Optimización de recursos para determinar la carga viral de HIV-1 en un país con pocos recursos / Optimizing resources to reduce costs to determine HIV viral load in limited resources settings

Resumen Introducción. Las metas globales para controlar la epidemia de HIV contemplan que la carga viral sea indetectable en 90 % de las personas en tratamiento. El costo de la medición de la carga viral en lotes de muestras puede reducirse y, así, aumentar la cobertura cuando los recursos son limitados; sin embargo, su eficacia disminuye al aumentar la prevalencia del fracaso del tratamiento antirretroviral. Objetivo. Evaluar estrategias para disminuir la proporción de pacientes con fracaso del tratamiento antirretroviral en los lotes de muestras y, de esta manera, aumentar el ahorro en las pruebas de carga viral. Materiales y métodos. Las estrategias evaluadas fueron: a) la organización de los lotes de muestras según el esquema de tratamiento antirretroviral, y b) la exclusión de aquellos pacientes con antecedente reciente de fracaso del tratamiento antirretroviral, aquellos con menos de 12 meses de tratamiento antirretroviral y aquellos sin tratamiento antirretroviral previo. Los resultados de los lotes se compararon con los resultados individuales. Resultados. El valor diagnóstico negativo fue similar para los pacientes con esquema de primera línea (100,0 %; IC95% 99,5-100,0) o de segunda línea de tratamiento (99,4 %; IC95% 96,9-99,9). La incidencia del fracaso del tratamiento antirretroviral fue menor en los pacientes con tratamiento de primera línea (p<0,01), lo cual permitió un mayor ahorro en las pruebas de laboratorio en este grupo (74,0 %; IC95% 71,0-76,7) que en los pacientes con tratamiento de segunda línea (50,9 %; IC95% 44,4-57,3) (p<0,01). Conclusión. La selección de las muestras que se incluyeron en los lotes para determinar la carga viral del HIV según el tipo de esquema de tratamiento, permitió maximizar el porcentaje de ahorro en pruebas de laboratorio.

Abstract Introduction: HIV viral load testing is a key factor to evaluate the accomplishment of the UNAIDS target of 90% of viral suppression among people receiving antiretroviral therapy. Pooled samples are a potentially accurate and economic approach in resource-constrained settings, but efficiency can be negatively affected by high prevalence rates of virological failure. Objective: Strategies were assessed to increase the relative efficiency of pooled HIV viral load testing in resource-constrained settings. Materials and methods: We evaluated two strategies: a) plasma samples were not included in pools if patients had <12 months on antiretroviral therapy, patients had previous viral load >1,000 copies/ml, or were antiretroviral therapy naïve patients, and b) plasma pools were organized separately for first and second-line antiretroviral therapy regimens. Individual viral load tests were used to compare pooled results. Results: Negative predictive values were similar for patients on first (100.0%; 95% CI 99.5 to 100.0) and second-line antiretroviral therapy regimens (99.4%; 95% CI 96.9 to 99.9). However, the incidence of virological failure among individuals on first-line antiretroviral therapy was lower than second-line antiretroviral therapypatients (p <0.01), resulting in greater savings in laboratory tests in patients on first-line antiretroviral therapy (74.0%; 95% CI 71.0 to 76.7) compared with the group of patients on second-line antiretroviral therapy (50.9%; 95% CI 44.4 to 57.3) (p<0.01). Conclusion: Selecting the samples to be included in the pools and selecting the pools according to ART regimens are criteria that could lead to decreased spending on laboratory tests for HIV viral load determination in resource-constrained settings.

Evaluation of HIV, STI and CD4 results among voluntary attendees at the HIV/AIDS program of Mexico City / Evaluación de resultados de VIH, ETS y CD4 en asistentes al programa de VIH/SIDA de la Ciudad de México

Abstract: Objective: To describe results of HIV, sexually transmitted diseases (STI) and CD4 counts at the HIV-specialized Condesa Clinic (CC) in Mexico City. Materials and methods: Individuals who requested voluntary counseling and testing at CC were studied. We identified antibodies against HIV, syphilis, hepatitis C, and hepatitis B HBsAg. CD4 cell counts and viral load of HIV positive individuals were also obtained. Late HIV infection diagnosis was established if CD4 counts were lower than 200 cells/μL. Results: Global seroprevalence of HIV, syphilis, HBsAg, and anti HCV markers was of 20.1, 6, 1 and 1, respectively. Men displayed higher seroprevalence of infection markers than women. Among men, HIV infection was related to age and with all STI markers. Late HIV diagnosis was 31.8%. The risk of late HIV diagnosis was higher among women and it increased as age increased. Conclusions: Differences between genders regarding HIV and STIs prevalence as well as risk factors for HIV infection and late HIV diagnosis were observed.

Resumen: Objetivo: Describir resultados del programa VIH/SIDA de la Clínica Especializada Condesa (CC). Material y métodos: Se identificaron anticuerpos contra VIH, sífilis y hepatitis C, así como HBsAg del virus de la hepatitis B. Se hizo un conteo de CD4 y carga viral en los positivos a VIH asistentes a la CC. El conteo CD4 menor a 200 células/μL definió el diagnóstico tardío de la infección por VIH. Resultados: La prevalencia de VIH, sífilis, HBsAg y virus de la hepatitis (HCV) fue de 20.1, 6, 1 y 1, respectivamente. Los hombres mostraron prevalencias mayores de infección que las mujeres y en ellos la infección por VIH estuvo relacionada con la edad y con los marcadores de ITS. El diagnóstico tardío de VIH fue de 31.8% y su riesgo fue mayor en las mujeres y se incrementó conforme la edad. Conclusión: Se encontraron diferencias de género en las prevalencias de VIH e ITS, en los riesgos de infección por VIH y en el diagnóstico tardío de esta infección.

Analysis of Immunoglobulin G subclasses in Hepatitis C virus patients: Correlation with disease activity and viraemia

Hepatitis C virus antibodies are found in the serum of most patients with chronic hepatitis C. However, the significance of the humoral response is still uncertain. There is no reliable and simple diagnostic marker available to diagnose recent hepatitis C virus [HCV] infection. It has been shown that the avidity of specific IgG antibody is low in primary viral infection and increases with time. In this study, in vitro IgG anti-hepatitis C virus secretion by peripheral blood mononuclear cells of patients with hepatitis C was analyzed. To assess the effect of elicited HCV antigen-specific IgG antibody subclasses on the clinical outcome of HCV infection. The study protocol was approved in Damnhour National Medical Institute, and patients provided written informed consent. Forty-five [45] hepatitis C patients [5ml blood for each pt.] were divided into 3 groups: the first HCV Patients with variable degrees of viraemia [low-moderate-high], second: HCV patients with end stage complications [cirrhosis and hepatocellular carcinoma] third: HCV sero-positive individuals without viraemia or complications; 15 patients [5ml blood for each pt.] were randomized into the normal group [HCV sero-negative could subjects]. Hepatitis C patients with [in vitro] IgG anti-hepatitis C virus secretion had higher AFP and IgG levels in serum than did patients without such secretion in vitro [p<0.002]. Furthermore, there is correlation between IgG antibody secretion levels and activity and degree of HCV patients [p<0.001]. The mean-counts of IgG plasma cells in 3 studied groups hepitits C patient were significantly correlated with activity of disease. Whereas IgG plasma cells resulted more correlated with normal group. IgG plasma cells on hepatitis C can be a valuable parameter for better diagnosis of degree of hepatitis C disease and also for follow up the patients

HLA-Bw4-B*57 and Cw*18 alleles are associated with plasma viral load modulation in HIV-1 infected individuals in Salvador, Brazil

Host genetic factors play an important role in mediating resistance to HIV-1 infection and may modify the course of infection. HLA-B alleles (Bw4 epitope; B*27 and B*57) as well as killer cell immunoglobulin-like receptors have been associated with slow progression of HIV-1 infection. OBJECTIVE: To evaluate the association between serological epitopes HLA-Bw4 and HLA-Bw6 and prognostic markers in AIDS. METHODS: 147 HIV-infected individuals in Bahia, Northeast Brazil, were genotyped for HLA class I locus. HLA class I genotyping was performed by hybridization with sequence-specific oligonucleotide probes following amplification of the corresponding HLA-A, HLA-B and HLA-C genes. Statistical analysis was performed using Fisher's exact and ANOVA tests for categorical and continuous variables, respectively. RESULTS: We detected a significant association (χ2 = 4.856; p = 0.018) between the presence of HLA-Bw4 and low levels of viremia. Eighteen out of the 147 HIV-infected individuals presented viremia <1,800 copies/mL and 129 presented viremia > 2,000 copies/mL. Ninety and four percent (17/18) of all individuals with viremia < 1,800 copies/mL carried HLA-Bw4, compared to 67.4 percent (87/129) of individuals with viremia > 2,000 copies/mL. Additionally, we found a significantly higher frequency of B*57 (OR = 13.94; 95 percent CI = 4.19-46.38; p < 0.0001) and Cw*18 (OR = 16.15; 95 percent CI = 3.46-75.43; p < 0.0001) alleles, favoring the group with lower viremia levels, in comparison with those with higher viral load. CONCLUSION: HLA-Bw4-B*57 and Cw*18 alleles are associated with lower level of viral load in HIV-infected Brazilian patients. These findings may help us in understanding the determinants of HIV evolution in Brazilian patients, as well as in providing important information on immune response correlates of protection for such population.

Defects in blood dendritic cell subsets in HIV-1 subtype C infected Indians.

Background & objective: DCs trigger both innate and adaptive immune responses to control HIV infection and represent a viral reservoir acting as target and HIV carriers for infection of permissive CD4+ T-cells. DCs thus form a very attractive study subject to further our existing knowledge of HIV induced immunopathogenesis due to its diverse and crucial role in HIV infection establishment, viral dissemination, immune evasion, viral persistence, etc. We aimed to characterize the effect of HIV infection on myeloid and plasmacytoid dendritic cell subsets in a group of HIV-1 subtype C infected treated or untreated Indian individuals. Methods: Blood DC subset numbers and immunophenotype were studied for 79 HIV infected subjects at various stages of disease and compared with 13 HIV-uninfected controls. Comparisons were also made between groups of subjects based on their CD4+ T cell counts and also experience of antiretrovirals. Results: Significant decreases were observed in blood DC counts and the two DC subsets in HIV infected individuals. Subjects with lowest CD4+ T cell counts also had a drastically reduced DC subset pool which correlated positively with plasma viraemia and negatively with CD4+ T cell counts. DC subsets from HIV infected subjects showed higher expression of co-stimulatory molecules CD40 and CD86, and HIV-1 co-receptors CXCR4 and CCR5 which correlated positively with HIV-1 plasma viraemia. The alterations in blood DCs were partly resolved in ART receiving study subjects. Interpretation & conclusions: Correlation between DC subset activation state and viraemia supports the role of DC activation on viral replication and CD4+ T cell depletion.

Impact of cytomegalovirus and grafts versus host disease on the dynamics of CD57+CD28-CD8+ T cells after bone marrow transplant

OBJECTIVES: The present study aimed to evaluate the dynamics of CD28 and CD57 expression in CD8+ T lymphocytes during cytomegalovirus viremia in bone marrow transplant recipients. METHODS: In a prospective study, blood samples were obtained once weekly once from 33 healthy volunteers and weekly from 33 patients. To evaluate the expression of CD57 and CD28 on CD8+ T lymphocytes, flow cytometry analysis was performed on blood samples for four months after bone marrow transplant, together with cytomegalovirus antigenemia assays. RESULTS: Compared to cytomegalovirus-seronegative healthy subjects, seropositive healthy subjects demonstrated a higher percentage of CD57+ and a lower percentage of CD28+ cells (p<0.05). A linear regression model demonstrated a continuous decrease in CD28+ expression and a continuous increase in CD57+ expression after bone marrow transplant. The occurrence of cytomegalovirus antigenemia was associated with a steep drop in the percentage of CD28+ cells (5.94 percent, p<0.01) and an increase in CD57+ lymphocytes (5.60 percent, p<0.01). This cytomegalovirus-dependent effect was for the most part concentrated in the allogeneic bone marrow transplant patients. The development of acute graft versus host disease, which occurred at an earlier time than antigenemia (day 26 vs. day 56 post- bone marrow transplant), also had an impact on the CD57+ subset, triggering an increase of 4.9 percent in CD57+ lymphocytes (p<0.05). CONCLUSION: We found continuous relative changes in the CD28+ and CD57+ subsets during the first 120 days post- bone marrow transplant, as part of immune system reconstitution and maturation. A clear correlation was observed between the expansion of the CD57+CD28-CD8+ T lymphocyte subpopulation and the occurrence of graft versus host disease and cytomegalovirus viremia.

Desarrollo del virus de la encefalitis equina venezolana en el tejido nervioso de ratones recién nacidos: ultraestructura e histoquímica / Ultrastructural and histochemical: study on the development of venezuelan equine encephalitis virus in the central nervous system of newborn mice

Se estudio la estructura del tejido nervioso de ratones recién nacidos inoculados con el virus de la EEV. Las células gliales mostraron hichamiento y aumento del glucógeno; las neuronas presentaron alteraciones en la estructura de las mitocondrias y en la organización de los ribosomas; el complejo de Golgi presentó gran desarrollo y tortuosidad de sus cisternas. La actividad de fosfatasa ácida en la cisterna perinuclear y en el retículoendoplasmático granular, sugiere un aumento de producción de esta enzima en las neuronas invadidas por el virus de la EEV. Se estudió la localización de las partículas virales en relación con la viremia y diseminación de la infección, destacándose la presencia de partículas virales en la luz de los vasos sanguíneos y alrededor de las vainas de mielina de algunos axones. El desarrollo de las partículas virales se comparó con las observaciones hechas en otras arbovirosis, proponiéndose que los precursores virales se originen en masas de viroplasma, desde donde migran a través del citoplasma, hacia las cisternas y vacuolas del complejo de Golgi o hacia la membrana plasmática, transformándose en partículas virales maduras por gemación