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Updated: Dec 31, 2025

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
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The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity

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Identification of an activation-related protein in B cells in the ABO incompatible condition

Jingsong Cao1,2,3, Cong Chen1, Luogen Liu2

  • 1Clinical Research Center, Institute of Pathogenic Biology, Medical College, Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control, University of South China, Hengyang, Hunan 421001, P.R. China.

Experimental and Therapeutic Medicine
|January 4, 2020
PubMed

In ABO-incompatible (ABOi) kidney transplantation (KT), antibodies can mediate immunological accommodation or immune rejection, but the mechanism by which B cells are induced to produce antibodies with different functions is still unclear. Previous research established an ABOi kidney cell model and identified that haptoglobin (HP) is associated with the activation of lymphocytes. In the present study, the results of a flow cytometric assay demonstrated that HP was expressed by B cells. Moreover, dot-ELISA and ELISA analyses showed that the concentrations of total IgG, blood group B antibody, IgG1, IgG2 and IgG4 were all significantly increased in the cell model. In addition, dot-ELISA and haptoglobin level analyses showed that HP protein expression was significantly increased, while RT-qPCR assay indicated that HP was significantly reduced at the mRNA level. Furthermore, bioinformatics analysis showed that HP could interact with Smad3, and the HP-Smad3 complex was detected in a peripheral blood mononuclear cell (PBMC) protein extract by a dot-ELISA method. This research revealed that HP was involved in the process of B-cell activation by interacting with Smad3, and the results will be helpful to reveal the mechanism of B-cell activation in ABOi-KT.

Keywords:
ABOi-KTB cellsSmad3haptoglobinimmunological accommodation

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