Boor Patrick P C, Ijzermans Jan N M, van der Molen Renate G, Binda Rekha, Mancham Shanta, Metselaar Herold J, Kusters Johannes G, de Jong Esther, Drexhage Hemmo A, Kwekkeboom Jaap
Department of Gastroenterology and Hepatology, Room L-455, Erasmus Medical Center, Rotterdam, The Netherlands.
Immunol Lett. 2005 Jul 15;99(2):162-8. doi: 10.1016/j.imlet.2005.02.013. Epub 2005 Mar 27.
It was investigated whether positive immunomagnetic selection with two novel DC-specific mAb allowed purification of functional myeloid dendritic cells (MDC) and plasmacytoid dendritic cells (PDC) from the human lymph nodes (LN). The results were compared with enrichment of DC by low-density Nycodenz gradient centrifugation followed by immunomagnetic depletion of residual B- and T-cells (Nycodenz method). MDC were selected from inguinal LN cell suspensions using CD1c mAb and PDC using anti-BDCA-4 mAb. Immunomagnetic selection with anti-CD1c mAb yielded highly pure MDC preparations (90 +/- 3% MDC; n = 7), provided that the B-cells were thoroughly depleted by using CD19 magnetic beads and Large Depletion (LD) columns prior to selection of MDC. The purified MDC comprised both mature and immature cells and were functional, secreting large amounts of cytokines upon stimulation and strongly stimulating allogeneic T-cell proliferation. Immunomagnetic selection with anti-BDCA-4 mAb enriched PDC 70-fold to a purity of 59 +/- 26% (n = 8). The contamination consisted mainly of BDCA-4(+) T- and NK-cells. The previously used Nycodenz method yielded mixtures of MDC and PDC, not allowing functional studies of MDC and PDC separately. In conclusion, positive immunomagnetic selection with CD1c mAb from human LN cell suspensions yields almost pure MDC preparations, which are, in contrast to those obtained by the Nycodenz method, not contaminated with PDC. Moreover, these MDC are functionally intact. Selection with anti-BDCA-4 mAb does enrich PDC from human LN, but the resulting preparations are contaminated with T- and NK-cells.
研究了使用两种新型树突状细胞（DC）特异性单克隆抗体（mAb）进行阳性免疫磁选是否能够从人淋巴结（LN）中纯化出功能性髓样树突状细胞（MDC）和浆细胞样树突状细胞（PDC）。将结果与通过低密度Nycodenz梯度离心富集DC，随后对残余B细胞和T细胞进行免疫磁耗竭的方法（Nycodenz法）进行比较。使用CD1c mAb从腹股沟LN细胞悬液中筛选MDC，使用抗BDCA-4 mAb筛选PDC。使用抗CD1c mAb进行免疫磁选可得到高度纯化的MDC制剂（90±3% MDC；n = 7），前提是在选择MDC之前，先用CD19磁珠和大耗竭（LD）柱彻底清除B细胞。纯化的MDC包括成熟和未成熟细胞，且具有功能，在受到刺激时分泌大量细胞因子，并强烈刺激同种异体T细胞增殖。使用抗BDCA-4 mAb进行免疫磁选可将PDC富集70倍，纯度达到59±26%（n = 8）。污染物主要由BDCA-阳性T细胞和NK细胞组成。先前使用的Nycodenz法产生了MDC和PDC的混合物，无法分别对MDC和PDC进行功能研究。总之，从人LN细胞悬液中使用CD1c mAb进行阳性免疫磁选可得到几乎纯的MDC制剂，与通过Nycodenz法获得的制剂不同，该制剂未被PDC污染。此外，这些MDC功能完整。使用抗BDCA-4 mAb进行筛选确实能从人LN中富集PDC，但所得制剂被T细胞和NK细胞污染。
