Prince A M, Horowitz B, Dichtelmueller H, Stephan W, Gallo R C
Cancer Res. 1985 Sep;45(9 Suppl):4592s-4594s.
Human T-lymphotropic retrovirus type III (HTLV-III) can be quantitatively assayed for infectivity by inoculation of serial dilutions into cultures of the H-9 cell line and testing for reverse transcriptase in the culture supernatants. Sequential harvests revealed that 14 days of incubation of cultures fed twice weekly was sufficient to reveal maximal titers. Stocks prepared from unconcentrated H9:HTLV-IIIb supernatants have contained from 10(4.5) to 10(6.0) (TCID50)/ml. Stocks prepared by 100-fold concentration of such fluids by pelleting or by polyethylene glycol precipitation followed by pelleting onto sucrose cushions contained 10(6.0)-10(6.5) TCID50/ml. Preliminary studies are under way to utilize this system for evaluation of sterilization processes which can be applied to blood derivatives. Exposure of HTLV-III suspended in Factor VIII preparations to 0.3% tri(n-butyl)phosphate-0.2% sodium cholate resulted in inactivation of greater than or equal to 10(4.5) TCID50 in 2.5 h at 27 degrees C. Exposure of HTLV-III suspended in 4 g of gamma-globulin per 100 ml to 0.14% beta-propiolactone for 4 h at room temperature at pH 8.0 inactivated greater than or equal to 10(4.5) TCID50. However, exposure to gamma-globulin alone inactivated about 99% of HTLV-III infectivity.
人类嗜T淋巴细胞逆转录病毒III型（HTLV - III）的感染性可通过将系列稀释液接种到H - 9细胞系培养物中，并检测培养上清液中的逆转录酶来进行定量测定。连续收获表明，每周喂食两次的培养物孵育14天足以显示最高滴度。由未浓缩的H9:HTLV - IIIb上清液制备的储备液每毫升含有10(4.5)至10(6.0)（半数组织培养感染剂量，TCID50）。通过沉淀或聚乙二醇沉淀然后在蔗糖垫层上沉淀将此类液体浓缩100倍制备的储备液每毫升含有10(6.0)-10(6.5) TCID50。正在进行初步研究以利用该系统评估可应用于血液制品的灭菌工艺。将悬浮在凝血因子VIII制剂中的HTLV - III在27℃下暴露于0.3%三（正丁基）磷酸酯 - 0.2%胆酸钠2.5小时，导致大于或等于10(4.5) TCID50失活。将每100毫升中悬浮有4克γ - 球蛋白的HTLV - III在室温下pH 8.0的条件下暴露于0.14%β - 丙内酯4小时，可使大于或等于10(4.5) TCID50失活。然而，仅暴露于γ - 球蛋白可使约99%的HTLV - III感染性失活。
