Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi · 2002
[Participation of bonth splenic CD(5)(+) and CD(5)(-) B lymphocytes in production of platelet glycoprotein-specific autoantibodies in chronic ITP].
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Lu Baojun, Hou Ming, Lu Lu, Shi Yan, He Qingsi, Ma Daoxin, Zhang Maohong
Аннотация
OBJECTIVE: To investigate the percentage of splenic CD(5)(+) B lymphocytes in chronic idiopathic thrombocytopenic purpura (IT) and the impact of splenic CD(5)(+) and CD(5)(-) B lymphocytes on the production of platelet glycoprotein (GP)-specific autoantibodies.
METHODS: Splenic CD(5)(+) B lymphocytes were identified by two-color flow cytometric analysis in eight patients. Four of the eight patients displayed plasma autoantibodies against both GPIIb/IIIa and GPIb/IX, and their splenic B lymphocytes were separated by Ficoll-Hypaque density gradient and sheep erythrocyte, and further purified by magnetic activate cell separation (MACS). Purified CD(5)(+) and CD(5)(-) B lymphocytes were cultured separately with or without staphylococcus aureus cowan I (SAC). GP specific autoantibodies in culture supernatants were measured by modified monoclonal antibody immobilization of platelet antigen assay (MAIPA).
RESULTS: The percentage of splenic CD(5)(+) B lymphocytes in ITP patients was slightly higher than that in control with no statistical significance. MACS purified splenic CD(5)(+) and CD(5)(-) B lymphocytes from three out of four ITP patients produced high levels of anti-GPIIb/IIIa and anti-GPIb/IX antibodies. Culture supernatants of CD(5)(+) B lymphocytes from the other patient showed positive reaction only in GPIb/IX MAIPA. Culture supernatant of CD(5)(-)B lymphocytes from the same patient were double positive in both GPIIb/IIIa and GPIb/IX MAIPA.
CONCLUSIONS: Both splenic CD(5)(+) and CD(5)(-) B lymphocytes produce platelet GP-specific autoantibodies in chronic ITP with similar antibody spectrum and titer, and may all play a role in the autoimmune pathogenesis of ITP.
METHODS: Splenic CD(5)(+) B lymphocytes were identified by two-color flow cytometric analysis in eight patients. Four of the eight patients displayed plasma autoantibodies against both GPIIb/IIIa and GPIb/IX, and their splenic B lymphocytes were separated by Ficoll-Hypaque density gradient and sheep erythrocyte, and further purified by magnetic activate cell separation (MACS). Purified CD(5)(+) and CD(5)(-) B lymphocytes were cultured separately with or without staphylococcus aureus cowan I (SAC). GP specific autoantibodies in culture supernatants were measured by modified monoclonal antibody immobilization of platelet antigen assay (MAIPA).
RESULTS: The percentage of splenic CD(5)(+) B lymphocytes in ITP patients was slightly higher than that in control with no statistical significance. MACS purified splenic CD(5)(+) and CD(5)(-) B lymphocytes from three out of four ITP patients produced high levels of anti-GPIIb/IIIa and anti-GPIb/IX antibodies. Culture supernatants of CD(5)(+) B lymphocytes from the other patient showed positive reaction only in GPIb/IX MAIPA. Culture supernatant of CD(5)(-)B lymphocytes from the same patient were double positive in both GPIIb/IIIa and GPIb/IX MAIPA.
CONCLUSIONS: Both splenic CD(5)(+) and CD(5)(-) B lymphocytes produce platelet GP-specific autoantibodies in chronic ITP with similar antibody spectrum and titer, and may all play a role in the autoimmune pathogenesis of ITP.