Onuma M, Tsukiyama K, Ohya K, Morishima Y, Ohno R
Microbiol Immunol. 1987;31(2):131-7. doi: 10.1111/j.1348-0421.1987.tb03076.x.
For detection of antibody to bovine leukemia virus (BLV) major core protein of p24 and cross-reactive antibody in human patients infected with human T cell leukemia virus type I (HTLV-I), monoclonal antibody, D432 against BLV p24 was used by competitive binding enzyme-linked immunoadsorbed assay (ELISA). In sera from cattle with enzootic bovine leukosis (EBL) which were positive for BLV antibodies by immunodiffusion test, 109 out of 112 (97.3%) were positive for BLV p24 antibody by competitive binding ELISA. By using the same procedures, 21 samples from adult T cell leukemia (ATL) patients and healthy carriers with HTLV-I were tested for cross-reactive antibody to BLV p24. All 21 samples were positive for HTLV-I antibodies by immunofluorescence test and/or ELISA. By competitive binding ELISA using non-treated BLV antigens, none of these 21 samples inhibited the binding of the D432. When the BLV antigen was treated by several different denaturation procedures, several HTLV-I positive samples showed the inhibition of the D432 binding and the most effective treatment was by 2-mercaptoethanol (2-ME). Sixteen out of 21 samples showed the presence of cross-reactive antibody against 2-ME-treated BLV antigens. The cross-reactivity of human sample to BLV p24 antigen was further confirmed by Western blotting of the 2-ME-treated BLV antigens. None of the 28 samples from leukemia patients other than ATL which were negative for HTLV-I antibodies showed inhibition of the D432 by the competitive binding ELISA.
为检测感染I型人类T细胞白血病病毒(HTLV-I)的人类患者体内针对牛白血病病毒(BLV)主要核心蛋白p24的抗体及交叉反应性抗体,采用竞争结合酶联免疫吸附测定(ELISA)法,使用抗BLV p24的单克隆抗体D432。在免疫扩散试验检测BLV抗体呈阳性的地方流行性牛白血病(EBL)牛血清中,112份血清中有109份(97.3%)通过竞争结合ELISA检测出BLV p24抗体呈阳性。采用相同程序,对21份来自成人T细胞白血病(ATL)患者及HTLV-I健康携带者的样本进行检测,以检测其对BLV p24的交叉反应性抗体。通过免疫荧光试验和/或ELISA检测,所有21份样本的HTLV-I抗体均呈阳性。使用未处理的BLV抗原进行竞争结合ELISA时,这21份样本均未抑制D432的结合。当BLV抗原经过几种不同的变性处理后,几份HTLV-I阳性样本显示出对D432结合的抑制作用,最有效的处理方法是用2-巯基乙醇(2-ME)处理。21份样本中有16份显示存在针对经2-ME处理的BLV抗原的交叉反应性抗体。通过对经2-ME处理的BLV抗原进行蛋白质印迹分析,进一步证实了人类样本与BLV p24抗原的交叉反应性。28份来自非ATL白血病患者且HTLV-I抗体呈阴性的样本,通过竞争结合ELISA检测均未显示对D432的抑制作用。