Lathey J L, Brambilla D, Goodenow M M, Nokta M, Rasheed S, Siwak E B, Bremer J W, Huang D D, Yi Y, Reichelderfer P S, Collman R G
Department of Pediatrics, University of California San Diego, La Jolla 92093-0672, USA.
J Leukoc Biol. 2000 Sep;68(3):324-30.
A monocyte-derived macrophage (MDM) culture assay was used to define the replication kinetics of HIV isolates. Ten-day-old MDMs were infected with HIV. Supernatants were collected and assayed for HIV p24 on days 3, 7, 10, and 14 post-infection (PI). In this assay, SF162 (macrophage tropic, NSI) produced increasing amounts of HIV p24 antigen with increasing time in culture. BRU (nonmacrophage tropic, SI) infection resulted in low levels of HIV p24 antigen with no increase in production during the culture period. A panel of 12 clinical isolates was evaluated. All isolates produced detectable levels of HIV p24 antigen in MDMs. However, the NSI viruses had significantly higher log10 HIV p24 antigen values at all times PI (P < 0.01). Co-receptor usage was determined for all 12 isolates (8 NSI and 4 SI). All SI isolates used CXCR4 for entry; two used CXCR4 only, one used CXCR4, CCR5, and CCR3, and one was a mixture of two isolates using CXCR4 and CCR5. None of the NSI viruses used CXCR4 for entry. All used CCR5 as their predominant co-receptor. Of the eight NSI isolates, three used CCR5 only, two used CCR5 and CCR2b, one used CCR5 and CCR3, and one used CCR5, CCR3, and CCR2b. Log10 HIV p24 antigen production on day 14 PI for viruses that used CCR5+CCR3 (3.79 + 1.40) was greater than for viruses that used CCR5+CCR2b (3.22 + 1.55) or CCR5 (3.32 + 1.49), and all were greater than those that used CXCR4 only (1.69 + 0.28), regardless of SI phenotype (P < 0.05). Thus, in these primary isolates, macrophage tropism and replication kinetics were closely linked to CCR5 utilization, whereas SI capacity was closely linked to CXCR4 utilization. Furthermore, viruses, which could use CCR5 and CCR3 for entry, had a replication advantage in macrophages, regardless of SI phenotype.
采用单核细胞衍生巨噬细胞（MDM）培养试验来确定HIV分离株的复制动力学。将10日龄的MDM用HIV感染。在感染后（PI）第3、7、10和14天收集上清液并检测HIV p24。在该试验中，SF162（巨噬细胞嗜性，非合胞体诱导型）在培养过程中随着时间推移产生的HIV p24抗原量不断增加。BRU（非巨噬细胞嗜性，合胞体诱导型）感染导致HIV p24抗原水平较低，且在培养期间产量没有增加。对一组12个临床分离株进行了评估。所有分离株在MDM中均产生可检测水平的HIV p24抗原。然而，非合胞体诱导型病毒在所有PI时间点的log10 HIV p24抗原值均显著更高（P<0.01）。确定了所有12个分离株（8个非合胞体诱导型和4个合胞体诱导型）的共受体使用情况。所有合胞体诱导型分离株利用CXCR4进入细胞；2个仅使用CXCR4，1个使用CXCR4、CCR5和CCR3，1个是使用CXCR4和CCR5的两个分离株的混合物。没有非合胞体诱导型病毒利用CXCR4进入细胞。所有病毒均以CCR5作为主要共受体。在8个非合胞体诱导型分离株中，3个仅使用CCR5，2个使用CCR5和CCR2b，1个使用CCR5和CCR3，1个使用CCR5、CCR3和CCR2b。对于使用CCR5 + CCR3的病毒，PI第14天的log10 HIV p24抗原产量（3.79 + 1.40）高于使用CCR5 + CCR2b的病毒（3.22 + 1.55）或仅使用CCR5的病毒（3.32 + 1.49），且所有这些产量均高于仅使用CXCR4的病毒（1.69 + 0.28），无论其合胞体诱导型表型如何（P<0.05）。因此，在这些原发性分离株中，巨噬细胞嗜性和复制动力学与CCR5的利用密切相关，而合胞体诱导型能力与CXCR4的利用密切相关。此外，无论合胞体诱导型表型如何，能够利用CCR5和CCR3进入细胞的病毒在巨噬细胞中具有复制优势。
