Nikolaeva A I
Biokhimiia. 1980 Jan;45(1):146-56.
An iodinated erythrocyte antigen isolated from sheep erythrocytes (125I-EAG) was injected to the BALB/c or CBA mice in a single dose of 20-25 mkg as for protein. The antigen determinants were detected by incorporation of a radioactive label into spleen B-lymphocytes on the 1sh-5th post-immunization days. It was shown that the T-cells contain no label and that 2/3 of the radioactivity of B-lymphocyte suspension of the 125I-EAG-immunized mice are detected in the cell nuclei within mRNP. During electrophoresis of the particles in 2,5% polyacrylamide gel nuclear mRNP isolated from B-lymphocytes of mice immunized with cold EAG are adsorbed by an immunoadsorbent containing antibodies against sheep erythrocytes. Fractionation of 125I-cytoplasmic extract obtained from 125I-EAG-immunized mouse B-lymphocytes in sucrose gradient revealed that the radioactive label was detected in the same two regions of the sucrose gradient, which is occupied by light RNP complexes containing mRNA and polyribosomes. Under experimental conditions allowing to detect the transitions of mRNP complexes into polyribosomes, a shift of the 125I-labelled material (probably together with the RNP particles) towards polyribosomes was observed. Thus, the antigen determinants are detected as part of nuclear and cytoplasmic mRNP of B-lymphocytes during the whole cycle of the primary immune response and are detected within the composition of cytoplasmic mRNP during the immunoglobulin polypeptide chain synthesis.
从绵羊红细胞中分离出的碘化红细胞抗原(125I-EAG),以20 - 25微克蛋白质的单剂量注射到BALB/c或CBA小鼠体内。在免疫后第1 - 5天,通过将放射性标记掺入脾脏B淋巴细胞中来检测抗原决定簇。结果表明,T细胞不含标记,并且在125I-EAG免疫小鼠的B淋巴细胞悬液中,2/3的放射性在细胞核内的mRNP中被检测到。在用冷EAG免疫的小鼠B淋巴细胞分离出的核mRNP在2.5%聚丙烯酰胺凝胶中进行颗粒电泳时,被含有抗绵羊红细胞抗体的免疫吸附剂吸附。对从125I-EAG免疫的小鼠B淋巴细胞获得的125I-细胞质提取物在蔗糖梯度中进行分级分离,结果显示放射性标记在蔗糖梯度的相同两个区域被检测到,这两个区域被含有mRNA和多核糖体的轻RNP复合物占据。在允许检测mRNP复合物向多核糖体转变的实验条件下,观察到125I标记的物质(可能与RNP颗粒一起)向多核糖体的转移。因此,在初次免疫反应的整个周期中,抗原决定簇被检测为B淋巴细胞的核和细胞质mRNP的一部分,并且在免疫球蛋白多肽链合成期间在细胞质mRNP的组成中被检测到。