HEMATOPOIETIC CYTOKINES EFFECT ON MODEL OF AIDS

SIV和HIV的血液学效应得到了很好的描述，但 发病机制尚不清楚。假设的机制包括 CD34+祖细胞生长调节异常，CD34+细胞生长调节失调 造血细胞因子，或存在一种抑制 造血术。无论机制如何，造血细胞因子都可以 用于改善外周血细胞计数，通过其对 病毒复制和负担问题尚未得到充分解决。至 研究潜在的机制和对病毒调控的影响，一组 用500只TCID50 SIVmac239感染27只恒河猴 在大约6个月(“早期”)使用细胞因子的意向 感染；n=12)，症状出现时(“晚期”感染； N=12)。在每组12只动物中，分配4个亚组(n=3 每个亚群)接受rh-Flt-3配体(干细胞 隔室特异性细胞因子)、rr-IL-3(已确定的祖细胞 隔室细胞因子)，rh-G-CSF/EPO/TPO组合(激活 成熟细胞系)，或rr-IL-12(作为免疫调节剂 细胞因子)。一个亚基接受了rh-flt-3配体和一个亚基 RR-IL-3(早期)联合G-CSF/EPO/TPO和IL-12(早期)亚组 马上开始。继rh-Flt-3配体后，CD34+祖细胞 细胞骨髓室未见明显扩张(1.03 +/-0.19%，第0天vs 1.48+/-0.8%，第15天)，尽管CD34+释放 进入外周血的细胞最高可达22.9×103/毫升 第8天。(这是与我们的SIV-猴子相比，在那里BM CD34+ 第0天细胞为0.75+/-0.22，第11天为2.11+/-1.14%[p&lt；0.05]， 第15天CD34+细胞达96.2×103/ml)。CFU-GM扩展和 定量聚合酶链式反应的病毒载量研究正在进行中。CD34+的聚合酶链式反应 经流式细胞仪分选的细胞不能扩增SIV基因组 元素。非黏附性外周血单核细胞的平行实验 来自该队列(使用和不使用5微克PHA刺激)和 随着细胞因子剂量的增加，&gt；2x的p27表达增加 由rr-IL-3控制，而不是rh-flt-3或EPO。饱满 描述了“早期”接受细胞因子的12只动物的特征， 包括CD34+细胞及其亚群、CD3、4、8、19、16/56、CFU-GM、 BFU-E、病毒共培养和病毒载量的聚合酶链式反应正在进行中。

The hematologic effects of SIV and HIV are well described, but the pathogenesis is poorly understood. Postulated mechanisms include abnormal regulation of CD34+ progenitor cell growth, dysregulation of hematopoietic cytokines, or the presence of an inhibitor of hematopoiesis. Regardless of mechanism, hematopoietic cytokines can be used to improve peripheral blood counts though their effect on viral replication and burden has not been adequately addressed. To study potential mechanisms and effect on viral regulation, a cohort of 27 rhesus macaques was infected with 500 TCID50 SIVmac239 with the intent of administering cytokines at approximately 6 months ("early" infection; n=12) and when symptomatic with disease ("late" infection; n=12). In each group of 12 animals, 4 subgroups are assigned (n=3 each subgroup) to receive either rh-flt-3 ligand (a stem cell compartment specific cytokine), rr-IL-3 (a committed progenitor cell compartment cytokine), combination rh-G-CSF/EPO/ TPO (active on maturing cell lineages), or rr-IL-12 (as an immunomodulatory cytokine). One subgroup has received rh-flt-3 ligand and one subgroup rr-IL-3 (early) with G-CSF/EPO/TPO and IL-12 (early) subgroups to commence presently. Following rh-flt-3 ligand, the CD34+ progenitor cell bone marrow compartment did not significantly expand (1.03 +/-0.19%, day 0 vs 1.48 +/- 0.8%, day 15), though release of CD34+ cells into the peripheral blood led to a maximum of 22.9 x 103/ml on day 8. (This is in comparison to our SIV- monkeys where BM CD34+ cells were 0.75 +/- 0.22, day 0 and 2.11 +/- 1.14% day 11 [p < .05], and day 15 CD34+ cells reached 96.2 x 103/ml ). CFU-GM expansion and viral burden studies by quantitative PCR are underway. PCR of CD34+ cells enriched by flow cytometric sorting did not amplify SIV genomic elements. Parallel experiments of non-adherent peripheral blood MNC from this cohort (with and without stimulation with 5 ug PHA) and with increasing doses of cytokines dem onstrated an increase in p27 of > 2x over control by rr-IL-3 but not rh-flt -3 or EPO. Full characterization of the 12 animals that received cytokines "early", including CD34+ cells and subsets, CD3, 4, 8, 19, 16/56, CFU-GM, BFU-E, viral coculture, and viral burden by PCR is underway.