Fibroblast-derived factors preserve viability in vitro of mononuclear cells isolated from subjects with HIV-1 infection.

OBJECTIVE: Peripheral blood mononuclear cells (PBMC) from HIV-infected subjects have an increased mortality rate (MR) when incubated in vitro for 3 days in a culture medium. We have previously shown that fibroblast-conditioned medium (FCM) can preserve viability, without significant activation, of human lymphocytes in vitro. We therefore tested the ability of two FCM and other factors to reduce spontaneous MR in HIV-positive PBMC. METHODS: PBMC were cultured for 3 days in control medium and medium supplemented with FCM or recombinant cytokines [interleukin (IL)-2, IL-6, IL-7, granulocyte macrophage colony-stimulating factor]. Cells viable at day 3 were counted in a cytofluorimeter after staining with ethidium bromide. DNA was extracted from the cultures and evaluated for the presence of low molecular weight fragmentation. RESULTS: The MR of PBMC from 51 HIV-positive subjects and from 21 healthy controls were 30.1 and 9.5%, respectively (P < 0.0001). The MR was higher in 40 patients with a CD4+ lymphocyte count < 400 x 10(6)/l than in subjects with a count > 400 x 10(6)/l (32.84 versus 20.96%; P = 0.047). IL-2 and FCM significantly reduced MR in HIV-positive subjects (MR: 17.8 and 20.4%; P: < 0.001 and 0.005, respectively). This effect was more evident in subjects with a CD4+ lymphocyte count < 400 x 10(6)/l and in subjects with negative p24 antigenaemia. Cellular proliferation accounts for increased survival in IL-2-supplemented cultures but not in those with FCM. DNA was extracted from fresh PBMC and cells cultured for 3 days for 22 HIV-positive cases. DNA degradation was documented and bands related to an apoptotic mechanism of death observed, especially in subjects with more advanced disease. CONCLUSIONS: Our data suggest that FCM inhibits accelerated cell death in vitro of PBMC isolated from HIV-positive patients.

Cross-cultural adaptation and validation of an Argentine Spanish Version of the Stanford Childhood Health Assessment Questionnaire.

OBJECTIVES: To translate into Argentine Spanish and cross-culturally adapt the Childhood Health Assessment Questionnaire (CHAQ) and validate the adapted instrument in Argentine patients with juvenile rheumatoid arthritis (JRA). METHODS: Five bilingual Argentine pediatric rheumatologists translated into Argentine Spanish and cross-culturally adapted the United States English CHAQ. Pretesting was done in a sample of 23 parents using a probe question technique. Parents of 70 patients with JRA and 21 healthy children (controls) participated in the validation phase. All were from Argentina. RESULTS: The mean disability index (DI) scores for patients with systemic, polyarticular, or pauciarticular onset JRA were 0.64, 0.32, and 0.1, respectively. Healthy controls averaged 0.2. Intercomponent correlations were between 0.4 and 0.9, suggesting internal consistency, but also some redundancy. Test-retest reliability, studied at a 1-week interval, was moderate (mean DI 0.44 [in clinic] and 0.29 [one week later], Pearson's correlation = 0.82). We compared CHAQ scores from 15 parents with those of their children > 10 years of age. Significantly higher DI scores were given by patients than their respective parents (P > 0.019), but the pairwise scores (parent-patient) were highly correlated (r = 0.986). CONCLUSIONS: Cross-cultural adaptation of the US CHAQ to Argentina required few changes. Although DI scores for all patient subgroups were higher than for controls subjects, the scores were low, particularly for those with pauciarticular disease. Prospective studies designed to examine the sensitivity to change and predictive validity will help to assess further the usefulness of the adapted CHAQ in the Argentine population.

The Argentinian version of the Childhood Health Assessment Questionnaire (CHAQ) and the Child Health Questionnaire (CHQ).

We report herein the results of the cross-cultural adaptation and validation into the Argentinian language of the parent's version of two health related quality of life instruments. The Childhood Health Assessment Questionnaire (CHAQ) is a disease specific health instrument that measures functional ability in daily living activities in children with juvenile idiopathic arthritis (JIA). The Child Health Questionnaire (CHQ) is a generic health instrument designed to capture the physical and psychosocial well-being of children independently from the underlying disease. The Argentinian CHAQ was already published and therefore it was revalidated while the Argentinian CHQ was derived from the European Spanish version by changing few words which use is different in the 2 countries. A total of 124 subjects were enrolled: 61 patients with JIA (29% systemic onset, 38% polyarticular onset, 7% extended oligoarticular subtype, and 26% persistent oligoarticular subtype) and 63 healthy children. The CHAQ clinically discriminated between healthy subjects and JIA patients, with the systemic onset, and polyarticular having a higher degree of disability, pain, and a lower overall well-being when compared to their healthy peers. Also the CHQ clinically discriminated between healthy subjects and JIA patients, with the systemic onset, polyarticular onset and extended oligoarticular subtypes having a lower physical and psychosocial well-being when compared to their healthy peers. In conclusion the Argentinian version of the CHAQ-CHQ is a reliable, and valid tool for the functional, physical and psychosocial assessment of children with JIA.

[HIV-1 virus transmission through maternal milk]. / Transmisión del HIV-1 a través de la leche materna.

Human immunodeficiency virus type 1 (HIV-1) may be vertically transmitted during the pre, peri or postpartum period. Postnatal transmission as well as an increased risk of vertical transmission with breastfeeding has been shown for HIV-1 in several reports. Breastfeeding was here analyzed as a risk of HIV-1 transmission in a group of infants born to HIV-1 infected mothers. Among the 215 children studied in our population a significant difference was detected between those who were breastfed vs those who were bottle fed and finally became infected (p < 0.000000, R.R. = 4.29). We also report the case of a postnatal infection in a baby born to an HIV-1 seropositive father and a seronegative mother. Due to the risk of infection of the mother she had been thoroughly controlled when pregnant and after delivery. Mother and child were negative when retested at delivery, and at 10 months post-partum. At the age of 32 months the child attended the outpatient clinic with generalized lymphadenopathy and right parotitis. HIV-1 infection was then confirmed in both mother and child. At that time it was discovered that the baby had been breastfed up to the age of 24 months. This is the first reported child in Argentina whose infection may undoubtedly be attributed to breastfeeding.

Human immunodeficiency virus bDNA assay for pediatric cases.

Techniques to quantify plasma HIV-1 RNA viral load (VL) are commercially available, and they are adequate for monitoring adults infected by HIV and treated with antiretroviral drugs. Little experience on HIV VL has been reported in pediatric cases. In Argentina, the evaluation of several assays for VL in pediatrics are now being considered. To evaluate the pediatric protocol for bDNA assay in HIV-infected children, 25 samples from HIV-infected children (according to CDC criteria for pediatric AIDS) were analyzed by using Quantiplex HIV RNA 2.0 Assay (Chiron Corporation) following the manufacturer's recommendations in a protocol that uses 50 microliters of patient's plasma (sensitivity: 10,000 copies/ml). When HIV-RNA was not detected, samples were run with the 1 ml standard bDNA protocol (sensitivity: 500 HIV-RNA c/ml). Nine samples belonged to infants under 12 months of age (group A) and 16 were over 12 months (group B). All infants under one year of age had high HIV-RNA copies in plasma. VL ranged from 30,800 to 2,560,000 RNA copies/ml (median = 362,000 c/ml) for group A and < 10,000 to 554,600 c/ml (median = < 10,000) for group B. Only 25% of children in group B had detectable HIV-RNA. By using the standard test of quantification, none of the patients had non detectable HIV-RNA, ranging between 950 and 226,200 c/ml for group B (median = 23,300 RNA c/ml). The suggested pediatric protocol could be useful in children under 12 months of age, but 1 ml standard protocol must be used for older children. Samples with undetectable results from children under one year of age should be repeated using the standard protocol.

Troublesome diagnosis in two children born to HIV-1 infected mothers.

In order to assess early HIV infection vertically transmitted in children it is necessary to use techniques that directly detect HIV. Positive results were found in some rare cases who later seemed to have cleared the infection. We communicate two cases of children with troublesome diagnosis. Two girls born to HIV-1 infected mothers were followed-up since birth up to 40 months old, with viral culture, polymerase chain reaction (PCR), p24 antigen detection and serologic techniques. PCRs were positive in three opportunities at 2, 8 and 30 months of age in the first child and confirmatory tests for specific antibodies remained indeterminate up to the age of 34 months. Positive viral DNAs were detected in two opportunities in the second child at 2 and 4 months of age. Western Blots were negative since 25 months. No virus was recovered nor was p24 antigen detected during the whole period of study in either child. Sequestration of the virus in lymphatic tissue, low replicative ability of the virus and/or immunological tolerance can be postulated in the first case. In patient 2, it could be hypothesized that infection, if any, had cleared up.

Diagnosis of perinatally acquired HIV-1 infection using an IgA ELISA test.

The clinical utility of the detection of anti-HIV-1 IgA antibodies using a modified commercial ELISA (EIA) test for the early diagnosis of perinatally acquired HIV-1 infection was evaluated. One hundred and seventeen sera were obtained from 86 infants born to HIV-1-infected mothers and tested for HIV IgA antibodies by an ELISA test (third generation) after removal of IgG with recombinant protein G. Infants were classified according to the Center for Disease Control and Prevention's (CDC) classification system after 15 months of age; 46 were classified as HIV-infected children and 40 as uninfected. HIV-IgA antibodies were detected in 53 of 64 serum samples from all infected children. No significant differences were observed in IgA detection among symptomatic or asymptomatic infected children. However, when analyzed by age a significant difference was observed in IgA detection when children who were over 6 months of age were compared with the younger group (Fisher exact test, p = 0.0000053). All 53 samples from 40 noninfected children were IgA-negative. Statistical analysis was assessed comparing IgA results with HIV infection status as the gold standard. Sensitivity (95%) and specificity (100%), positive predictive value (100%), and negative predictive value (94%) of IgA antibody determination were analyzed taking into account only one sample per child and only children older than 6 months. Positive likelihood ratio was 95.9% and negative likelihood ratio was 94%. Test efficiency was 97%. The detection of IgA HIV antibodies using EIA is an effective method for early diagnosis of HIV-infected infants in comparison with conventional IgG HIV antibody tests. It is a simple and inexpensive method that could be used in both developed and developing countries.

Specific IgA detection can be used for perinatal diagnosis of HIV in children under protocol ACTG 076.

Detection of anti-HIV-1 IgA antibodies using a modified ELISA test for the early diagnosis of perinatally acquired HIV-1 infection in children treated with protocol ACTG 076 was evaluated. A total of 177 sera were obtained from 141 infants between 1 and 12 months of age (46 were treated and 95 were non-treated with protocol ACTG 076) and tested for HIV IgA antibodies by an ELISA test after removal of IgG with recombinant protein G. Infants were classified according to CDC's classification system after a follow-up until 20 months of age. Of the 46 treated children 22 turned out to be infected and in the group of 95 untreated children, 52 were infected. All 81 samples from uninfected children treated or untreated with protocol ACTG 076 were persistently IgA-negative. HIV IgA antibodies were detected in 14 of 25 plasma samples from infected children with treatment, and in 58 of 71 samples in infected children without treatment. Considering that the sensitivity of this test is lower in children younger than 6 months the population of children studied was divided into two groups; those under and those over 6 months of age. No significant differences were observed in the detection of IgA in treated or untreated children in both age groups. The overall specificity of the test was 100 per cent; sensitivity in children older than 6 months was 76.92 per cent in treated children and 93.10 per cent in untreated children. In spite of the small number of samples studied it could be demonstrated that treatment with zidovudine does not affect the detection of IgA antibodies. This is a simple and inexpensive method that could be used for diagnosis of treated and untreated children in developing countries.

HIV-1 and hepatitis B virus infections in adolescents lodged in security institutes of Buenos Aires.

The goal of this study was to determine the prevalence of human immunodeficiency virus type 1 (HIV-1) and hepatitis B virus (HBV) infections in street youth lodged in security institutes, from February 1992 to March 1995, to correlate these infections with nontherapeutic drug use, and to compare these results with a previous study done in a similar population. A total of 1460 white adolescents, 276 females and 1184 males, were enrolled (mean age 16.6 years). Prevalence of HIV-1 was 4.58% and of HBV was 6.78%. The prevalence of dual HIV-1/HBV infection was 1.91%; the prevalence of HBV infection was significantly higher in HIV-positive subjects (p < 0.0000000, chi 2 = 136.17, OR = 13.37) than in those not infected with HIV-1. Prevalences were higher in males. Intravenous drug addiction proved to be a significant risk factor for both viruses (HIV-1, p < 0.0000000, chi 2 = 171.34, OR = 16.84; HBV, p = 0.000044, chi 2 = 16.67, OR = 3.17); 6.43% of the total population were intravenous drug users. Comparison of the current results with our previous study (1989-1992) showed that the prevalence of HIV-1, HBV, and concurrent HIV/HBV as well as intravenous drug addiction has decreased significantly in our current cohort (chi 2 = 134.85, p < 0.0000000; chi 2 = 126.62, p < 0.0000000; chi 2 = 110.05, p < 0.0000000; and chi 2 = 158.3, p < 0.0000000) respectively. Progress appears to have been made in the fight against acquired immunodeficiency syndrome (AIDS), and promising results have been obtained. However, if further viral spread is to be avoided, the emphasis on prevention should be energetically maintained.

Infantile-onset multisystem inflammatory disease: a differential diagnosis of systemic juvenile rheumatoid arthritis.

We describe four unrelated children with neonatal maculopapular rash, fever, arthritis, hepatosplenomegaly, lymphadenopathy, eye involvement, and neurologic symptoms. Radiographs of the joints were surprisingly similar, showing an abnormal epiphyseal and metaphyseal appearance. These clinical and radiologic findings allowed us to include these children in a very peculiar syndrome described as infantile-onset multisystemic inflammatory disease. A chondrosarcoma developed in one of our patients.

Retrospective study of children born to HIV-1-infected mothers in a pediatric hospital in Argentina.

The aim of this retrospective study, which included 103 children born to human immunodeficiency virus type 1 (HIV-1)- infected mothers, is to initiate a database on HIV-infected children, which has to date been unavailable in Argentina. All HIV-1 seropositive children admitted to the Pedro de Elizalde Children's Hospital in Buenos Aires from March 1, 1987, to December 31, 1992, were enrolled in this study. The number of patients enrolled dramatically increased each year during the period of study. Of the 60 infected children, 22 (36.66%) have died with a clinical diagnosis of HIV-1 infection; in 10 of those children HIV infection was also confirmed by P24 antigenemia and/or polymerase chain reaction (PCR): 20 qualified for the Centers for Disease Control and Prevention (CDC) P2D class (P2D1 = 7, P2D2 = 10, P2D3 = 3), 1 for P2C, and 1 for P2A, whose cause of death was pneumonia. The mean age of death was 14.8 months, 18 (82%) died before 18 months. When immunoglobulin G (IgG), IgM, and IgA levels were determined according to age and clinical status, significant differences (P < 0.005) were observed when both asymptomatic and symptomatic infected children (P1, P2) were compared with noninfected children (P3). A significant difference was also obtained between those children who qualified for P2 classification prior to 12 months of age who died early (at or prior to 25 months) and those who reached stage P2 after 12 months of age and have survived to date (X2 = 24.73, p < 0.0001; RR = 5.83, 2.52 < RR < 13.49).

Troublesome diagnosis in two children born to HIV-1 infected mothers

Para determinar la infección vertical por HIV-1 en niños en forma temprana es necesario el uso de técnicas de diagnóstico virológico directo tales como la detección del ADN proviral por la reacción en cadena de la polimerasa (PCR), el aislamiento viral por cocultivo y la detección de antígeno p24 en plasma. Con el uso de estos métodos se describieron casos con resultados positivos en niños que finalmente resultaron no estar infectados. En este trabajo se describen dos casos de niñas hijas de madres HIV-1 positivas que presentaron dificultad para su diagnóstico. Se realizó el seguimiento clínico y de laboratorio de las dos pacientes hasta los 40 meses de edad. Las dos pacientes presentaron un desarrollo pondoestatural y madurativo normal, sin síntomas de enfermedad relacionados con el HIV. En ambas pacientes se encontró, como único hallazgo, la detección del genoma proviral por PCR en más de una muestra. Sin embargo la pérdida de anticuerpos específicos en pacientes con función inmune normal, clínicamente sanas, descartaría la infección por HIV-1. Cabría plantearse la infección con una variante defectuosa, una infección silente o la expresión de tolerancia inmunológica. Además podría plantearse como hipótesis el clearence viral

Human immunodeficiency virus bDNA assay for pediatric cases

Techniques to quantify plasma HIV-1 RNA viral load (VL) are commercially available, and they are adequate for monitoring adults infected by HIV and treated with antiretroviral drugs. Little experience on HIV VL has been reported in pediatric cases. In Argentina, the evaluation of several assays for VL in pediatrics are now being considered. To evaluate the pediatric protocol for bDNA assay in HIV-infected children, 25 samples from HIV-infected children (according to CDC criteria for pediatric AIDS) were analyzed by using Quantiplex HIV RNA 2.0 Assay (Chiron Corporation) following the manufacturer's recommendations in a protocol that uses 50 microliters of patient's plasma (sensitivity: 10,000 copies/ml). When HIV-RNA was not detected, samples were run with the 1 ml standard bDNA protocol (sensitivity: 500 HIV-RNA c/ml). Nine samples belonged to infants under 12 months of age (group A) and 16 were over 12 months (group B). All infants under one year of age had high HIV-RNA copies in plasma. VL ranged from 30,800 to 2,560,000 RNA copies/ml (median = 362,000 c/ml) for group A and < 10,000 to 554,600 c/ml (median = < 10,000) for group B. Only 25 of children in group B had detectable HIV-RNA. By using the standard test of quantification, none of the patients had non detectable HIV-RNA, ranging between 950 and 226,200 c/ml for group B (median = 23,300 RNA c/ml). The suggested pediatric protocol could be useful in children under 12 months of age, but 1 ml standard protocol must be used for older children. Samples with undetectable results from children under one year of age should be repeated using the standard protocol.