Because helminth infections and human immunodeficiency virus (HIV) coexist in areas where the spread of AIDS is most dramatic, their in vitro interaction was explored. Cryopreserved peripheral blood mononuclear cells (PBMC) from patients with filarial infections (n=24) and from unexposed control subjects (n=12) were depleted of CD8 T cells and were infected with macrophage (M)- and T cell-tropic viruses. A trend toward increased HIV replication in PBMC from filaria-infected patients was observed. Furthermore, PBMC from 6 filaria-infected patients before antifilarial treatment were significantly more susceptible to replication of M-tropic virus than their posttreatment PBMC (P=.03). No intergroup differences were found in the surface expression of HLA-DR, CD25, CCR5, CXCR4, CCR3 on CD4 T cells, or monocytes before infection. PBMC from filaria-infected patients produced less RANTES (P=.02) but more intracellular interleukin-4 than those of control subjects. Thus, PBMC from persons with filarial infections appear to have enhanced susceptibility to HIV-1 infection mediated by an undetermined mechanism.
Early and accurate diagnosis and prognosis of patients with fulminant liver failure is of critical importance for optimum management. We investigated the role of transjugular liver biopsy in the management of patients with fulminant liver failure and assessed its value in comparison with the recently proposed King's College criteria. Sixty-one patients with fulminant liver failure, ages 2 to 82 yr, were retrospectively analyzed. The main outcome measures were survival vs. death or progression to orthotopic liver transplantation. Transjugular liver biopsy was successful in 60 of 61 patients, with a mean core tissue length of 2.1 cm. There were eight minor complications, all of which were managed conservatively. Biopsy specimens were evaluated for degree of fibrosis, percentage of hepatocellular necrosis and presence of bile duct proliferation, hepatocellular mitotic figures and binucleate hepatocytes for each of the 54 specimens available for analysis. In 34 of 54 patients (63%), the presumed clinical diagnosis was confirmed by transjugular liver biopsy. In 11 patients the procedure served to clarify clinical uncertainty, whereas in 9 of 54 (16.7%) the diagnosis was altered after transjugular liver biopsy. The percentage of necrosis was the only histological parameter that appeared to have significant discriminatory prognostic value, with only 2 of 19 survivors having greater than 70% necrosis. Twenty-one of these biopsy specimens were reviewed by two pathologists, and their degree of correlation for the various features was assessed. Almost perfect concordance was found between the two pathologists on the percentage of hepatocellular necrosis.(ABSTRACT TRUNCATED AT 250 WORDS)
To assess the utility of a polymerase chain reaction (PCR)-based method for diagnosis of Wuchereria bancrofti infection, blood, plasma, and paraffin-embedded tissue samples were tested using a PCR-based assay that detects a W. bancrofti-specific repetitive DNA sequence. The assay was positive in 100 microL of blood from 40 of 42 microfilaria-positive subjects, the 2 subjects with negative assays having microfilarial counts of 1. Samples from 127 uninfected subjects were PCR-negative. The assay was also positive in 7 of 10 daytime samples in regions where infection is nocturnally periodic; PCR amplification from paraffin-embedded sections established the diagnosis of W. bancrofti infection in another 2 cases. A microtiter ELISA plate-based method was developed for rapid evaluation of large numbers of samples. These results suggest that this PCR-based assay will be useful in diagnosis of W. bancrofti infection in a variety of clinical settings.
Treatment of patients with patent Wuchereria bancrofti infection results in an acute clinical reaction and peripheral eosinophilia. To investigate the dynamics of the eosinophil response, changes in eosinophil activation and degranulation and plasma levels of eosinophil-active chemokines and cytokines were studied in 15 microfilaremic individuals in south India by sequential blood sampling before and after administration of 300 mg of diethylcarbamazine (DEC). Clinical symptoms occurred within 24 h. Plasma interleukin-5 (IL-5) and RANTES levels peaked 1 to 2 days posttreatment, preceding a peak peripheral eosinophil count at day 4. Major basic protein secretion from eosinophils paralleled IL-5 secretion, while levels of eosinophil-derived neurotoxin peaked at day 13 after treatment. Expression of the activation markers HLA-DR and CD25 on eosinophils rose markedly immediately after treatment, while expression of VLA-4 and ␣47 showed an early peak within 24 h and a second peak at day 13. Thus, the posttreatment reactions seen in filarial infections can be divided into an early phase with killing of microfilariae, clinical symptomatology, increases in plasma IL-5 and RANTES levels, and eosinophil activation and degranulation and a later phase with expression of surface integrins on eosinophils, recruitment of eosinophils from the bone marrow to tissues, and clearance of parasite antigen.
Two variants of the circumsporozoite (CS) protein of Plasmodium vivax (VK210 and VK247) have been identified. Recently, a putative third CS variant of P. vivax, referred to as causing P. vivax-like malaria, has been reported from Papua New Guinea. The objective of this study was to confirm and extend findings on the global distribution of the P. vivax-like parasite. Blood samples were obtained from 126 untreated patients with P. vivax infection acquired in Central America, South America, Africa, Southeast Asia, and the Indian subcontinent. The P. vivax CS gene was amplified by polymerase chain reaction and hybridized with probes specific for the VK210, VK247, and P. vivax-like CS variants. All samples were positive for VK210, VK247, or both. No sample was positive for P. vivax-like DNA. Therefore, the existence of a third variant of P. vivax cannot be confirmed in the geographic areas studied.
The disease course of human immunodeficiency virus (HIV) is often altered by existing or newly acquired coinfections. Treatment or prevention of these concomitant infections often improves the quality and duration of life of HIV-infected persons. The impact of helminth infections on infections with HIV is less clear. However, HIV is frequently most problematic in areas where helminth infections are common. In advance of the widespread distribution of drugs for elimination of lymphatic filariasis, we assessed the prevalence of active Wuchereria bancrofti infection among HIV-positive patients in Chennai, India at two time points separated by four years. We found that the overall prevalence of W. bancrofti infections among HIV-positive persons was 5-9.5%, and there were no quantitative differences in circulating filarial antigen levels between HIV-positive and HIV-negative filarial-infected patients.
The identification of eosinophils in lysed whole blood by flow cytometry can be problematic, since these cells overlap significantly with the neutrophil cluster on forward scatter versus side scatter plots of whole blood samples. Current methods can be time‐consuming when running multiple samples or may compromise yield in the interests of greater accuracy. The use of eosinophil purification techniques prior to FACS analysis or sorting as a way of ensuring purity may have unpredictable effects on eosinophil activation, leading to questionable data interpretation. Here we describe a simple, single‐step method for definition of eosinophils utilizing their high side scatter and CD16 fluorescence negativity to differentiate them from neutrophils. The purity of the neutrophil and eosinophil populations sorted with this gate is close to 100% regardless of the peripheral blood eosinophil count, while the population obtained by sorting on a plot of FSC/SSC was a mixture of eosinophils and neutrophils. We suggest this method as a simple, reproducible, and accurate way of defining eosinophils by flow cytometry for analysis or sorting. Cytometry 30:313–316, 1997. © 1997 Wiley‐Liss, Inc.
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