Phenotypic and functional analysis of the HLA-class I-specific inhibitory receptors of natural killer cells isolated from peripheral blood of patients undergoing bone marrow transplantation from matched unrelated donors.
BACKGROUND
Limited information is available on the natural killer cell reconstitution after bone marrow transplantation and on the possible role of these cells in graft-versus-host-disease.


METHODS
Blood samples were collected at different time intervals after transplantation. Lymphocytes were analyzed for informative markers by immunocytofluorimetric analysis. Natural killer cells derived from patients undergoing matched unrelated donor transplant were cloned by limiting dilution in the presence of phytohemoagglutinin and IL2. The natural killer cell clones were analyzed for cytolytic activity.


RESULTS
In the nine patients analyzed undergoing transplantation from sibling donors, the majority of peripheral blood lymphocytes during the first 80 d after BMT were represented by T lymphocytes, while in the 15 patients undergoing matched unrelated donor transplant natural killer cells consistently outnumbered T lymphocytes. During the early phases after transplantation, most CD56+CD3- natural killer cells did not express CD16 which was expressed at later intervals. Analysis of the inhibitory receptors specific for HLA-class I molecules showed that CD94/NKG2A, specific for HLA-E, unlike normal donors, was expressed by all natural killer cells including the early appearing CD16-negative ones. Killer inhibitory receptors of the Ig superfamily were expressed late and in low percentages after transplantation and were always coexpressed with CD94/NKG2A. Natural killer-cell clones efficiently lysed the HLA-class I-negative cell lines K562 and 721-221. Natural killer-cell populations or clones isolated from patients with graft-versus-host-disease, failed to lyse donor or recipient derived phytohemoagglutinin-induced lymphoblasts.


CONCLUSIONS
Our analysis shows that (1) recipients of matched unrelated donors transplants exhibit a high proportion of NK cells (2) all NK cells express CD94/NKG2A while the HLA-class I-specific inhibitory receptors of Ig superfamily appear at later stages and (3) donor NK cells do not lysed donor or recipient target cells.
Vitale
C
C
Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy, and Dipartimento di Medicina Sperimentale Università di Genova, Italy.
Pitto
A
A
Benvenuto
F
F
Ponte
M
M
Bellomo
R
R
Frassoni
F
F
Mingari
M C
MC
Bacigalupo
A
A
Moretta
L
L
D000293:Adolescent; D000328:Adult; D000483:Alleles; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology*; D016026:Bone Marrow Transplantation / Q000276:immunology*; D018929:Cell Culture Techniques / Q000379:methods; D002999:Clone Cells; D005260:Female; D015235:HLA-B Antigens / Q000235:genetics / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000235:genetics / Q000276:immunology*; D006650:Histocompatibility Testing; D006801:Humans; D016130:Immunophenotyping; D007694:Killer Cells, Natural / Q000276:immunology*; D007938:Leukemia / Q000097:blood / Q000276:immunology / Q000628:therapy; D008297:Male; D008875:Middle Aged; D009190:Myelodysplastic Syndromes / Q000097:blood / Q000276:immunology / Q000628:therapy; D009679:Nuclear Family; D013601:T-Lymphocytes / Q000276:immunology; D013997:Time Factors; D014019:Tissue Donors
D015703:Antigens, CD; D015235:HLA-B Antigens; D015395:Histocompatibility Antigens Class I