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Biomedical subjects

R F Hector

Publications and source records attributed to R F Hector.

30 records · Page 2Linked to original sources

Innate and acquired immune responses against Candida albicans in congenic B10.D2 mice with deficiency of the C5 complement component.

Congenic mice, sufficient or deficient with respect to the C5 component of complement, were evaluated for their innate and acquired immune responses to Candida albicans. When unimmunized mice were challenged intravenously and sacrificed at intervals for cultural analyses of kidneys, it was clear that C5-sufficient mice were able to deal more effectively with C. albicans during the first week after challenge than C5-deficient mice. When immunized mice were challenged intravenously to assess the development of protective responses, an intact complement cascade appeared to contribute to the more rapid clearance of fungi during the first few weeks following challenge, but by the fourth week after challenge, the numbers of fungi had decreased significantly in both types of mice and were at levels which were not significantly different. No significant differences were detected in the development of delayed hypersensitivity or Candida-specific antibody between C5-sufficient and C5-deficient mice either. C5-deficient mice did have slightly elevated levels over the C5+ mice, but this may simply reflect the prolonged antigenic load during the first 3 weeks following intravenous challenge in both immune and nonimmune animals. The later-acting complement components, while appearing to contribute to the early inhibition of the growth of C. albicans in the nonimmune animal, had no adverse effect on the development of specific immune responses, in that delayed hypersensitive responses were equivalent between the two groups, antibody response was not significantly altered and the ultimate outcome of challenge in immunized animals was not affected.

Animals↗

Synergistic action of nikkomycins X and Z with papulacandin B on whole cells and regenerating protoplasts of Candida albicans.

Combinations of nikkomycin X (NX) or nikkomycin Z (NZ), known inhibitors of chitin synthesis in fungi, together with papulacandin B (PB), an inhibitor of beta-glucan synthesis, were tested for synergistic activity against four isolates of Candida albicans by using the broth microdilution checkerboard technique and a method to assess the regeneration of cell wall material in protoplasts. The construction of isobolograms from the data generated by the checkerboard determinations revealed a synergistic effect for the two classes of compounds against all strains. The combination of NX and PB was more effective than the combination of NZ and PB, perhaps reflecting the lower Ki value of NX. While the presence of NX and NZ reduced chitin synthesis, as determined by staining with calcofluor white and assaying with a microfluorimeter, cells treated with PB demonstrated an increased synthesis of chitin. Protoplast regeneration experiments using similar concentrations of the two classes of compounds resulted in comparable findings. The combination of NX and PB resulted in a greater inhibition of chitin synthesis than did equivalent combinations of NZ and PB. These data suggest that combinations of agents active against cell wall synthesis in fungi may prove more useful as chemotherapeutic agents than such compounds used singly.

Aminoglycosides↗

A 96-well epifluorescence assay for rapid assessment of compounds inhibitory to Candida spp.

A rapid method for the screening of potential antifungal compounds was developed. A variety of compounds were tested against regenerating protoplasts of Candida spp. in a microtiter format. The degree of cell wall formation was assessed by staining with Cellufluor (Polysciences, Inc., Warrington, Pa.), a fluorochrome with known affinity for chitin, followed by determination of fluorescence by using a Dynatech Microfluor reader (Dynatech Laboratories, Inc., Alexandria, Va.). Compounds with known activity against the cell wall or cytoplasmic membrane of fungi inhibited wall synthesis in a concentration-dependent fashion. Treatment with 5-fluorocytosine, however, resulted in no inhibition. In general, protoplasts of C. albicans regenerated more quickly and were more sensitive to the compounds tested than protoplasts of C. tropicalis and C. parapsilosis. While the described method is not specific for a given class of antifungal agents, it may prove useful for testing large numbers of compounds quickly.

Antifungal Agents↗

Immunodeficient CBA/N mice respond effectively to Candida albicans.

An immune defect in CBA/N mice diminishes their ability to respond adequately to certain well-defined antigens. Since the contribution of T and B cells to immunity in candidiasis has not been clearly defined, it was hoped that CBA/N mice might prove a useful model for the study of specific responses to Candida albicans. Therefore, immunodeficient CBA/N and immunocompetent CBA/J mice were immunized by two cutaneous inoculations of viable C. albicans B311 given 2 weeks apart and challenged iv 14 days after the second inoculation. Delayed-type hypersensitivity (DTH) was tested with a membrane-derived antigen (B-HEX) 7 days following the second inoculation, and lymphocyte stimulation with B-HEX, a cytoplasmic antigen (SCS), and mitogens was done at 12 days. Antibody to SCS was determined by ELISA 2 days after DTH testing and 28 days after iv challenge, at which time the animals were sacrificed for quantitative culture of kidneys and brains. Naive CBA/N mice were no more susceptible to challenge than CBA/J mice in that the mean log colony-forming units (CFU) were 3.79 and 5.48, respectively. Both strains responded to immunization by a similar reduction in CFU, a marked DTH response (e.g., reactions at 24 hr were 1.12 mm for CBA/N and 1.34 mm for CBA/J), and significant and similar quantities of antibody. The immune defect in CBA/N mice had no demonstrable effect on the development of immune responses to infection with C. albicans.

Animals↗

High-pressure liquid chromatographic assay of Bay n 7133 in human serum.

A high-pressure liquid chromatographic method that includes a Sep-Pak (Waters Associates, Inc., Milford , Mass.) preparation of human serum was employed for the quantitative assay of Bay n 7133. Drug levels of 0.1 to 20 micrograms/ml could be detected. No interference from amphotericin B was found in the chromatographic analysis of Bay n 7133.

Amphotericin B↗

Control of systemic spread of Candida albicans with ketoconazole in the stomachs of mice treated with cytarabine.

Attempts were made to colonize the gastrointestinal tract of outbred CD-1 mice with Candida albicans or Candida tropicalis before assessing the effects of cytarabine and ketoconazole on colonization. Stable populations of C. albicans but not C. tropicalis could be maintained in the stomachs of mice infected as adults if they were maintained on penicillin and tetracycline, or in untreated mice if the fungus was administered while they were infants. Treatment with cytarabine of mice colonized with C. albicans resulted in increased levels of yeast in the stomach, and small numbers of yeasts were recovered from the livers, kidneys, and spleens. Ketoconazole limited the multiplication of the yeast in the stomachs of colonized mice, although the effect was somewhat variable. Systemic spread of the fungus was essentially eliminated in animals receiving both ketoconazole and cytarabine, suggesting that ketoconazole may be an appropriate prophylactic agent for individuals undergoing therapy with cytarabine.

Animals↗

Inhibition of chitin synthesis in the cell wall of Coccidioides immitis by polyoxin D.

The cell walls of both growth phases of Coccidioides immitis were studied by light and electron microscopy and biochemical procedures in an effort to assess the role of chitin in the fungus. Inhibition of normal chitin synthesis in the spherule by exposure to several concentrations of polyoxin D (PD) led to multiple morphological effects. Exposure of the mycelial phase to significantly higher levels of the compound had no morphological effect, as determined by autoradiography and light and electron microscopy. However, when equal masses of both morphological phases were treated with PD and pulsed with labeled N-acetylglucosamine, there was a greater relative (percent) reduction of incorporation of label in PD-treated mycelia compared with that of spherules. Nevertheless, the treated and untreated mycelia incorporated severalfold more counts than did corresponding spherules. The results suggest that chitin is important in maintaining the structural integrity of the spherule phase, but the role of chitin in the mycelial phase is less clear.

Autoradiography↗

Mammary gland contamination as a means of establishing long-term gastrointestinal colonization of infant mice with Candida albicans.

Infant outbred CD-1 mice were infected intragastrically with Candida albicans by inoculating the mammary glands of the lactating mothers with viable blastospores and allowing the infants to suckle. Levels of colonization were determined by quantitative cultures of stomachs and selected organs at various intervals up to 6 weeks after infection. The results demonstrate that a high percentage of infant mice can be colonized in this manner and that the colonization is of long duration. Although systemic spread of the yeast to other visceral organs did occur, the numbers of yeasts recovered were minimal.

Animals↗

Immune responses to Candida albicans in genetically distinct mice.

Mice from six genetically distinct strains were examined for their immune responses to Candida albicans in in vitro and in vivo assays, and naive mice and mice immunized with the fungus were challenged intravenously with three different doses of C. albicans to determine differences in susceptibility. Naive mice from the six groups showed substantial differences in resistance to challenge based on mortalities and quantitative cultures of kidneys, with mice from strains C57BL/6J and BALB/cByJ showing the most resistance; mice from strains A/J, C3H/HeJ, and CBA/J showing moderate susceptibility; and mice from strain DBA/2J showing the highest degree of susceptibility to challenge. Unimmunized mice from strains C57BL/6J and BALB/cByJ did not produce detectable levels of Candida-specific antibody by the end of the 28-day observation period when challenged intravenously, but the other strains did. Immunized mice showed a degree of protection to challenge, with all groups except mice from strain BALB/cByJ showing a reduction of two to three log units in the level of colonization in their kidneys and all strains producing significant levels of antibody. Additionally, the immunized mice of all strains developed substantial levels of delayed-type hypersensitivity and demonstrated nearly identical lymphocyte proliferative responses to Candida antigens. The results indicate that resistance to systemic candidiasis is dependent upon a combination of innate factors, predominately an intact complement system, and the acquisition of an immune response, most likely of a cell-mediated type. Additionally, the findings suggest that genetic control of acquired resistance to C. albicans may not be associated with the H-2 complex.

Animals↗

Immunological relatedness among Candida albicans and other pathogenic Candida species.

Membrane-mitochondrial (butanol-hot phosphate-buffered saline) and cytosol (soluble cytoplasmic substances) extracts from seven pathogenic species of Candida were used in in vivo and in vitro immunological assays to study antigenic similarities among the strains with respect to C. albicans. Mice were sensitized with C. albicans serotype A for footpad testing or to provide cells for lymphocyte stimulation assays, and guinea pigs were immunized with whole cells or butanol-hot phosphate-buffered saline extracts of C. albicans to obtain antisera for immunodiffusion assays. When extracts from each of the seven species were used in the assays, they consistently segregated, as determined by statistical or subjective analyses, into three groups. Extracts of C. albicans serotype A or B and C. stellatoidea were the most immunologically reactive in all assays, indicating close similarities between those two species, whereas extracts of C. tropicalis and C. parapsilosis elicited only moderate responses. Extracts from C. krusei, C. guilliermondii, and C. pseudotropicalis were hypo- or nonreactive in the assays, indicating a low level of antigenic relatedness to C. albicans.

Animals↗

[Prevention of gram-negative and gram-positive infections using 3 intravenous immunoglobulin preparations and therapy of experimental polymicrobial burn infection using intravenous Pseudomonas immunoglobulin G and ciprofloxacin in an animal model].

Three immunoglobulin preparations for intravenous infusion were compared in vivo to determine their relative protective capacity against several gram-negative and gram-positive pathogens. Polyglobin N is a conventional IgG concentrate. Psomaglobin N is identical in formulation to Polyglobin N but is prepared from the plasma of donors who have naturally high levels of antibody to lipopolysaccharide antigens of Pseudomonas aeruginosa. IgGMA is a conventional IgG concentrate containing 12% IgG and 16% IgA. In a murine model of burn wound sepsis the three IgG preparations were similarly protective against three or ten strains of P. aeruginosa. Psomaglobin N and Polyglobin N were significantly (p less than or equal to 0.015) more protective than IgG-MA against six of ten and three of ten strains of P. aeruginosa, respectively. In a murine model of Streptococcus pneumoniae type 3 pneumonia, the three Ig preparations were similarly protective. IgG-MA was significantly more protective (p less than or equal to 0.025) than Psomaglobin N and Polyglobin N against Salmonella typhimurium in murine peritonitis. However, the mean protective dose (PD50) of the two later preparations was less than or equal to 20 mg/kg body weight. In models of peritonitis both Psomaglobin N and Polyglobin N were more protective than IgGMA (p less than or equal to 0.004) against Haemophilus influenzae b, Klebsiella pneumoniae, Serratia marcescens 06:H3 and group B Streptococcus types 1b and 1c. Psomaglobin N and ciprofloxacin were employed to treat established polymicrobial murine burn wound sepsis resulting from contamination of the burn site with mixtures of P. aeruginosa and Staphylococcus aureus. Psomaglobin N or albumin was given once 16 h after challenge.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Immunoglobulin G: potentiation of tobramycin and azlocillin in the treatment of Pseudomonas aeruginosa sepsis in neutropenic mice and neutralization of exotoxin A in vivo.

Mice with cyclophosphamide-induced neutropenia were challenged with four immunotypes of Pseudomonas aeruginosa by contamination of a small dorsal surface wound. The infections were lethal; 100% of control animals (n = 80) treated only with albumin died. Administration of an immunoglobulin G intravenous preparation (IGIV) and/or therapy with tobramycin or azlocillin was begun 16 hr after challenge. Mortality among mice (n = 120) treated only with an antibiotic was 75.0%, while that among mice (n = 80) treated only with IGIV was 78.8%. Combination therapy with IGIV and an antibiotic (n = 120) resulted in mortality of 38.3%. The protection afforded by IGIV may have resulted in part from neutralization of exotoxin A, as mice treated with IGIV before challenge with exotoxin A were subject to lower mortality and had lower levels of serum aspartate and alanine aminotransferases than controls.

ADP Ribose Transferases↗