Hodgkin病单个H/R-S细胞Ig基因重排的研究

Hodgkin′s disease: Ig gene rearrangements in single H/Rs cells

目的探讨Ｈｏｄｇｋｉｎ病（ＨＤ）Ｈｏｄｇｋｉｎ／ＲｅｅｄＳｔｅｒｎｂｅｒｇ（Ｈ／ＲＳ）细胞的性质和来源。方法从８例ＨＤ冰冻切片上共提取Ｈ／ＲＳ细胞６８个，用ＩｇＨ通用引物ＦＲⅢａ／ＪＨ和κ、λ轻链家族性特异性引物行ＰＣＲ检测。结果１例淋巴细胞为主型（ＬＰ）ＨＤ的Ｈ／ＲＳ细胞重复出现ＩｇＨ和Ｖκ４家族重排；２例结节硬化型ＨＤ（ＮＳＨＤ）中，１例Ｈ／ＲＳ细胞有单次的ＩｇＨ、Ｖκ４和Ｖλ３重排；１例有重复的Ｖκ４重排，单次的ＩｇＨ和Ｖκ２、４重排；５例混合细胞型（ＭＣ）ＨＤ中，１例有重复的ＩｇＨ重排，单次的Ｖκ３和Ｖλ４重排；１例有重复的Ｖκ４重排，单次的Ｖλ３和ＩｇＨ重排；２例有重复的ＩｇＨ重排和单次的ＩｇＨ重排；１例有重复的ＩｇＨ和Ｖκ３重排，单次的Ｖλ２、４重排。结论（１）ＬＰＨＤ的Ｈ／ＲＳ细胞具有克隆性ＩｇＨ和Ｖκ４基因重排，支持ＬＰＨＤ为Ｂ细胞克隆性增生的结论；（２）ＩｇＨ和κ、λ基因重排的出现，提示ＮＳＨＤ和ＭＣＨＤ的Ｈ／ＲＳ细胞来源于不同分化阶段的Ｂ细胞，可能为单克隆性，也有可能为寡克隆性增生。本研究的新颖之处在于：（１）改进后的单个细胞ＰＣＲ方法和Ｋ¨ｕｐｐｅｒｓ等人的原方法相比较，?

Objective To investigate the origin and cell lineage of Hodgkin/ReedSternberg (H/RS) cells from various subtypes of Hodgkin′s disease (HD). Methods 68 single H/RS cells were taken from frozen section of 8 cases of HD. The DNA from these single cells was amplified by polymerase chain reaction (PCR) with immunoglobulin heavy chain gene FRⅢa/JH primers and light chain familyspecific primers. Results (1) IgH and Vκ4 rearrangements were repeatedly found in H/RS cells from one case of lymphocytic predominant HD (LPHD). (2) Repeated and unique IgH and Vκ2,4 rearrangements were observed in 1 case of nodular sclerosing HD (NSHD) and unique IgH, Vλ3/Vκ4 rearrangements in another case of NSHD. (3) Repeated IgH/Vλ3 and unique Vλ2,4 rearrangements, repeated Vκ4 and unique IgH/Vκ3 rearrangements, plus repeated IgH and unique Vκ3/Vλ4 rearrangements were detected respectively in 3 cases of mixed cellularity HD (MCHD). Repeated and unique rearrangements were found respectively in 2 cases of MCHD. Conclusions The H/RS cells isolated from LPHD had monoclonal Igh and Vκ4 gene rearrangements, which supports the conclusion that LPHD is a monoclonal proliferation of B cells. The appearance of IgH and κ,λ gene rearrangement suggest that the H/RS cells isolated from classical HD (MCHD and NSHD) originate from B lineage cells at various stages of differentiation.