pmc logo imageJournal ListSearchpmc logo image
Logo of iaiInfect Immun SubscriptionsInfect Immun Web Site
Infect Immun. 1991 November; 59(11): 3909–3916.
PMCID: PMC258976
Antibody recognition and isoelectrofocusing of antigens of the malaria parasite Plasmodium yoelii.
F A Kironde, A Kumar, A R Nayak, and J L Kraikov
Malaria Group, International Centre for Genetic Engineering and Biotechnology, New Delhi, India.
Abstract
Inbred BALB/c mice were either immunized with Triton X-100-extracted antigens of blood-stage Plasmodium yoelii or infected with P. yoelii and cured in three successive schedules. Whereas the immunized BALB/c became only partially protected from subsequent challenge infection with blood-stage P. yoelii, the convalescent mice acquired total immunity. When total P. yoelii antigen extract was resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and immunoblotted with anti-P. yoelii serum, five major protein bands of 150, 84, 40, 19, and 16 kDa were recognized by the sera of fully protected convalescent mice but not by the sera of partially protected mice. The utility of comparing reactivities of sera from fully protected and partially protected malaria hosts and the possibility that antigens uniquely recognized by the convalescent mouse sera may contribute to immunity against P. yoelii infection are discussed. Although previously reported to be an effective adjuvant for immunization against P. yoelii infection in (BALB/c x C57BL)F1 hybrid mice, saponin did not promote protection any better than did Freund adjuvant in BALB/c mice immunized with detergent-extracted P. yoelii antigen. Most of the P. yoelii proteins (14 to 250 kDa) found in Triton X-100 extracts of P. yoelii-parasitized erythrocytes isoelectrofocused as a single peak in the pH region 4.4 to 4.6, suggesting a rationale for previous findings that the most anti-P. yoelii protective and T-helper activities are induced by antigens isoelectrically focused in a fraction of similar pH.
Full text
Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.
Images in this article
Click on the image to see a larger version.
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
  • Anders, RF; Coppel, RL; Brown, GV; Kemp, DJ. Antigens with repeated amino acid sequences from the asexual blood stages of Plasmodium falciparum. Prog Allergy. 1988;41:148–172. [PubMed]
  • Bouharoun-Tayoun, H; Attanath, P; Sabchareon, A; Chongsuphajaisiddhi, T; Druilhe, P. Antibodies that protect humans against Plasmodium falciparum blood stages do not on their own inhibit parasite growth and invasion in vitro, but act in cooperation with monocytes. J Exp Med. 1990 Dec 1;172(6):1633–1641. [PubMed]
  • Certa, U; Ghersa, P; Döbeli, H; Matile, H; Kocher, HP; Shrivastava, IK; Shaw, AR; Perrin, LH. Aldolase activity of a Plasmodium falciparum protein with protective properties. Science. 1988 May 20;240(4855):1036–1038. [PubMed]
  • De Souza, JB; Playfair, JH. Immunization of mice against blood-stage Plasmodium yoelii malaria with isoelectrically focused antigens and correlation of immunity with T-cell priming in vivo. Infect Immun. 1988 Jan;56(1):88–91. [PubMed]
  • Fenton, B; Walker, A; Walliker, D. Protein variation in clones of Plasmodium falciparum detected by two dimensional electrophoresis. Mol Biochem Parasitol. 1985 Aug;16(2):173–183. [PubMed]
  • FULTON, JD; GRANT, PT. The sulphur requirements of the erythrocytic from of Plasmodium knowlesi. Biochem J. 1956 Jun;63(2):274–282. [PubMed]
  • Groux, H; Gysin, J. Opsonization as an effector mechanism in human protection against asexual blood stages of Plasmodium falciparum: functional role of IgG subclasses. Res Immunol. 1990 141(6):529–542.Jul–Aug; [PubMed]
  • Herrera, S; Herrera, MA; Perlaza, BL; Burki, Y; Caspers, P; Döbeli, H; Rotmann, D; Certa, U. Immunization of Aotus monkeys with Plasmodium falciparum blood-stage recombinant proteins. Proc Natl Acad Sci U S A. 1990 May;87(10):4017–4021. [PubMed]
  • Holder, AA. The precursor to major merozoite surface antigens: structure and role in immunity. Prog Allergy. 1988;41:72–97. [PubMed]
  • Homewood, CA; Neame, KD. A comparison of methods used for the removal of white cells from malaria-infected blood. Ann Trop Med Parasitol. 1976 Jun;70(2):249–251. [PubMed]
  • Jayawardena, AN; Targett, GA; Leuchars, E; Davies, AJ. The immunological response of CBA mice to P. yoelii. II. The passive transfer of immunity with serum and cells. Immunology. 1978 Jan;34(1):157–165. [PubMed]
  • Kemp, DJ; Coppel, RL; Stahl, HD; Bianco, AE; Corcoran, LM; McIntyre, P; Langford, CJ; Favaloro, JM; Crewther, PE; Brown, GV, et al. The Wellcome Trust lecture. Genes for antigens of Plasmodium falciparum. Parasitology. 1986;92 Suppl:S83–108. [PubMed]
  • Laemmli, UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • LaRochelle, WJ; Froehner, SC. Immunochemical detection of proteins biotinylated on nitrocellulose replicas. J Immunol Methods. 1986 Aug 21;92(1):65–71. [PubMed]
  • Miller, LH; Howard, RJ; Carter, R; Good, MF; Nussenzweig, V; Nussenzweig, RS. Research toward malaria vaccines. Science. 1986 Dec 12;234(4782):1349–1356. [PubMed]
  • Mitchell, GH. An update on candidate malaria vaccines. Parasitology. 1989;98 Suppl:S29–S47. [PubMed]
  • Ozaki, LS; Mattei, D; Jendoubi, M; Druihle, P; Blisnick, T; Guillotte, M; Puijalon, O; Da Silva, LP. Plaque antibody selection: rapid immunological analysis of a large number of recombinant phage clones positive to sera raised against Plasmodium falciparum antigens. J Immunol Methods. 1986 May 22;89(2):213–219. [PubMed]
  • Perrin, LH; Merkli, B; Gabra, MS; Stocker, JW; Chizzolini, C; Richle, R. Immunization with a Plasmodium falciparum merozoite surface antigen induces a partial immunity in monkeys. J Clin Invest. 1985 May;75(5):1718–1721. [PubMed]
  • Peterson, GL. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. [PubMed]
  • Playfair, JH; De Souza, JB. Vaccination of mice against malaria with soluble antigens. I. The effect of detergent, route of injection, and adjuvant. Parasite Immunol. 1986 Sep;8(5):409–414. [PubMed]
  • Saul, A. Kinetic constraints on the development of a malaria vaccine. Parasite Immunol. 1987 Jan;9(1):1–9. [PubMed]
  • Tait, A. Analysis of protein variation in Plasmodium falciparum by two-dimensional gel electrophoresis. Mol Biochem Parasitol. 1981 Feb;2(3-4):205–218. [PubMed]
  • Towbin, H; Staehelin, T; Gordon, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Troye-Blomberg, M; Perlmann, P. T cell functions in Plasmodium falciparum and other malarias. Prog Allergy. 1988;41:253–287. [PubMed]
  • Yoeli, M; Hargreaves, B; Carter, R; Walliker, D. Sudden increase in virulence in a strain of Plasmodium berghei yoelii. Ann Trop Med Parasitol. 1975 Jun;69(2):173–178. [PubMed]