Skip to main content
Bulletin of the World Health Organization logoLink to Bulletin of the World Health Organization
. 1977;55(2-3):317–331.

The isolation and fractionation of malaria-infected cells*

Julius P Kreier
PMCID: PMC2366739  PMID: 338186

Abstract

This paper is a critical review of procedures for the isolation of malarial parasites from host cells and their fractionation. The procedures are grouped according to the stage of parasite being isolated, and the procedures for isolation of the erythrocytic stages are further grouped by techniques used. Some types of procedure are described for isolation of all stages of the parasite, both those in the invertebrate and vertebrate hosts. The uses and limitations of the various procedures are described. It is concluded that all the procedures are useful for some purposes, but that from a morphological standpoint only natural release in culture and continuous flow oscillation provide large yields of intact erythrocytic parasites free of host cell membranes.

Full text

PDF
325

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aikawa M., Cook R. T. Plasmodium: electron microscopy of antigen preparations. Exp Parasitol. 1972 Feb;31(1):67–74. doi: 10.1016/0014-4894(72)90048-3. [DOI] [PubMed] [Google Scholar]
  2. Alger N. E. In vitro development of Plasmodium berghei ookinetes. Nature. 1968 May 25;218(5143):774–774. doi: 10.1038/218774a0. [DOI] [PubMed] [Google Scholar]
  3. Ball G. H., Chao J. The cultivation of Plasmodium relictum in mosquito cell lines. J Parasitol. 1971 Apr;57(2):391–395. [PubMed] [Google Scholar]
  4. Brown I. N., Brown K. N., Hills L. A. Immunity to malaria: the antibody response to antigenic variation by Plasmodium knowlesi. Immunology. 1968 Jan;14(1):127–138. [PMC free article] [PubMed] [Google Scholar]
  5. Carter R., Chen D. H. Malaria transmission blocked by immunisation with gametes of the malaria parasite. Nature. 1976 Sep 2;263(5572):57–60. doi: 10.1038/263057a0. [DOI] [PubMed] [Google Scholar]
  6. Chow J. S., Kreier J. P. Plasmodium berghei: adherence and phagocytosis by rat macrophages in vitro. Exp Parasitol. 1972 Feb;31(1):13–18. doi: 10.1016/0014-4894(72)90042-2. [DOI] [PubMed] [Google Scholar]
  7. Coggeshall L. T., Eaton M. D. THE COMPLEMENT FIXATION REACTION IN MONKEY MALARIA. J Exp Med. 1938 May 31;67(6):871–882. doi: 10.1084/jem.67.6.871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cook R. T., Aikawa M., D'Antonio L. E., Silverman P. H. Malaria resistance: artificial induction with a partially purified plasmodial fraction. Science. 1971 Mar 19;171(3976):1175–1176. doi: 10.1126/science.171.3976.1175. [DOI] [PubMed] [Google Scholar]
  9. Cook R. T., Aikawa M., D'Antonio L. E., Silverman P. H. Malaria resistance: artificial induction with a partially purified plasmodial fraction. Science. 1971 Mar 19;171(3976):1175–1176. doi: 10.1126/science.171.3976.1175. [DOI] [PubMed] [Google Scholar]
  10. Cook R. T., Aikawa M., Rock R. C., Little W., Sprinz H. The isolation and fractionation of Plasmodium knowlesi. Mil Med. 1969 Sep;134(10):866–883. [PubMed] [Google Scholar]
  11. Corradetti A., Verolini F., Ilardi A., Bucci A. Immunoelectrophoretic analysis of water-soluble antigens extracted from parasitic bodies of Plasmodium berghei separated from the blood. Bull World Health Organ. 1966;35(5):802–805. [PMC free article] [PubMed] [Google Scholar]
  12. D'Antonio L. E. Plasmodium: a resumé of the isolation of the vaccine fraction by the French pressure cell technique. Exp Parasitol. 1972 Feb;31(1):75–81. doi: 10.1016/0014-4894(72)90049-5. [DOI] [PubMed] [Google Scholar]
  13. D'Antonio L. E., Von Doenhoff A. E., Jr, Fife E. H., Jr Serological evaluation of the specificity and sensitivity of purified malaria antigens prepared by a new method. Mil Med. 1966 Sep;131(9):1152–1156. [PubMed] [Google Scholar]
  14. Dennis E. D., Mitchell G. H., Butcher G. A., Cohen S. In vitro isolation of Plasmodium knowlesi merozoites using polycarbonate sieves. Parasitology. 1975 Dec;71(3):475–481. doi: 10.1017/s0031182000047235. [DOI] [PubMed] [Google Scholar]
  15. Diggs C. L. Immunodiffusion studies of Plasmodium berghei: interactions of an extract of the erythrocytic forms with rabbit antisera. Exp Parasitol. 1966 Dec;19(3):237–248. doi: 10.1016/0014-4894(66)90073-7. [DOI] [PubMed] [Google Scholar]
  16. Doby J. M., Barker R. Essais d'obtention in vitro des formes pré-érythrocytaires de Plasmodium vivax en cultures de cellules hépatiques humanines inoculées par sporozoïtes. C R Seances Soc Biol Fil. 1976 Oct;170(3):661–665. [PubMed] [Google Scholar]
  17. FULTON J. D., GRANT P. T. The sulphur requirements of the erythrocytic from of Plasmodium knowlesi. Biochem J. 1956 Jun;63(2):274–282. doi: 10.1042/bj0630274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fife E. H., Jr Advances in methodology for immunodiagnosis of parasitic diseases. Exp Parasitol. 1971 Aug;30(1):132–163. doi: 10.1016/0014-4894(71)90079-8. [DOI] [PubMed] [Google Scholar]
  19. Fife E. H., Jr Current state of serological tests used to detect blood parasite infections. Exp Parasitol. 1972 Feb;31(1):136–152. doi: 10.1016/0014-4894(72)90056-2. [DOI] [PubMed] [Google Scholar]
  20. Geiman Q. M., Meagher M. J. Susceptibility of a New World monkey to Plasmodium falciparum from man. Nature. 1967 Jul 22;215(5099):437–439. doi: 10.1038/215437a0. [DOI] [PubMed] [Google Scholar]
  21. Gravely S. M., Kreier J. P. Babesia microti (gray strain): removal from infected hamster erythrocytes by continuous-flow ultrasonication. Tropenmed Parasitol. 1974 Jun;25(2):198–206. [PubMed] [Google Scholar]
  22. Gwadz R. W. Successful immunization against the sexual stages of Plasmodium gallinaceum. Science. 1976 Sep 17;193(4258):1150–1151. doi: 10.1126/science.959832. [DOI] [PubMed] [Google Scholar]
  23. Hamburger J., Kreier J. P. Antibody-mediated elimination of malaria parasites (plasmodium berghei) in vivo. Infect Immun. 1975 Aug;12(2):339–345. doi: 10.1128/iai.12.2.339-345.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hamburger J., Kreier J. P. Interaction between protective antibodies and malaria parasites (Plasmodium berghei): involvement of low avidity antibodies. Tropenmed Parasitol. 1976 Sep;27(3):385–390. [PubMed] [Google Scholar]
  25. Hamburger J., Kreier J. P. Plasmodium berghei: use of free blood stage parasites to demonstrate protective humoral activity in the serum of recovered rats. Exp Parasitol. 1976 Oct;40(2):158–169. doi: 10.1016/0014-4894(76)90078-3. [DOI] [PubMed] [Google Scholar]
  26. Hamburger J., Zuckerman A. Plasmodium berghei: I. Immunochemical properties of fractions of a soluble extract. Exp Parasitol. 1976 Jun;39(3):460–478. doi: 10.1016/0014-4894(76)90050-3. [DOI] [PubMed] [Google Scholar]
  27. Hamburger J., Zuckerman A. Plasmodium berghei: II. Immunobiological properties of fractions of a soluble extract. Exp Parasitol. 1976 Jun;39(3):479–495. doi: 10.1016/0014-4894(76)90051-5. [DOI] [PubMed] [Google Scholar]
  28. Haynes J. D., Diggs C. L., Hines F. A., Desjardins R. E. Culture of human malaria parasites Plasmodium falciparum. Nature. 1976 Oct 28;263(5580):767–769. doi: 10.1038/263767a0. [DOI] [PubMed] [Google Scholar]
  29. Heidelberger M., Mayer M. M. NORMAL HUMAN STROMATA AS ANTIGENS FOR COMPLEMENT FIXATION IN THE SERA OF PATIENTS WITH RELAPSING VIVAX MALARIA. Science. 1944 Oct 20;100(2599):359–360. doi: 10.1126/science.100.2599.359. [DOI] [PubMed] [Google Scholar]
  30. Jerusalem C., Eling W. Active immunization against Plasmodium berghei malaria in mice, using different preparations of plasmodial antigen and different pathways of administration. Bull World Health Organ. 1969;40(6):807–818. [PMC free article] [PubMed] [Google Scholar]
  31. Krettli A., Chen D. H., Nussenzweig R. S. Immunogenicity and infectivity of sporozoites of mammalian malaria isolated by density-gradient centrifugation. J Protozool. 1973 Nov;20(5):662–665. doi: 10.1111/j.1550-7408.1973.tb03594.x. [DOI] [PubMed] [Google Scholar]
  32. Langreth S. G., Trager W. Fine structure of the malaria parasite Plasmodium lophurae developing extracellularly in vitro. J Protozool. 1973 Nov;20(5):606–613. doi: 10.1111/j.1550-7408.1973.tb03584.x. [DOI] [PubMed] [Google Scholar]
  33. Mannweiler E., Oelerich S. Untersuchungen über die antigenen Eigenschaften von Plasmodium berghei. Z Tropenmed Parasitol. 1969 Sep;20(3):265–279. [PubMed] [Google Scholar]
  34. Martin W. J., Finerty J., Rosenthal A. Isolation of Plasmodium berghei (malaria) parasites by ammonium chloride lysis of infected erythrocytes. Nat New Biol. 1971 Oct 27;233(43):260–261. doi: 10.1038/newbio233260a0. [DOI] [PubMed] [Google Scholar]
  35. McAlister R. O., Gordon D. M. Schizont-infected cell enrichment in rodent malaria. J Parasitol. 1976 Oct;62(5):664–669. [PubMed] [Google Scholar]
  36. Miller L. H., Chien S. Density distribution of red cells infected by Plasmodium knowlesi and plasmodium coatneyi. Exp Parasitol. 1971 Jun;29(3):451–456. doi: 10.1016/0014-4894(71)90054-3. [DOI] [PubMed] [Google Scholar]
  37. Mitchell G. H., Butcher G. A., Cohen S. A merozoite vaccine effective against Plasmodium knowlesi malaria. Nature. 1974 Nov 22;252(5481):311–313. doi: 10.1038/252311a0. [DOI] [PubMed] [Google Scholar]
  38. Mitchell G. H., Butcher G. A., Cohen S. Merozoite vaccination against Plasmodium knowlesi malaria. Immunology. 1975 Aug;29(2):397–407. [PMC free article] [PubMed] [Google Scholar]
  39. Prior R. B., Kreier J. P. Plasmodium berghei freed from host erythrocytes by a continuous-flow ultrasonic system. Exp Parasitol. 1972 Oct;32(2):239–243. doi: 10.1016/0014-4894(72)90030-6. [DOI] [PubMed] [Google Scholar]
  40. Prior R. B., Smucker R. A., Kreier J. P., Pfister R. M. A comparison by electron microscopy of Plasmodium berghei freed by ammonium chloride lysis to P. berghei freed by ultrasound in a continuous-flow system. J Parasitol. 1973 Feb;59(1):200–201. [PubMed] [Google Scholar]
  41. Richards W. H., Williams S. G. The removal of leucocytes from malaria infected blood. Ann Trop Med Parasitol. 1973 Jun;67(2):249–250. doi: 10.1080/00034983.1973.11686885. [DOI] [PubMed] [Google Scholar]
  42. Rosales-Ronquillo M. C., Nienaber G., Silverman P. H. Plasmodium berghei ookinete formation in a nonvector cell line. J Parasitol. 1974 Dec;60(6):1039–1040. [PubMed] [Google Scholar]
  43. Rosales-Ronquillo M. C., Silverman P. H. In vitro ookinete development of the rodent malarial parasite, Plasmodium berghei. J Parasitol. 1974 Oct;60(5):819–824. [PubMed] [Google Scholar]
  44. Rutledge L. C., Ward R. A. Effects of ultrasound on Plasmodium gallinaceum-infected chick blood. Exp Parasitol. 1967 Apr;20(2):167–176. doi: 10.1016/0014-4894(67)90036-7. [DOI] [PubMed] [Google Scholar]
  45. SHERMAN I. W. ANTIGENS OF PLASMODIUM LOPHURAE. J Protozool. 1964 Aug;11:409–417. doi: 10.1111/j.1550-7408.1964.tb01772.x. [DOI] [PubMed] [Google Scholar]
  46. SPIRAD, ZUCKERMAN A. Antigenic structure of Plasmodium vinckei. Science. 1962 Aug 17;137(3529):536–537. doi: 10.1126/science.137.3529.536. [DOI] [PubMed] [Google Scholar]
  47. STEIN B., DESOWITZ R. S. THE MEASUREMENT OF ANTIBODY IN HUMAN MALARIA BY A FORMOLIZED TANNED SHEEP CELL HAEMAGGLUTINATION TEST. Bull World Health Organ. 1964;30:45–49. [PMC free article] [PubMed] [Google Scholar]
  48. Sadun E. H., Gore R. W. Mass diagnostic test using Plasmodium falciparum and chimpanzee erythrocyte lysate. Exp Parasitol. 1968 Oct;23(2):277–285. doi: 10.1016/0014-4894(68)90069-6. [DOI] [PubMed] [Google Scholar]
  49. Scheibel L. W., Miller J. Cytochrome oxidase activity in platelet-free preparations of Plasmodium knowlesi. J Parasitol. 1969 Aug;55(4):825–829. [PubMed] [Google Scholar]
  50. Schenkel R. H., Simpson G. L., Silverman P. H. Vaccination of Rhesus monkeys (Macaca mulatta) against Plasmodium knowlesi by the use of nonviable antigen. Bull World Health Organ. 1973 May;48(5):597–604. [PMC free article] [PubMed] [Google Scholar]
  51. Seed T. M., Aikawa M., Sterling C. R. An electron microscope-cytochemical method for differentiating membranes of host red cells and malaria parasites. J Protozool. 1973 Nov;20(5):603–605. doi: 10.1111/j.1550-7408.1973.tb03583.x. [DOI] [PubMed] [Google Scholar]
  52. Seed T. M., Aikawa M., Sterling C., Rabbege J. Surface properties of extracellular malaria parasites: morphological and cytochemical study. Infect Immun. 1974 Apr;9(4):750–761. doi: 10.1128/iai.9.4.750-761.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Seed T. M., Kreier J. P. Autoimmune reactions in chickens with Plasmodium gallinaceum infection: the isolation and characterization of a lipid from trypsinized erythrocytes which reacts with serum from acutely infected chickens. Mil Med. 1969 Sep;134(10):1220–1227. [PubMed] [Google Scholar]
  54. Seed T. M., Kreier J. P. Surface properties of extracellular malaria parasites: electrophoretic and lectin-binding characteristics. Infect Immun. 1976 Dec;14(6):1339–1347. doi: 10.1128/iai.14.6.1339-1347.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Shapiro M., Espinal-Tejada C., Nussenzweig R. S. Evaluation of a method for in vitro ookinete development of the rodent malarial parasite, Plasmodium berghei. J Parasitol. 1975 Dec;61(6):1105–1106. [PubMed] [Google Scholar]
  56. Simpson G. L., Schenkel R. H., Silverman P. H. Vaccination of rhesus monkeys against malaria by use of sucrose density gradient fractions of Plasmodium knowlesi antigens. Nature. 1974 Feb 1;247(5439):304–305. doi: 10.1038/247304a0. [DOI] [PubMed] [Google Scholar]
  57. Smalley M. E., Butcher G. A. The in vitro culture of the blood stages of Plasmodium berghei. Int J Parasitol. 1975 Apr;5(2):131–132. doi: 10.1016/0020-7519(75)90018-1. [DOI] [PubMed] [Google Scholar]
  58. Smalley M. E. Plasmodium falciparum gametocytogenesis in vitro. Nature. 1976 Nov 18;264(5583):271–272. doi: 10.1038/264271a0. [DOI] [PubMed] [Google Scholar]
  59. Sodeman W. A., Jr, Meuwissen H. E. Disk electrophoresis of Plasmodium berghei. J Parasitol. 1966 Feb;52(1):23–25. [PubMed] [Google Scholar]
  60. Speer C. A., Silverman P. H., Barr M. L. Ultrastructural study of Plasmodium knowlesi antigen used in vaccination of rhesus monkeys. J Protozool. 1976 Aug;23(3):437–442. doi: 10.1111/j.1550-7408.1976.tb03806.x. [DOI] [PubMed] [Google Scholar]
  61. Speer C. A., Silverman P. H., Schiewe S. G. Cultivation of the erythrocytic stages of Plasmodium berghei in primary bone marrow cells. J Parasitol. 1976 Oct;62(5):657–663. [PubMed] [Google Scholar]
  62. Spira D., Zuckerman A. Recent advances in the antigenic analysis of plasmodia. Mil Med. 1966 Sep;131(9 Suppl):1117–1123. [PubMed] [Google Scholar]
  63. TORREY J. C., KAHN M. C. Precipitin-tests in Plasmodium lophurae malaria. J Immunol. 1949 May;62(1):41–48. [PubMed] [Google Scholar]
  64. Trager W., Jensen J. B. Human malaria parasites in continuous culture. Science. 1976 Aug 20;193(4254):673–675. doi: 10.1126/science.781840. [DOI] [PubMed] [Google Scholar]
  65. Trigg P. I., Shakespeare P. G., Burt S. J., Kyd S. I. Ribonucleic acid synthesis in Plasmodium knowlesi maintained both in vivo and in vitro. Parasitology. 1975 Oct;71(2):199–209. doi: 10.1017/s0031182000046655. [DOI] [PubMed] [Google Scholar]
  66. Walter R. D. Untersuchungen über die Entwicklung freier, erythroxytärer Schizonten bei Infektionen mit Plasmodium berghei und Plasmodium chabaudi. Z Tropenmed Parasitol. 1968 Dec;19(4):415–426. [PubMed] [Google Scholar]
  67. Ward P. A. Conran PB: Immunopathologic studies of simian malaria. Mil Med. 1966 Sep;131(9 Suppl):1225–1232. [PubMed] [Google Scholar]
  68. Weiss M. M., Vanderberg J. P. Studies on Plasmodium ookinetes. 1. Isolation and concentration from mosquito midguts. J Protozool. 1976 Nov;23(4):547–551. doi: 10.1111/j.1550-7408.1976.tb03837.x. [DOI] [PubMed] [Google Scholar]
  69. Wilson R. J., Bartholomew R. K. The release of antigens by Plasmodium falciparum. Parasitology. 1975 Oct;71(2):183–192. doi: 10.1017/s0031182000046631. [DOI] [PubMed] [Google Scholar]
  70. Yu D. T., Peter B. J., Paulus H. E., Machleder H. I. Lymphocyte populations: separation by discontinuous density gradient centrifugation. J Immunol. 1973 Jun;110(6):1615–1622. [PubMed] [Google Scholar]

Articles from Bulletin of the World Health Organization are provided here courtesy of World Health Organization

RESOURCES