Influence of chloroquine or acid treatment of human platelets on the antigenicity of HLA and the 'thrombocyte-specific' glycoproteins Ia/IIa, IIb, and IIb/IIIa.

The influence of treatment of platelets with citrate buffer (pH 7.2), chloroquine, or citric acid at pH 3 on the expression of HLA class I antigens and 'thrombocyte-specific' glycoproteins was investigated by means of flow cytometry. After treatment with citric acid at pH 3 and chloroquine, the expression of HLA class I was significantly reduced, while the density of the molecules GPIa/IIa, GPIIb, and GPIIb/IIIa (GP = glycoprotein) carrying 'thrombocyte-specific' antigens was not or only weakly decreased on the surface of the platelets. The use of two monoclonal antibodies (HC-10 and HC-A2) against the native heavy chain of the HLA class I molecule revealed that 'antigen stripping' with chloroquine or citric acid does not affect the entire molecule: only the beta 2-microglobulin is cleaved, or only some epitopes on the heavy chain are altered by this procedure. The treatment with citric acid yielded better results with respect to the removal of HLA class I activity and the preservation of 'thrombocyte-specific' glycoproteins. The presence of the heavy chain of HLA class I molecules on the surface of platelets after treatment with citric acid and chloroquine confirms the hypothesis that platelets--like nucleated cells--bear HLA class I antigens inserted in the cell by a cytoplasmic and a transmembrane domain.