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. 2017 Dec 16;18(1):49.
doi: 10.1186/s12865-017-0233-9.

The influence of HK2 blood group antigen on human B cell activation for ABOi-KT conditions

Jingsong Cao  1   2 , Luogen Liu  2 , Yunsheng Zhang  2 , Jianhua Xiao  3   4 , Yi Wang  5   6
Affiliations

The influence of HK2 blood group antigen on human B cell activation for ABOi-KT conditions

Jingsong Cao et al. BMC Immunol. .

Abstract

Background: It is well known that ABO blood group system incompatible kidney transplantation (ABOi-KT) is an effective strategy for end-stage renal disease. The main barrier for ABOi-KT is how to keep host B cell activation and blood group antibody titer in low levels. Moreover, the mechanism of B cell activation induced by blood group antigen was unclear in ABOi-KT.

Results: In this study, HK2 cells were identified to express blood group B antigen when cocultured with lymphocytes of blood group A. Optical microscope observation demonstrated that HK2 cells in coculture group gradually decreased. Furthermore, flow cytometer assay identified that T cell phenotypes (CD3+, CD3+CD4+ and CD3+CD8+) had no significant change and B cell phenotypes (CD19+ and CD138+) were all significantly enhanced (3.07 and 3.02 folds) at day 4. In addition, immunoturbidimetry analysis demonstrated that blood group B antibody was significantly increased to 2.35 fold at day 4, IgG was significantly increased to 3.60 and 2.81 folds at days 4 and 8 respectively, while IgM had no significant change at the measured time points.

Conclusions: Taken together, B cells were activated and secreted blood group B antibody after treatment with HK2 expressing blood group B antigen. The results of this study maybe useful for further determination of the mechanism of B cell activation after ABO incompatible kidney endothelial cells stimulation.

Keywords: ABOi-KT; B cells activation; Blood group B antibody; Blood group B antigen; HK2.

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Conflict of interest statement

Ethics approval and consent to participate

Written consent was obtained from the participants. The experiments were approved by the Animal Welfare and Research Ethics Committee of the Institute of University of South China.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interest.

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Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Blood group antigen identified by immunohistochemistry. a PB group, b Blood group A antigen analysis, c Blood group B antigen analysis. All groups were repeated for 3 times
Fig. 2
Fig. 2
Optical microscope analyzed for HK2 cells. a PB group, b HK2 group, c Coculture group. d the analysis of histogram for HK2 cell quantity. Bars represent mean ± standard deviation (n = 3). Significant differences between the PB and the coculture groups were indicated with one (p < 0.05) or two (p < 0.01) asterisks
Fig. 3
Fig. 3
The T cells and B cells subsets analyzed by flow cytometry. a CD3+ phenotype T cells, b CD3+CD8+ phenotype T cells, c CD3+CD4+ phenotype T cells, d CD19+ phenotype B cells, e CD138+ phenotype B cells. f The flow cytometry scatter plot of CD19+ and CD138+ phenotype B cells. Bars represent mean ± standard deviation (n = 3). Significant differences between the PB and the coculture groups were indicated with one (p < 0.05) asterisks
Fig. 4
Fig. 4
The analysis of western-blotting for mouse monoclonal to anti-blood group B antibody in reagent. Lane 1. PAGE assay, lane 2. Western-blotting assay, lane 3. protein mark
Fig. 5
Fig. 5
Immunoturbidimetry analysis for blood group B antibody, IgG and IgM. a Blood group B antibody concentration assay; b IgG concentration assay; c IgM concentration assay. Bars represent mean ± standard deviation (n = 3). Significant differences between the PB and the coculture group were indicated with one (p < 0.05) or two (p < 0.01) asterisks. Conc.: concentration
Fig. 6
Fig. 6
The activation of T follicular helper cells for B cells*. Tfh: T follicular helper, TCR: T cell receptor, MHC II: major histocompatibility complex class II, ICOS: Inducible costimulator, ICOSL: ICOS ligand, CD40L: CD40 ligand, IL: interleukin, IL-R: interleukin receptor, CSR: class switch recombination, SHM: somatic hypermutation, AID: activation-induced cytidine deaminase, UNG: uracil-N-glycosylase. * The figure was drawn as Durandy er al [35] reported with some modification
See this image and copyright information in PMC

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