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Biomedical subjects

D O Freedman

Publications and source records attributed to D O Freedman.

At least 37 records · Page 2Linked to original sources
PubMed1995

Abnormal lymphatic function in presymptomatic bancroftian filariasis.

Despite the common association of filarial infection with elephantiasis, the great majority of those infected are in fact clinically asymptomatic microfilariae carriers. The assumption has been that infection but not disease exists in these presymptomatic persons. In an area Brazil where Wuchereria bancrofti is endemic, flow studies done with dynamic radionuclide lymphoscintigraphy were used to compare 30 limbs from asymptomatic microfilaremic subjects with 16 control limbs. Geometric mean values for T1/2 (19.8 vs. 37.7 min; P < .001), appearance time (7.9 vs. 27.9 min; P < .001), percent uptake at the region of interest (0.67% vs. 0.14%; P < .001), and peak activity (62.6 vs. 2.6 cps; P < .001) each indicated an enhanced pattern of rapid, increased lymph flow in asymptomatic microfilaremic subjects. The abnormal lymphatic function in these subjects indicates that current passive intervention strategies may need to change if the debilitating sequelae of this parasitic infection are to be avoided.

Adolescent↗
PubMed1995

Lymphoscintigraphic assessment of the effect of diethylcarbamazine treatment on lymphatic damage in human bancroftian filariasis.

Despite many millions of doses administered over the past 40 years, basic and crucial issues regarding the use, mode of action, and effectiveness of diethylcarbamazine (DEC) in many clinical situations remain unresolved. To directly investigate whether the well-known microfilaricidal and macrofilaricidal actions of DEC actually result in subsequent improvement in existing damage to lymphatic vessels or lymph nodes, 29 study subjects in Recife, Brazil were stratified into three groups according to the severity of clinical manifestations of lymphatic insufficiency. After baseline radionuclide lymphoscintigraphy was performed, subjects were treated with two courses of DEC separated by at least a six-month interval and then rescanned one year after the baseline scan. A side-by-side comparison of images obtained at baseline with those obtained at follow-up in 13 asymptomatic microfilaremic individuals, six individuals with filarial fever, and in 10 individuals with chronic pathology demonstrated essentially unchanged lymphatic morphology in all but one individual whose disease actually progressed in the face of therapy. We conclude that two 12-day treatment courses of DEC did not have a demonstrable direct or indirect effect on existing structural damage to the lymphatic system even in those individuals with preclinical disease.

Acute Disease↗
PubMed1995

Predominant CD8+ infiltrate in limb biopsies of individuals with filarial lymphedema and elephantiasis.

In 34 individuals with a spectrum of clinical manifestations of Bancroftian filariasis, we investigated whether immunoperoxidase-stained, random, superficial dermal biopsies could further elucidate the nature of the diffuse damage to superficial lymphatics that had been recently demonstrated by radionuclide lymphoscintigraphy. A total of 78% and 68% of limbs from patients with clinical disease and asymptomatic microfilaremia, respectively, contained EN4+PAL-E- lymphatic vessels that were abnormally dilated. The majority of subjects, regardless of clinical classification, had a CD3+ perivascular but not a perilymphatic infiltrate in tissues and no parasites were present. In contrast to those individuals with asymptomatic infection, a striking predominance of CD8+ T cells was found in the tissue of individuals with clinical disease. Tissue pathology consistent with cutaneous bacterial infection was not observed. The prominent perivenular and pericapillary mononuclear infiltrates likely indicate, in light of current understanding of lymphocyte recirculation, the extravasation of lymphocytes from the vascular circulation into the inflamed filarial tissue.

Animals↗
PubMed1994

The Onchocerca volvulus homologue of the multifunctional polypeptide protein disulfide isomerase.

Protein disulfide isomerase (PDI) functions to catalyze the formation of correct disulfide bonds in nascent proteins, and also acts as one of the subunits of prolyl-4 hydroxylase, the enzyme responsible for the oxidative maturation of procollagen. Since the cuticle of parasitic nematodes consists primarily of a network of collagen molecules which are connected through intermolecular disulfide bonds, PDI might be expected to be involved in the process of cuticle biosynthesis. The isolation and characterization of a cDNA encoding the PDI homologue of Onchocerca volvulus is described. This cDNA contains a single, long open reading frame that encodes sequence motifs identical to the two known active sites of PDI for isomerase activity. The O. volvulus PDI appears to be encoded by a single copy gene. Both in situ hybridization and immunolocalization data suggest that PDI is both spatially and temporally regulated in O. volvulus. The pattern of spatial and temporal regulation is consistent with the involvement of PDI in the biosynthesis of the parasite cuticle. The parasite protein appears to be an antigen recognized by a minority of individuals exposed to O. volvulus.

Amino Acid Sequence↗
PubMed1994

Lymphoscintigraphic analysis of lymphatic abnormalities in symptomatic and asymptomatic human filariasis.

To obtain high-resolution radionuclide lymphoscintigraphic images of affected limbs in persons with both symptomatic and asymptomatic filarial infection, 36 volunteers were recruited from a Wuchereria bancrofti-endemic area of Recife, Brazil, for a prospective, controlled analysis. Subjects were stratified after determination of serologic and clinical determinants of filarial infection status. Widespread lymphatic abnormalities were found in clinically asymptomatic microfilaremic persons, who had been assumed to have infection but not disease. All patients with clinical manifestations of lymphatic pathology and marked abnormalities. No correlation was found between clinical findings and actual lymphatic function as demonstrated by lymphoscintigraphy. The initial diagnosis of lymphatic filariasis, whether asymptomatic or symptomatic, is based on nonimaging laboratory criteria. After diagnosis, lymphoscintigraphy is a valuable tool for initial assessment of any lymphatic damage. Changes in strategies for therapeutic interventions in asymptomatic microfilaremic persons, who are not usually aggressively treated, may be warranted.

Adolescent↗
PubMed1994

Cloning and characterization of a potentially protective chitinase-like recombinant antigen from Wuchereria bancrofti.

While there is no direct evidence demonstrating the existence of protective immunity to Wuchereria bancrofti infection in humans, the presence of individuals, in populations in areas where infection is endemic, with no clinical evidence of past or current infection despite appreciable exposure to the infective larvae, suggests that protective immunity to filarial parasites may occur naturally. Earlier work indicated that such putatively immune individuals generated antibodies to a 43-kDa antigen from larval extracts of the related filarial parasite Brugia malayi that was recognized by only 8% of the infected population. With rabbit antiserum raised against this 43-kDa antigen, this current study identified a recombinant clone, WbN43, with an insert size of 2.3 kb, from a W. bancrofti genomic expression library. The recombinant fusion protein was differentially recognized by the putatively immune individuals but not by the infected patients. The coding sequence (684 bp) from the 5' end had significant sequence similarity to chitinases from Serratia marcescens, Bacillus circulans, Streptomyces plicatus, and B. malayi. Peptide sequencing of the expressed product also defined a chitinase-like sequence. Molecular characterization indicated WbN43 to be a low-copy-number gene, with expression predominantly in infective larvae and microfilariae but not in adult parasites.

Adult↗
PubMed1992

Rapid diagnosis of Bancroftian filariasis by acridine orange staining of centrifuged parasites.

A rapid diagnostic test for detection of microfilaremia using a microhematocrit tube precoated with acridine orange (the Quantitative Buffy Coat [QBC] tube) was compared with a conventional 50 microliters-thick blood film (TF) in 119 volunteers in an area of Recife, Brazil that was endemic for Wuchereria bancrofti. Both the QBC and the TF techniques were 100% specific, and thus positive predictive values were equal at 100% for each technique in all subjects studied. Both techniques had equal negative predictive values of 100% in subjects with microfilarial counts > 100 per milliliter (mf/ml). Counts < 20 mf/ml are below a cutoff equal to the calculated limit of sensitivity of each of the two techniques. For those individuals with counts between 20 and 99 mf/ml, negative predictive values were, for practical diagnostic purposes, equivalent at 97.5% for the QBC technique and 99.0% for the TF. Because the QBC technique has predictive values as high as conventional TF, the convenience and rapidity of the technique will make the QBC technique a desirable alternative diagnostic method in those clinical settings where the equipment is available. A positive result will be available in less than six min after obtaining the specimen in individuals with counts < 100 mf/ml, and individuals with lower or no microfilaremia will have a result within 6-12 min.

Acridine Orange↗
PubMed1991

Immunoregulation in onchocerciasis. Functional and phenotypic abnormalities of lymphocyte subsets and changes with therapy.

To help define the immunoregulatory defects in patients with onchocerciasis, flow cytometric analysis of circulating lymphocyte subpopulations was performed in parallel with functional assays. No significant differences in CD4/CD8 ratios were seen when microfilariae-positive individuals from Guatemala were compared with Guatemalan controls. However, the infected individuals had significantly increased numbers of circulating CD4+CD45RA+ lymphocytes (mean 38.3%) when compared with controls (mean 16.0%). Coexpression of the activation marker HLA-DR was significantly increased on CD4+ cells from infected individuals. In contrast, no up-regulation of HLA-DR was seen on CD8+ or CD19+ cells. At 1 year after initiation of treatment with semiannual doses of the microfilaricide ivermectin, there were significant increases (P less than 0.05) in the percentage of CD4+CD45RA- cells, the percentage of CD4+HLA-DR+ cells, and mitogen-induced lymphokine production (IL-2, IL-4). Despite these changes, parasite-specific IL-2 and IL-4 production which had been undetectable before treatment did not manifest itself even by the 2-yr follow-up. Defects in the T-cell activation pathway in Onchocerca volvulus-infected individuals may thus exist at several independent points; a state of parasite antigen-specific tolerance appears to remain even after the relative reversal of other generalized immunoregulatory defects.

Adolescent↗
PubMed1989

Selective up-regulation of endothelial cell class I MHC expression by cytokines from patients with lymphatic filariasis.

Local host immune responses to the lymphatic-dwelling filarial parasite Wuchereria bancrofti are important in the pathogenesis of the lymphangitis that leads to filarial elephantiasis. That the lymphatic endothelial cells may be important in this inflammatory process was shown by the ability of supernatants generated from filarial Ag-driven PBMC of individuals with filarial elephantiasis caused by W. bancrofti infection to up-regulate class I MHC expression on human umbilical vein endothelial cells when compared to unstimulated control supernatants from the same individual (relative fluorescence intensity = 159% +/- 13.5; p less than 0.001). In contrast, individuals with the same filarial infection but manifesting no lymphatic disease were unable to generate, in response to filarial Ag the cytokines required for this activation (relative fluorescence intensity = 93% +/- 2.6). Supernatants induced by a non-filarial Ag (purified protein derivative) were able to effect class I MHC up-regulation in both patient groups. The same filarial Ag-driven supernatants did not cause detectable class II MHC staining on human umbilical vein endothelial cells. These results suggest a likely role for parasite Ag-driven, cytokine-mediated endothelial cell activation in the pathogenesis of lymphatic inflammatory/obstructive filarial disease.

Adult↗
PubMed1989

Protective immunity in bancroftian filariasis. Selective recognition of a 43-kD larval stage antigen by infection-free individuals in an endemic area.

There is little information about naturally occurring protective immunity in individuals living in areas endemic for lymphatic filariasis, though an immunologically hyperresponsive, uninfected group of "endemic normal" individuals that may be immune has been previously recognized. To analyze the nature of the hyperresponsiveness and its potential relation to a state of protective immunity in such individuals, strict clinical, parasitological, and serological criteria were applied to select seven "infection-free" endemic normal individuals (ENs) from a population of 459 persons resident in an area heavily endemic for bancroftian filariasis. Immunoblot analysis was used to compare the qualitative antigen recognition patterns of these endemic normal individuals to those of a group of 12 clearly infected microfilaremic individuals (MFs) from the same endemic area. Though immunoblot analysis using microfilarial and adult stage filarial antigens revealed no distinct differences in antigen recognition patterns between the two groups, when responses to infective larval stage antigens were assessed, 7/7 (100%) of the ENs were found to recognize a 43-kD antigen that was recognized by only 1/12 (8%) of the MFs. These findings are consistent with the concept that recognition of unique larval antigens may induce protective immunity to human filarial parasites and they identify a candidate immunogen for further functional assessment.

Adult↗
PubMed1988

Enhanced solubilization of immunoreactive proteins from Brugia malayi adult parasites using cetyltrimethylammonium bromide.

To determine an optimal method for extracting immunoreactive proteins from filarial parasites, we have subjected Brugia malayi adult worms to a variety of solubilization regimens and compared the results. The parasites were extracted in one of seven detergents (including anionic, cationic, nonionic, and zwitterionic compounds) under varying conditions of pH, detergent concentrations, and incubation time. The individual antigen preparations were then compared both by one-dimensional SDS-PAGE and by immunoblotting analysis using a serum pool from individuals resident in an area endemic for lymphatic filariasis. The cationic detergent cetyltrimethylammonium bromide (CTAB) at 1.0% concentration, pH 7.2, consistently solubilized more proteins immunoreactive with the sera tested. Additionally, CTAB never failed to solubilize immunoreactive proteins solubilized by those other detergents or combinations of detergents studied.

Animals↗
PubMed1988

Eggs of Schistosoma mansoni stimulate endothelial cell proliferation in vitro.

In humans chronically infected with Schistosoma mansoni, total hepatic blood flow and normal hepatic function appear to be maintained by neovascularization of the periportal fibrotic tissue. To identify possible stimuli for this neovascularization, we evaluated parasite-derived products for their ability to activate endothelial cells in vitro. Soluble egg antigen (SEA) of S. mansoni at concentrations as low as 30 ng/mL induced a marked proliferation of human umbilical vein endothelial cells (HUVE) that increased in a dose-dependent fashion; equivalent effects were seen with bovine adrenal and murine lung endothelial cells. Live, intact S. mansoni eggs secreted a soluble factor that stimulated HUVE in a manner similar to crude SEA. In contrast, adult worm extracts of S. mansoni had no significant effect on endothelial cell proliferation in any of the three cell lines tested. Preliminary biochemical characterization showed the activity to be protease sensitive and heat stable. S. mansoni SEA, in the absence of inflammatory cells, can stimulate endothelial cell proliferation in vitro.

Animals↗
PubMed1987

Loiasis in an expatriate American child: diagnostic and treatment difficulties.

On the basis of this experience, we recommend the following when faced with possible filariasis in an expatriate from Western Central Africa: (1) Attempt a clinical differentiation between L loa and other filarial infections present in West Africa. It is important to suspect loiasis because this is the only filarial infection that is readily curable; (2) ophthalmologic assessment to diagnose onchocerciasis; (3) if L loa is suspected, thick blood smears should be obtained from midmorning to midafternoon and stained with Giemsa or hematoxylin stains, after a concentration technique is used. Nighttime blood specimens should be obtained if the patient has been in an area where W. bancrofti is prevalent; (4) skin snip biopsies prepared as follows: Bilateral symmetrical skin snips should be taken. In the case of suspected West African filariasis, the pelvic girdle, iliac crest, and back of scapula are thought to have the highest yield. One snip from each of six different sites should be obtained. Each skin snip should be approximately 2 to 3 mm (a cornealoscleral biopsy forceps can be used). Each skin snip is placed in 100 microL (approximately one drop) of normal saline in a flat-bottomed microtiter plate. The plate is incubated at room temperature and checked periodically for 24 hours under a dissecting microscope (X20 to X40). If present, the small worms will be seen wiggling and squirming in the drop of saline; (5) serologic diagnostic methods are most efficient if human filarial antigens are used; (6) if treatment is with diethylcarbamazine, the initial dose should be small.(ABSTRACT TRUNCATED AT 250 WORDS)

Cameroon↗