Gessain A, Saal F, Giron M L, Lasneret J, Lagaye S, Gout O, De Thé G, Sigaux F, Peries J
UPR A0043 CNRS Rétrovirus et Rétrotransposons des Vertébrés, Hôpital Saint-Louis, Paris, France.
J Gen Virol. 1990 Feb;71 ( Pt 2):333-41. doi: 10.1099/0022-1317-71-2-333.
Twelve long-term cell lines were established from peripheral blood mononuclear cells (PBMC) or cerebrospinal fluid cells of patients with human T lymphotropic virus type I (HTLV-1) seropositive tropical spastic paraparesis (TSP) originating from the French West Indies, French Guyana or the Central African Republic. Most of these long-term interleukin-2-dependent cell lines exhibited a pattern characteristic of CD4(+)-activated T cells with high expression of CD2, CD3 and CD4 antigens, associated with a strong density of TAC and DR molecules. Nevertheless, in five cases CD8 expression was present at a significant level. HTLV-I antigens were never detected in uncultured PBMC, but they were expressed in a few cells after short-term culture and after 4 months the majority of the cells were HTLV-I positive, as demonstrated by indirect immunofluorescence (IF) using polyclonal or monoclonal anti-p19 and anti-p24 antibodies. Low and variable levels of reverse transcriptase activity were detected in supernatant fluids of these cell lines only after 4 months of culture, when at least 50% of the cells exhibited HTLV-I antigens by IF. However, numerous type C HTLV-I-like viral particles were detected, mostly in the extracellular spaces, with rare budding particles. Similar findings were found in three T cell lines derived from West Indian and African patients with adult T-cell leukaemia/lymphoma (ATLL). Differences in high Mr polypeptides were detected by Western blot in cell lysates when comparing TSP- or ATLL-derived T cell lines. Thus a signal of 62K was easily detectable in all the TSP lines, but not in the ATLL lines. In all cell lines bands corresponding to p53, p24 and p19 viral core polypeptides were present, as was the env gene-coded protein p46.
从来自法属西印度群岛、法属圭亚那或中非共和国的人类嗜T淋巴细胞病毒I型(HTLV-1)血清阳性热带痉挛性截瘫(TSP)患者的外周血单核细胞(PBMC)或脑脊液细胞中建立了12个长期细胞系。这些长期依赖白细胞介素-2的细胞系大多表现出CD4(+)活化T细胞的特征模式,CD2、CD3和CD4抗原高表达,伴有TAC和DR分子的强密度。然而,在5例中CD8表达水平显著。未培养的PBMC中从未检测到HTLV-I抗原,但短期培养后在少数细胞中表达,4个月后大多数细胞为HTLV-I阳性,这通过使用多克隆或单克隆抗p19和抗p24抗体的间接免疫荧光(IF)得以证明。仅在培养4个月后,当至少50%的细胞通过IF显示出HTLV-I抗原时,在这些细胞系的上清液中检测到低水平且变化不定的逆转录酶活性。然而,检测到大量C型HTLV-I样病毒颗粒,大多在细胞外空间,出芽颗粒很少。在源自西印度和非洲成年T细胞白血病/淋巴瘤(ATLL)患者的三个T细胞系中也发现了类似结果。比较TSP或ATLL来源的T细胞系时,通过蛋白质印迹法在细胞裂解物中检测到高分子量多肽的差异。因此,在所有TSP系中都很容易检测到62K的信号,但在ATLL系中未检测到。在所有细胞系中都存在与p53、p24和p19病毒核心多肽相对应的条带,env基因编码的蛋白p46也存在。