Burger D R, Vandenbark A A, Dunnick W, Kraybill W, Daves G D, Vetto R M
J Immunol. 1979 Mar;122(3):1091-8.
Leukocyte extracts containing human transfer factor (TF) were fractionated by exclusion chromatography, and the active fraction (Sephadex G25, Fraction IIIa) was subjected to high pressure, reverse phase (HPRP) chromatography and enzymatic degradation. TF activity was assessed by the systemic transfer of dermal skin test reactivity from KLH-immunized donors to naive recipients. Preparative HPRP chromatography resolved Fraction IIIa into multiple chromophoric regions, two of which demonstrated transfer of KLH reactivity. Alkaline phosphatase treatment of Fraction IIIa converted the major ultraviolet-absorbing component, 5'-inosine monophosphate, to inosine and resulted in TF activity being restricted to one region. This HPRP region (R1A) contained less than 1% of the UV254 active material in Fraction IIIa but greater than 90% of the reactivity. The sensitivity of TF to pronase, proteinase K, phosphodiesterase I, and phosphodiesterase II was evaluated by inhibition of systemic transfer of KLH reactivity. Pronase and proteinase K destroyed systemic transfer activity and the pronase destruction could be inhibited with traysylol. Phosphodiesterase I, a 3' exonuclease, destroyed activity, whereas phosphodiesterase II, a 5' exonuclease, did not. The data are consistent with a phosphodiester-containing polypeptide in the structure of human TF for KLH reactivity.
含人转移因子（TF）的白细胞提取物通过排阻色谱法进行分级分离，活性级分（葡聚糖凝胶G25，IIIa级分）进行高压反相（HPRP）色谱分析和酶降解。通过将来自钥孔血蓝蛋白（KLH）免疫供体的皮肤试验反应性全身性转移至未免疫受体来评估TF活性。制备性HPRP色谱将IIIa级分分离为多个发色区，其中两个显示出KLH反应性的转移。用碱性磷酸酶处理IIIa级分可将主要的紫外线吸收成分5'-肌苷单磷酸转化为肌苷，并使TF活性局限于一个区域。这个HPRP区域（R1A）在IIIa级分中所含的254nm紫外线活性物质不到1%，但反应性超过90%。通过抑制KLH反应性的全身性转移来评估TF对链霉蛋白酶、蛋白酶K、磷酸二酯酶I和磷酸二酯酶II的敏感性。链霉蛋白酶和蛋白酶K破坏全身性转移活性，而链霉蛋白酶的破坏作用可用抑肽酶抑制。3'外切核酸酶磷酸二酯酶I破坏活性，而5'外切核酸酶磷酸二酯酶II则不破坏。这些数据与含磷酸二酯的多肽在人TF对KLH反应性结构中的存在情况一致。
