Human cytomegalovirus (HCMV)-specific CD4+ T lymphocyte response in AIDS patients with no past or current HCMV disease following HAART.

BACKGROUND: The incidence of Human Cytomegalovirus (HCMV) end-organ disease has dramatically decreased since the implementation of highly active antiretroviral therapies (HAARTs), but the precise immune mechanism whereby HCMV is controlled remains to be elucidated. OBJECTIVES: To investigate the effect of (HAART) on CD4+ T-cell immunity to HCMV in AIDS patients with no past or current HCMV disease. STUDY DESIGN: Seventeen patients were prospectively examined for CD4+ (CD45RO+ and CD45 RA+) T-cell counts (flow cytometry), HIV RNA load (Amplicor HIV test), HCMV leukoDNAemia and HCMV DNA in urine (nested PCR), lymphoproliferative response (LPR) to HCMV, phytohemagglutinin (PHA) and purified protein derived from Mycobacterium tuberculosis (PPD) by measurement of 5-bromo-2'-deoxyuridine incorporation to DNA (ELISA) and cytokine secretion (IFN-gamma, IL-4 and IL-10) by HCMV-stimulated peripheral blood mononuclear cell (PBMC) cultures (ELISA). RESULTS: Fifteen patients responded favorably to HAART (virologically, immunologically, or both). Of these, six patients presented LPR to HCMV at least once during follow-up, whereas most displayed detectable LPRs to PHA. IFN-gamma was detected at least once in supernatants of HCMV-stimulated PBMC cultures from 14 of the 17 patients. All but one patient tested negative for HCMV leukoDNAemia and HCMV DNA in urine, and none developed HCMV disease during the observation period. CONCLUSIONS: Control of HCMV replication and the absence of HCMV disease are not consistently associated with recovery and/or maintenance of LPR to HCMV in AIDS patients under HAART and with no prior HCMV disease. Whether detection of IFN-gamma by PBMCs upon HCMV antigenic stimulation may serve as a surrogate marker for protection against HCMV disease requires further investigation.

Application of a 5-bromo-2'-deoxyuridine ELISA for measuring the lymphoproliferative response to human cytomegalovirus in HIV-1-infected patients.

Assessment of the lymphoproliferative response to human cytomegalovirus (HCMV) may help to identify human immunodeficiency virus (HIV)-1-infected patients at high risk of developing HCMV end-organ disease. The tritiated thymidine ([3H]-TdR)-incorporation assay is the gold standard for measuring lymphoproliferative responses, though it is unsuitable as a routine laboratory procedure. An alternative non-radioactive technique, a 5-bromo-2'-deoxyuridine (BrdU) enzyme-linked immunosorbent assay, was applied for measuring T-cell proliferation in response to HCMV. Stimulation of either 1 x 10(5) or 5 x 10(4) peripheral blood mononuclear cells (PBMCs)/well with 10 PFU/well (before inactivation) of inactivated HCMV (AD169 strain) virions during 5 days, followed by an 18 h period of pulsing with BrdU (10 microM) proved to be the optimal laboratory conditions. The assay is simple, economical and feasible for monitoring the lymphoproliferative response to HCMV in HIV-1-infected patients.

[Clinico-radiological profile of acute disseminated encephalomyelitis in the childhood population. A retrospective analysis of a series of 20 patients in a tertiary hospital]. / Perfil clinicorradiológico de la encefalomielitis aguda diseminada en la población infantil. Análisis retrospectivo de una serie de 20 pacientes de un hospital terciario.

INTRODUCTION: Acute disseminated encephalomyelitis (ADE) is an inflammatory disorder of the central nervous system that is mediated immunologically and of unknown pathogenesis. It can present at any age, but is much more frequent in children. ADE has no specific biological marker and diagnosis is based on findings from clinical and neuroimaging studies. AIM: To enhance our knowledge of the clinico-radiological profile of this disease. PATIENTS AND METHODS: This retrospective study involved patients under 14 years of age who were admitted to a tertiary hospital over the last 15 years with a diagnosis of ADE. History, clinical presenting symptoms, lab findings from blood/cerebrospinal fluid analyses and radiological semiology were reviewed. In 16 cases an average follow-up of 25 months was performed. RESULTS: The study examined 20 patients, 70% children, with a mean age of 4.4 years. Forty per cent had a previous febrile episode. Eighty-five per cent presented fever or vomiting, and 70% had altered states of mind. Motor deficits (45%), convulsions (35%) and involvement of the cranial nerves (30%) were predominant. Three children progressed with relapses and three others were left with motor sequelae. Magnetic resonance imaging showed hyperintense lesions in T2, with a pattern of scarce/no enhancement, which were predominantly located in the thalamus (70%), the spinal cord (67%) and the white matter of the sub-cortex (50%). Haemorrhagic ADE was diagnosed in two patients. CONCLUSIONS: ADE is a condition with an important degree of general involvement and neurological repercussions, as well as considerable potential to leave the patient with sequelae. Clinico-analytical data and magnetic resonance scans of the head and spinal cord are relevant for the initial diagnosis and follow-up of patients with ADE.

TITLE: Perfil clinicorradiologico de la encefalomielitis aguda diseminada en la poblacion infantil. Analisis retrospectivo de una serie de 20 pacientes de un hospital terciario.Introduccion. La encefalomielitis aguda diseminada (EAD) es un trastorno inflamatorio del sistema nervioso central mediado inmunologicamente y de patogenia desconocida. Puede presentarse en cualquier edad, pero es mucho mas frecuente en niños. La EAD no tiene marcador biologico especifico y el diagnostico se basa en hallazgos clinicos y neurorradiologicos. Objetivo. Mejorar el conocimiento del perfil clinicorradiologico de esta enfermedad. Pacientes y metodos. Estudio retrospectivo con inclusion de pacientes menores de 14 años ingresados en un hospital terciario en los ultimos 15 años con el diagnostico de EAD. Se revisaron antecedentes, signos clinicos de presentacion, datos analiticos en sangre/liquido cefalorraquideo y la semiologia radiologica. En 16 casos se realizo un seguimiento medio de 25 meses. Resultados. Se revisaron 20 pacientes, un 70% niños, con una edad media de 4,4 años. El 40% tuvo un episodio febril previo. El 85% presento fiebre o vomitos, y el 70%, afectacion del estado de consciencia. Predominaron los deficits motores (45%), las convulsiones (35%) y la afectacion de pares craneales (30%). Tres niños presentaron una evolucion recidivante, y otros tres, secuelas motoras. Los estudios de resonancia magnetica mostraron lesiones hiperintensas en secuencias T2, con patron de realce escaso o nulo, que predominaron en los talamos (70%), la medula (67%) y la sustancia blanca subcortical (50%). En dos pacientes se diagnostico EAD hemorragica. Conclusiones. La EAD representa una entidad con importante afectacion general y repercusion neurologica, que muestra un potencial secuelar considerable. Los datos clinicoanaliticos y la resonancia magnetica cerebral y medular son relevantes para el diagnostico inicial y seguimiento de pacientes con EAD.

Assessment of human cytomegalovirus specific T cell immunity in human immunodeficiency virus infected patients in different disease stages following HAART and in long-term non-progressors.

T cell immunity to human cytomegalovirus (HCMV) was assessed in HAART-treated HIV-1 infected patients (9 asymptomatic, CDC group A; and 22 symptomatic, CDC group B), and in eight HIV-1 long term non-progressors. Patients were either prospectively or cross-sectionally examined for CD4(+) T cell counts, HIV RNA load, HCMV leukoDNAemia, HCMV DNA in urine, lymphoproliferative response (LPR) to HCMV and phytohemagglutinin (PHA), and cytokine secretion (IFN-gamma and IL-4) by HCMV-stimulated peripheral blood mononuclear cell (PBMC) cultures. No patient either progressed to clinical AIDS or developed HCMV active infection during the study period. Twenty-nine patients responded to HAART, though 12 patients failed to recover the LPR to HCMV over the study period (three from CDC group A and nine from CDC group B). In contrast to healthy control individuals, most patients displaying positive LPRs LPRs to HCMV had unstable responses. Sustained LPRs to HCMV were significantly associated with high pre-HAART nadir CD4(+) T cell counts. Long-term suppression of HIV viremia correlated with recovery of LPR to HCMV. Sequential PBMC cultures from most patients secreted IFN-gamma (but not IL-4) at normal levels upon HCMV stimulation, irrespective of the pre-HAART nadir CD4(+) T cell counts and CDC group to which patients belonged. Failure to reconstitute IFN-gamma response was associated with very low pre-HAART nadir CD4(+) T cell counts. Control of HCMV infection in the cohort was associated with either recovery or maintenance of IFN-gamma response rather than with reconstitution of LPR to HCMV. A LPR to HCMV was absent in three out of eight long term non-progressors; contrarily, all patients showed preserved IFN-gamma responses.