Serum-free media were used to culture Plasmodium falciparum. A commercial preparation, Daigo's GF21 developed as a growth-promoting factor for many kinds of mammalian cells, and consisting of the 55–70% ammonium sulphate fraction of adult bovine serum, insulin, transferrin, ethanolamine and sodium selenite, was found to sustain growth of the parasite when Daigo's T was employed as a basal medium. The optimal Daigo's GF21 concentration for parasite growth was between 5 and 20% (v/v), with the best results at 10%. Differential counts indicated that Daigo's GF21 is essential for schizogony. Established serum-free medium, GIT, consisting of Daigo's T basal medium and Daigo's GF21, yielded good parasite growth without any supplementation. Growth-promoting factor derived from adult bovine serum in Daigo's GF21 was shown to be crucial to parasite growth. The results presented here will not only be of practical value, but will provide important information about the developmental requirements for the parasite.

The purpose of this work is to review the progress towards malaria vaccination that has been made over the last four or so years, and the prospects and difficulties as they now appear. Although some of the older literature will be referred to as necessary background, it is not treated here in any detail. The reader who wishes for a fuller historical perspective should see, for instance, Brown (1969), Cohen & Mitchell (1978), Desowitz & Miller (1980), Mitchell (1984), Miller, David & Hadley (1984), Heidrich (1986) and, specifically for a consideration of sporozoite vaccination, Nussenzweig & Nussenzweig (1984, 1986). Naturally, any summary of vaccination will lean on the immunology of the disease, but malaria immunology is not reviewed here in its own right. The reader requiring the most recent attempt to cover this field should see the work edited by Perlmann & Wigzell (1988); some individual chapters of that volume will be referred to below.

Murine monoclonal antibodies (Mabs) were produced by vaccination of Balb/c mice with live Plasmodium falciparum infected red cells (iRBC). The iRBC Mabs recognized altered forms of the human erythrocyte membrane protein band 3; however, these Mabs did not recognize the band 3 molecule in uninfected or ring-infected red cells. The location of epitopes was determined by studying the binding of the iRBC Mabs after selective proteolysis of band 3 as well as by the reactivity of these Mabs to synthetic peptides that corresponded to putative exofacial regions of band 3. Treatment of uninfected red cell membranes with trypsin under low ionic strength conditions resulted in exposure of cryptic epitopes of band 3 which were recognized by the iRBC Mabs. Several of the anti-iRBC Mabs (two of which were described previously) inhibited the in vitro adherence of infected erythrocytes to C32 amelanotic melanoma cells. A mouse polyclonal serum against a synthetic peptide based on an amino acid sequence motif of band 3 reacted (by immunostaining) only with the surface of iRBC and blocked adhesion. Thus, it appears that cryptic residues of the band 3 protein become exposed upon parasitization, and their presence contributes to the increased adhesiveness of the P. falciparum-infected red cell.

A low molecular weight factor in a basal medium essential for erythrocytic Plasmodium falciparum development in a serum-free medium using a cell growth-promoting factor derived from adult bovine serum was detected. The factor was hypoxanthine. The optimal hypoxanthine concentration for parasite growth was between 15 and 120 μM. The contribution of hypoxanthine to increased parasite growth was clearly evident in cultures on day 4. Among various low molecular weight supplements tested, adenine, adenosine, AMP, ATP, cyclic AMP, guanine, guanosine, inosine, inosine mono-phosphate, xanthine, NAD, NADH, NADP, NADPH and deoxyguanosine triphosphate showed a similar effect to that of hypoxanthine in the serum-free culture system. On the other hand, the addition of uric acid, FAD, thymidine, uridine, orotic acid, deoxythymidine triphosphate, deoxycytidine triphosphate, deoxyadenosine triphosphate, ribose-1-phosphate, or ethanolamine was not beneficial to the parasite growth. The results presented here will not only be of practical value, but will provide important information about the developmental requirements of the parasite.