Ganciclovir, when combined with high-dose intravenous immune globulin, appears to have significantly altered the outcome of patients with cytomegalovirus pneumonia after allogeneic bone marrow transplantation.
Eighteen episodes of community-acquired bacterial pneumonia were diagnosed in 13 patients among 336 with the acquired immunodeficiency syndrome (AIDS) cared for at Memorial Sloan-Kettering Cancer Center since 1979. Bacterial pathogens isolated in 16 of 18 episodes were Haemophilus influenzae in 8, Streptococcus pneumoniae in 6, group B streptococcus in 1, and Branhamella catarrhalis in 1. Eight episodes were presumed Pneumocystis carinii pneumonia until cultures obtained at bronchoscopy confirmed a bacterial cause. Specific antibacterial therapy was curative in 16 of 18 episodes; 2 patients died. Given an estimated yearly incidence of pneumococcal pneumonia in the general population of 2.6/1000, 1.09 cases were expected in our patients with AIDS; we saw 6 (p = 0.001), for an attack rate of 17.9/1000. Bacteria associated with B-cell defects should be anticipated when formulating empiric antibiotic therapy, pending a definitive diagnosis, for pulmonary infiltrates in patients with AIDS.
Factors that influence viability and function of cryopreserved peripheral blood mononuclear cells (PBMC)were identified on 54 samples from 27 AIDS Clinical Trial Units. PBMC viability ranged from 1 to 96% with a median of 70%, was higher in laboratories with experienced staff, and was not significantly associated with CD4 cell number. Function of cryopreserved PBMC, measured by lymphocyte proliferation, was associated with viability. Preparations with viability greater than or equal to 70% had consistent proliferative responses and were suitable for functional analyses.Highly active antiretroviral therapy often results in potent and durable suppression of human immunodeficiency virus (HIV) replication, elevation of CD4 cell counts, decreased incidence of opportunistic infections, and increased survival in HIV-infected patients, including those with advanced disease (6, 7). Immunologic studies in these patients (1, 4) show improvement of some immune functions, but not others, raising the question of whether restoration of this immune function will be complete. Such studies are sometimes performed on cryopreserved peripheral blood mononuclear cells (PBMC). Our understanding of the extent, timing, and determinants of the immune reconstitution will be expanded if assays can be reliably performed on frozen and thawed cells from well-characterized patients.To identify biological and/or technical factors that influence functional assays performed on cryopreserved PBMC, we analyzed the results of the cryopreservation quality control (QC) program established for the immunology component of protocol AIDS Clinical Trial Group (ACTG) 360, "A Longitudinal Study of the Predictive Value of Quantitative CMV Viremia Assays for CMV Disease in Persons with AIDS". This is an ongoing study which enrolled 403 patients at 27 AIDS Clinical Trials Units (ACTU). PBMC from all subjects were cryopreserved every 4 months at ACTU sites. Personnel at the ACTU were asked to freeze the cells following the ACTG cryopreservation protocol, which included separation of PBMC on FicollHypaque gradients, washing the cells, and resuspending them at 10 7 PBMC/ml in cold fetal calf serum with 10% dimethyl sulfoxide. Working on ice, we aliquoted the cell suspension into cryovials at 0.5 ml/vial (5 ϫ 10 6 cells/vial), gradually brought the suspension to a temperature less than or equal to Ϫ70°C over 24 h by using Mr. Frosty devices (Curtis Matheson Scientific) or controlled-rate freezers, and then transferred the frozen aliquots to liquid nitrogen tanks. For testing, the cryovials were shipped every 6 months on dry ice to the University of Colorado Health Sciences Center. The QC protocol was performed on randomly selected samples from each participating ACTU. Cells were thawed by quickly bringing them to 4°C followed by slow addition of cold RPMI medium containing 10% human AB serum. Cells were washed and counted in 0.5% trypan blue to assess numbers of viable cells. The total number of cells in each vial and the percentage and absolute number of viable ce...
We undertook a prospective study to analyze cytomegalovirus (CMV) end-organ disease (EOD) in subjects with advanced human immunodeficiency virus (HIV) infection. Of 403 individuals without prior CMV EOD who were followed up for a median of 151 weeks, 56 died and 21 developed CMV EOD. Twenty of the subjects with CMV EOD had CD4 cell counts of < or =50 cells/mm3 and HIV RNA level of >10,000 copies/mL of plasma at baseline; in these 20 subjects, an increase of CMV DNA level to greater than the quantification limits was associated with CMV EOD. A CD4 cell count of < or =100 cells/mm3 and an HIV RNA level of >10,000 copies/mL of plasma at baseline, a CMV DNA level of >200 copies/mL of blood during follow-up, or development of CMV EOD were all associated with decreased survival. HIV-infected subjects with CD4 cell counts of < or =50 cells/mm3 and HIV RNA levels of >10,000 copies/mL of plasma should have blood fractions screened for CMV DNA; if CMV DNA is detected, CMV prophylaxis might be considered.
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