Nuclear matrix protein is released from apoptotic white cells during cold (1-6 degrees C) storage of concentrated red cell units and might induce antibody response in multiply transfused patients.
BACKGROUND
A previous study showed that white cells in blood units undergo apoptosis during storage.


METHODS
The present study attempts to show the release of nuclear matrix protein (NMP) in the supernatants of red cell units and to determine whether antibodies against nuclear components may be present in multiply transfused patients; the methods employed were enzyme-linked immunosorbent assay, flow cytometry, microscopy, immunoblotting, immunofluorescence, and confocal laser-scanning microscopy.


RESULTS
NMP is released from white cells in the supernatant of packed red cell units upon cold storage (1-6 degrees C). The concentration of NMP correlates well with the degree of apoptosis, as analyzed by flow cytometry, nuclear dye staining, and DNA gel electrophoresis. Immunofluorescence also shows that white cells undergoing apoptosis (pre-G(1) peak, as seen by propidium iodide staining and flow cytometry) have an NMP content lower than control cells, which confirms an actual release of NMP. Moreover, immunoblotting analysis and immunofluorescent staining showed that, in 4 of 38 multiply transfused patients, autoantibodies against NMPs were present without any clinical or laboratory sign of autoimmune disease. One of the sera, recognizing a 64-kDa NMP, immunostained nuclear dots that were identified as coiled bodies because of their colocalization with p 80 coilin.


CONCLUSIONS
NMP is released in the supernatant of red cell units. The results obtained from patients suggest that nuclear proteins released during apoptosis, once transfused, may induce an immune response in multiply transfused patients.
Martelli
A M
AM
Department of Normal Human Morphology, University of Trieste, Trieste, Italy.
Tazzari
P L
PL
Bortul
R
R
Riccio
M
M
Tabellini
G
G
Santi
S
S
Frabetti
F
F
Musiani
D
D
Bareggi
R
R
Conte
R
R
D000906:Antibodies / Q000097:blood; D000917:Antibody Formation; D034961:Antigens, Nuclear; D017209:Apoptosis / Q000276:immunology; D001793:Blood Preservation / Q000379:methods; D001803:Blood Transfusion; D003080:Cold Temperature; D004396:Coloring Agents; D004912:Erythrocytes / Q000737:chemistry*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting; D007962:Leukocytes / Q000166:cytology*; D018613:Microscopy, Confocal; D009687:Nuclear Proteins / Q000276:immunology* / Q000378:metabolism*
D000906:Antibodies; D034961:Antigens, Nuclear; D004396:Coloring Agents; D009687:Nuclear Proteins