HEMATOPOIETIC CYTOKINES EFFECT ON MODEL OF AIDS

SIV和HIV的血液学效应已得到充分描述，但 发病机制知之甚少。 假设的机制包括 CD 34+祖细胞生长调节异常， 造血细胞因子，或造血细胞因子抑制剂的存在， 造血 无论机制如何，造血细胞因子可以 可用于改善外周血细胞计数， 病毒复制和负荷尚未得到充分解决。 到 研究潜在的机制和对病毒调节的影响， 用500 TCID 50 SIVmac 239感染27只恒河猴， 在大约6个月时（“早期”）施用细胞因子的意图 感染; n=12）和有症状的疾病（“晚期”感染; n=12）。 在每组12只动物中，分配4个亚组（n=3 每个亚组）接受rh-FLT-3配体（干细胞 区室特异性细胞因子）、rr-IL-3（定向祖细胞 隔室细胞因子）、rh-G-CSF/EPO/ TPO组合（对 成熟细胞谱系），或rr-IL-12（作为免疫调节剂 细胞因子）。 一个亚组接受了rh-FLT-3配体， rr-IL-3（早期）与G-CSF/EPO/TPO和IL-12（早期）亚组 马上开始。 在rh-flt-3配体后，CD 34+祖细胞 细胞骨髓室没有显著扩大（1.03 +/-0.19%，第0天vs 1.48 +/-0.8%，第15天），但释放CD 34 + 细胞进入外周血导致22.9 × 103/ml的最大值。 第八天。 (This与我们的SIV-猴相比， 细胞为0.75 +/-0.22，0天和2.11 +/-1.14%，11天[p <0.05]， 第15天CD 34+细胞达到96.2 × 103/ml）。CFU-GM扩增和 定量聚合酶链反应的病毒负荷研究正在进行中。 CD 34+的PCR 流式细胞仪分选富集的细胞不能扩增SIV基因组 元素 非粘附性外周血单个核细胞的平行实验 从这个队列（用和不用5 μ g PHA刺激）和用 增加细胞因子剂量dem显示p27增加> 2x rr-IL-3而不是rh-flt-3或EPO的过度控制。 充分 表征“早期”接受细胞因子的12只动物， 包括CD 34+细胞和亚群，CD 3，4，8，19，16/56，CFU-GM， 正在进行BFU-E、病毒共培养和通过PCR测定病毒负荷。

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.