Current HIV Research (v.9, #5)

Differential Innate Immune Responses to Low or High Dose Oral SIV Challenge in Rhesus Macaques by Andre Durudas, Hui-Ling Chen, Melanie A. Gasper, Vasudha Sundaravaradan, Jeffrey M. Milush, Guido Silvestri, Welkin Johnson, Luis D. Giavedoni, Donald L. Sodora (276-288).
Mucosal transmission of HIV predominately occurs during sexual intercourse or breast-feeding and generallyresults in a successful infection from just one or few founder virions. Here we assessed the impact of viral inoculum sizeon both viral and immune events within two groups of Rhesus macaques that were non-traumatically, orally inoculatedwith either multiple low (1000 to 4000 TCID50) or high (100,000 TCID50) doses of SIV. In agreement with previousstudies, more diverse SIV variants were observed in macaques following infection with high dose oral SIV compared to alow dose challenge. In peripheral blood cells, the immune gene transcript levels of CXCL9, IFN.., TNF.. and IL10remained similar to uninfected macaques. In contrast, OAS and CXCL10 were upregulated following SIV infection inboth the high and low dosed macaques, with a more rapid kinetics (detectable by 7 days) following the high SIV dosechallenge. In peripheral lymph nodes, an increase in CXCL10 was observed irrespective of viral dose while CXCL9 andOAS were differentially regulated in the two SIV dosed groups. Magnetic bead sorting of CD3+, CD14+ and CD3-/CD14- cells from peripheral blood identified the increase in OAS expression primarily within CD14+ monocytes,whereas the CXCL10 expression was primarily in CD3+ T cells. These findings provide insights into the impact of SIVchallenge dose on viral and innate immune factors, which has the potential to inform future SIV/HIV vaccine efficacytrials in which vaccinated hosts have the potential to be infected with a range of viral challenge doses.

HIV Infection-Related Premature Immunosenescence: High Rates of Immune Exhaustion After Short Time of Infection by Sara Ferrando-Martinez, Ezequiel Ruiz-Mateos, Maria Concepcion Romero-Sanchez, Mª Angeles Munoz-Fernandez, Pompeyo Viciana, Miguel Genebat, Manuel Leal (289-294).
Premature immunosenescence has been reported in different HIV scenarios. However, how premature is theHIV-related immunosenescent phenotype is still unknown. Thus, the aim of this study was to analyze theimmunosenescent status of young viraemic naive HIV-infected individuals, with less than four years from infection. Tothis end, replicative senescence, activation and proliferation T-cell levels were analyzed in chronically HIV-infectedyoung individuals and both, elderly and young healthy controls. We show that young HIV-infected viraemic patients, withless than four years from infection, have early immune exhaustion leading to a premature immunosenescence comparableto healthy people 40 years elder. In addition, memory T-cell subsets showed greater alterations than elder healthy controlsand, in patients with high viral loads, CD57 expression at the memory T-cell subsets was correlated with lower viralincreases but higher CD4 T-cell lost during follow up.

The Vpr Gene Polymorphism of Human Immunodeficiency Virus Type 1 in China and its Clinical Significance by Xia Chen, Yuhuang Zheng, Hui Li, Diallo Mamadou, Chunying Zhang, Yan He, Huaying Zhou, Zi Chen, Meng Liu (295-299).
Recent studies have explored that mutated Human immunodeficiency virus type 1(HIV-1) Vpr genes likelyinfluence clinical manifestations of HIV infected patients. However, the relationship between the mutation sites on HIVVpr gene and subsequent function changes is still not clear. In this study we investigated such relationship in analyzingthe Vpr genes of HIV-1 viruses isolated from 208 HIV-1 infected patients from different regions in China. Reversetranscription polymerase chain reaction (RT-PCR) and nested PCR were used to amplify HIV-1 Vpr gene extracted fromplasma of 208 HIV-1 infected patients and 153 isolates displayed the target gene sequences. Biological analysis softwareanalyzed the deduced amino acid sequence, and identified the characteristics of the polymorphism of HIV-1 Vpr gene andits clinical significance. Results show the sequence subtypes as follows: CRF01-AE is 51.63%, subtype C is 24.84%,subtype B is 17.65%, CRF03-AB is 3.92% and CRF08-BC is 1.31%. This paper revealed for the first time the HIV-1 Vprgene polymorphism in HIV-1 positive individuals in China.: the subtype CRF01-AE is the main Vpr gene subtype in thisregion. The mutations in the C-terminal were more obvious than those observed in the N-terminal. It was also discoveredthat in the 77th position, 84.3% of the 153 amino acid sequences were glutamine (Q), which differ from overseas reports.Our data showed that the mutations 63, 70, 85, 86, 89 and 94 of the Vpr gene were possibly correlated with the clinicalmanifestations of the HIV-1 infected individuals.

The relationship between chemical structure and CCR5 anti HIV-1 activity was investigated in the series of1-[N-(Methyl)-N-(phenylsulfonyl)amino]-2-(phenyl)-4-[4-(substituted) piperidin-1-yl]butanes derivatives including 114molecules by using the Electron-Topological Method (ETM). In the frameworks of this approach, its input data weretaken as the results of conformational and quantum-mechanical calculations. Conformational analysis and quantumchemicalcalculations were carried out for each molecule. Then molecular fragments being specific for active moleculesand non-active molecules were revealed by using ETM. The result of testing showed the high ability of ETM in predictingthe activity and inactivity investigated series. In order to classify and to develop a model for those molecules, cluster anddiscriminant analyses are conducted. First, cluster analysis is implemented in order to classify similar molecules into thegroups. Then, discriminant analysis is used to construct models including descriptors. By doing so, two obtaineddiscriminant functions segregate those molecules into three different groups by using the descriptors called EHOMO, DipoleMoment and SEZPE.

HIV Shedding in Cervico-Vaginal Secretions in Pregnant Women by Barbara Gardella, Marianna Roccio, Anna Maccabruni, Bianca Mariani, Lucia Panzeri, Francesca Zara, Arsenio Spinillo (313-320).
Objective: The purpose of this study was to evaluate the presence of HIV-1 in cervico-vaginal secretions ofpregnant as compared to non-pregnant HIV-seropositive women.Patients and Methods: We compared 43 known HIV seropositive pregnant patients versus 241 age-matched (± 2 years)control non-pregnant HIV-seropositive subjects. In pregnant patients blood and cervico-vaginal samples were obtainedduring each trimester of pregnancy. In control subjects the same samples were obtained at enrolment. HIV-1 RNA wasmeasured in plasma; proviral HIV-1 DNA, cell-associated and cell-free HIV-1 RNA in cervico-vaginal secretion bycompetitive polymerase chain reaction (cRT-PCR) and reverse transcriptase PCR.Results: The genital shedding of HIV-DNA (22/43 as compared to 79/241, p = 0.02), and cell-free HIV-RNA detection(26/43 as compared to 72/241, p <.001) was more common in first-trimester pregnant than in non pregnant women.Pregnancy correlated with a significant positive trend in the cervico-vaginal load of HIV-DNA (Spearman Rho= 0.149, p=0.012), and cell-free HIV-RNA (Spearman Rho= 0.253, p<.001), but not of HIV-RNA transcripts (Spearman Rho = 0.06,p= 0.31). After correction for potential confounders, first trimester pregnant women had increased rates of genital HIVDNA(odds ratio = 1.94, 95% confidence interval = 1.01 - 3.78) and cell-free HIV-RNA (odds ratio = 4.07, 95%confidence interval = 1.97 - 8.41) detection compared to nonpregnant controls.Conclusion: The shedding of genital HIV was increased in pregnant compared to non pregnant subjects, even in patientswith undetectable viremia. In this low-risk HIV-positive population the risks of vertical or horizontal transmissions shouldnot be underestimated.

Lipodystrophy, Insulin Resistance, and Adiponectin Concentration in HIV-Infected Children and Adolescents by Alessandra Vigano, Gian Vincenzo Zuccotti, Chiara Cerini, Sara Stucchi, Maria Puzzovio, Vania Giacomet, Stefano Mora (321-326).
Alterations of fat distribution and insulin resistance are associated with increased risk of metabolicderangements and cardiovascular disease. HIV-infected adult patients on antiretroviral treatment often showlipodystrophy, insulin resistance and hypoadiponectinemia, but data in children are controversial. We investigated serumadiponectin concentration in a cohort of HIV-infected youths, and we assessed the relationships with lipodystrophy andinsulin resistance. We studied 36 HIV-infected patients (aged 5.0 - 19.4 years), and 171 healthy subjects (aged 4.9 - 17.9years) for adiponectin measurements. All patients underwent body composition assessment by dual-energy x-rayabsorptiometry, and an oral glucose tolerance test to determine the fasting insulin concentration, the insulin area under thecurve (AUC), and the HOMA index. Adiponectin serum concentration was measured by an immunoenzymatic assay.Sixteen patients had central fat accumulation, 6 had peripheral lipoatrophy, 5 had a mixed phenotype, and the remaining 9were non-lipodystrophic. Fasting insulin, insulin AUC, and HOMA index were significantly higher in patients withcentral fat adiposity and mixed phenotype than in the other two groups. The patients of the former two groups hadadiponectin concentration much lower than healthy controls, and patients with peripheral lipoatrophy or normal phenotypehad normal concentration. Low adiponectin concentration is associated to central fat and mixed lipohypertrophy, and tosigns of insulin resistance in HIV-infected youths. Strict monitoring of metabolic and cardiovascular evolution should beperformed in these patients.

Accuracy of Ultrasound Imaging Technique for Assessing Lipoatrophy in HIV-Infected Subjects by Giovanna Ferraioli, Carmine Tinelli, Luigia Scudeller, Gaetano Filice, Carlo Filice (327-333).
Objective: To compare the accuracy of ultrasound imaging technique to that of clinical diagnosis in evaluatingsubcutaneous fat changes in HIV-infected subjects.Methods: HIV-uninfected control subjects (Group A), HIV-infected subjects with clinically assessed lipoatrophy (GroupB), and HIV-infected subjects without clinical lipoatrophy (Group C) underwent ultrasound measurements ofsubcutaneous fat thickness at facial, brachial and thigh regions. ROC curve analyses were used to estimate ultrasoundprediction accuracy and cut-off values of subcutaneous fat thickness.Results: 228 subjects were enrolled: 78 in Group A, 73 in Group B, and 77 in Group C. Facial lipoatrophy: ROC curveanalysis identified optimal cut-off value of 13.3 mm [sensitivity, 96.0%; specificity, 76.9% AUC 0.92], 5.0 mm[sensitivity, 71.4%; specificity, 92.3%; AUC 0.90] and 11.2 mm [sensitivity, 95.8%; specificity, 89.7%; AUC 0.97] forfemales and 12.05 mm [sensitivity, 51.2%; specificity, 87.2%; AUC 0.74], 4.1 mm [sensitivity, 76.2%; specificity, 89.7%;AUC 0.85] and 4.35 mm [sensitivity, 60.0%; specificity, 89.7%; AUC 0.82] for males in assessing facial, brachial andcrural lipoatrophy respectively. Using this cut-off values, 12/25 (48%) females and 17/49 (34.7%) males, 12/28 (42.9%)females and 23/49 (46.9%) males, 19/28 (67.9%) females and 12/49 (24.5%) males in Group C would be classified assub-clinical facial, brachial and crural lipoatrophy respectively.Conclusions: The results of our study show that in the assessment of subtle subcutaneous fat changes ultrasound is moreaccurate than clinical evaluation and confirm the usefulness of ultrasound imaging technique in identifying lipoatrophy atan early stage.

Itraconazole vs Fluconazole as a Primary Prophylaxis for Fungal Infections in HIV-Infected Patients in Thailand by Romanee Chaiwarith, Apinya Fakthongyoo, Jutarat Praparattanapan, Darakorn Boonmee, Thira Sirisanthana, Khuanchai Supparatpinyo (334-338).
Background: Disseminated fungal infections are common presenting opportunistic infections among AIDSpatients in developing countries. Primary prophylaxis with itraconazole has been shown to be effective in northernThailand. This study aimed to compare the efficacy of fluconazole vs itraconazole as primary prophylaxis for fungalinfections in HIV-infected patients.Methods: A retrospective cohort study was conducted among HIV-infected patients who received primary prophylaxiswith fluconazole 400 mg once weekly or itraconazole 200 mg once daily at Chiang Mai University Hospital. Wecompared the incidence of systemic fungal infections and the probability of disease-free survival between groups.Results: From January 2000 to June 2010, 308 HIV-infected patients who received primary fungal prophylaxis wereenrolled; 148 were male (48.1%) and the mean age was 38.2 ± 8.0 years. 276 patients received fluconazole and 32received itraconazole. Baseline CD4+ cell count was 35 (IQR 15, 70) and 50 (IQR 21,75) cells/mm3 in fluconazole anditraconazole groups, respectively (p=0.159). The median follow-up time was 12 months (IQR 7, 19) in fluconazole groupand 15.5 months (IQR 9, 21.5) in itraconazole group. Seven patients (2.5%) who received fluconazole and 2 patients(6.3%) who received itraconazole developed systemic fungal infections, giving the incidence of 17.0 and 34.8/10000person-months, respectively (p=0.261). The probability of developing any systemic fungal infections or death did notdiffer between groups.Conclusions: Although P. marneffei has a reduced susceptibility in in vitro to fluconazole, our study has demonstrated thatonce-weekly fluconazole is at least as effective as once-daily itraconazole as primary prophylaxis for systemic fungalinfections in AIDS patients in northern Thailand.

Objectives: To evaluate gender differences in liver fibrosis and hepatitis C virus-related parameters in patientscoinfected with human immunodeficiency virus.Methods: Transversal study of 782 patients who underwent a complete clinical and laboratory evaluation. Fibrosis wasmeasured by transient elastometry (TE) and by commonly used laboratory-derived fibrosis indexes.Results: Men were older, had higher rates of alcohol abuse, higher HCV viral load and liver tests, lower platelet values,poorer CDC clinical stages, longer duration of HCV infection, shorter time on successful antiretroviral therapy (ART) andhad appreciably more advanced fibrosis than women. Multivariate analysis revealed that male gender (P<0.0001), longertime since HCV acquisition (P<0.0001), alcohol abuse (P<0.0001), HCV genotype 3 (P=0.01), shorter time on successfulART (P=0.005) and worse CDC clinical stages (P=0.03) were independently associated with significant or higher stagesof fibrosis. Male gender was also independently predictive of advanced or higher stages of fibrosis (P=0.06) or cirrhosis(P=0.02). In patients with no alcohol abuse, men had worse fibrosis parameters than women (P<0.01 for each), but thesedifferences decreased in patients with alcohol abuse and became non-significant.Conclusions: HIV-HCV-coinfected women have more favorable HCV virological and clinical profile than men and,particularly, lower degrees of fibrosis. Alcohol abuse seemed to result more deleterious in women than in men. Thereportedly poorer outcomes of liver disease in HIV-HCV-coinfected patients, as compared with their HCV-monoinfectedcounterparts, could be ameliorated by addressing these cofactors, some of them preventable or treatable.

Women's Desire for Children in an HIV Population: A Clinical Pilot Study in Burkina Faso by Lemoine Caroline, Willems Nathalie, Hien Alain Diedon, Karama Robert, Somda Idovide, Englert Yvon (346-351).
Introduction: Today, thanks to the HAART, HIV has become a chronic disease. In most cases, HIV positive women are ofreproductive age and at present, the vertical transmission rate is around 0.1% for women with an undetectable viral load. So, it is normalthat the question of seropositive women's desire to have children is on the table.Methods: In this experimental study, 50 HIV-seropositive and 44 seronegative women were interviewed about their desire to havechildren. Some of the questions asked were: How many children did you want to have before you got married/at 15 years of age if youaren't married? How many children would you like to have today, considering your present situation?. In case of a difference betweenbefore and now, we asked them: What are the reasons for this difference?. This study was performed in Burkina Faso.Results: The positive women tend to desire more children before and fewer children now than negative women (OR: 1.33; C.I. 95%:0.86-2; p= 0.19 vs OR: 0.78; C.I. 95%: 0.51-1.21; p= 0.27). 62% of HIV positive women mention multiple reasons directly linked totheir seropositivity to explain the difference between before and now. 70% of HIV positive women still want to have children. Wehave noted that the positive women who still want children are more likely to be younger (p<0.05 by Two-Sample T tests), in arelationship (p<0.01 by Chi-Square Test) and to have been diagnosed earlier than the positive women who don't want any children(p=0.01 by Wilcoxon Rank Sum Test).Conclusion: Even if the results of this pilot study are preliminary, they show that HIV positive and negative women have a relativelysimilar desire for children, even though seropositive women seem to want fewer children than their uninfected counterparts. Most of thereasons which reduce HIV positive women's desire to have children are directly linked to HIV. This is why getting these womeninformed about materno-fetal transmission risks and existing treatments is really important to give them the opportunity to make aconscious choice.