Volume 19, Issue 4 (winter 2022)
Sci J Iran Blood Transfus Organ 2022, 19(4): 257-269 |
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Amrovani M, Yari F, Milani S, Amoohossein B. Evaluation of effective factors in the optimization of immunization to achieve antibodies against RhD antigen in culture medium. Sci J Iran Blood Transfus Organ 2022; 19 (4) :257-269
URL: http://bloodjournal.ir/article-1-1457-en.html
URL: http://bloodjournal.ir/article-1-1457-en.html
Abstract: (843 Views)
Abstract
Background and Objectives
Antibody against RhD antigen is important in blood transfusion and clinical medicine. Therefore, in this study, the effective factors in the optimization of in vitro immunization to achieve immunized lymphocytes that produce antibodies were evaluated to obtain antibodies against RhD antigen in the culture medium.
Materials and Methods
In an experimental study peripheral blood mononuclear cells were isolated from O-Negative whole blood bags using the Ficoll method. These cells were treated with L-leucyl-L-leucine methyl ester (LLME) and were exposed to soluble and particulate RhD antigen, interferon-gamma (IFN-γ), interleukin-4 (IL-4) and CpGODN in a culture plate at 37°C for one week. After this period of incubation, in order to evaluate the production of antibodies from lymphocytes on the supernatant obtained from the culture medium, ELISA assay was performed.
Results
The use of IFN-γ, IFN-γ + IL-4 and IFN-γ + CpGODN was identified as the most optimal factors for the production of antibodies in an in vitro immunization (by producing antibodies with concentrations of 75.851 ± 36.831, 82.195 ± 31.293, and 66.134 ± 24.814
Conclusions
The use of immunization optimization agents, has an effective role in the production of antibodies against RhD antigen in the in vitro culture.
Background and Objectives
Antibody against RhD antigen is important in blood transfusion and clinical medicine. Therefore, in this study, the effective factors in the optimization of in vitro immunization to achieve immunized lymphocytes that produce antibodies were evaluated to obtain antibodies against RhD antigen in the culture medium.
Materials and Methods
In an experimental study peripheral blood mononuclear cells were isolated from O-Negative whole blood bags using the Ficoll method. These cells were treated with L-leucyl-L-leucine methyl ester (LLME) and were exposed to soluble and particulate RhD antigen, interferon-gamma (IFN-γ), interleukin-4 (IL-4) and CpGODN in a culture plate at 37°C for one week. After this period of incubation, in order to evaluate the production of antibodies from lymphocytes on the supernatant obtained from the culture medium, ELISA assay was performed.
Results
The use of IFN-γ, IFN-γ + IL-4 and IFN-γ + CpGODN was identified as the most optimal factors for the production of antibodies in an in vitro immunization (by producing antibodies with concentrations of 75.851 ± 36.831, 82.195 ± 31.293, and 66.134 ± 24.814
Conclusions
The use of immunization optimization agents, has an effective role in the production of antibodies against RhD antigen in the in vitro culture.
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