Volume 19, Issue 4 (winter 2022)
Sci J Iran Blood Transfus Organ 2022, 19(4): 292-300 |
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Shekeft M, Milani S, Sharifi Z, Oodi A. Establishment of lymphoblastoid cell line producing Anti-RhE antibody using alloimmune thalassemia patient's blood. Sci J Iran Blood Transfus Organ 2022; 19 (4) :292-300
URL: http://bloodjournal.ir/article-1-1454-en.html
URL: http://bloodjournal.ir/article-1-1454-en.html
Abstract: (886 Views)
Abstract
Background and Objectives
One of the applications of Epstein-Barr virus in research is the immortalization of B lymphocytes and the creation of the lymphoblastoid cell line. People with thalassemia usually need blood transfusion, the main complication of which is alloimmunization. Antibody against RhE antigen causes hemolytic reactions. The aim of this study is to create a lymphoblastoid cell line that produces anti-E using peripheral blood mononuclear cells of alloimmune thalassemia patients against E antigen when exposed to Epstein Bar Virus (EBV).
Materials and Methods
In an experimental study, the mononuclear cells of 3 patients were isolated by Ficol and mixed with EBV. The transformed cells were cultured in RPMI medium containing cyclosporine. After 2-3 weeks, the formation of clones in the wells was checked and the hemagglutination test was performed for the wells containing the growing cells; the total amount of antibody in positive wells was measured by ELISA method.
Results
Exposure of mononuclear cells of patients with EBV led to cell transformation and creation of clones in some wells. The results of hemagglutination and ELISA methods showed that the clones were able to produce antibodies against the RhE antigen.
Conclusions
Anti-E antibody-producing lymphoblastoid cell line can be created by using sensitized cells of thalassemia patients against RhE antigen and its immortalization by EBV transformation method.
Background and Objectives
One of the applications of Epstein-Barr virus in research is the immortalization of B lymphocytes and the creation of the lymphoblastoid cell line. People with thalassemia usually need blood transfusion, the main complication of which is alloimmunization. Antibody against RhE antigen causes hemolytic reactions. The aim of this study is to create a lymphoblastoid cell line that produces anti-E using peripheral blood mononuclear cells of alloimmune thalassemia patients against E antigen when exposed to Epstein Bar Virus (EBV).
Materials and Methods
In an experimental study, the mononuclear cells of 3 patients were isolated by Ficol and mixed with EBV. The transformed cells were cultured in RPMI medium containing cyclosporine. After 2-3 weeks, the formation of clones in the wells was checked and the hemagglutination test was performed for the wells containing the growing cells; the total amount of antibody in positive wells was measured by ELISA method.
Results
Exposure of mononuclear cells of patients with EBV led to cell transformation and creation of clones in some wells. The results of hemagglutination and ELISA methods showed that the clones were able to produce antibodies against the RhE antigen.
Conclusions
Anti-E antibody-producing lymphoblastoid cell line can be created by using sensitized cells of thalassemia patients against RhE antigen and its immortalization by EBV transformation method.
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