蛋白和基因水平探索H/RS细胞的起源

目的：从蛋白、组织和单细胞的基因水平进一步探索H／RS细胞的来源及其克隆性。方法首先对33例经典型霍奇金淋巴瘤(cHL)标本进行B细胞特异性激活蛋白(BSAP)和CD20的检测，然后对33例cHL患的石蜡刮片组织和部分阳性病例的微切割细胞进行免疫球蛋白重链基因克隆性重排检测，并对同一病例的石蜡刮片组织和微切割细胞的扩增产物进行测序分析比较。结果33例中30例的H／RS细胞表达BSAP，10例表达CD20，BSAP和CD20的表达率差异有显性(P=0.000)，对照病例中反应性增生淋巴结的B淋巴细胞和B细胞淋巴瘤肿瘤细胞的BSAP和CD20表达均为100％，T细胞淋巴瘤的肿瘤细胞无表达；33例中的16例石蜡刮片组织IgH基因重排阳性，微切割的19管H／RS细胞有14管出现重排阳性，细胞数目不同的各管阳性率差异无显性(P=0．280)；同一病例的石蜡刮片组织和微切割细胞的PCR产物测序结果均为IgH可变区片段，但是碱基序列并不完全相同。结论：进一步支持cHL中大多数H／RS细胞为B细胞起源，且可能来源于其不同的分化阶段。

Explore the origin of H/RS cells on protein and gene

Objective To explore the origin and clonality of H/RS cells. Methods Immunohistochemical method was used to detect the expression of B-cell-specific activator protein (BSAP) and CD 20 in 33 paraffin-embedded tissues of classical Hodgkin lymphoma(cHL). IgH gene rearrangement was detected in 33 paraffin-embedded cHL tissue and 6 microsectioned H/RS cell samples. The PCR products of a case of cHL and its microsectioned cells were sequenced. Results H/RS cells were positive for BSAP in 30 of 33(90.91%) cHL cases and positive for CD 20 in 10/33(30.30%) cases. There was a significant difference between the expression of BSAP and CD 20 in H/RS cells(P=0.000). BSAP and CD 20 were positive in almost all B cells of lymph node reactive hyperplasia and malignant cells in B-cell lymphomas while were negative in all maliganant cells of T-cell lymphomas. 16 of 33 cHL were positive for gene rearrangement,and microsectioned H/RS cells in 14 of 19 tubes displayed clonal bands of rearrangement.There was no significant difference among the rearrangement rates in tubes containing different numbers of H/RS cells(P=0.280). Sequencing analyses of the PCR products from both paraffin-embedded tissue and microsection of the same patient revealed the rearranged V segments,but the sequences were not identical. Conclusion H/RS cells were originated from B cells of different differentiation stage.