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Evaluation of effective factors in the optimization of immunization to achieve antibodies against RhD antigen in culture medium
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M. Amrovani   , F. Yari  , S. Milani , B. Amoohossein |
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| Keywords: Key words: Immunization, In Vitro Techniques, Antibodies |
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| Abstract (HTML) (482 Views)
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Type of Study: Research |
Subject:
Imunohematology
Published: 2022/12/31
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Full-Text: (336 Views) |
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Sci J Iran Blood Transfus Organ 2022;19 (4): 257-269
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Evaluation of effective factors in the optimization of immunization to achieve antibodies against RhD antigen
in culture medium
Amrovani M.1, Yari F.1, Milani S.1, Amoohossein B.1
1Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran
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 ngrml in 10 exposures).
Conclusions
The use of immunization optimization agents, has an effective role in the production of antibodies against RhD antigen in the in vitro culture.
Key words: Immunization, In Vitro Techniques, Antibodies
Received: 6 Aug 2022
Accepted: 6 Sep 2022
Correspondence: Yari F., PhD of Immunology. Professor of Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine.
P.O.Box: 14665-1157, Tehran, Iran. Tel: (+9821) 82052237; Fax: (+9821) 88601555
E-mail: f.yari@ibto.ir
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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
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