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Plasmodium falciparum (human malaria) infections are characterized by the attachment of erythrocytes infected with mature stage parasites to endothelial cells lining the post-capillary venules, a phenomenon known as sequestration. In the human body, the microvessels of the heart, lungs, kidneys, small intestine, and liver are the principal sites of sequestration. Sequestered cells that clog the brain capillaries may reduce blood flow sufficiently so that there is confusion, lethargy, and unarousable coma—cerebral malaria. This review considers what is known about the molecular characteristics of the surface proteins, that is, the red cell receptors and the endothelial cell ligands, involved in sequestration. Recent work from our laboratory on the characterization of the adhesive proteins on the surface of the P falciparum-infected red cell, and the ligands to which they bind on human brain endothelial cells is also discussed. Finally, consideration is given to the multifactor processes involved in sequestration and cerebral malaria, as well as the possible role of ‘anti-adhesion therapy’ in the management of severe malaria.

Ribosomes were isolated from Plasmodium lophurae by Triton X-100 lysis and ultracentrifugation. Plasmodium lophurae ribosomal subparticles, produced by treatment of ribosomes with puromycin, were separated by sucrose gradient centrifugation. The nucleotide base composition of P. lophurae ribosomal RNA (rRNA) from intact ribosomes and subparticels was low in guanine-cytosine content (approximately 37% G + C) whereas the rRNA of the host cell (duck reticulocyte) was considerably higher (64% G + C). The rRNA of the malaria parasite was typically protozoan, with sedimentation values of 25S and 17S. In vitro, free parasites and malaria-infected red cells incorporated radioactive adenosine into both the large and the small plasmodial subparticles; label was also recovered in the 25S and 17S RNAs. These results suggest that host ribosomes do not contribute to ribosome biogenesis in this malaria, rather parasite rRNA is the transcription product of plasmodial cistrons.

SYNOPSIS Plasmodium lophurae-infected red blood cells utilized considerably greater quantities of glucose than did uninfected duckling red cells. Kinetic analysis of glucose transport showed: (A). Below a concentration of 2 mM in the medium the uptake process followed Michaelis-Menten kinetics (carrier-mediated facilitated diffusion) whereas at concentrations greater than this simple diffusion became the main mode of entry. (B). The apparent transport constants, Kt, for normal and infected cells were similar. However there was an 8-fold increase in the maximal velocity, Vmax, for infected cells. (C). “Free” malaria parasites had a significantly lower Kt and a higher Vmax than did normal or infected red cells. Entry and exit studies with the nonmetabolizable sugar analog, 3-0-methyl glucose, demonstrated that the enhanced rate of uptake by infected cells involved an increase in the simple diffusion component and the degree of enhancement was correlated with the size of the intracellular parasite. Competition experiments suggested that in the malaria-infected cell one locus is involved in the carrier-mediated transport of glucose, mannose and galactose whereas another locus transports fructose and/or glycerol. These results indicate that the enhanced entry of glucose into the malaria-infected red cell is a consequence of factors other than increased glucose catabolism by the host-parasite complex, and the host cell's capacity to take up greater quantities of sugar directly involves the growing intracellular plasmodium.

SYNOPSIS. Plasma membranes of normal duckling erythrocytes were prepared by blender homogenization and nitrogen decompression. Surface membrane vesicles of red cells infected with the avian malaria Plasmodium lophurae were produced by nitrogen decompression. Membranes of erythrocyte-free malaria parasites were removed from cytoplasmic constituents by Dounce homogenization. These membranes were collected by centrifugation in a sucrose step gradient and purified on a linear sucrose gradient. Red cell membranes had a buoyant density of 1.159 g/cm3, whereas plasmodial membranes banded at 2 densities: 1.110 g/cm3 and 1.158 g/cm3. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the isolated red cell membranes revealed 7 major protein bands with molecular weights (MW) ranging from 230,000 to 22,000, and 3 glycoprotein bands with MW of 160,000, 88,000 and 37,000. Parasite membranes also had 7 major bands with MW ranging from 100,000 to 22,000. No glycoproteins were identifiable in these membranes. The proteins of the surface membranes from infected red cells had MW similar to those from normal red cells; however, there was some evidence of a reduction in the amount of the high MW polypeptides. The red cell membrane contained 79 nmoles sialic acid/mg membrane protein, whereas plasmodial membranes had 8 nmoles sialic acid/mg membrane protein. The sialic acid content of the surface membranes of infected red cells was significantly smaller than that of normal cells. Lactoperoxidase-glucose oxidase-catalyzed iodination of intact normal and malaria-infected erythrocytes labeled 7 surface components. Although no observable differences in iodinatable proteins were seen in these preparations, there was a striking reduction in the iodinatability of erythrocytic membranes obtained from P. lophurae-infected cells. Erythrocyte-free plasmodia bound very little radioactive iodine; the small amount of radioactivity was distributed among 3 major bands with MW of 42,000, 32,000 and 28,000. It is suggested that the alterations of the surface of the P. lophurae-infected erythrocyte do not occur by a wholesale insertion of plasmodial membrane proteins into the red cell plasma membrane, but rather that there are parasite-mediated modifications of existing membrane polypeptides.

SYNOPSIS. The use of saponin hemolysis, buffer washings and deoxyribonuclease yields quantities of erythrocyte-free plasmodia sufficiently pure for physico-chemical studies. The pigment produced by Plasmodium lophurae, unlike the pigments of the simian and human malarias, is of such low solubility in buffer solutions that urea is necessary as an additive to the buffer for adequate solubilization. On the basis of spectral and electrophoretic qualities, the pigment is a protein-porphyrin complex closely resembling methemoglobin but clearly distinct from hematin. Extraction of the pigment in solvents which degrade hemoglobin, e.g. phenol, 0.1 N NaOH, cannot be used to ascertain the properties of hemozoin, for the pigment in such solvents shows a spectrum identical to hemoglobin. The soluble parasite proteins which constitute approximately 55% of the totaI volume of the parasite as revealed by electrophoretic and ultracentrifugal analysis appear to be homogeneous. They show a marked similarity to the host hemoglobin, but differ sufficiently to verify their integrity and individuality.

SYNOPSIS. Plasmodium lophurae hemozoin (malaria pigment) is a heme-containing protein which is distinctly different from hemoglobin and hematin by immunologic, spectrophotometric, fingerprint, heme-iron, gel filtration, and starch gel electrophoretic analyses. The calculated average molecular weight of P. lophurae hemozoin is ca. 40,000. Hemozoin contains at least 3 antigenic components and shows some indication of cross-reaction with hemoglobin.

SYNOPSIS. Molecular heterogeneity of malic dehydrogenase (MDH) in malaria was shown by zone electrophoresis in potato starch, starch gel, and by enzymatic activity with analogs of the coenzyme diphosphopyridine nucletide. A single anodal peak of MDH characterized the normal duck red blood cell whereas P. lophurae free of the host cell had a cathodal form of the enzyme. Infected duck erythrocytes had a combination of these electrophoretic forms. The isolated enzymes had different pH optima with oxaloacetate as substrate: pH 7.4 for the duck red cell and 6.4 for the plasmodial enzyme. The Km of each enzyme for oxaloacetate varied with the pH. The Km at pH 7.4 was 4.1 and 4.4 × 105 M for parasite and host, respectively, whereas at 6.4 it was 2.0 × 105 M for P. lophurae and 6.3 × 105M for the duck erythrocyte. At pH 7.4 both enzymes were inhibited by oxaloacetate concentrations greater than 10−4 M. P. berghei MDH also had a different electrophoretic character from that of the mouse red blood cell. Quantitatively, MDH activity increased with parasitization, and erythrocyte-free P. lophurae contained approximately twice the activity found in the uninfected duck erythrocyte. The quantity of MDH activity of the infected cell was ca. 50% less than the sum of the activities of the parasite and the uninfected cell. It is suggested that these properties of the parasite MDH may give it a physiologic advantage over the red cell under the conditions which prevail intraerythrocytically.

SYNOPSIS. A study by means of paper electrophoresis of the scrum.proteins and lipoproteins during the course of infection of chicks with Plasmodium lophurae, reveals a qualitative change in the gamma-globulin and demonstrates: a significant crisis period depression of the albumin; no alteration of the alpha-globulin or alpha-lipoprotein; a significant crisis period rise in the beta- and gamma-g'obuiins with the former persisting in higher amounts well after the crisis of the infection. The significance of such changes with reference to previous work and the induced pathology is discussed.