RESUMO
OBJECTIVE: Immunofluorenscence methods to detect pp65 antigenemia were implemented for identifying the circulating virus-infected cells in individuals known to have cytomegalovirus infection and disease symptoms. MATERIAL AND METHODS: Between December-2002 and July-2003, 110 peripheral blood samples were obtained from 46 immunosuppressed patients. pp65 antigenemia and the presence of circulating cells were determined by indirect immunofluorescence using a commercial kit to detect CMV pp65 antigen in peripheral blood leukocytes. Antigenemia was positive when one or more cells was observed with multilobulated, homogenous fluorescent stain in the nucleus. The presence of infected circulating cells (peripheral blood mononuclear cells) was determined when an extended pattern of fluorescent stain was observed throughout the cytoplasm in cells of 35 to 50 microm. RESULTS: Eight antigenemias from 7 patients (15%) were positive. Of these, 4 (57%) were also positive for circulating infected cells and consisted of 3 kidney transplant recipients and 1 liver transplant recipient. The number of positive cells in antigenemia was greater in kidney-transplant recipients than in the rest of immunosupressed patients (457 vs. 1.96, p < 0.005). No association was seen between the presence of infected circulating cells, morbidity, mortality or the development of GVHD (p < 0.001). CONCLUSION: No correlation was observed between the presence of infected cells, antigenemia and mortality. To substantiate the lack of correlation amongst these factors, prospective studies with larger sample sizes are necessary. These studies will aid in better defining the clinical application in immunosupressed patients.
Assuntos
Infecções por Citomegalovirus/sangue , Citomegalovirus/isolamento & purificação , Hospedeiro Imunocomprometido/imunologia , Fosfoproteínas/sangue , Proteínas da Matriz Viral/sangue , Antígenos Virais/sangue , Antígenos Virais/imunologia , Citomegalovirus/imunologia , Infecções por Citomegalovirus/imunologia , Humanos , Fosfoproteínas/imunologia , Proteínas da Matriz Viral/imunologiaRESUMO
The incidence of maternal-to-fetal human immunodeficiency virus type 1 (HIV-1) transmission is 25-30% in absence of antiretroviral therapy, and is inversely associated with Human leukocyte antigens (HLA) class-I discordance. Based on our earlier report that mixed lymphocyte reactions (MLR) induce a ribonuclease (RNase) that inhibits HIV-1 replication, we proposed that maternal-fetal alloantigen stimulation activates factors that protect the fetus against vertically-transmitted infections. We investigate here whether the degree of mother-infant HLA discordance associates with the ability to produce anti-HIV-1 alloantigen-stimulated factor (ASF), and affects placental RNases. We also determine whether such HLA association is influenced by the mother's HIV-1 status. Paired maternal and cord blood leukocytes were tested for the induction of ASF by MLR, and typed for HLA-A and -B. The placentas were tested for mRNA expression of three RNases. Neonate anti-mother, but not mother anti-neonate MLR generated supernatants with anti-HIV-1 activity, that was associated with HLA class I discordance. This HLA association was not seen in the HIV-infected cohort. HLA class I discordance was also associated with expression of placental RNase 1. Our findings are consistent with the hypothesis that HLA class I discordance induces expression of RNases in the placenta that contribute to innate host resistance to HIV-1 and other viral infections.
Assuntos
Fármacos Anti-HIV/metabolismo , Infecções por HIV/transmissão , Histocompatibilidade Materno-Fetal/imunologia , Transmissão Vertical de Doenças Infecciosas/prevenção & controle , Placenta/enzimologia , Ribonucleases/metabolismo , Adolescente , Adulto , Fármacos Anti-HIV/farmacologia , Feminino , Infecções por HIV/imunologia , Infecções por HIV/virologia , HIV-1/efeitos dos fármacos , Antígenos HLA-A/imunologia , Antígenos HLA-A/metabolismo , Antígenos HLA-B/imunologia , Antígenos HLA-B/metabolismo , Humanos , Lactente , Recém-Nascido , Isoantígenos/imunologia , Teste de Cultura Mista de Linfócitos , Gravidez , Complicações Infecciosas na Gravidez/imunologia , Complicações Infecciosas na Gravidez/virologia , Ribonucleases/farmacologia , Replicação Viral/efeitos dos fármacosRESUMO
Objetivo. Buscar la correlación entre la presencia de células citomegálicas circulantes con la antigenemia pp65 para citomegalovirus y el desarrollo de complicaciones clínicas inherentes a infección y enfermedad por citomegalovirus. Materiales y métodos. Entre diciembre de 2002 y julio de 2003 se procesaron 110 muestras de sangre periférica, obtenidas de 46 pacientes inmunosuprimidos. La antigenemia pp65 y la presencia de células citomegálicas circulantes se determinaron mediante inmuno-fluorescencia indirecta utilizando un estuche comercial para la detección del antígeno pp65 de citomegalovirus. En leucocitos de sangre periférica, la antigenemia fue positiva cuando se encontró una o más células con tinción fluorescente, multilobulada y homogénea en el núcleo celular. Se determinó la presencia de células citomegálicas circulantes cuando se observaron células de gran tamaño (35 a 50 mm) con un patrón de tinción fluorescente extendido en todo el citoplasma celular en células mononucleares de sangre periférica. Resultados. Se encontraron ocho pruebas con antigenemia positiva, procedentes de siete pacientes (15 por ciento). De éstos, cuatro (57 por ciento) también presentaron células citomegálicas circulantes, tres habían recibido un trasplante renal y uno un trasplante hepático. El número de células positivas en la antigenemia fue mayor en los pacientes con trasplante renal que en el resto de pacientes inmunosuprimidos (457 vs.1,96; p<0,005). No se encontró asociación entre la presencia de células citomegálicas circulantes, la morbilidad, la mortalidad y el desarrollo de enfermedad injerto contra huésped (GVHD) (p<0,001). Discusión. No encontramos correlación entre la detección de células citomegálicas circulantes, la antigenemia y la mortalidad en estos pacientes. Sin embargo, es necesario realizar estudios prospectivos con un mayor número de individuos para determinar mejor dicha correlación y definir su utilidad clínica en pacientes inmunosuprimidos