A rapid and efficient strategy to generate allele-specific anti-HLA monoclonal antibodies.
That generation of allele-specific anti-human leukocyte antigen (HLA) monoclonal antibodies (ASHmAb) is very difficult is well known. This is thought to be due to the unique epitope structure, an assemblage of amino acid residues that lie separately in the amino acid sequence of human HLA, and to its low antigenicity compared with that of common epitopes recognized as xenogeneic determinants by mice. Here we report a rapid and efficient strategy to generate ASHmAb. Different from usual immunization methods is that we suppressed the production of non-allele-specific anti-HLA antibodies against xenogeneic determinants of HLA molecules by immunizing human HLA-B51 transgenic mice against non-HLA-B51 HLA tetramers. In addition, HLA-coated beads enabled rapid and efficient screening for ASHmAb. ASHmAb generated by this strategy will be useful for HLA typing and for clinical diagnosis, such as flow cytometry-based chimerism analysis for early detection of graft failure and relapse of leukemia after HLA-mismatched hematopoietic stem cell transplantation.
Yamazaki
Satoshi
S
Division of Stem Cell Therapy, Center for Stem Cell Biology and Medicine, The Institute of Medical Science, The University of Tokyo, Minato-ku, Tokyo, Japan.
Suzuki
Nao
N
Saito
Tsuneyoshi
T
Ishii
Yumiko
Y
Takiguchi
Masafumi
M
Nakauchi
Hiromitsu
H
Watanabe
Nobukazu
N
D000483:Alleles; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000918:Antibody Specificity; D002459:Cell Fusion; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D015235:HLA-B Antigens / Q000235:genetics / Q000276:immunology*; D059949:HLA-B51 Antigen; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015235:HLA-B Antigens; D059949:HLA-B51 Antigen