Ethics code: IR.TMI.REC.1403.004
Abstract: (27 Views)
Background and Objectives
The Rh blood group system is one of the most significant and complex antigenic systems of blood group. The production of specific antibodies against RhD is of great importance for diagnostic and therapeutic applications. This study aims to evaluate the immune response generated in peipheral blood mononuclear cells (PBMCs) by red blood cells containing the RhD antigen and their membrane extracts in a cell culture medium.
Materials and Methods
In this experimental study, PBMCs were isolated from two whole blood units with O- blood type using a density gradient method and Ficoll reagent. After a 40-minute exposure to L-leucine methyl ester (2.5 mM) and subsequent washing, the cells were exposed to red blood cells containing the RhD antigen or their membrane extract in a culture medium. Their ability to induce the production of specific antibodies was assessed using the ELISA method.
Results
Following immunization of PBMCs with O+ red blood cells or their membrane extract, antibody production was quantitatively assessed using ELISA method. The mean concentration of total antibodies was 159.3±5.19 ng/mL in response to membrane extract (n=4) and 136.5±38.9 ng/mL following stimulation with O+ RBC (n=2). In addition, the concentration of RhD-specific antibodies was measured as 38.17±10.58 ng/mL in response to membrane extract and 18.75±2.95 ng/mL in response to O+ RBC cells. The data indicate that both antigenic forms are capable of inducing an immune response. Wilcoxon method showed that the differences between two groups of antigens in both evaluated total antibodies and RhD-specific antibodies were not significant.
Conclusions
Based on the findings of this study, it seems that the membrane extract of RBC plays a more effective role in the immunogenicity against the RhD antigen in compare to O+ RBC cells but the difference were not significant. Based on the data, the use of both membrane extracts and the whole red blood cells could be helpful in the context of immunization strategies in culture conditions.
The Rh blood group system is one of the most significant and complex antigenic systems of blood group. The production of specific antibodies against RhD is of great importance for diagnostic and therapeutic applications. This study aims to evaluate the immune response generated in peipheral blood mononuclear cells (PBMCs) by red blood cells containing the RhD antigen and their membrane extracts in a cell culture medium.
Materials and Methods
In this experimental study, PBMCs were isolated from two whole blood units with O- blood type using a density gradient method and Ficoll reagent. After a 40-minute exposure to L-leucine methyl ester (2.5 mM) and subsequent washing, the cells were exposed to red blood cells containing the RhD antigen or their membrane extract in a culture medium. Their ability to induce the production of specific antibodies was assessed using the ELISA method.
Results
Following immunization of PBMCs with O+ red blood cells or their membrane extract, antibody production was quantitatively assessed using ELISA method. The mean concentration of total antibodies was 159.3±5.19 ng/mL in response to membrane extract (n=4) and 136.5±38.9 ng/mL following stimulation with O+ RBC (n=2). In addition, the concentration of RhD-specific antibodies was measured as 38.17±10.58 ng/mL in response to membrane extract and 18.75±2.95 ng/mL in response to O+ RBC cells. The data indicate that both antigenic forms are capable of inducing an immune response. Wilcoxon method showed that the differences between two groups of antigens in both evaluated total antibodies and RhD-specific antibodies were not significant.
Conclusions
Based on the findings of this study, it seems that the membrane extract of RBC plays a more effective role in the immunogenicity against the RhD antigen in compare to O+ RBC cells but the difference were not significant. Based on the data, the use of both membrane extracts and the whole red blood cells could be helpful in the context of immunization strategies in culture conditions.
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