New strategies for selection of unrelated bone marrow donors.
An important problem in the selection of unrelated donors for bone marrow transplantation (UD-BMT), is HLA matching, between selected donor and recipient. Serological screening, mixed lymphocyte culture (MLC), and sequence specific oligonucleotide genotyping (PCR-SSO) are the methods commonly used for typing of HLA-genes. These ways to select donor candidates are time-expensive. We set up new applications of the "fingerprinting-PCR" technique, to analyse the polymorphic second exon of DRB, DQB, DQA, DPB of HLA Class II and second exon A, B, C HLA-Class I genes, and to search for identity between patient and serologically selected unrelated donors. In an assessment of the technique, 50 normal samples, and 4 unrelated HLA-A and HLA-B serological matched patient-donor pairs were analysed for HLA polymorphic regions. In 3 of the 4 cases (UD-BMT) at least HLA-DRB mismatched different donor-transplanted patterns were identified. In all cases PCR-SSO analysis was performed as control. Based on our data, we suggest that identification of UD for allogeneic BMT should follow these steps: 1) serological HLA-Class I and II genes screening; 2) HLA-Class II DRB gene PCR fingerprinting; 3) confirmation by SSO analysis in case of fingerprinting identity. 4) HLA-Class II DQA, DQB, DPB PCR fingerprinting. Moreover, confirmation by PCR fingerprinting or protein isoelectrofocusing of HLA-Class I identity is recommended. This "strategy" may contribute to rapid and specific selection of unrelated marrow donors.
Martinelli
G
G
Institute of Hematology L. e A. Seragnoli, University of Bologna, Italy.
Buzzi
M
M
Farabegoli
P
P
Testoni
N
N
Mantovani
V
V
Calori
E
E
Rosti
G
G
Bandini
G
G
Bragliani
M
M
Panzica
G
G
D001483:Base Sequence; D001853:Bone Marrow / Q000276:immunology; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation / Q000276:immunology* / Q000379:methods; D002985:Clinical Protocols; D004247:DNA / Q000235:genetics; D016172:DNA Fingerprinting; D006682:HLA-DP Antigens / Q000032:analysis / Q000235:genetics / Q000276:immunology; D006683:HLA-DQ Antigens / Q000032:analysis / Q000235:genetics / Q000276:immunology; D059848:HLA-DQ alpha-Chains; D059866:HLA-DQ beta-Chains; D006684:HLA-DR Antigens / Q000032:analysis / Q000235:genetics / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000032:analysis / Q000235:genetics / Q000276:immunology; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000235:genetics / Q000276:immunology; D006801:Humans; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D014019:Tissue Donors
D006682:HLA-DP Antigens; D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; D059866:HLA-DQ beta-Chains; C069050:HLA-DQA1 antigen; C057615:HLA-DQbeta antigen; D006684:HLA-DR Antigens; D015395:Histocompatibility Antigens Class I; D000949:Histocompatibility Antigens Class II; D004247:DNA