Two populations of mouse lymphokine-activated killer cells separated by use of soybean agglutinin.
We separated lymphokine-activated killer (LAK) cells induced from spleen cells of BALB/c mice by culturing with recombinant interleukin-2 (rIL-2) into soybean agglutinin-positive (SBA+) and soybean agglutinin-negative (SBA-) fractions with a cell sorter at the time when LAK activity reached a maximum (day 3). We found that the cells with LAK activity were enriched in the SBA+ fraction. Analysis of cell surface phenotypes revealed that the SBA+ cells are of non-T cell origin, while the SBA- fraction consists of T cells. We also found that the large granular lymphocyte (LGL) fraction of spleen cells obtained with a Percoll gradient became SBA+ after culture for 3 days with rIL-2, whereas cells of high density did not. The change in SBA binding sites was also examined on C57BL/6 mouse spleen cells and we found that NK1.1+ non-T cells selectively acquire SBA binding sites at an early stage of activation with rIL-2. On the other hand, sorted SBA- cells gradually acquired SBA binding sites on extended culturing with rIL-2. These results suggest that the expression of SBA binding sites is related to the stage of cell activation with rIL-2, though cells of the NK-lineage became SBA+ at an earlier period of a culture with rIL-2 than cells of the T-lineage. Utilizing these findings, we separated NK-derived LAK cells by means of SBA-binding, and used the separated cells for adoptive immunotherapy for experimental pulmonary metastasis. We found that SBA(+)-NK cell-derived LAK cells showed stronger activity for the inhibition of experimental pulmonary metastasis than SBA(-)-T cell-derived LAK cells.
Takano
M
M
Division of Chemical Toxicology and Immunochemistry, Faculty of Pharmaceutical Sciences, University of Tokyo.
Okada
T
T
Maruyama
T
T
Harada
K
K
Imai
Y
Y
Osawa
T
T
D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D002469:Cell Separation / Q000379:methods*; D005260:Female; D007376:Interleukin-2 / Q000494:pharmacology; D015979:Killer Cells, Lymphokine-Activated / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D037102:Lectins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D037121:Plant Lectins; D011975:Receptors, Mitogen / Q000032:analysis; D011994:Recombinant Proteins / Q000494:pharmacology; D030262:Soybean Proteins; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D010835:Phytohemagglutinins
D000945:Antigens, Differentiation, T-Lymphocyte; D007376:Interleukin-2; D037102:Lectins; D037121:Plant Lectins; D011975:Receptors, Mitogen; D011994:Recombinant Proteins; D030262:Soybean Proteins; C028824:soybean lectin; D010835:Phytohemagglutinins; C046772:soybean lectin receptor