Ann Lab Med.  2020 Jan;40(1):48-56. 10.3343/alm.2020.40.1.48.

Human B1 Cells are the Main Blood Group A-Specific B Cells That Have a Moderate Correlation With Anti-A Antibody Titer

• Xu Y
• Lee JG
• Yan JJ
• Ryu JH
• Xu S
• Yang J

• KMID: 2457493
• DOI: http://doi.org/10.3343/alm.2020.40.1.48

Abstract

BACKGROUND
Anti-carbohydrate antibody responses, including those of anti-blood group ABO antibodies, are yet to be thoroughly studied in humans. Because anti-ABO antibody-mediated rejection is a key hurdle in ABO-incompatible transplantation, it is important to understand the cellular mechanism of anti-ABO responses. We aimed to identify the main human B cell subsets that produce anti-ABO antibodies by analyzing the correlation between B cell subsets and anti-ABO antibody titers.
METHODS
Blood group A-binding B cells were analyzed in peritoneal fluid and peripheral blood samples from 43 patients undergoing peritoneal dialysis and 18 healthy volunteers with blood group B or O. The correlation between each blood group A-specific B cell subset and anti-A antibody titer was then analyzed using Pearson's correlation analysis.
RESULTS
Blood group A-binding B cells were enriched in CD27⁺CD43⁺CD1c− B1, CD5⁺ B1, CD11b⁺ B1, and CD27⁺CD43⁺CD1c+ marginal zone-B1 cells in peripheral blood. Blood group A-specific B1 cells (P=0.029 and R=0.356 for IgM; P=0.049 and R=0.325 for IgG) and marginal zone-B1 cells (P=0.011 and R=0.410 for IgM) were positively correlated with anti-A antibody titer. Further analysis of peritoneal B cells confirmed B1 cell enrichment in the peritoneal cavity but showed no difference in blood group A-specific B1 cell enrichment between the peritoneal cavity and peripheral blood.
CONCLUSIONS
Human B1 cells are the key blood group A-specific B cells that have a moderate correlation with anti-A antibody titer and therefore constitute a potential therapeutic target for successful ABO-incompatible transplantation.

Keyword

Anti-ABO antibodies; Blood group A antigen; Human B1 cells; Human marginal zone B cells; Peritoneal B cells

MeSH Terms

Antibodies
Antibody Formation
Ascitic Fluid
B-Lymphocyte Subsets
B-Lymphocytes*
Healthy Volunteers
Humans*
Immunoglobulin M
Peritoneal Cavity
Peritoneal Dialysis
Antibodies
Immunoglobulin M

Figure

• 

  Fig. 1 Gating strategy for naïve, memory, marginal zone, B1 and marginal zone-B1 B cells B1 subpopulations. (A) For analysis of B cells in peripheral blood, singlet blood lymphocytes were gated for total CD20+ B cells or ‘A’ antigen-capturing, blood group A-specific CD20+ B cells. Next, the CD20+ B cells were divided into various subsets. (B) For analysis of B1 cell subpopulations in peripheral blood, CD27+CD43+ B1 cells were subdivided into various subpopulations based on CD5 and CD11b expressions.Abbreviations: MZB, marginal zone B cells; MZ-B1, marginal zone-B1 cells.

• 

  Fig. 2 The frequency of each blood group A-specific B cell subset. (A) The proportion of each B cell subset among the total CD20+ B cells (left panel) and A-specific CD20+ B cells (right panel). (B) The proportion of blood group A-specific B cells among each B cell subset. *P<0.05, **P<0.01 compared with the naïve B2 cell group. ‡P<0.01 compared with the memory B2 cell group. (C) The proportions of each B1 cell subset among the total CD20+ B cells (left panel) and A-specific CD20+ B cells (right panel). (D) The proportions of blood group A-specific B cells among each B1 cell subset. *P<0.05, **P<0.01 compared with the naïve and memory B2 cell group. †P<0.05, ‡P<0.01 compared with the B1 cell group. The results are expressed as mean±SEM; N=30–45 per group.Abbreviations: FITC, fluorescein isothiocyanate; FSC, forward scatter; MZB, marginal zone B cells; MZ-B1, marginal zone-B1 cells; SEM, standard error of the mean.

• 

  Fig. 3 The correlation between the frequency of each B cell subset and anti-blood group A IgM (A) and IgG (B) titer in the peripheral blood. The anti-A IgM titer was positively correlated with the frequency of CD20+CD27+CD43+CD1c− B1 cells and CD20+CD27+CD43+CD1c+ MZ-B1 cells (B1 cells, P=0.029, R=0.356; MZ-B1 cells, P=0.011, R=0.410). The anti-A IgG titer was positively correlated with the frequency of CD20+CD27+CD43+CD1c− B1 cells (P=0.049; R=0.325).Abbreviations: MZB, marginal zone B cells; MZ-B1, marginal zone-B1 cells; R, Pearson correlation coefficient.

• 

  Fig. 4 The correlation between the frequency of each B cell subset and anti-blood group A IgM (A) and IgG (B) titer in the peripheral blood. The frequency of each B1 subpopulation showed a positive correlation tendency with the anti-A IgM titer.Abbreviation: R, Pearson correlation coefficient.

• 

  Fig. 5 B cell subsets and blood group A-specific B cells analyzed using the (A) CD27, CD43, CD1c and (B) CD43, CD5, CD11b classification methods. The proportion of each B cell subset within the total CD20+ B cell population (left panel), A-specific CD20+ B cells (middle panel), and blood group A-specific B cells among each B cell subset (right panel) were compared between peripheral blood and peritoneal cavity. The results are expressed as mean±SEM; N=15–30 per group.*P<0.05, **P<0.01 comparing between peripheral blood and peritoneal cavity.Abbreviations: MZB, marginal zone B cells; MZ-B1, marginal zone-B1 cells; SEM, standard error of the mean.

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