Metabolism of benzo[a]pyrene by murine splenic cell types.
Metabolism of benzo[a]pyrene by murine splenic cell types.
复制标题
小鼠脾细胞类型对苯并[a]芘的代谢。
DOI：
10.1016/0041-008x(92)90304-b
复制
发表时间：
1992
影响因子：
3.8
通讯作者：
WhiteJr,KL
中科院分区：
文献类型：
--
作者：
Ladics,GS;Kawabata,TT;Munson,AE;WhiteJr,KL
本研究的目的是确定哪种B6 C3 F1小鼠脾细胞类型能够代谢B（a）P。通过不连续Percoll梯度离心分离脾细胞，沿着结合免疫磁性阴性选择或抗体介导的补体裂解，获得高度富集的脾细胞群，用于B（a）P代谢研究。免疫荧光细胞染色结合流式细胞术和Giemsa染色的细胞离心涂片细胞制备物的检查表明，常规获得纯度大于95%的B-或T-细胞群和80-90%纯度的脾巨噬细胞群。将脾细胞群与[3 H]B（a）P孵育24小时。使用高压液相色谱法分离和定量富集的脾细胞群产生的B（a）P代谢物。这些研究的结果表明，巨噬细胞是负责脾内B（a）P代谢的细胞类型。产生的B（a）P的主要代谢产物如下：含有多羟基化代谢产物、B（a）P-9，10-二羟基-9，10-二氢二醇和B（a）P-7，8-二羟基-7，8-二氢二醇的极性代谢产物的未鉴别峰。检查的其他脾细胞类型，包括B和T细胞、多形核细胞或脾被膜未产生显著高于背景水平的B（a）P代谢物。基于这些发现，巨噬细胞是代谢B（a）P的脾细胞类型。因此，巨噬细胞可能是B（a）P靶向的细胞类型，导致脾体液免疫应答抑制。
The objective of the present study was to determine which splenic cell type(s) of B6C3F1 mice was capable of metabolizing B(a)P. Separation of splenocytes based on density by centrifugation through discontinuous Percoll gradients along with immunomagnetic negative selection or antibody-mediated complement lysis was utilized to obtain highly enriched populations of splenocytes for B(a)P metabolism studies. Immunofluorescent cell staining in conjunction with flow cytometry and examination of Giemsa-stained cytospin cell preparations indicated that B- or T-cell populations of greater than 95% purity and an 80–90% pure population of splenic macrophages were routinely attained. Splenic cell populations were incubated with [3H]B(a)P for 24 hr. High-pressure liquid chromatography was used to separate and quantitate the B(a)P metabolites generated by the enriched splenic cell populations. The results of these studies demonstrate that the macrophage is the cell type responsible for the metabolism of B(a)P within the spleen. The major metabolites of B(a)P produced were as follows: an unidentified peak of polar metabolites containing polyhydroxylated metabolites, B(a)P-9,10-dihydroxy-9,10-dihydrodiol, and B(a)P-7,8-dihydroxy-7,8-dihydrodiol. Other splenic cell types examined, including B and T cells, polymorphonuclear cells, or the spleen capsule did not produce amounts of B(a)P metabolites significantly above background levels. Based on these findings, macrophages are the splenic cell types which metabolize B(a)P. As a result, macrophages may be the cell type targeted by B(a)P resulting in suppression of splenic humoral immune responses.