Shen Y, Rudnik J, Cassol S, Drouin J, Cameron W, Izaguirre C A, Filion L G
Department of Microbiology, University of Ottawa General Hospital, Ontario, Canada.
Clin Diagn Lab Immunol. 1994 Sep;1(5):531-7. doi: 10.1128/cdli.1.5.531-537.1994.
In blood, the CD4+ T cells of patients with human immunodeficiency virus type 1 (HIV-1) harbor HIV-1; however, whether the CD4+ blood monocytes carry the virus is controversial. Tissue macrophages are known to be infected. To determine in blood monocytes from HIV-1-seropositive patients contain HIV-1, we separated monocytes and T-cell subsets by using monoclonal antibodies bound to magnetic beads and by monocyte adherence to glass. Monocytes were cultured with macrophage colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and interleukin-3. After 14 days in culture, cells were analyzed for the presence of HIV-1 antigen and multinucleated giant cells (MGCs). Freshly isolated cell subsets were analyzed for HIV-1 proviral DNA by PCR with modified env (SK68i and SK69i2) and gag (SK145i and SK150) primers. We found that (i) monocytes cultured without depletion of CD4+ T cells (11 of 11 patients) were HIV-1 antigen positive and showed dramatically increased spontaneous formation of MGCs (ii) monocytes cultured after depletion of CD4+ T cells (three experiments) were HIV-1 antigen negative and showed markedly decreased MGC formation, and (iii) in specimens from 14 patients subsequently analyzed by PCR, purified CD4+ T cells were positive for HIV-1 proviral DNA in all patients. In 11 of 14 patients (79%), the monocyte fractions were HIV-1 proviral DNA negative, while in the remaining 3 patients, the monocytes were positive for HIV-1 proviral DNA. In conclusion, the major reservoir for HIV-1 infection in human peripheral blood is the CD4+ T cell (14 of 14 cases).(ABSTRACT TRUNCATED AT 250 WORDS)
在血液中，1型人类免疫缺陷病毒（HIV-1）患者的CD4+ T细胞携带HIV-1；然而，CD4+血液单核细胞是否携带该病毒仍存在争议。已知组织巨噬细胞会被感染。为了确定HIV-1血清阳性患者的血液单核细胞中是否含有HIV-1，我们通过使用与磁珠结合的单克隆抗体以及单核细胞对玻璃的黏附作用来分离单核细胞和T细胞亚群。将单核细胞与巨噬细胞集落刺激因子、粒细胞-巨噬细胞集落刺激因子和白细胞介素-3一起培养。培养14天后，分析细胞中HIV-1抗原和多核巨细胞（MGCs）的存在情况。通过使用修饰的env（SK68i和SK69i2）和gag（SK145i和SK150）引物进行PCR，对新鲜分离的细胞亚群进行HIV-1前病毒DNA分析。我们发现：（i）在未去除CD4+ T细胞的情况下培养的单核细胞（11例患者中的11例）HIV-1抗原呈阳性，且MGCs的自发形成显著增加；（ii）在去除CD4+ T细胞后培养的单核细胞（三个实验）HIV-1抗原呈阴性，且MGC形成明显减少；（iii）在随后通过PCR分析的14例患者的样本中，所有患者的纯化CD4+ T细胞HIV-1前病毒DNA均为阳性。在14例患者中的11例（79%）中，单核细胞部分HIV-1前病毒DNA为阴性，而在其余3例患者中，单核细胞HIV-1前病毒DNA为阳性。总之，人类外周血中HIV-1感染的主要储存库是CD4+ T细胞（14例中的14例）。（摘要截短于250字）
