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Repeat sequences from the Plasmodium falciparum blood stage antigen Pf332 frequently comprise the pentapeptide VTEEI, an epitope recognized by certain parasite neutralizing antibodies. This B-cell epitope was assembled in an octavalent multiple antigen peptide (MAP) system either as trimers (VTEEI)3 (MAP1) or as an integral part of a naturally occurring Pf332 undecamer repeat sequence SVTEEIAEEDK (MAP2). Characteristics of the immunogenicity of these subunit constructs were evaluated in H-2 congenic mice. MAP1 generated antibody responses in mice of the H-2d, H-2k and H-2q haplotypes, but not in H-2b or H-2s mice, whereas MAP2 only induced antibodies in mice of H-2k haplotype. When analysing T-cell responses induced by the MAP, lymph node cells from responder strains primed in vivo with MAPI proliferated in response to restimulation with both MAPI and the peptide (VTEEI)3. MAP2, however, did not induce a detectable T-cell proliferation. Additionally, the lack of antibody response to MAP1 in H-2b mice could be circumvented by combining the MAP1 peptide and a H-2b-restricted T-cell epitope in a diepitope MAP construct. Despite the fact that the motif VTEEI has not been identified in Pf332 sequences in the form of a trimer, MAP1 did induce Pf332 protein-reactive antibodies. Assembly of multimers of short defined epitopes in MAP constitutes an interesting approach for the design of polyvalent subunit immunogens. [ABSTRACT FROM AUTHOR]

An enzyme linked immunosorbent assay (ELISA) has been developed to estimate disease related antibodies in sera from malaria patients or individuals living in malaria endemic areas. As antigen, Percoll enriched fractions (mainly late trophozoites, schizonts) from Plasmodium falciparum in vitro cultures were used. An ELISA with ghosts from normal human red blood cells (RBC) was performed in parallel. One hundred and seventy-five sera were tested for their reactivity with either one of the two antigens. Seven sera from patients with acute P.falciparum infection were negative. Most of these had been taken very early in infection and consecutive samples taken later usually were positive. The antibodies reacting with the P. falciparum antigen had a high parasite specificity as indicated by inhibition and absorption experiments. Many sera also had elevated levels of antibodies specific for RBC antigens. A correlation, most pronounced in the IgM system, was also seen between the anti-RBC and the -duti-P. falciparum antibody levels. [ABSTRACT FROM AUTHOR]

The IgG subclass levels of anti-Plasmodium falciparum antibodies in human sera were determined in ELISA with monoclonal mouse antibodies specific for the human IgG subclasses as analytical reagents. The parasite antigen was a trophozoite/schizont enriched preparation of in vitro cultivated P. falciparum. Serum samples were from Swedish malaria patients and adult Liberians. Parasite specific antibodies were found in all four subclasses. Relatively elevated levels of IgGl antibodies were found both in Swedish patients and in the Liberians. Relative to the Liberians, high IgG2 antibody levels were seen in most Swedish patients. In Liberian sera but not in those from Swedish patients elevated IgG3 levels were found. In two Swedish patients followed consecutively for a period of 15 weeks, elevated levels of IgG1 and IgG3 antibodies were seen after relapse. No differences between the groups in regard to the levels origG4 antibodies were noticed. The differences between the Swedish patients and the Liberians were also in part reflected by differences in the total amounts of the four IgG subclasses. [ABSTRACT FROM AUTHOR]

The antigen-induced DNA synthesis in vitro in lymphocytes from patients with acute Plasmodium falciparum malaria was investigated. The patients and healthy controls from Sweden or Colombia were the same as those studied in the accompanying paper (Troye-Blomberg el al., 1983). The malarial antigens used were sonicated membrane preparations or purified and concentrated supernatants from in vitro cultures of P. facliparum, similar preparations derived from normal human erythrocytes served as control antigen. In the patients' lymphocytes P. falciparum antigens induced a weak or moderate but significant stimulation of DNA synthesis, peaking after 3-4 days of incubation. This early response was specific for P. faleiparum since it was not obtained with lymphocytes from healthy donors nor with those from patients with acute P. vivax or P. ovale malaria. No antigen-induced response was seen in about half of the P.falciparum patients. However in a few negative eases, available for consecutive testing, positive reactions were seen with lymphocytes taken 2 weeks after infection when the blood of these patients was free of parasites. The early response induced in patients' lymphocytes to the P. falciparum antigens was not obtained with RBC antigen, however, these preparations frequently induced a response rising to significant levels later during incubation (day 5-6)- Similar delayed responses were obtained when either- patients' or control donors' lymphocytes were exposed to the P. falciparum antigens. This indicates that both the RBC and the parasite preparations contained mitogenic substances affecting human lymphocytes in general and easily obscuring the P, falciparum specific responses seen only in the patients. This latter response was relatively low and short lived, suggesting that it reflected a secondary in vitro stimulation of in vivo primed lymphocytes and that it was regulated by suppressor mechanisms. [ABSTRACT FROM AUTHOR]