concanavalin-a and Candidiasis

In a previous study, our group verified that 100% of mice survived to a lethal dose of Candida albicans following pretreatment with concanavalin-A (Con-A) for 3 days. This work proposed to investigate whether treatment could mediate an adaptative immune response involving T(H) 17 cells. A significant increase in IL-17 levels at 6 h postinfection was observed and was maintained up to 18 h in the Con-A group, whereas in control mice, a reduction in this cytokine was verified. In addition, T(H) 17 cells develop in the presence of TGF-β, IL-1 β, and IL-6 that were increased significantly 2 h postinfection in Con-A-treated mice. Macrophages were involved in the process, engulfing greater numbers of yeast cells, and were activated through TNF-α and interferon-γ produced at significant levels at 2 h postinfection. A significant increase in IL-12 levels was also observed at 2 h postinfection. Thus, activated macrophages were probably more capable of killing and processing Candida antigens, signalizing an adaptative immune response. Macrophages from controls did not prevent yeast-to-hyphae transition and were partially destroyed, as shown in scanning microscopy. These results suggest that treatment with Con-A facilitated the triggering of T(H) 17 and T(H) 1 responses via IL-17 and IFN-γ production, leading to the resolution of C. albicans infection.

Topics: Animals; Ascitic Fluid; Candida albicans; Candidiasis; Concanavalin A; Cytokines; Interleukin-17; Macrophages; Male; Mice; Phagocytosis; Th1 Cells; Th17 Cells

As of late, numerous reports have demonstrated the multiple biological activities of polyphenolic flavonoids. Amongst these reports, some indicate that the flavonoids play an important role in inflammation therapy. In this present study, we investigated the effect of rutin, a polyphenolic flavonoid, on septic arthritis due to Candida albicans, a major etiological agent that causes fungal arthritis. To induce septic arthritis, an emulsified mixture of C. albicans cell wall and Complete Freund's Adjuvant (CACW/CFA) was injected into BALB/c mice via hind footpad route once a day, everyday, for three days. In order to determine the effect of rutin, twenty-four hours after the final injection, mice having the swollen footpad were given the flavonoid (1 mg/dose/mouse) intraperitoneally every other day for three times. The footpad-edema was measured for a period of 17 days. Results showed that the rutin treatment reduced app. 45% of the edema at the peak day (day 11) of septic arthritis (P<0.05). In addition, 6 days after the peak, there was an app. 35% additional reduction of the edema (P<0.05). We found that this anti-arthritic activity was mediated by rutin's ability to inhibit nitric oxide production from macrophages and T-cells proliferation. Furthermore, this flavonoid also inhibited the growth of C. albicans yeast cells (P<0.01) and resulted in no hemolysis. These data indicate that rutin, which has both anti-arthritic and antifungal effects, can safely be administered into the blood circulation for treatment of septic arthritis caused by C. albicans. Ultimately, it can be suggested that the dual effects of rutin, anti-arthritic and anti-candidal may be helpful as an all-in-one treatment for septic arthritis.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Antifungal Agents; Arthritis, Experimental; Arthritis, Infectious; Candida albicans; Candidiasis; Cell Proliferation; Concanavalin A; Female; Hemolysis; Lipopolysaccharides; Macrophages; Mice; Mice, Inbred BALB C; Mitogens; Nitric Oxide; Rutin; T-Lymphocytes

The mechanisms through which Candida albicans is recognized by immune cells and how it triggers host defence are not completely understood. In this study, we evaluated the effect of Concanavalin-A on the clearance of C. albicans by infected mice and their production of proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha). Subgroups of 5 animals were pretreated with Con-A (250 mug mL(-1) PBS) and after 96 h were infected intraperitoneally with 10(7) cells of C. albicans CR15 (an isolate from a HIV+ person); 30 min, 2, 6, 24 or 72 h after infection the mice were sacrificed. Phagocytosis of C. albicans by peritoneal macrophages increased 30 min after infection in mice pretreated with Con-A. The liver presented the greatest number of CFUs, and this number was reduced by pretreatment with Con-A. Control animals infected with C. albicans presented a significant increase in plasmatic alanine aminotransferase, which was not observed in mice treated with Con-A. Two hours after infection the production of TNF-alpha in the liver of mice pretreated with Con-A was significantly increased. These results suggest that a single dose of Con-A caused a beneficial modulating action of the inflammatory response during infection with C. albicans.

Topics: Alanine Transaminase; Animals; Candida albicans; Candidiasis; Colony Count, Microbial; Concanavalin A; Disease Models, Animal; Histocytochemistry; Immunologic Factors; Liver; Macrophages, Peritoneal; Mice; Phagocytosis; Survival Analysis; Tumor Necrosis Factor-alpha

Specific epitope (DEPAGE) of Candida albicans heat shock protein 90 (SE-CA-HSP90) was successfully expressed on the surface of filamentous phage fd, fused to the major coat protein pVIII. Protective immune responses mediated by hybrid-phage expressing SE-CA-HSP90 in C57BL/6J mice were evaluated in this paper. The results showed that hybrid-phage particles induced the specific antibody response against SE-CA-HSP90, enhanced delayed-type hypersensitivity (DTH) response, natural killer (NK) cell activity and concanavalin A (ConA)-induced splenocyte proliferation. Furthermore, this study demonstrated that hybrid-phage-immunized mice had fewer colony forming unites (CFU) in the kidneys compared with wild-type phage-immunized mice and TE (1.0mM EDTA, 0.01M Tris-HCl, pH 8.0)-injected mice and showed statistically significant survival advantage over TE-injected group. In conclusion, the results suggest that the hybrid-phage displaying SE-CA-HSP90 may be considered as a potential candidate for producing vaccines against systemic candidiasis.

Topics: Animals; Antibody Specificity; Bacteriophage M13; Candida albicans; Candidiasis; Cell Proliferation; Concanavalin A; Epitopes, B-Lymphocyte; HSP90 Heat-Shock Proteins; Hypersensitivity, Delayed; Immunity, Cellular; Killer Cells, Natural; Mice; Mice, Inbred C57BL; Spleen

In this study we tested the hypothesis that after administration of a single intraperitoneal dose of concanavalin A (Con-A) to mice, the proportion of neutrophils and macrophages in the peritoneal exudate and their phagocytic and candidacidal activities should change with time. The number of neutrophils in the peritoneal exudate was greatly increased 6 h after administration of Con-A, and those cells were able to kill both intracellular and extracellular yeast and germ tube forms of Candida albicans. Addition of catalase to the culture medium reduced the killing of C. albicans, suggesting that the candidacidal activity depended on the myeloperoxidase system. The survival of mice pretreated with Con-A and submitted to an inoculum of C. albicans 6 h afterwards was twice higher than that of controls, which suggests that neutrophils were able to clear the experimental infection. One day after the treatment, the population of neutrophils in the exudate was about 45%, but after 2 days it was reduced to only 5% and the candidacidal activity was also reduced. After 4 days the exudate contained over 95% of macrophages, the candidacidal activity reached a maximum, and the phagocytosis mediated by both complement receptors and mannose receptors was increased. Uptake of FITC-mannose-BSA by macrophages was maximal on about the 4th day and was inhibited by mannan, suggesting that treatment with Con-A increased the activity of mannose receptors. These results support the hypothesis that activation of cellular immunity by Con-A occurred in two phases, one dominated by neutrophils, and the other by macrophages expressing increased activity of mannose receptors.

Topics: Animals; Candida albicans; Candidiasis; Concanavalin A; Lectins, C-Type; Macrophage Activation; Macrophages, Peritoneal; Male; Mannose Receptor; Mannose-Binding Lectins; Mice; Neutrophil Infiltration; Neutrophils; Phagocytosis; Receptors, Cell Surface

The lectin-mediated agglutination kinetics of Candida albicans, Candida tropicalis, Candida glabrata, Candida krusei, Candida kefyr, and Candida parapsilosis strains isolated from immunocompromised patients was investigated. The rate of the lectin-induced cell agglutination depends on the physiological state of the yeast cell population. Therefore, the Candida strains have to be cultivated and investigated under identical conditions. Lentil lectin (prepared from Lens culinaris), castor lectin, and concanavalin A were used. Different yeast species showed different agglutination behaviour. Furthermore, the lectin-mediated rate of agglutination is a strain-specific property which makes it possible to distinguish between different yeast strains of the same species. It is concluded that the lectin-mediated agglutination kinetics allows reproducible differentiation of yeast strains of the same species.

Topics: Agglutination; Agglutination Tests; Candida; Candidiasis; Concanavalin A; Humans; Immunocompromised Host; Lectins; Plant Lectins; Species Specificity; Time Factors

Germ-free C57BL/6 x 129 interferon-gamma knockout (IFN-gamma(-/-)) mice and their immunocompetent (+/-, +/+) counterparts were colonized with a pure culture of Candida albicans to assess their natural susceptibility to mucosal and systemic candidiasis of endogenous origin. Colonization with a pure culture of C. albicans was not lethal for adult or neonatal IFN-gamma(-/-) gnotobiotic mice over the 15-week study. The IFN-gamma(-/-) mice were more susceptible to gastric (cardia-antrum section), anorectal, and acute systemic (intravenous challenge) candidiasis than immunocompetent controls, and some IFN-gamma(-/-) mice developed intestinal adenomas after colonization with C. albicans. The enhanced susceptibility of IFN-gamma(-/-) mice, compared with immunocompetent controls, may be associated with a poor proliferative response of spleen cells to C. albicans antigens and a T helper 2 (IgG1) serum antibody response to C. albicans antigens. Thus, IFN-gamma is important for murine resistance to gastric, anorectal, and acute systemic candidiasis.

Topics: Adenoma; Administration, Oral; Animals; Antibodies, Fungal; Candidiasis; Cell Division; Concanavalin A; Disease Models, Animal; Disease Susceptibility; Female; Genotype; Germ-Free Life; Interferon-gamma; Intestinal Neoplasms; Lipopolysaccharides; Lymphocytes; Mice; Mice, Inbred C57BL; Mice, Knockout; Mitogens

In vitro studies have suggested a role for interferon gamma (IFN-gamma) in host defense against disseminated candidiasis, but in vivo studies are inconclusive. We utilized homozygous IFN-gamma knockout (GKO) mice to determine if the cytokine is essential in host defense against this disease. Genotypes of mice were determined by PCR with specific primers for the normal or disrupted IFN-gamma gene. The GKO status of the mice was confirmed by an enzyme-linked immunosorbent assay, which showed no detectable IFN-gamma produced by their splenocytes stimulated by concanavalin A. To test the susceptibility of GKO mice to candidiasis, the animals were infected either intravenously (i.v.) or intragastrically (i.g.) with Candida albicans. GKO mice infected i.v. survived as long as wild-type (WT) mice and showed no difference in Candida CFU counts in liver, spleen, or kidneys compared to those for WT mice. When animals were given Candida i.g., at 3 h or at 10 or 21 days after infection, there was no dissemination of Candida to the lung, liver, spleen, or kidneys for either GKO or WT mice. There was no difference in Candida CFU counts recovered from the stomach or intestines between GKO and WT mice. Histological examination of the stomach cardial-atrium fold, where the fungus was located, showed that GKO mice did not have evidence of more tissue damage or fungal invasion than WT mice. Finally, the jejunum for both types of mice showed no evidence of tissue damage or fungal invasion. These studies indicate that IFN-gamma is not essential in host defense against C. albicans that originates from a mucosal site or that is given directly into the bloodstream in a mouse model.

Topics: Animals; B-Lymphocytes; Candidiasis; Colony Count, Microbial; Concanavalin A; Enzyme-Linked Immunosorbent Assay; Interferon-gamma; Intestines; Jejunum; Kidney; Liver; Mice; Mice, Inbred BALB C; Mice, Knockout; Polymerase Chain Reaction; Spleen; Stomach

Surfactant protein A (SP-A) contributes to host defense by opsonizing microbial organisms for phagocytosis by alveolar macrophages (AM). The role of SP-A as opsonin for phagocytosis of Candida albicans was analyzed. AM in suspension exhibited no phagocytosis of nonopsonized yeast. This was not increased by SP-A, whether provided for preincubation of AM or yeast or present during coincubation. However, the engulfment of serum-opsonized yeast by AM in suspension was inhibited by SP-A. This inhibitory effect was mimicked by complement subcomponent C1q and concanavalin A but not by type IV collagen. SP-A did not interfere with phagocytosis of serum-opsonized yeast by adherent AM, monocytes, neutrophils, or peritoneal macrophages. SP-A lacks function as an opsonin for the phagocytosis of C. albicans by AM but interferes with binding of yeast to AM, inhibiting subsequent ingestion. The role of SP-A as an alveolar space opsonin may thus critically depend on the microbial species involved.

Topics: Animals; Candidiasis; Collagen; Complement C1q; Concanavalin A; Female; Flow Cytometry; Macrophages, Alveolar; Macrophages, Peritoneal; Male; Monocytes; Neutrophils; Opsonin Proteins; Phagocytosis; Proteolipids; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants; Rabbits

The effect of the cilofungin, a beta 1-3 glucan synthase inhibitor, on the incorporation of the glucan associated proteins (GAP) into the mycelial wall of Candida albicans was investigated. For this study sub-inhibitory (< 2 micrograms/ml) doses of cilofungin were employed during the yeast to mycelial transition in a defined chemical medium, at 37 degrees C for 24 hours. Under these conditions, and particularly at the dose of 0.50 micrograms/ml cilofungin exerted a marked effect on GAP incorporation into the mycelial cell wall. The changes were essentially the absence of the two prominent bands of 46 and 31 kDa of the untreated cell wall coupled with an apparent increase in the amount of 55-56 kDa constituent, as well as of a minor constituent of 27-28 kDa. Radiolabel incorporation experiments demonstrated increased synthesis of a 34 kDa GAP, in addition to confirming the absence of the 46 kDa constituent, in mycelial cells under cilofunging treatment. Thus, sub-inhibitory doses of cilofungin may greatly alter the pattern of essential cell wall constituents such as the glucan-associated proteins, suggesting that this drug also has important effects on cell wall structure and fine organization, independent of, or prior to, its principal lytic effect on the fungal organism.

Topics: Blotting, Western; Candida albicans; Candidiasis; Cell Wall; Concanavalin A; Echinocandins; Electrophoresis, Polyacrylamide Gel; Fungal Proteins; Glucans; Peptides, Cyclic

The complement C3d-binding protein (CR2) of Candida albicans has been purified by immunoaffinity chromatography, and its specificity has been characterized by immunoblotting with monoclonal antibodies to the C. albicans CR2 and the mammalian CR2. Recent studies with immunoelectron microscopy indicated that the CR2 was expressed during a systemic infection in a murine model of candidiasis. As a continuation of these observations, the immunogenicity of the C. albicans CR2 was investigated in a lymphoblastogenesis assay. Lymph node cells as well as splenic lymphocytes from mice infected subcutaneously with viable blastoconidia of C. albicans reacted to the C. albicans CR2 to a significantly greater extent than did lymphocytes from uninfected mice (P less than 0.01). The maximum stimulation of splenic lymphocytes by the purified receptor occurred at a concentration of 0.54 micrograms of protein per ml after 72 h of incubation of lymphocytes and receptor. Also, splenocytes from infected or CR2-immunized mice exhibited significantly reduced responses to the T-cell-dependent mitogen phytohemagglutinin (P less than 0.01). These data indicate that lymphocytes from infected mice respond to the C. albicans CR2 in a lymphoproliferation assay to a greater extent than do lymphocytes from uninfected mice, indicating that the CR2 is expressed in vivo.

Topics: Animals; Antibodies, Monoclonal; Antigens, Differentiation, B-Lymphocyte; Blotting, Western; Candida albicans; Candidiasis; Chromatography, Affinity; Concanavalin A; Disease Models, Animal; Epitopes; Immunization; Lipopolysaccharides; Lymph Nodes; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Phytohemagglutinins; Pokeweed Mitogens; Proteins; Receptors, Complement; Receptors, Complement 3d; Spleen

Cell walls were isolated from stationary-phase cultures of Candida albicans grown at 25 degrees C or 37 degrees C, in iron-depleted and iron-sufficient conditions. Proteins solubilised from cell-wall fractions were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Approximately 40 protein bands were detected by Coomassie blue staining in all wall extracts, regardless of temperature or other growth condition. Sera from patients with oral or systemic candidosis, from whom the isolates were obtained, and pooled normal human serum were examined for the presence of IgG and IgM antibodies to cell-wall proteins by Western blotting. Patient sera recognised more antigens than pooled normal human serum. In particular, an antigen of 44 kda was detected by IgG antibodies in the sera of patients and two antigens of 41 and 14 kda were detected by their IgM antibodies when the sera were used as probes against walls from iron-depleted cells, but not from iron-sufficient cells, grown at 25 degrees C. Two antigens of 45 and 40 kda were detected by IgM antibodies in the sera of patients tested against walls from iron-depleted but not from iron-sufficient cells grown at 37 degrees C. IgG antibodies did not distinguish between these wall preparations from cells grown at 37 degrees C. These results suggest that the specific cell-wall proteins induced during growth in iron-depleted conditions, as well as other proteins, were immunogenic and were recognised by the patients' antibodies.

Topics: Antibodies, Fungal; Antigens, Fungal; Antigens, Surface; Blotting, Western; Candida albicans; Candidiasis; Cell Wall; Concanavalin A; Electrophoresis, Polyacrylamide Gel; Humans; Iron; Temperature

The ability of murine recombinant gamma interferon (IFN) or lymphokines to enhance the fungicidal activity of murine pulmonary macrophages (PuM) was studied in in vitro. PuM monolayers were incubated overnight with IFN, lymph node cells (LNC) plus concanavalin A, supernatants from Con A stimulated LNC or spleen cell cultures (Con A Sup), or tissue culture medium (TCM) +/- Con A (5 micrograms/ml) or +/- lipopolysaccharide (LPS, 10 ng to 10 micrograms/ml). After treatment, culture fluids were removed and PuM were challenged for 4 h with the yeast-form Blastomyces dermatitidis or 2 h with Candida albicans. Inoculum colony forming units (CFU) of B. dermatitidis were significantly reduced by PuM treated with 1000 U/ml of IFN (25 +/- 3%), Con A Sup (25 +/- 3%) or LNC plus Con A (37-44%), but not by TCM, ConA or LPS. Candida albicans was killed by PuM treated with Con A Sup (33 +/- 8%) or LNC plus Con A (30-43%), but not by TCM, Con A, or LPS, and the activity of Con A Sup was neutralized by anti-IFN antibody. Candida albicans was not significantly killed by PuM treated with IFN doses ranging from 1 to 10(5) U/ml; nor did addition of LPS to IFN, or prolonged (3 day) treatment with IFN, result in significant killing of C. albicans by PuM. However, IFN (100 U/ml) could activate resident peritoneal macrophages for significant candidacidal activity (63%). These data indicate that PuM can be activated for fungicidal activity, and that PuM differ from resident peritoneal macrophages with regard to induction of candidacidal activity by recombinant gamma-IFN.

Topics: Animals; Blastomycosis; Candidiasis; Concanavalin A; Interferon-gamma; Lipopolysaccharides; Lung; Lymph Nodes; Lymphokines; Macrophage Activation; Male; Mice; Mice, Inbred BALB C; Mycoses; Spleen

To investigate whether there was an immunologic basis for recurrent Candida albicans-induced vaginitis, peripheral blood lymphocytes were isolated from six women with this disorder and from six healthy control women. No differences were observed in the proliferative responses of peripheral blood lymphocytes to mitogens. However, only the control peripheral blood lymphocytes proliferated in response to a C. albicans extract. Furthermore, patients' lymphocytes or serum suppressed the proliferative response of control lymphocytes to C. albicans but not to mitogens. Women with recurrent C. albicans vaginitis appear to produce Candida-specific suppressor lymphocytes which block the cellular immune response to this organism.

Topics: Blood; Candida albicans; Candidiasis; Concanavalin A; Female; Humans; Immune Tolerance; Lymphocyte Activation; Phytohemagglutinins; Recurrence; T-Lymphocytes, Regulatory; Vaginitis

Four paediatrics patients affected by Chronic Mucocutaneous Candidiasis associated with hypoparathyroidism have been studied. In vivo and in vitro responses to Candida albicans antigens and to other common antigens (Dermatophytin T, Varidase), the percentage of T and B cells and the response to the selective polyclonal phytomitogens (PHA, ConA, PWM) have been investigated in order to quantitate and functionally characterize the B and T lymphocyte classes. The HLA A an B typing has been also performed. Our results show a remarkable and most likely primitive impairment of the T cells, unresponsive either to the C. albicans and the two other skin test antigens or to the selective polyclonal mitogens in vitro. The infectious disease (Mucocandidiasis) is most likely secondary to the T-cell failure. The association between the Mucocandidiasis, hypoparathyroidism and the T-cell deficiency is probably due to a common primitive disembryogenetic defect.

Topics: Antigens, Fungal; B-Lymphocytes; Candida albicans; Candidiasis; Candidiasis, Chronic Mucocutaneous; Child; Concanavalin A; Female; HLA Antigens; HLA-A Antigens; HLA-B Antigens; Humans; Hypoparathyroidism; Lymphocyte Activation; Male; Phytohemagglutinins; Pokeweed Mitogens; T-Lymphocytes

Topics: Animals; Antigens, Fungal; Candida albicans; Candidiasis; Concanavalin A; Endocarditis; Female; Heart Valves; Lymphocyte Activation; Rabbits

Three hundred fifty human sera were tested by double immunodiffusion, crossed-line electrophoresis, and crossed immuno-affinoelectrophoresis with a concanavalin A intermediate gel for precipitating antibodies to antigens present in cytoplasmic extracts of Candida albicans. Sera from 48 of 287 hospitalized patients at risk of invasive candidiasis contained precipitating antibodies to Candida antigens. Of these 48 sera, 27 had precipitating antibodies only to cell-wall antigens present in the cytoplasmic extract, and 21 sera had precipitating antibodies to both cytoplasmic and cell-wall antigens. The latter sera came from patients who were 2.5 times as likely to have deep-seated candidiasis as those patients with precipitins exclusively to cell-wall antigens. Sera from seven of 22 patients with vaginal candidiasis and 10 of 41 patients with other fungal infections had precipitating antibodies to C. albicans cell-wall antigens; only two of these sera also contained precipitating antibodies to the cytoplasmic antigens. Crossed immunoaffinoelectrophoresis with concanavalin A reduced the number of false-positive results and increased the predictive value positive of the precipitin test for deep-seated candidiasis from 31% to 71%.

Topics: Antigens, Fungal; Candidiasis; Concanavalin A; Evaluation Studies as Topic; Female; Humans; Immunodiffusion; Immunoelectrophoresis; Precipitin Tests

Topics: Bone Marrow Examination; Candidiasis; Concanavalin A; Cytomegalovirus Infections; Fluorescent Antibody Technique; Histocompatibility Antigens; Humans; Hypersensitivity, Delayed; Immune Sera; Immunity, Maternally-Acquired; Immunoglobulins; Immunologic Deficiency Syndromes; Infant; Lectins; Lymph Nodes; Lymphocyte Activation; Male; Mitogens; Palatine Tonsil; Pedigree; Pneumonia, Pneumocystis; Thymidine; Tritium