Yamada K, Akiyoshi K, Murakami H, Sugahara T, Ikeda I, Toyoda K, Omura H
Department of Food Science and Technology, Faculty of Agriculture, Kyushu University, Fukuoka, Japan.
In Vitro Cell Dev Biol. 1989 Mar;25(3 Pt 1):243-7. doi: 10.1007/BF02628461.
We screened for immunoglobulin (Ig) production stimulating factor (IPSF) which enhanced Ig production of human-to-human hybridomas in serum-free culture, and found that culture supernatant and lysate of human lymphoblastoid Namalwa cells stimulated proliferation and Ig production of human-to-human hybridoma HB4C5 cells. The IPSF in Namalwa lysate was partially purified with DEAE-Toyopearl 650M, hydroxylapatite and Superose 6HR 10/30 column chromatographies. The partially purified IPSF was a macromolecule of about 500,000 dalton containing 72,000 dalton protein as a major component. The activity was stable at pH 6 to 12, but inactivated partially by heating over 40 degrees C (60% decrease) and completely by trypsin digestion. These results suggest that the IPSF activity is due to its protein and heat-stable components. The Namalwa IPSF stimulated proliferation of human-to-human hybridomas but not that of mouse-to-mouse hybridomas. The IPSF also stimulated Ig production of human-to-human hybridomas derived from NAT-30 cells, but not that of other human-to-human or mouse-to-mouse hybridomas. NAT-30 is a human fusion partner derived from Namalwa cells. These results suggest that the Namalwa IPSF is an autocrine factor that stimulates proliferation and Ig production of hybridomas derived from NAT-30 cells.
我们筛选了可在无血清培养中增强人源杂交瘤免疫球蛋白（Ig）产生的免疫球蛋白（Ig）产生刺激因子（IPSF），发现人淋巴母细胞样Namalwa细胞的培养上清液和裂解物可刺激人源杂交瘤HB4C5细胞的增殖和Ig产生。用DEAE-Toyopearl 650M、羟基磷灰石和Superose 6HR 10/30柱色谱对Namalwa裂解物中的IPSF进行了部分纯化。部分纯化的IPSF是一种约500,000道尔顿的大分子，主要成分是72,000道尔顿的蛋白质。该活性在pH 6至12时稳定，但在40℃以上加热会部分失活（降低60%），经胰蛋白酶消化则完全失活。这些结果表明IPSF活性归因于其蛋白质和热稳定成分。Namalwa IPSF刺激人源杂交瘤的增殖，但不刺激鼠源杂交瘤的增殖。该IPSF还刺激源自NAT-30细胞的人源杂交瘤的Ig产生，但不刺激其他任何来源的人源或鼠源杂交瘤的Ig产生。NAT-30是源自Namalwa细胞的人融合伴侣。这些结果表明Namalwa IPSF是一种自分泌因子，可刺激源自NAT-30细胞的杂交瘤的增殖和Ig产生。
