Evolution of proviral gp120 over the first year of HIV-1 subtype C infection

The evolution of proviral gp120 during the first year after seroconversion in HIV-1 subtype C infection was addressed in a case series of eight subjects. Multiple viral variants were found in two out of eight cases. Slow rate of viral RNA decline and high early viral RNA set point were associated with a higher level of proviral diversity from 0 to 200 days after seroconversion. Proviral divergence from MRCA over the same period also differed between subjects with slow and fast decline of viral RNA, suggesting that evolution of proviral gp120 early in infection may be linked to the level of viral RNA replication. Changes in the length of variable loops were minimal, and length reduction was more common than length increase. Potential N-linked glycosylation sites ranged ± one site, showing common fluctuations in the V4 and V5 loops. These results highlight the role of proviral gp120 diversity and diversification in the pathogenesis of acute HIV-1 subtype C infection.     

差别聚合酶链反应定量检测艾滋病患者外周血前病毒DNA水平

目的　为了评价艾滋病的治疗效果、筛选有效治疗药物 ,了解患者体内病情变化 ,常常需要对治疗前后患者体内病毒水平进行客观的定量。方法　采用差别聚合酶链反应 ,共同扩增待测靶基因HIV 1 gag和参照模板 β globin基因 ,建立起检测HIV 1复制水平的定量方法 ,并对 8例艾滋病患者外周血前病毒DNA含量进行定量。结果　 8例患者的前病毒DNA水平介于 0 5 0 8～ 2 2 1 0拷贝数 /μgDNA之间 ,其复制和整合水平存在差异。结论　该方法具有快速、可靠、重复性好等优点 ,非常适合于典型的临床标本的检测 ,为艾滋病疗效判定提供客观依据     

Provirus Load in Patients with Human T-Cell Leukemia Virus Type 1 Uveitis Correlates with Precedent Graves' Disease and Disease Activities

We previously demonstrated the increased provirus load in the peripheral blood of patients with human T-cell leukemia virus type 1 (HTLV-1) uveitis (HU). To delineate the relevance of the increased provirus load to clinical and immunologic parameters, we studied the correlation between them. Seventy-nine HU patients (24 male and 55 female) were included in the study, with their informed consent. Plasma samples and genomic DNA of the peripheral blood mononuclear cells were isolated and the provirus load was estimated by semi-quantitative polymerase chain reaction of the gag region sequence. Serum levels of anti-HTLV-1 antibodies and soluble IL-2R were determined by electrochemiluminescence immuno assay and by ELISA, respectively. Disease activities were assessed and graded 0 to 4 according to the evaluation system. Recurrence of the disease during the follow-up period was diagnosed ophthalmologically. The provirus load was significantly higher in the HU patients after Graves' disease (GD) than in those without GD ( P <0.05). It correlated with disease activities assessed in terms of vitreous inflammation and interval to recurrence (both P <0.05). In the HU patients without GD, it correlated with the serum levels of soluble IL-2 receptor ( P <0.01), and nearly with those of HTLV-1 antibody ( P =0.063). These correlations were not found in the HU patients after GD under methimazole treatment. The results suggested a direct involvement of HTLV-1-infected cells in the pathogenesis of uveitis, and raise the possibility that hyperthyroidism may contribute to the clonal expansion of HTLV-1-infected cells.     

Tumor model-specific proviral insertional mutagenesis of the Fos/Jdp2/Batf locus

Retroviral activation of the AP-1/ATF super family member Jdp2 was recently reported to be a common event in M-MLV-induced T cell lymphoma in p27-null C57x129 mice as compared to wild type-inoculated mice but has not been found important in other models. On the basis of retroviral tag retrieval from 1190 individual Akv- and SL3-3-induced lymphomas, we here report that insertional mutagenesis into the 250-kb Fos/Jdp2/Batf locus is associated with SL3-3 MLV-induced T but not Akv-induced B cell lymphomas of NMRI and SWR mice. Integration pattern and clonality analyses suggest that Jdp2 participates in SL3-3-induced tumorigenesis distinctly as compared to the M-MLV setting. Northern blot analysis showed Jdp2 to be alternatively spliced in various normal tissues as well as MLV-induced lymphomas. Interestingly, in some tumors, proviral insertion seems to activate different mRNA sub-species. Whereas elevated mRNA levels of the Fos gene could not be correlated with provirus presence, in one case, Northern blot analysis as well as quantitative real-time PCR indicated proviral activation of the AP-1 super family member Batf, a gene not previously reported to be a target of insertional mutagenesis. A novel integration cluster between Jdp2 and Batf apparently did not influence the expression level of either gene, underscoring the importance of addressing expression effects to identify target genes of insertion. Altogether, such distinct insertion patterns point to different mechanism of activation of specific proto-oncogenes and are consequently of importance for the understanding of proviral activation mechanisms as well as the specific role of individual oncogenes in tumor development.