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Abstract
BackgroundTo date, human peripheral blood mononuclear cells (PBMCs) have been used mainly in immune stimulation assays and the interpretation of data can be influenced by the previous immunological history of donors and cross reactivity with other infectious agents. Resolving these limitations requires an alternative in vitro model to uncover the primary response profiles.MethodsA novel in vitro model of mononuclear cells (MNCs) generated from haematopoietic stem cells (HSCs) was developed and these cells were then co-cultured with various antigens from Plasmodium falciparum and Plasmodium vivax to investigate the response of naïve immune cells to malaria antigens by flow cytometry.ResultsIn vitro stimulation of naïve lymphocytes showed that CD4+ and CD8+ T lymphocytes were significantly reduced (P < 0.01) by exposure to lysates of infected erythrocytes or intact erythrocytes infected with P. falciparum. The depletion was associated with the expression of CD95 (Fas receptor) on the surface of T lymphocytes. Maturation of T lymphocytes was affected differently, showing elevated CD3+CD4+CD8+ and CD3+CD4−CD8− T lymphocytes after stimulation with cell lysates of P. falciparum and P. vivax, respectively. In addition, antigen presenting monocytes and dendritic cells derived from haematopoietic stem cells showed impaired HLA-DR expression as a consequence of exposure to different species of malaria parasites.ConclusionThese results suggest that naïve mononuclear cells differentiated in vitro from HSCs could provide a valid model for the assessment of immunity. P. falciparum and P. vivax malaria parasites could modulate various populations of immune cells starting from newly differentiated mononuclear cells.
Highlights
To date, human peripheral blood mononuclear cells (PBMCs) have been used mainly in immune stimulation assays and the interpretation of data can be influenced by the previous immunological history of donors and cross reactivity with other infectious agents
Taken together (Fig. 1d), mononuclear cells (52.4% ±2.26) were the majority of immune cells in this haematopoietic stem cells (HSCs) culture giving twofold higher numbers than that of the polymorphonuclear cells (25.6% ±1.94)
Results obtained from the in vitro model presented in this study showed that intact Plasmodium falciparum-infected erythrocytes (PfIEs) significantly increased the CD34+ HSCs remaining in the culture, resulting in a 1.5 ± 0.24-fold higher abundance of these cells when compared to the control (Fig. 6)
Summary
Human peripheral blood mononuclear cells (PBMCs) have been used mainly in immune stimulation assays and the interpretation of data can be influenced by the previous immunological history of donors and cross reactivity with other infectious agents. Resolving these limitations requires an alternative in vitro model to uncover the primary response profiles. Several diseases have shown cross reactivity with malaria, i.e. schistosomiasis [4, 5], leishmaniasis, toxoplasmosis, and Chagas’ disease [6] Resolving these limitations requires an alternative in vitro model such as newly produced immune cells to uncover primary response profiles. The system uses newly differentiated immune cells derived from cord blood-HSCs to verify the modulatory roles of Plasmodium falciparum and Plasmodium vivax in the manipulation of naïve immune cells without the interference of prior donor immunological exposure
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