Distribution of cyclophilin B-binding sites in the subsets of human peripheral blood lymphocytes.
Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein, mainly associated with the secretory pathway and released in biological fluids. We have recently demonstrated that both free CyPB and CyPB-CsA complex specifically bind to peripheral blood T lymphocytes and are internalized. These results suggest that CyPB might promote the targeting of the drug into sensitive cells. Peripheral blood lymphocytes are subdivided in several populations according to their biological functions and sensitivity to CsA. We have investigated the binding of CyPB to these different subsets using a CyPB derivatized by fluorescein through its single cysteine which retains its binding properties. We have confirmed that only T cells were involved in the interaction with CyPB. The ligand binding was found to be heterogeneously distributed on the different T-cell subsets and surface-bound CyPB was mainly associated with the CD4-positive cells. No significant difference was noted between the CD45RA and CD45RO subsets, demonstrating that CyPB-binding sites were equally distributed between native and memory T cells. CD3 stimulation of T lymphocytes led to a decrease in the CyPB-binding capacity, that may be explained by a down-regulation of the CyPB-receptor expression upon T-cell activation. Finally, we demonstrated that CyPB-receptor-positive cells, isolated on CyPB sulphydryl-coupled affinity matrices, are more sensitive to CyPB-complexed CsA than mixed peripheral blood lymphocytes, suggesting that CyPB potentiates CsA activity through the binding of the complex. Taken together, our results demonstrate that CyPB-binding sites are mainly associated with resting cells of the helper T lymphocyte, and that CyPB might modulate the distribution of CsA through the drug targeting to sensitive cells.
Denys
A
A
Laboratoire de Chimie Biologique, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Allain
F
F
Foxwell
B
B
Spik
G
G
D000591:Amino Acid Isomerases / Q000378:metabolism*; D001665:Binding Sites; D001667:Binding, Competitive; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D018929:Cell Culture Techniques; D021983:Cyclophilins; D016572:Cyclosporine / Q000494:pharmacology; D005452:Fluoresceins; D006801:Humans; D016130:Immunophenotyping; D007166:Immunosuppressive Agents / Q000494:pharmacology; D008213:Lymphocyte Activation; D019696:Peptidylprolyl Isomerase; D011994:Recombinant Proteins / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000187:drug effects / Q000378:metabolism*
D002352:Carrier Proteins; D005452:Fluoresceins; D007166:Immunosuppressive Agents; D011994:Recombinant Proteins; C081786:cyclophilin B; D016572:Cyclosporine; D000591:Amino Acid Isomerases; D021983:Cyclophilins; D019696:Peptidylprolyl Isomerase