure 1
Figure 1
VH/κ library construction. A schematic diagram of the steps involved in constructing a VH/κ library from mRNA isolated from PBL.
Figure 2
Figure 2
Illustration of vectors pORFES, JC-M13-88 and pAbHIS.
Figure 3
Figure 3
Indirect immunofluorescence assay (IFA) on Plasmodium falciparum sporozoites. Panel (A) PfNPNA-1 VH/κ, (B) 2A10 MAb.
Figure 4
Figure 4
Competition ELISA.
Figure 5
Figure 5
Determination of specificity of PfNPNA-1. The binding of immune serum (WR5), non-immune serum and PfNPNA-1 VH/κ to R32tet32, recombinant hepatitis core containing (NANP)4 peptide sequence and (NPNA)3C-BSA conjugate coated microtiter plate well was determined by ELISA essentially as described in Figure 4. The serum(s) and the recombinant PfNPNA-1 VH/κ were diluted 1/16 and 1/10 respectively.

References

    1. Breman JG, Egan A, Keusch GT. The intolerable burden of malaria: a new look at the numbers. Am J Trop Med Hyg. 2001;64:iv–vii.
    1. Sachs J, Malaney P. The economic and social burden of malaria. Nature. 2002;415:680–685. doi: 10.1038/415680a.
    1. Nussenzweig RS, Vanderberg J, Most H, Orton C. Protective immunity produced by the injection of x-irradiated sporozoites of Plasmodium berghei. Nature. 1967;216:160–162.
    1. Hoffman SL, Goh LM, Luke TC, Schneider I, Le TP, Doolan DL, Sacci J, de la Vega P, Dowler M, Paul C, Gordon DM, Stoute JA, Church LW, Sedegah M, Heppner DG, Ballou WR, Richie TL. Protection of humans against malaria by immunization with radiation-attenuated Plasmodium falciparum sporozoites. J Infect Dis. 2002;185:1155–1164. doi: 10.1086/339409.
    1. Potocnjak P, Yoshida N, Nussenzweig RS, Nussenzweig V. Monovalent fragments (Fab) of monoclonal antibodies to a sporozoite surface antigen (Pb44) protect mice against malarial infection. J Exp Med. 1980;151:1504–1513. doi: 10.1084/jem.151.6.1504.
    1. Dyson HJ, Satterthwait AC, Lerner RA, Wright PE. Conformational preferences of synthetic peptides derived from the immunodominant site of the circumsporozoite protein of Plasmodium falciparum by 1H NMR. Biochemistry. 1990;29:7828–7837.
    1. Egan JE, Hoffman SL, Haynes JD, Sadoff JC, Schneider I, Grau GE, Hollingdale MR, Ballou WR, Gordon DM. Humoral immune responses in volunteers immunized with irradiated Plasmodium falciparum sporozoites. Am J Trop Med Hyg. 1993;49:166–173.
    1. Huse WD, Sastry L, Iverson SA, Kang AS, Alting-Mees M, Burton DR, Benkovic SJ, Lerner RA. Generation of a large combinatorial library of the immunoglobulin repertoire in phage lambda. Science. 1989;246:1275–81.
    1. Kang A, Burton D, Lerner RA. Combinatorial immunoglobulin libraries in phage lambda. Methods: A companion to Methods in Enzymology. 1991;2:111–118.
    1. McCafferty J, Griffiths AD, Winter G, Chiswell DJ. Phage antibodies: filamentous phage displaying antibody variable domains. Nature. 1990;348:552–554. doi: 10.1038/348552a0.
    1. Kang AS, Barbas CF, Janda KD, Benkovic SJ, Lerner RA. Linkage of recognition and replication functions by assembling combinatorial antibody Fab libraries along phage surfaces. Proc Natl Acad Sci U S A. 1991;88:4363–4366.
    1. Barbas CF, 3rd, Kang AS, Lerner RA, Benkovic SJ. Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Proc Natl Acad Sci U S A. 1991;88:7978–7982.
    1. Winter G, Griffiths AD, Hawkins RE, Hoogenboom HR. Making antibodies by phage display technology. Annu Rev Immunol. 1994;12:433–455. doi: 10.1146/annurev.iy.12.040194.002245.
    1. Kang A. Modulation of polypeptide display on modified filamentous phage use: 2003. US 6,586,236 B2.
    1. Chappel JA, He M, Kang AS. Modulation of antibody display on M13 filamentous phage. J Immunol Methods. 1998;221:25–34. doi: 10.1016/S0022-1759(98)00094-5.
    1. He M, Kang AS, Hamon M, Humphreys AS, Gani M, Taussig MJ. Characterization of a progesterone-binding, three-domain antibody fragment (VH/K) expressed in Escherichia coli. Immunology. 1995;84:662–668.
    1. Nardin EH, Nussenzweig V, Nussenzweig RS, Collins WE, Harinasuta KT, Tapchaisri P, Chomcharn Y. Circumsporozoite proteins of human malaria parasites Plasmodium falciparum and Plasmodium vivax. J Exp Med. 1982;156:20–30. doi: 10.1084/jem.156.1.20.
    1. Zavala F, Tam JP, Hollingdale MR, Cochrane AH, Quakyi I, Nussenzweig RS, Nussenzweig V. Rationale for development of a synthetic vaccine against Plasmodium falciparum malaria. Science. 1985;228:1436–1440.
    1. Young JF, Hockmeyer WT, Gross M, Ballou WR, Wirtz RA, Trosper JH, Beaudoin RL, Hollingdale MR, Miller LH, Diggs CL, et al. Expression of Plasmodium falciparum Circumsporozoite proteins in Escherichia coli for potential use in a human malaria vaccine. Science. 1985;228:958–962.
    1. Schodel F, Wirtz R, Peterson D, Hughes J, Warren R, Sadoff J, Milich D. Immunity to malaria elicited by hybrid hepatitis B virus core particles carrying Circumsporozoite protein epitopes. J Exp Med. 1994;180:1037–1046. doi: 10.1084/jem.180.3.1037.
    1. Seehaus T, Breitling F, Dubel S, Klewinghaus I, Little M. A vector for the removal of deletion mutants from antibody libraries. Gene. 1992;114:235–237. doi: 10.1016/0378-1119(92)90580-I.
    1. Edelman R, Hoffman SL, Davis JR, Beier M, Sztein MB, Losonsky G, Herrington DA, Eddy HA, Hollingdale MR, Gordon DM, Clyde DF. Long-term persistence of sterile immunity in a volunteer immunized with X-irradiated Plasmodium falciparum sporozoites. J Infect Dis. 1993;168:1066–1070.
    1. Hoffman SL, Franke ED, Hollingdale MR, Druilhe P. Attacking the infected hepatocyte. In: Hoffman SL, editor. in Malaria Vaccine Development: A Multi-Immune Response Approach. ASM Press: Washington, DC; 1996. pp. 35–75.
    1. Ballou WR, Sherwood JA, Neva FA, Gordon DM, Wirtz RA, Wasserman GF, Diggs CL, Hoffman SL, Hollingdale MR, Hockmeyer WT, Schneider I, Young JF, Reeve P, Chulay JD. Safety and efficacy of a recombinant DNA Plasmodium falciparum sporozoite vaccine. Lancet. 1987;i:1277–1281.
    1. Herrington DA, Clyde DF, Losonsky G, Cortesia M, Murphy JR, Davis J, Baqar S, Felix AM, Heimer EP, Gillessen D, Nardin E, Nussenzweig RS, Nussenzweig V, Hollingdale MR, Levine MM. Safety and immunogenicity in man of a synthetic peptide malaria vaccine against Plasmodium falciparum sporozoites. Nature. 1987;328:257–259. doi: 10.1038/328257a0.
    1. Marsh K, Hayes RH, Carson DC, Otoo L, Shenton F, Byass P, Zavala F, Greenwood BM. Anti-sporozoite antibodies and immunity to malaria in a rural Gambian population. Trans R Soc Trop Med Hyg. 1988;82:532–537. doi: 10.1016/0035-9203(88)90495-6.
    1. Webster HK, Brown AE, Chuenchitra C, Permpanich B, Pipithkul J. Characterization of antibodies to sporozoites in Plasmodium falciparum malaria and correlation with protection. J Clin Microbiol. 1988;26:923–927.
    1. Hoffman SL, Wistar R, Jr, Ballou WR, Hollingdale MR, Wirtz RA, Schneider I, Marwoto HA, Hockmeyer WT. Immunity to malaria and naturally acquired antibodies to the circumsporozoite protein of Plasmodium falciparum. N Engl J Med. 1986;315:601–606.
    1. Hoffman SL, Oster CN, Plowe CV, Woollett GR, Beier JC, Chulay JD, Wirtz RA, Hollingdale MR, Mugambi M. Naturally acquired antibodies to sporozoites do not prevent malaria: vaccine development implications. Science. 1987;237:639–642.
    1. Luke TC, Hoffman SL. Rationale and plans for developing a non-replicating, metabolically active, radiation-attenuated Plasmodium falciparum sporozoite vaccine. J Exp Biol. 2003;206:3803–3808. doi: 10.1242/jeb.00644.
    1. Clyde DF, McCarthy VC, Miller RM, Hornick RB. Specificity of protection of man immunized against sporozoite-induced falciparum malaria. Am J Med Sci. 1973;266:398–403.
    1. Clyde DF, Most H, McCarthy VC, Vanderberg JP. Immunization of man against sporozite-induced falciparum malaria. Am J Med Sci. 1973;266:169–177.
    1. Rieckmann KH, Carson PE, Beaudoin RL, Cassells JS, Sell KW. Letter: Sporozoite induced immunity in man against an Ethiopian strain of Plasmodium falciparum. Trans R Soc Trop Med Hyg. 1974;68:258–259. doi: 10.1016/0035-9203(74)90129-1.
    1. Clyde DF. Immunization of man against falciparum and vivax malaria by use of attenuated sporozoites. Am J Trop Med Hyg. 1975;24:397–401.
    1. McCarthy VC, Clyde DF. Plasmodium vivax : correlation of circumsporozoite precipitation (CSP) reaction with sporozoite-induced protective immunity in man. Exp Parasitol. 1977;41:167–171. doi: 10.1016/0014-4894(77)90142-4.
    1. Rieckmann KH, Beaudoin RL, Cassells JS, Sell KW. Use of attenuated sporozoites in the immunization of human volunteers against falciparum malaria. Bull World Health Organ. 1979;57:261–265.
    1. Clyde DF. Immunity to falciparum and vivax malaria induced by irradiated sporozoites: a review of the University of Maryland studies 1971–75. Bull World Health Organ. 1990;68:9–12.
    1. Rieckmann KH. Human immunization with attenuated sporozoites. Bull World Health Organ. 1990;68:13.
    1. Nguyen TV, Fujioka H, Kang AS, Rogers WO, Fidock DA, James AA. Stage-dependent localization of a novel gene product of the malaria parasite, Plasmodium falciparum. J Biol Chem. 2001;276:26724–26731. doi: 10.1074/jbc.M103375200.
    1. Saul A. Kinetic constraints on the development of a malaria vaccine. Parasite Immunol. 1987;9:1–9.
    1. Sawada-Hirai R, Jiang I, Wang F, Sun SM, Nedellec R, Ruther P, Alvarez A, Millis D, Morrow PR, Kang AS. Human anti-anthrax protective antigen neutralizing monoclonal antibodies derived from donors vaccinated with anthrax vaccine adsorbed. J Immune Based Ther Vaccines. 2004;2:5. doi: 10.1186/1476-8518-2-5.
    1. Wild MA, Xin H, Maruyama T, Nolan MJ, Calveley PM, Malone JD, Wallace MR, Bowdish KS. Human antibodies from immunized donors are protective against anthrax toxin in vivo. Nat Biotechnol. 2003;21:1305–1306. doi: 10.1038/nbt891.
    1. Gilbert SC, Plebanski M, Gupta S, Morris J, Cox M, Aidoo M, Kwiatkowski D, Greenwood BM, Whittle HC, Hill AV. Association of malaria parasite population structure, HLA, and immunological antagonism. Science. 1998;279:1173–1177. doi: 10.1126/science.279.5354.1173.
    1. Hill AV, Allsopp CE, Kwiatkowski D, Anstey NM, Twumasi P, Rowe PA, Bennett S, Brewster D, McMichael AJ, Greenwood BM. Common west African HLA antigens are associated with protection from severe malaria. Nature. 1991;352:595–600. doi: 10.1038/352595a0.
    1. Zavala F, Masuda A, Graves PM, Nussenzweig V, Nussenzweig RS. Ubiquity of the repetitive epitope of the CS protein in different isolates of human malaria parasites. J Immunol. 1985;135:2790–2793.
    1. Charoenvit Y, Collins WE, Jones TR, Millet P, Yuan L, Campbell GH, Beaudoin RL, Broderson JR, Hoffman SL. Inability of malaria vaccine to induce antibodies to a protective epitope within its sequence. Science. 1991;251:668–671.
    1. Hockmeyer W, Ballou W. In: in Progress in Allergy. Perlmann P, Wigzell H, editor. Karger: Basel; 1988. pp. 1–14.
    1. Simmons LC, Reilly D, Klimowski L, Raju TS, Meng G, Sims P, Hong K, Shields RL, Damico LA, Rancatore P, Yansura DG. Expression of full-length immunoglobulins in Escherichia coli: rapid and efficient production of aglycosylated antibodies. J Immunol Methods. 2002;263:133–147. doi: 10.1016/S0022-1759(02)00036-4.
    1. Gasdaska J, Spencer D, Dickey L. Advantages of Therapeutic Protein Production in the Aquatic Plant Lemna. BioProcessing Journal. 2003;2:49–56.

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