Chaturvedi P, Mukherjee R, Chaturvedi U C, Mathur A
Postgraduate Department of Microbiology, KG Medical College, Lucknow, India.
Int J Exp Pathol. 1992 Jun;73(3):263-72.
Dengue type 2 virus (DV) induces a subpopulation of T lymphocytes of mouse spleen to secrete a soluble helper cytokine (HF) which enhances the DV-specific IgM antibody plaque forming cells (PFC). The present study undertaken to purify and characterize HF shows that it can be purified by low pressure liquid chromatography (LPLC) using Sephacryl S-200 column. HF consisted of two subunits, having a M(r) of 65-68 kDa on SDS-PAGE, and both had similar activity. The isoelectric point of HF was 6.5. HF-specific antisera (HFAS) raised in mice neutralized the activity of HF in mice, reacted with it in a Western blot assay, and bound HF in an immunosorbent column. HF bound to DV-antigen in an immunosorbent column and enhanced only the DV-specific PFC. HF had no effect on PFC against heterologous antigens such as Japanese encephalitis virus, Coxsackie B4 virus or sheep red blood cells. HF generated in mice of H-2k haplotype, enhanced DV-specific PFC in the same strain of mice but had no effect on that in the H-2d or H-2q haplotype strains of mice. Thus, DV-induced HF with a M(r) of 65-68 kDa, antigen-specificity and genetic-restriction differs from most of the similarly acting cytokines but appears similar to the cell-free form of T cell receptor alpha beta dimer.
2型登革病毒(DV)可诱导小鼠脾脏中的一个T淋巴细胞亚群分泌一种可溶性辅助细胞因子(HF),该因子可增强DV特异性IgM抗体空斑形成细胞(PFC)。本研究旨在纯化和鉴定HF,结果表明它可以通过使用Sephacryl S - 200柱的低压液相色谱(LPLC)进行纯化。HF由两个亚基组成,在SDS - PAGE上的分子量为65 - 68 kDa,且两者具有相似的活性。HF的等电点为6.5。在小鼠中产生的HF特异性抗血清(HFAS)可中和小鼠体内HF的活性,在蛋白质免疫印迹分析中与HF发生反应,并在免疫吸附柱中结合HF。HF在免疫吸附柱中与DV抗原结合,且仅增强DV特异性PFC。HF对针对异源抗原(如日本脑炎病毒、柯萨奇B4病毒或绵羊红细胞)的PFC没有影响。在H - 2k单倍型小鼠中产生的HF可增强同品系小鼠中的DV特异性PFC,但对H - 2d或H - 2q单倍型品系小鼠中的PFC没有影响。因此,DV诱导产生的分子量为65 - 68 kDa、具有抗原特异性和遗传限制性的HF不同于大多数具有类似作用的细胞因子,但似乎与无细胞形式的T细胞受体αβ二聚体相似。