A single VH gene is utilized predominantly in anti-BrMRBC hybridomas derived from purified Ly-1 B cells. Definition of the VH11 family.
A single VH gene is utilized predominantly in anti-BrMRBC hybridomas derived from purified Ly-1 B cells. Definition of the VH11 family.
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DOI：
10.4049/jimmunol.142.10.3643
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发表时间：
1989-05
影响因子：
4.4
通讯作者：
Richard R. Hardy;C. Carmack;S. Shinton;R. Riblet;Kyoko Hayakawa
中科院分区：
文献类型：
--
作者：
Richard R. Hardy;C. Carmack;S. Shinton;R. Riblet;Kyoko Hayakawa
通过从两个不同的表面 IgM+ 脾 B 细胞群、Ly-1 B 细胞和“传统”(Ly-1-) B 细胞建立杂交瘤，我们发现与传统 B 细胞 (0.03%) 相比，Ly-1 B 细胞群包含对菠萝蛋白酶处理的自体红细胞 (抗 BrMRBC) 具有特异性的细胞频率高出 30 至 70 倍 (1 至 2%)。我们从用从 SM/J 小鼠脾脏分选的 Ly-1 B 细胞制成的两个杂交瘤中克隆并测序了编码抗 BrMRBC 抗体的 V 基因。 VH 序列（两者）与之前报道的与该特异性相关的序列相同，并且属于新的 VH 基因家族。该基因家族在此定义为 VH11，只有两个成员，是 Ly-1 B 衍生的抗 BrMRBC 杂交瘤集合中重排的主要 VH，始终与单个 VL 基因（V kappa 9 家族的成员）相关。此外，对从腹膜分选的 Ly-1 B 细胞制成的杂交瘤的分析显示，VH11 编码的抗 BrMRBC 特异性的频率更高（30%）。 Ly-1 群体中抗 BrMRBC 频率的这种变化取决于位置，再加上 VH11 与特定 V kappa 基因的重复关联，表明抗原驱动的选择（至少部分）是该群体中看到的偏向 V 基因表达的原因。此外，通过将 VH11 5' 定位到几个更接近 J 的家族（Q52，7183），排除了可能导致偏向表达的机制，即由于与 J 非常接近（如前 B 系中所见）而导致的优先重排。
By establishing hybridomas from two distinct surface IgM+ splenic B cell populations, Ly-1 B cells and "conventional" (Ly-1-) B cells, we found that the Ly-1 B population includes a 30 to 70 times higher frequency (1 to 2%) of cells with specificity for bromelain treated autologous red blood cells (anti-BrMRBC) when compared with conventional B cells (0.03%). We cloned and sequenced the V genes encoding anti-BrMRBC antibody from two hybridomas made with Ly-1 B cells sorted from the spleen of SM/J mice. The VH sequence (for both) is identical with the previously reported sequence associated with this specificity and belongs to a new VH gene family. This gene family, defined here as VH11, has only two members and is the predominant VH rearranged in a collection of Ly-1 B derived anti-BrMRBC hybridomas, always in association with a single VL gene (a member of the V kappa 9 family). Furthermore, analysis of hybridomas made with Ly-1 B cells sorted from the peritoneum reveals a yet higher increased frequency of VH11-encoded anti-BrMRBC specificity (30%). This variation in frequency of anti-BrMRBC in the Ly-1 population depending on location, together with the repeated association of VH11 with a particular V kappa gene suggest that antigen driven selection is (at least in part) responsible for the biased V gene expression seen in this population. Furthermore, a mechanism that might contribute to biased expression, preferential rearrangement due to close proximity to J (as seen in pre-B lines), is excluded by localization of VH11 5' to several of the more J-proximal families (Q52, 7183).