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Biomedical subjects

M C Healey

Publications and source records attributed to M C Healey.

52 records · Page 3Linked to original sources
PubMed1987

PR 879-317A enhances in vitro immune activity of peripheral blood mononuclear cells from patients with Down syndrome.

Down syndrome is associated with immune deficiencies which result in increased incidences of respiratory infections and lymphocytic leukemia. Peripheral blood mononuclear cells (PBMC) from patients with Down Syndrome were assayed in several in vitro assays following incubation in medium or various concentrations of PR 879-317A (2,3,5,6,7,8-hexahydro-2-phenyl-8,8-dimethoxyimidazo (1,2a) pyridine), a selective immunorestorative agent. PBMC of the patients, incubated in medium, exhibited significantly reduced activities in the natural killer (NK) cell, antibody-dependent cell-mediated cytotoxicity (ADCC), T-cell blastogenesis and leukocyte-inhibition factor (LIF) assays. Incubation in PR 879-317A significantly increased the NK and ADCC activities of PBMC from both patients and healthy subjects. However, the effect was much more pronounced on the patients' cells increasing their NK and ADCC activities to normal levels. Incubation in PR 879-317A augmented to normal levels the responses of patients' cells in various assessments of T-cell immunity including blastogenic responses to phytohemagglutinin and concanavalin A and production of LIF. In addition, the number of patients' cells forming spontaneous rosettes with sheep red blood cells was increased following incubation in PR 879-317A. In contrast this compound did not significantly modify the T-cell responses of cells from the healthy subjects suggesting that this compound does not affect normal T-cell function.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1987

Production of monoclonal antibodies specific for Eimeria tenella microgametocytes.

A panel of 4 monoclonal antibodies specific for Eimeria tenella, the causative agent of cecal coccidiosis of birds in the genus Gallus, was produced by standard techniques. The indirect fluorescent antibody (IFA) test demonstrated specificity of these 4 antibodies for the microgametocytes. Hybridoma TIA3B9 secreted a monoclonal antibody of subisotype IgG2b that was used throughout the course of this study. Immunologic potency of this antibody was demonstrated by in vitro experiments that revealed a greater than 50% reduction in oocyst production, indicating an apparent inhibition of fertilization.

Animals↗
PubMed1986

Use of monoclonal antibodies to identify outer membrane antigens of Actinobacillus species.

Three monoclonal antibodies (LG17, LG30, and LG33) were used to identify outer membrane antigens of Actinobacillus sp (As8C isolate) cultured from the epididymides of an infected ram lamb. Specificity of the 3 antibodies to As8C antigens was determined by use of bacterial agglutination, the enzyme-linked immunosorbent assay, and the indirect fluorescent antibody test. Results of immunoelectron microscopy confirmed that each antibody was specific for epitopes on As8C outer membrane antigens. Evaluation by use of enzyme-linked immunoelectrotransfer blot indicated that target antigens for LG17 and LG33 antibodies had molecular weights of 10 kilodaltons and 43 kilodaltons, respectively. Multiple-band staining was observed with the LG33 antibody. The target antigen for the LG30 antibody could not be discerned by use of enzyme-linked immunoelectrotransfer blot. For each of the 3 monoclonal antibodies, enzyme-linked immunosorbent assay titers were obtained for Actinobacillus seminis, A actinomycetemcomitans, and 10 field isolates of Actinobacillus spp. Target antigens for LG17 and LG30 antibodies occurred infrequently or were absent on these bacteria. However, the target antigen for the LG33 antibody was shared by Actinobacillus seminis, A actinomycetemcomitans, and the 10 field isolates of Actinobacillus spp, indicating some diversity of outer membrane antigens between isolates.

Actinobacillus↗
PubMed1985

Production and preliminary characterization of monoclonal antibodies to Actinobacillus sp isolated from epididymitis lesions in a ram.

A panel of 55 monoclonal antibodies to an Actinobacillus sp isolate (As8C strain) cultured from the epididymides of an infected ram was produced. Cell lines producing 5 of these antibodies were cloned and expanded by hybridoma tumor production in Balb/c mice. An isotype profile revealed that 1 cloned antibody belonged to the immunoglobulin (Ig) M class and the 4 remaining antibodies belonged to the IgG class. Within the IgG class, 1 clone produced IgG1, 1 clone produced IgG2a, and 2 clones produced IgG2b. Ascites fluid antibody titers from the cloned hybridomas ranged from 6,400 to 51,200, as determined by enzyme-linked immunosorbent assay. Specificity of the antibodies to target As8C antigens could be demonstrated by enzyme-linked immunosorbent assay inhibition. Ascites fluid from 2 clones contained antibodies that agglutinated As8C. Two additional clones produced antibodies capable of only partial agglutination, whereas 1 clone produced antibody that did not agglutinate As8C. The indirect fluorescent antibody test revealed that target antigens for at least 4 of the 5 monoclonal antibodies were most likely located on the bacterial cell surface. Antigens were extracted from As8C, using 5 surface active chemicals. An attempt to immunoprecipitate these antigens in agarose by reacting individual extracts with each of the antibodies was unsuccessful.

Actinobacillus↗
PubMed1984

The effect of erythrosin B on infective larvae of bovine gastrointestinal nematodes.

The effect of erythrosin B and visible light on 3rd stage larvae of gastrointestinal nematodes of naturally-infected cattle was studied. Cattle were treated orally with erythrosin B at dosages of 30 and 40 mg kg-1 day-1 for as many as 17 days. Feces from treated and untreated animals were collected and prepared for culture. Third stage larvae were then collected by Baermannization and exposed to light. Both sunlight and artificial fluorescent light were shown to be toxic to 3rd-stage-larvae after treatment with erythrosin B. This toxic reaction was significant after only 2 consecutive daily treatments.

Animals↗
PubMed1980

Ophthalmooestriasis externa.

In this case of ophthalmooestriasis externa in a 20-year-old white male student attending Mississippi State University, three first instar larvae from the sheep nasal bot fly, Oestrus ovis, were removed. The patient responded well to application of sulfa eye drops. The exact means by which the patient acquired the larvae was not determined.

Adult↗
PubMedDate not supplied

Comparison of the host ranges and antigenicity of Cryptosporidium parvum and Cryptosporidium wrairi from guinea pigs.

Oocysts of a Cryptosporidium isolate from guinea pigs were not infectious for adult mice, but were infectious for two of three newborn calves and for suckling mice. However, oocysts isolated from calves or mice infected with guinea pig Cryptosporidium were not infectious for guinea pigs. Four isolates of C. parvum from calves were incapable of infecting weanling guinea pigs. Microscopic examination of tissue from the colon and cecum of suckling guinea pigs inoculated with C. parvum revealed sparse infection of some pups. These host range studies and previously described differences in 125I-labeled oocyst surface protein profiles between Cryptosporidium sp. from guinea pigs and C. parvum suggest they are distinct species. We propose the name Cryptosporidium wrairi be retained. Studies with monoclonal antibodies indicate that C. wrairi and C. parvum are antigenically related.

Animals↗
PubMedDate not supplied

Hemolytic properties of lytic peptides active against the sporozoites of Cryptosporidium parvum.

Cryptosporidium parvum is a protozoan parasite that causes mildto-severe diarrheal disease in animals and humans. There are currently no effective chemotherapeutic agents available for the treatment of cryptosporidiosis. Recently, small, naturally occurring antimicrobial lytic peptides with anti-protozoal activities have been described. In the present study, we compare the in vitro anti-cryptosporidial activities of synthetic lytic peptides and their corresponding hemolytic activities after a 30 min incubation at 37 degrees C. Sporozoite viability was assessed microscopically by the uptake of the vital dyes fluorescein diacetate (FDA) and propidium iodide (PI). Hemolysis was assessed spectrophotometrically by the release of soluble hemoglobulin. The most active peptide, Hecate-1, reduced sporozoite viability by 85.5% with a corresponding hemolytic activity of 21.5% at a concentration of 10 microM.

Amino Acid Sequence↗
PubMedDate not supplied

Cryptosporidial infections in SCID mice reconstituted with human or murine lymphocytes.

Severe combined immunodeficient (SCID) mice were experimentally infected with Cryptosporidium parvum. Adoptive transfer of BALB/c thymocytes, spleen and bone marrow cells resulted in functional immunologic reconstitution followed by complete eradication of the cryptosporidial infection. Additional SCID mice were injected with human blood peripheral blood lymphocytes and were subsequently infected with C. parvum. The latter mice (SCID-hu-PBL) were at least partially reconstituted with human lymphoid tissues, as evidenced by flow cytometric identification of human cell populations in the SCID mouse spleens and the response of these cells to the T-cell mitogen phytohemagglutinin. The SCID-hu-PBL mice did not resolve the cryptosporidial infections, although a transient reduction in parasitemia was noted 4-6 wk post-reconstitution.

Animals↗
PubMedDate not supplied

Effects of dexamethasone and dehydroepiandrosterone in immunosuppressed rats infected with Cryptosporidium parvum.

Treatment of rats immunosuppressed with dexamethasone and inoculated with Cryptosporidium parvum oocysts were treated with dehydroepiandrosterone. A significant reduction in cryptosporidial activity was observed as determined by oocyst shedding and colonization of host tissue by parasites. Dexamethasone treatment alone resulted in decreases in T-, B- and natural killer (NK) cell responses and antibody production that, with the exception of NK-cell activity, were all reversed after administration of dehydroepiandrosterone.

Animals↗
PubMedDate not supplied

Increasing the yield of Eimeria tenella oocysts in primary chicken kidney cells.

The present study was undertaken to increase the yield of Eimeria tenella oocysts in primary chicken kidney cells (PCKCs) using a comparatively inexpensive cell-culture system. PCKCs growing on coverslips positioned on the bottoms of individual wells in 24-well tissue-culture plates were infected with sporozoites of E. tenella. The effects of changing the culture medium (RPMI 1640), medium pH, serum type, and serum concentration in the wells were determined by counting newly produced oocysts at 7 days postinoculation. There were significantly more (P < 0.01) oocysts produced when the medium was supplemented with 10% fetal bovine serum (FBS) and changed either daily or every other day compared with not changing the medium. When the same medium was changed daily, significantly more (P < 0.05) oocysts were produced at pH 7.4 than at pH 8.0 but not at pH 6.0. If the medium was changed daily, significantly more (P < 0.05) oocysts were produced when medium was supplemented with 10% FBS only rather than 5% and 10% chicken serum. The cell-culture system described in this study offers a convenient and efficient method for investigating the biological, biochemical, and immunological relationships between E. tenella and the host cell.

Animals↗
PubMedDate not supplied

Adapting Eimeria tenella to grow in primary chicken kidney cells following repeated passages between cell culture and chickens.

The present study was undertaken to adapt a field isolate (F0) of Eimeria tenella to grow in primary chicken kidney cells (PCKCs) by selecting for characteristics of the parasite rather than modifying the culture and/or environmental conditions. Fourteen generations (F1 to F14) of E. tenella were produced following repeated passages between PCKCs and chickens. Although F1 yielded only a 28% increase in oocysts, in PCKCs compared with F0, F2 to F5 produced from 259% to 277% more oocysts, respectively. There was no significant increase in the percentage of oocysts produced in PCKCs by F6 to F14 compared with F5. Generations F1 to F14 demonstrated a greater propensity for multiple infections within the same host cell than did F0. For example, it was not uncommon to observe two, three, and occasionally four oocysts within a single PCKC. Chickens inoculated with F0 oocysts generally experienced greater pathogenesis by day 7 postinoculation than chickens inoculated with F14 oocysts as measured by decreased body weights, increased cecal lesions, and a higher mortality rate.

Animals↗