GENETIC-DIFFERENCES BETWEEN BLOOD-DERIVED AND BRAIN-DERIVED VIRAL SEQUENCES FROM HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1-INFECTED PATIENTS - EVIDENCE OF CONSERVED ELEMENTS IN THE V3-REGION OF THE ENVELOPE PROTEIN OF BRAIN-DERIVED SEQUENCES
GENETIC-DIFFERENCES BETWEEN BLOOD-DERIVED AND BRAIN-DERIVED VIRAL SEQUENCES FROM HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1-INFECTED PATIENTS - EVIDENCE OF CONSERVED ELEMENTS IN THE V3-REGION OF THE ENVELOPE PROTEIN OF BRAIN-DERIVED SEQUENCES
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DOI：
10.1128/jvi.68.11.7467-7481.1994
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发表时间：
1994-11-01
影响因子：
5.4
通讯作者：
WOLINSKY, SM
中科院分区：
文献类型：
--
作者：
KORBER, BTM;KUNSTMAN, KJ;WOLINSKY, SM
人类免疫缺陷病毒 1 型 (HIV-1) 序列是从六名接受诊断性神经外科手术的患者的血液和通过立体定向活检获得的脑组织中生成的。通过巢式 PCR 直接扩增原病毒 DNA，并对每个样本中的 8 至 36 个克隆进行测序。对患者体内包膜 V3-V5 区 HIV-1 DNA 序列集的系统发育分析显示，脑病毒序列相对于血液病毒序列聚集在一起，这表明在 6 例病例中的 4 例中存在病毒的组织特异性区室化。在另外两个案例中，血液和脑病毒序列在系统发育分析中混合在一起，表明病毒在这两种组织之间运输。对两名患者脑活检样本进行基于载玻片的 PCR 驱动的原位杂交证实了我们对患者内系统发育分析的解释。患者间 V3 区脑源序列距离显着小于血液源序列距离。相对于环的尖端，与血液源性病毒序列组相比，脑源性病毒序列组具有带负电荷或中性电荷的趋势。熵计算被用来衡量血液和脑病毒序列比对中每个位置的变异性。发现了一组相对保守的位置，大脑中的熵明显低于血液来源的病毒序列。这些位点构成了大脑的“特征模式”，或者是 V3 区中一组不连续的氨基酸，这些氨基酸在源自脑组织的病毒序列中保守。这种源自大脑的特征模式在之前体外鉴定为巨噬细胞趋向性的分离株中也得到了很好的保存。巨噬细胞单核细胞的趋向性可能是导致病毒脑特征模式保存的生物学限制。
Human immunodeficiency virus type 1 (HIV-1) sequences were generated from blood and from brain tissue obtained by stereotactic biopsy from six patients undergoing a diagnostic neurosurgical procedure. Proviral DNA was directly amplified by nested PCR, and 8 to 36 clones from each sample were sequenced. Phylogenetic analysis of intrapatient envelope V3-V5 region HIV-1 DNA sequence sets revealed that brain viral sequences were clustered relative to the blood viral sequences, suggestive of tissue-specific compartmentalization of the virus in four of the six cases. In the other two cases, the blood and brain virus sequences were intermingled in the phylogenetic analyses, suggesting trafficking of virus between the two tissues. Slide-based PCR-driven in situ hybridization of two of the patients' brain biopsy samples confirmed our interpretation of the intrapatient phylogenetic analyses. Interpatient V3 region brain-derived sequence distances were significantly less than blood-derived sequence distances. Relative to the tip of the loop, the set of brain-derived viral sequences had a tendency towards negative or neutral charge compared with the set of blood-derived viral sequences. Entropy calculations were used as a measure of the variability at each position in alignments of blood and brain viral sequences. A relatively conserved set of positions were found, with a significantly lower entropy in the brain than in the blood-derived viral sequences. These sites constitute a brain ''signature pattern,'' or a noncontiguous set of amino acids in the V3 region conserved in viral sequences derived from brain tissue. This brain-derived signature pattern was also well preserved among isolates previously characterized in vitro as macrophage tropic. Macrophage monocyte tropism may be the biological constraint that results in the conservation of the viral brain signature pattern.