Sequential analysis of anti-alpha Gal natural antibody-producing B cells in GalT knockout mice in cyclophosphamide-induced tolerance.

Publisher: Blackwell Scientific Publications Country of Publication: England NLM ID: 0323767 Publication Model: Print Cited Medium: Print ISSN: 0300-9475 (Print) Linking ISSN: 03009475 NLM ISO Abbreviation: Scand J Immunol Subsets: MEDLINE

Publication: Oxford : Blackwell Scientific Publications
Original Publication: Oslo, Universitetsforlaget.

B-Lymphocyte Subsets/*drug effects ; B-Lymphocyte Subsets/*immunology ; Clonal Anergy/*drug effects ; Cyclophosphamide/*pharmacology ; Galactosyltransferases/*deficiency ; Galactosyltransferases/*genetics ; Isoantibodies/*biosynthesis ; Trisaccharides/*immunology; Animals ; B-Lymphocyte Subsets/metabolism ; Bone Marrow Transplantation/immunology ; Busulfan/administration & dosage ; Busulfan/pharmacology ; Cyclophosphamide/administration & dosage ; Erythrocyte Transfusion ; Female ; Galactosyltransferases/metabolism ; Gene Deletion ; Graft Survival/genetics ; Graft Survival/immunology ; Heart Transplantation/immunology ; Mice ; Mice, Knockout ; Mice, SCID ; Protein Binding/immunology ; Rabbits ; Skin Transplantation/immunology ; Spleen/cytology ; Spleen/transplantation ; Transplantation Conditioning ; Trisaccharides/metabolism

Previously, we have shown that cyclophosphamide (CP)-induced tolerance, marked by permanent acceptance of donor skin graft and establishment of donor mixed chimerism, was readily induced with treatment with donor spleen cells (SC), CP, busulfan (BU) and donor bone marrow cells (BMC). Here, we investigated the mechanism of anti-donor natural antibody (nAb) producing B-cell tolerance in our CP-induced tolerance systems in alpha1,3-galactosyltransferase-deficient knockout mice (GalT KO; GalT-/-, H-2(b/d)). After induction of tolerance using donor AKR SC and BMC, survival of donor heart and skin grafts and production of anti-Galalpha1-3Galbeta1-4GlcNAc (anti-alphaGal) Ab in recipient GalT KO mice were analyzed. In addition, the production of anti-alphaGal Ab and the presence of Gal-BSA binding B cells in GalT KO mice were analyzed by flow cytometry (FCM) after treatments with rabbit red blood cells (RRBC) and CP. Permanent acceptance of donor skin and heart grafts and abrogation of anti-alphaGal Ab were achieved in GalT KO mice treated with donor SC + CP/BU + BMC. However, in the GalT KO mice treated with donor SC and CP, donor skin grafts were acutely rejected, even though anti-alphaGal Ab was undetectable. Similarly, anti-alphaGal Ab was undetectable in GalT KO mice treated with RRBC and CP. Our data strongly indicated the following mechanisms: the clonal destruction in the early stage and the clonal anergy or ignorance in the late stage after conventional conditioning with RRBC and CP. In conclusion, our drug-induced tolerance protocols are effective to induce tolerance in recipients that produce anti-donor nAb.

0 (Isoantibodies)
0 (Trisaccharides)
0 (alpha-galactosyl epitope)
8N3DW7272P (Cyclophosphamide)
EC 2.4.1.- (Galactosyltransferases)
EC 2.4.1.133 (alpha 1,3 galactosyltransferase, mouse)
G1LN9045DK (Busulfan)

Date Created: 20060613 Date Completed: 20060725 Latest Revision: 20131121

20221213

10.1111/j.1365-3083.2006.001763.x

16764697