Vujanovic N L, Rabinowich H, Lee Y J, Jost L, Herberman R B, Whiteside T L
Department of Pathology, University of Pittsburgh School of Medicine, Pennsylvania.
Cell Immunol. 1993 Oct 1;151(1):133-57. doi: 10.1006/cimm.1993.1227.
Human natural killer (NK) cells can be functionally subdivided into adherent (A) and non-adherent (NA) subpopulations. In the presence of 22 nM of interleukin 2(IL2), a substantial proportion of resting (R)-NK cells developed adherence to plastic as early as after 5 min of IL2 incubation, and by 1-5 hr of IL2 induction, 16% (range, 4-30%) of NK cells were adherent. Optimal concentration of IL2 for adherence of NK cells was 2-22 nM. This adherence was blocked completely by antibody to IL2 receptor (IL2R)-beta and, partially, by antibodies to beta 1 or beta 2 integrins, ICAM-1, CD2 or LFA3, but not by antibodies to the IL2R-alpha or CD56 antigen. A-NK cells separated from NA-NK cells after 5 hr of incubation in the presence of IL2 were significantly (P < 0.05) enriched in CD56dimCD16dim or -IL2Rp55+ and IL2Rp75+ cells, but were depleted of CD56bright CD16- cells. While surface density of CD56 and CD16 antigens was lower, that of beta 2 integrins (CD18, CD11a, CD11b) was higher on A-NK than on NA-NK cells. In a single-cell cytotoxicity assay, 61% of A-NK vs 37% of NA-NK cells bound, and 24% of A-NK vs 11% of NA-NK cells killed, K562 targets. In 4-day cultures with 0.02 or 2.2 nM of IL2, A-NK cells developed lymphokine-activated killer (LAK) activity later than NA-NK cells. By autoradiography, three to eight times more A-NK than NA-NK cells incorporated [3H]TdR into cell nuclei between 48 and 96 hr of IL2 incubation. In 14-day cultures in the presence of 22 nM of IL2, A-NK cells, which were initially adherent but later grew as single-cell suspensions, proliferated better (30-fold; P < 0.03) and expressed lower membrane density of CD56 than NA-NK cells. In culture, A-NK cells had consistently higher cytotoxicity against K562 targets than NA-NK cells, but cytotoxicity against Daudi was similar for both subsets. The data indicate that short incubation (1-5 hr) of human NK cells in the presence of 22 nM of IL2 allows for selection of a subpopulation which differs from the rest of NK cells not only by properties of rapid adherence to plastic, but also by a characteristic phenotype (CD3-CD56dim or -CD16dim or -beta 2integrinsbrightIL2Rp75+), rapid expression of IL2R-alpha, higher NK activity, delayed development of LAK activity, and ability to respond optimally in the presence of 22 nM of IL2.(ABSTRACT TRUNCATED AT 400 WORDS)
人类自然杀伤(NK)细胞在功能上可细分为黏附性(A)和非黏附性(NA)亚群。在存在22 nM白细胞介素2(IL2)的情况下,相当一部分静息(R)-NK细胞早在IL2孵育5分钟后就开始黏附于塑料,到IL2诱导1-5小时时,16%(范围为4%-30%)的NK细胞具有黏附性。NK细胞黏附的最佳IL2浓度为2-22 nM。这种黏附被抗IL2受体(IL2R)-β抗体完全阻断,部分被抗β1或β2整合素、细胞间黏附分子-1(ICAM-1)、CD2或淋巴细胞功能相关抗原3(LFA3)抗体阻断,但不被抗IL2R-α或CD56抗原抗体阻断。在IL2存在下孵育5小时后从NA-NK细胞中分离出的A-NK细胞,在CD56dimCD16dim或-IL2Rp55+和IL2Rp75+细胞中显著富集(P<0.05),但CD56bright CD16-细胞减少。虽然A-NK细胞上CD56和CD16抗原的表面密度较低,但β2整合素(CD18、CD11a、CD11b)的表面密度高于NA-NK细胞。在单细胞细胞毒性试验中,61%的A-NK细胞与37%的NA-NK细胞结合,24%的A-NK细胞与11%的NA-NK细胞杀伤K562靶细胞。在含有0.02或2.2 nM IL2的4天培养中,A-NK细胞比NA-NK细胞更晚产生淋巴因子激活的杀伤(LAK)活性。通过放射自显影,在IL2孵育48至96小时之间,掺入[3H]TdR进入细胞核的A-NK细胞比NA-NK细胞多三至八倍。在存在22 nM IL2的14天培养中,最初黏附但后来以单细胞悬液形式生长的A-NK细胞增殖更好(30倍;P<0.03),且其CD56的膜密度低于NA-NK细胞。在培养中,A-NK细胞对K562靶细胞的细胞毒性始终高于NA-NK细胞,但两个亚群对Daudi细胞的细胞毒性相似。数据表明,在22 nM IL2存在下对人类NK细胞进行短时间孵育(1-5小时),可选择出一个亚群,该亚群不仅具有快速黏附于塑料的特性,还具有独特的表型(CD3-CD56dim或-CD16dim或-β2整合素brightIL2Rp75+)、快速表达IL2R-α、更高的NK活性、LAK活性发育延迟以及在22 nM IL2存在下能最佳应答的能力。(摘要截断于400字)