Polliack A, Lampen N, Clarkson B D, De Harven E, Bentwich Z, Siegal F P, Kunkel H G
J Exp Med. 1973 Sep 1;138(3):607-24. doi: 10.1084/jem.138.3.607.
In this study a variety of human lymphocytes of known B or T cell type, obtained from multiple sources, were prepared for scanning electron microscopy (SEM) by the critical point drying method. Distinction between normal B and T lymphocytes was relatively easy in most instances, on the basis of their surface architecture. Using immunological methods, between 20 and 30% of normal peripheral blood lymphocytes (PBL) were identified as B cells and from 69 to 82% as T cells. SEM results showed that 20% of the PBL had a complex villous surface and approximately 80% of cells were smaller and had a relatively smooth surface. Comparison of the above data and enrichment of B cells from PBL, by centrifugation after T cell rosettes had formed, indicated that the "villous" cells were B lymphocytes and the "relatively smooth" cells were T lymphocytes. T cells obtained from two human thymuses were also of the generally smooth cell type. Further evidence for the distinction of B and T lymphocytes, on the basis of surface morphology, was obtained from the examination of cultured lymphoid cell lines of known B or T cell derivation. Cells from cases of chronic lymphocytic leukemia also provided support for the above interpretations. Five of six untreated cases were clearly of B cell type by immunologic and SEM criteria. One unusual case showed the presence of T and B lymphocytes in almost equal numbers by SEM and a mixture of B and T cells by immunologic markers. An additional case that had received chemotherapy showed numerous atypical cells that were difficult to classify by SEM. Detailed examination of the smoother T cells showed that at least half of them had a moderate number of surface digitations and a small proportion had an intermediate surface morphology with a relatively large number of surface digitations. The latter presented difficulties in classification and may correspond to different stages of differentiation and represent subpopulations of lymphocytes. The distinction between human B and T lymphocytes on the basis of their surface architecture can be made by SEM of critical point dried samples, with relative ease in most but not all instances. The effects of stimulation, cell cycle, differentiation, intercellular contact, and density of cell population, on the surface architecture of lymphoid cells, remain to be determined.
在本研究中，通过临界点干燥法，对从多个来源获取的多种已知B细胞或T细胞类型的人淋巴细胞进行扫描电子显微镜（SEM）制备。在大多数情况下，根据其表面结构，正常B淋巴细胞和T淋巴细胞之间的区分相对容易。采用免疫学方法，正常外周血淋巴细胞（PBL）中20%至30%被鉴定为B细胞，69%至82%为T细胞。SEM结果显示，20%的PBL具有复杂的绒毛状表面，约80%的细胞较小且表面相对光滑。上述数据与通过T细胞花环形成后离心从PBL中富集B细胞的比较表明，“绒毛状”细胞是B淋巴细胞，“相对光滑”的细胞是T淋巴细胞。从两个人类胸腺获得的T细胞也属于一般的光滑细胞类型。通过对已知B细胞或T细胞来源的培养淋巴细胞系的检查，获得了基于表面形态区分B淋巴细胞和T淋巴细胞的进一步证据。慢性淋巴细胞白血病病例的细胞也为上述解释提供了支持。六例未经治疗的病例中有五例根据免疫学和SEM标准明确为B细胞类型。一个不寻常的病例通过SEM显示T淋巴细胞和B淋巴细胞数量几乎相等，通过免疫标记显示为B细胞和T细胞的混合物。另一例接受化疗的病例显示出许多非典型细胞，通过SEM难以分类。对较光滑的T细胞的详细检查表明，其中至少一半具有中等数量的表面指状突起，一小部分具有中等表面形态，表面指状突起数量相对较多。后者在分类上存在困难，可能对应于不同的分化阶段，代表淋巴细胞亚群。通过临界点干燥样品的SEM，可以根据其表面结构区分人B淋巴细胞和T淋巴细胞，在大多数但并非所有情况下相对容易。刺激、细胞周期、分化、细胞间接触和细胞群体密度对淋巴细胞表面结构的影响仍有待确定。
