Binns R M, Duncan I A, Powis S J, Hutchings A, Butcher G W
Department of Immunology, AFRC Institute of Animal Physiology and Genetics Research, Babraham, Cambridge, U.K.
Immunology. 1992 Oct;77(2):219-27.
Rat monoclonal antibodies (mAb) against isolated pig Null T cells were derived using a novel two-colour cytofluorometric assay. One (MAC320) identified all blood CD2-sIg- 'Null' cells (present at up to approximately 6 x 10(6)/ml). Another type (MAC319 and MAC318) identified a subset comprising approximately 60% or approximately 30% of the Null cell population. This percentage appears genetically determined. This subset partially overlapped with a gamma delta T-cell receptor+ (TcR+) population which consisted of approximately 40% of Null T cells. The antibodies did not react with other leucocyte or lymphocyte populations. In non-reducing conditions, MAC320 precipitated two molecules at approximately 270,000-280,000 MW in SDS-PAGE; the larger of which was also precipitated by MAC319 (and MAC318, which binds to the same epitope). Under reducing conditions, MAC320 immunoprecipitated two or three polypeptide chains at approximately 130,000-160,000 MW; MAC319 precipitated only the largest of these polypeptides. The large MAC319+ MAC320+ molecule on one subset is removed by bromelain treatment; the smaller MAC319- MAC320+ molecule on the remaining Null cells is not bromelain sensitive. Several properties of this new antigen complex specific to pig Null T cells show that it is distinct from the ruminant T19 complex.
利用一种新型双色细胞荧光分析方法获得了针对分离出的猪Null T细胞的大鼠单克隆抗体(mAb)。其中一种(MAC320)可识别所有血液中的CD2-sIg-“Null”细胞(含量高达约6×10⁶/ml)。另一种类型(MAC319和MAC318)识别出一个亚群,该亚群约占Null细胞群体的60%或约30%。这个百分比似乎由基因决定。该亚群与γδT细胞受体阳性(TcR+)群体部分重叠,后者约占Null T细胞的40%。这些抗体不与其他白细胞或淋巴细胞群体发生反应。在非还原条件下,MAC320在SDS-PAGE中沉淀出两个分子量约为270,000 - 280,000的分子;其中较大的分子也可被MAC319(以及与相同表位结合的MAC318)沉淀。在还原条件下,MAC320免疫沉淀出两条或三条分子量约为130,000 - 160,000的多肽链;MAC319仅沉淀出这些多肽中最大的一条。一个亚群上的大的MAC319 + MAC320 +分子可通过菠萝蛋白酶处理去除;其余Null细胞上较小的MAC319 - MAC320 +分子对菠萝蛋白酶不敏感。这种猪Null T细胞特有的新抗原复合物的几个特性表明它与反刍动物的T19复合物不同。