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Updated: Aug 25, 2026

Isolation, Cultivation, and Transient Transfection of Primary Human T Cells to Generate Chimeric Antigen Receptor (CAR) T Cells
Published on: March 27, 2026
Two different forms of human CTLA-4 proteins following peripheral T cell activation
Taehoon Chun1, Hak-Jong Choi, Yong-Hoon Chung
1Department of Microbiology and Immunology, School of Medicine, Hanyang University, Haeng-Dang Dong, Sung-Dong Ku, Seoul 133-791, South Korea.
Abstract:
To study expression of human CTLA-4 in peripheral T cells, we developed a new anti-peptide (anti-CTLA4pB) antibody. Both denatured and native forms of CTLA-4 can be recognized by anti-CTLA4pB antibody. After peripheral T cell activation, we observed that most of CTLA-4 is localized primarily to intracellular compartment rather than to the cell surface. Biochemical study using anti-CTLA4pB antibody was revealed two forms of CTLA-4 on activated peripheral T cells. The majority of CTLA-4 was detected as a 34 kDa moiety in cytosolic fraction under reducing conditions. The presence of a 30 kDa CTLA-4 moiety was observed simultaneously in membrane fraction under reducing condition. The post-induction ratio of cytosolic:membrane-bound forms of CTLA-4 was approximately 50:1. Further analysis was indicated that the 34 kDa moiety in cytosolic fraction is part of a 250 kDa multi-protein complex. Antibodies raised against this complex recognized 54 and 75 kDa proteins together with a 34 kDa form of CTLA-4. In contrast, membrane-bound forms of CTLA-4 were 60 and 120 kDa under non-reducing condition. The molar ratio of 60 and 120 kDa oligomers was approximately 1:1. These results may provide evidence for post-translational modification of human CTLA-4 after T cell activation.
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