TY - JOUR
T1 - Enumeration of human lymphocyte subsets by monoclonal antibodies and flow cytometry
T2 - a comparative study using whole blood or mononuclear cells separated by density gradient centrifugation
AU - De Paoli, Paolo
AU - Reitano, Michele
AU - Battistin, Sandro
AU - Castiglia, Carla
AU - Santini, Gianfranco
PY - 1984/9/4
Y1 - 1984/9/4
N2 - Enumeration of lymphocyte subpopulations by combined use of flow cytometry and monoclonal antibodies is influenced by the separation method used. T or B lymphocyte antigen frequencies do not differ in samples of whole blood or after separation on Ficoll-Paque or Percoll, but there is a significant increase of Leu7+ Leu11+ lymphocytes showing strong natural killer activity at the sample-separation medium interface. At the bottom of the tubes a selective loss of OKT8+, Leu11- cells is found; at this level cytotoxicity is very low. Our data suggest that Leu7+ cells could be subdivided into 2 subpopulations differing in reactivity with the monoclonal antibody to Leu11, natural killer activity and density. Differences observed in the percentages of Leu7+, Leu11+ lymphocytes between whole blood and separated cells could be due to an enrichment produced by centrifugation or to the presence in whole blood of a factor interfering with antibody binding.
AB - Enumeration of lymphocyte subpopulations by combined use of flow cytometry and monoclonal antibodies is influenced by the separation method used. T or B lymphocyte antigen frequencies do not differ in samples of whole blood or after separation on Ficoll-Paque or Percoll, but there is a significant increase of Leu7+ Leu11+ lymphocytes showing strong natural killer activity at the sample-separation medium interface. At the bottom of the tubes a selective loss of OKT8+, Leu11- cells is found; at this level cytotoxicity is very low. Our data suggest that Leu7+ cells could be subdivided into 2 subpopulations differing in reactivity with the monoclonal antibody to Leu11, natural killer activity and density. Differences observed in the percentages of Leu7+, Leu11+ lymphocytes between whole blood and separated cells could be due to an enrichment produced by centrifugation or to the presence in whole blood of a factor interfering with antibody binding.
KW - density gradient centrifugation
KW - lymphocytes
KW - monoclonal antibodies
KW - whole blood
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U2 - 10.1016/0022-1759(84)90003-6
DO - 10.1016/0022-1759(84)90003-6
M3 - Article
C2 - 6332151
AN - SCOPUS:0021227952
VL - 72
SP - 349
EP - 353
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
SN - 0022-1759
IS - 2
ER -