Plasma IP-10 Is Associated with Rapid Disease Progression in Early HIV-1 Infection

Abstract Cytokines play key roles in modulating disease progression in simian immunodeficiency virus/human immunodeficiency virus (SIV/HIV) infection. There are a few studies on the relationship between early cytokines and HIV disease prognosis. In this study, we first report the relationship based on two groups with clearly different disease progression. We found that IP-10 was the only cytokine among the 26 cytokines tested that was always positively correlated with disease progression, and was associated with the time for CD4 counts to fall below 200 cells/μL during Fiebig stages III-V in HIV-1 infection. This suggests that high IP-10 levels in the blood are associated with rapid disease progression during Fiebig stages III-V in HIV-1 infection.     

Rapid Disease Progression in HIV-1-Infected Men Who Have Sex with Men Is Negatively Correlated with Peripheral Plasmacytoid Dendritic Cell Counts at the Early Stage of Primary Infection

To explore the relationship between absolute dendritic cell (DC) counts at the early stage of primary human immunodeficiency virus type 1 (HIV-1) infection (PHI) and subsequent disease progression, we performed a prospective study of 16 rapid progressors (RPs) and 12 typical progressors (TPs) from a PHI cohort of men who have sex with men. Significantly decreased plasmacytoid DC (pDC) and myeloid DC (mDC) counts in the blood of RPs were observed at study entry as compared with TPs and healthy HIV-1-negative subjects. Low baseline pDC counts were significantly associated with rapid disease progression after adjustment for baseline CD4(+) T cell counts, mDC counts, and HIV-1 load. Kaplan-Meier survival analysis showed that low pDC counts were strongly associated with rapid disease progression. Our findings demonstrated the predictive value of blood absolute pDC counts at baseline in PHI for HIV-1 disease progression. Further studies are required to confirm this notion.     

Low Frequency of CXCR4-Using Viruses in Patients at the Time of Primary Non-Subtype-B HIV-1 Infection

We used genotypic and phenotypic assays to estimate the frequency of X4/DM viruses in 131 patients infected with non-subtype-B viruses at the time of primary HIV-1 infection (PHI). All patients were enrolled in the French PRIMO Cohort from 1996 to 2007. Most strains belonged to CRF02_AG (51.1%) and subtype A (14.5%). Sixteen viruses (12.2%) were classified as CXCR4 tropic (“X4 strains”) by the combined criteria of amino acids 11 and 25 of the V3 loop (11/25) and net charge rules and/or the SVMgeno2pheno_10% algorithm: 6 strains by the combined genotypic rule, 7 by the SVMgeno2pheno_10% algorithm, and 3, clustering in subtype D, by both algorithms. However, only one strain (0.8%), belonging to subtype A, was defined as a dual-tropic (DM) virus by the phenotypic assay. The 67 CRF02_AG strains included 2 classified as X4 strains by the combined genotypic rule (3%) and 2 others classified as X4 strains by SVMgeno2pheno_10% (3%), but none of these 4 strains was an X4 or DM strain according to the phenotypic assay. These results suggest that the cellular virus reservoir was established with X4 strains in very few non-subtype-B-infected patients at the time of PHI. Genotypic predictions can overestimate the proportion of non-subtype-B X4 viruses at PHI.Human immunodeficiency virus type 1 (HIV-1) can be characterized by the host chemokine coreceptor that it uses to enter CD4-expressing cells. HIV-1 variants usually bind to the CCR5 chemokine coreceptor early in the course of disease. These are “R5” viruses (3, 31, 48). Viruses that use another chemokine coreceptor, CXCR4, are “X4” viruses, and they emerge later in HIV infection. They can account for up to 40 to 50% of all viruses in heavily treated patients with advanced disease (1, 32). The presence of X4 viruses has been associated with accelerated disease progression and a precipitous loss of CD4 T cells (27, 29, 40). A recent Swiss study suggested that the presence of X4 strains and the X4-specific virus load strongly predict clinical disease progression during combined antiretroviral therapy (cART), in addition to the CD4 T-cell count or viral load (44). This potential correlation between virus tropism and disease progression has important clinical implications. The development of coreceptor CCR5 antagonists for treating retroviruses and the lack of a virological response by patients infected with X4 or dual/mixed (X4/DM) viruses have increased the need to determine HIV-1 tropism.Recent studies have found the frequency of X4/DM dual-tropic strains in plasma samples from recently infected patients in the United States and Spain to be from 3.2% to 17.5% (14, 15, 16). Similarly, we found 15.9% (95% confidence interval [CI], 12.3% to 19.5%) strains of X4/DM viruses in 390 HIV-1 subtype B-infected patients diagnosed at the time of primary HIV-1 infection (PHI) in France from 1996 to 2007 (18).One of the major challenges of determining tropism is to select the best method for identifying coreceptor usage. HIV coreceptor usage is most commonly determined with a recombinant phenotype assay in clinical studies (28, 45). Bioinformatic tools based on the virus genotype may also be able to predict coreceptor usage. They are faster, less expensive, and more suitable for studies of a large number of patients than are phenotypic recombinant assays. Each available genotypic test is adequately specific but not very sensitive for detecting X4/DM or X4 variants. An overall concordance of 71.2 to 92% between genotypic and phenotypic assays has been reported (8, 15, 37, 41). However, most of these studies included HIV-1 subtype B strains. Genotypic algorithms may not be suitable for predicting the tropism of non-subtype-B HIV-1 strains (20). Two recent studies demonstrated that genotypic tests performed well for predicting the coreceptor usage of CRF02_AG and subtype C strains (36, 38), but no study has examined the correlation between genotypic and phenotypic tests for predicting the tropism of non-subtype-B HIV-1 at the time of PHI. The French PRIMO Cohort contained a large proportion of patients infected with a non-subtype-B virus (25.5% in 2005 to 2006) (6).We have therefore estimated the frequency of X4/DM viruses in 131 patients infected with non-subtype-B viruses at the time of PHI. All of them were enrolled in the French PRIMO Cohort from 1996 to 2007. We also studied the concordance between genotypic and phenotypic assays for predicting the tropism of non-subtype-B viruses in these patients.     

Serum Cytokine Profiles in Acute Primary HIV-1 Infection and in Infectious Mononucleosis

Serum cytokine profiles, T-cell subsets, and general parameters of immune activation were evaluated in 15 patients with acute primary HIV-1 infection, and compared with those obtained from 18 patients with acute primary Epstein-Barr virus (EBV) infection and from 18 control subjects in order to elucidate possible defects of immune response to HIV in early phases of virus-host interaction. Mean CD4⁺cell count, serum concentrations of interleukin (IL)-2, IL-4, soluble IL-2 receptor (sIL-2R), tumor necrosis factor (TNF)-α,5′-neopterin, and β₂-microglobulin were significantly lower in acute HIV-1 infection than in EBV infection. Both acute HIV-1 and EBV infections were characterized by significantly higher mean CD8⁺cell count and soluble CD8 antigen (sCD8) levels compared to control subjects, while acute HIV-1 infection was accompanied by the highest interferon (IFN)-γ serum concentrations. In primary HIV-1 infection, significant impairment of CD4⁺-mediated T-helper function may lead to viral escape and persistence of infection despite an early and vigorous CD8⁺T-lymphocyte activation.     

Broad neutralizing antibody response and genetic variation in HIV-1 env genes in Koreans with primary HIV-1 infections

To determine the neutralization profiles induced by HIV-1 Korean clade B, which has a monophyletic lineage and relative limited genetic diversity, we investigated the ability of HIV variants to elicit neutralizing antibodies in the immune response to primary infection. We selected seven Korean drug-na?ve subjects with an HIV-1 primary infection and did pseudovirion-based neutralization assays using env genes of Korean HIV origin. The neutralizing antibody responses to the Korean clade B showed broad reactivity to subtype B but a highly subtype-specific pattern. The lengths of the amino acid sequences and the PNGS numbers in the V1-V5 region were positively correlated with neutralization. These results imply that the genetic characteristics of HIV-1 env may affect neutralizing antibody responses in HIV-1-infected individuals. This is the first report describing the relationship between neutralizing antibody responses and HIV-1 genetic characteristics in Korean subjects. It can be useful for developing AIDS vaccines against HIV-1 subtype B strains.     

HIV-Ⅰ慢病毒载体的研究进展与应用

基因治疗是治疗肿瘤、遗传病等难治性疾病的最有效手段之一,但目前的基因转移方法存在一定的局限性,成为基因治疗和广泛应用的障碍.以1型人类免疫缺陷病毒(HIV-1)为基础构建的慢病毒载体(LV)具有感染分裂及非分裂细胞、使目的 基因产物表达稳定、表达时间长、载体自身免疫原性小等优点,尤其是LV能有效诱导免疫应答,抑制肿瘤生长,诱导移植免疫耐受等,是很有发展潜力的体内基因治疗新载体.     

Primary CXCR4 Co-receptor Use in Acute HIV Infection Leads to Rapid Disease Progression in the AE Subtype

Abstract This is a comparative study of HIV co-receptor usage in the early stages of HIV infection between two distinct patient groups, one with a low CD4 count (group 1), and the other with a high CD4 count (group 2). Group 1 progressed to a CD4 count below 200 cells/μL within 2 y, while group 2 had a CD4 count above 500 cells/μL within 2 y. Viral RNA was extracted from the plasma of these patients, and the C2-V5 region of the HIV-1 env genes were cloned and sequenced. The co-receptor usage was predicated based on V3 loop amino acid sequences using Geno2pheno and PSSM programs. Our results indicate that in acute HIV infection of rapid progressors (low CD4 count; group 1), the primary co-receptor usage is CXCR4, while in the high CD4 count group (group 2), the co-receptor usage is predominantly CCR5. One-year follow-up data from these patients showed no obvious change in HIV co-receptor usage in either group. Sequence analysis of patients from both study groups showed prevalence of the AE subtype, and therefore we can speculate that the CXCR4 co-receptor may be the primary HIV-1 co-receptor used in the HIV-1 AE subtype, and may be responsible for rapid HIV-1 disease progression in the MSM cohort.     

Validation of the Gen-Probe Aptima Qualitative HIV-1 RNA Assay for Diagnosis of Human Immunodeficiency Virus Infection in Infants

The qualitative Roche HIV-1 DNA Amplicor assay has been used for the past 20 years to diagnose HIV infection in infants and young children but is being phased out; hence, alternative assays must be found. The Gen-Probe Aptima qualitative HIV-1 RNA assay is currently the only FDA-cleared HIV-1 nucleic acid assay approved for diagnosis, but data on the use of this assay with infant plasma are limited. We assessed Aptima''s performance using control material for reproducibility and limit of detection and 394 plasma samples (0.2 to 0.5 ml) from HIV-exposed infected and uninfected infants and children for analytical sensitivity and specificity. Assays to assess within-run repeatability and between-run reproducibility indicated that the controls with 10,000 (5 of 5), 200 (5 of 5), 100 (16 of 16), 50 (12 of 12), and 25 (20 of 20) HIV-1 RNA copies/ml (cp/ml) were always positive, and negatives were always negative (20 of 20). The limit of detection was 14 cp/ml, as determined by probit analysis. The analytic sensitivity of the assay was 99.5% (189/190 samples; 95% confidence interval [CI], 97.1 to 99.9%) and specificity was 99.5% (199/200 samples; 95% CI, 97.2 to 99.9%). These results suggest that the assay is suitable for early infant diagnosis of HIV-1.     

Impact of Proficiency Testing Program for Laboratories Conducting Early Diagnosis of HIV-1 Infection in Infants in Low- to Middle-Income Countries

A voluntary, cost-free external quality assessment (EQA) program established by the U.S. Centers for Disease Control and Prevention (CDC) was implemented to primarily monitor the performance of laboratories conducting HIV Early Infant Diagnosis (EID) from dried blood spots (DBS) in low- to middle-income countries since 2006. Ten blind DBS proficiency test (PT) specimens and 100 known HIV-positive and -negative DBS specimens (to be used as internal controls) were shipped triannually to participating laboratories with reports for the PT specimens due within 30 days. The participant''s results and a summary of the performance of all participating laboratories and each diagnostic method were provided after each test cycle. Enrollment in the CDC PT program expanded progressively from 17 laboratories from 11 countries in 2006 to include 136 laboratories from 41 countries at the end of 2012. Despite external pressures to test and treat more children while expanding EID programs, mean PT test scores significantly improved over time as demonstrated by the upward trend from mid-2006 to the end of 2012 (P = 0.001) and the increase in the percentage of laboratories scoring 100% (P = 0.003). The mean test scores plateaued over the past 10 testing cycles, ranging between 98.2% and 99.7%, and discordant test results still occur but at a rate of no higher than 2.6%. Analysis of these test results suggests a positive impact of proficiency testing on the testing performance of the participating laboratories, and a continuous training program and proficiency testing participation may translate into laboratories improving their testing accuracy.     

Involvement of Envelope-Glycoprotein Glycans in HIV-1 Biology and Infection

Infection of host cells with HIV-1 depends on a highly glycosylated virus envelope glycoprotein (Env) and host-cell receptors. Glycans participate substantially in Env folding and in the binding of virions to the host-cell surface and indirectly affect cellular uptake of HIV-1. Moreover, Env glycans could protect HIV-1 from host’s neutralizing antibodies, but some glycans, on the other hand, represent targets for neutralizing antibodies. Variability of Env and its glycans in the HIV-1 strains from around the world as well as in patients during disease progression contributes substantially to further HIV-1 spreading in spite of the progress in basic HIV-1 research, vaccine development, and highly active antiretroviral therapy of HIV-1 infections.     

Infection with retroviruses other than HIV-1 in Spain: a retrospective analysis for HIV-2, HTLV-I,and/or HTLV-II

HIV-2, human T-cell lymphotropic virus (HTLV)-I, and HTLV-II infections are currently circulating in Spain with no evidence of an increase in the number of reported cases over time. Up to June 2002, a total of 106, 53, and 460 cases of HIV-2, HTLV-I, and HTLV-II infection, respectively, have been identified in Spain. Most HIV-2-infected and HTLV-I-infected individuals are immigrants who come from endemic areas or are Spaniards with a past history of travel to or sexual contacts with persons originating in those areas. In contrast, HTLV-II infection is mainly limited to native intravenous drug users who are frequently coinfected with HIV-1.     

Infection with Retroviruses Other Than HIV-1 in Spain: A Retrospective Analysis for HIV-2, HTLV-I,and/or HTLV-II

Abstract

HIV-2, human T-cell lymphotropic virus (HTLV)-I, and HTLV-II infections are currently circulating in Spain with no evidence of an increase in the number of reported cases over time. Up to June 2002, a total of 106, 53, and 460 cases of HIV-2, HTLV-I, and HTLV-II infection, respectively, have been identified in Spain. Most HIV-2-infected and HTLV-I-infected individuals are immigrants who come from endemic areas or are Spaniards with a past history of travel to or sexual contacts with persons originating in those areas. In contrast, HTLV-II infection is mainly limited to native intravenous drug users who are frequently coinfected with HIV-1.     

阴性和阳性症状为主型精神分裂症患者脑部质子波谱与症状相关性

目的探讨阴性和阳性症状为主型的精神分裂症患者脑部磁共振质子波谱(1H-MRS)与临床症状之间的相关性。方法对住院诊治的89例精神分裂症患者采用阳性与阴性症状量表(PANSS)评定症状,用1H-MRS测定患者前额叶及丘脑生化代谢物N-乙酰基天门冬氨酸(NAA)、肌酸复合物(Cr),计算NAA/Cr值。结果阴性组阴性症状(29.03±7.13)分、阳性症状(20.43±4.46)分、一般病理(40.21±5.43)分、总分(90.24±8.87)分;阳性组阴性症状(20.12±3.64)分、阳性症状(30.31±3.55)分、一般病理(40.76±5.54)分、总分(89.98±9.23)分,两组患者阴性症状及阳性症状分值之间比较存在显著性差异(P0.05);NAA/Cr值阴性组前额叶左侧(1.34±0.35)、右侧(1.38±0.37),丘脑左侧(1.45±0.36)、右侧(1.46±0.37);阳性组前额叶左侧(1.37±0.36)、右侧(1.40±0.37),丘脑左侧(1.49±0.36)、右侧(1.64±0.39),丘脑右侧NAA/Cr值在两组间比较存在显著性差异(P0.05);阴性组患者左侧前额叶NAA/Cr值与PANSS总分、阴性症状总分之间均具有负相关(r=-0.34,-0.36;P0.05),阳性组患者左侧前额叶NAA/Cr值与PANSS总分之间具有负相关(r=-0.35,P0.05)。结论精神分裂症阳性与阴性为主型的患者前额叶及丘脑均存在NAA/Cr值变化,前额叶NAA/Cr值与阴性症状的严重程度密切相关。     

Detection and enumeration of circulating HIV-1-specific memory B cells in HIV-1-infected patients

Memory B cells are long-living cells that circulate throughout the body and differentiate into plasma cells after stimulation by antigens, cytokines, and direct cell-to-cell interaction in lymphoid tissues. For HIV-1-infected patients, we assessed whether in vitro polyclonal B cell activation that induces immunoglobulin secreting cells (SCs) also generates HIV-1-specific resting B cells to synthesize antibodies specific to HIV-1. To this end, highly purified B cells from 10 HIV-1-untreated patients were cultured with or without mouse fibroblastic cells expressing the CD40 ligand in the presence of IL-2 and IL-10. The percentage of immunoglobulin SCs we obtained by using the B cell-CD40L stimulation system was equal to 55% to 98% of the circulating memory B cells. Moreover, the anti-HIV-1 IgG, IgA, or IgM antibody SCs represented 1 x 10-2 to 1 x 10-3 of the total immunoglobulin SCs. The anti-HIV-1-specific antibodies detected in cell culture supernatants were directed to gag-, pol-, and env-encoded viral proteins. We found that in AIDS patients, HIV-1-specific resting memory B cells circulate in the blood and can be quantified by their anti-HIV-1 antibody secretion after strong B cell polyclonal stimulation.