Influence of Candida albicans glycoprotein on 3-methylcholanthrene induced malignancy in newborn rodents

In the experimental conditions reportedCandida albicans glycoprotein has a stimulating effect on the course of 3-methylcholanthrene carcinogenesis in newborn rodents. Stimulation of 3-methylcholanthrene carcinogenesis in newborn rats was found following injections of 9.5 µg/g and 18.5 µg/g ofCandida albicans glycoprotein. Subcutaneous tumors occurred in the experimental animals earlier, with higher frequency and attained larger dimensions than in the control animals. Similar effects were observed in mice. In the experimental mice there was also a significantly higher number of thymomas. From the evidence presentedCandida albicans glycoprotein appears to act as a cocarcinogenic substance to 3-methylcholanthrene.This work was supported partially byDora Kaplan, Joan Sloan, Cathy Cooper Memorial Funds and the Leo Roon Foundation.     

Seizure induction by pentylenetetrazol in animals experimentally infected with Candida albicans

Summary The prolonged presence ofCandida albicans in the spleen of experimental rats and mice predisposes them to pentylenetetrazol induced seizures. The seizure threshold of the experimental animals is lowered. This becomes statistically significant at ten months after the operation. In the experimental animals the latent period was sometimes reduced. The convulsions in experimental rats and mice were prolonged, sometimes repeated, and on three occasions (2 rats and 1 mouse) ended in death. Electroencephalographic studies confirmed the increased proneness to seizures in the animals to whichCandida albicans was introduced to the spleen.This work was supported by the Damon Runyon Grant no 720, and Joan Sloan Memorial Fund.     

Effects of surface active agents on the susceptibility of Swiss mice to Candida albicans

Summary The LD50's ofCandida albicans combined with Plurafac B26, Mulsor 224, Pluronic L62, and Polyethylene Glycol 400 Mono Laurate injected intraperitoneally into female white Swiss mice were calculated by the method ofReed &Muench. Each of the previously mentioned surface-active agent —Candida albicans combinations had a lower LD50 than theCandida albicans control.Plurafac B26 was found to intially decrease the number of leukocytes in the peritoneal cavity and thus enhance the invasiveness ofCandida albicans.     

Occurrence of atypical lymphocytes following intravenous injection of Candida albicans in mice

The occurrence of atypical lymphocytes has been observed in the course of experimental acute infection withCandida albicans in mice. In animals injected intravenously with 2.5 × 10⁶ ofCandida albicans cells, an increased number of monocytes was seen in 24 hours. Monocytes showed toxic vacuolisation in most instances in protoplasm and sometimes in the nuclei. Only a few atypical lymphocytes could be seen at that time. In the following days the number of monocytes diminished and the number of atypical lymphocytes increased. After four days atypical lymphocytes constituted frequently over 20 % of white cells. The autopsy of sacrificed or dead animals with the presence of such elevated percentages of atypical lymphocytes showed enlargement of cervical lymphnodes in all animals. In mice infected with 1.4 × 10³ ofCandida albicans cells, no level higher than 12% of atypical lymphocytes was seen. Pictures were returning to normal with only a few atypical lymphocytes present among the animals which survived for two months after infection withCandida albicans.This work was supported partially byDora Kaplan, Joan Sloan, Cathy Cooper. Memorial Funds and the Roon Foundation.     

Production of glycoprotein by Candida albicans in a synthetic medium and its influence on the growth of newborn mice

Summary Growth ofCandida albicans on a synthetic medium for a period of 6 weeks produces glycoprotein substance in the approximate amount of 150 mg per liter. The polysaccharide component is formed by glucose and mannose in the approximate ratio 3:1. The protein component is composed of at least 15 different amino acids. Half percent of glucosamine was also found. The glycoprotein substance is water soluble and toxic to Swiss mice. LD50 is 0.75 mg/g of body weight when injected intravenously. Subcutaneous injection to newborn Swiss mice produced inhibition of growth. The degree of inhibition varied with the dosage. On one occasion when using a small amount of compound, stimulation of growth was also seen.This work was supported by Damon Runyon Memorial Fund Grant 720 and Joan Sloan Memorial Fund.     

Increased carbohydrate content in the thymus and spleen in mice receiving Candida albicans orally (occurence of lymphomas)

Following oral administration ofCancida albicans to non — inbred albino mice in the experimental conditions reported, an increased level of carbohydrate was found in the spleen and thymus. Significant increase of carbohydrate content in these organs was found to occur in the third month of the experiment and remained so, during the ten months of its duration with the exception of the fourth and fifth month. The carbohydrate content in other essential organs was in a similar range in the control and experimental animals. No significant differences in protein level in the control and experimental animals were found. This was regarded as an indication that the incresaed carbohydrate level in the thymus and spleen of experimental animals represents their reaction to the pathological influences of metabolic products ofCandida albicans.Appearance of lymphomas was observed between the seventh and tenth month. Three lymphomas were found in sixty four animals which were autopsied. Previously reported occurrence of lymphomas in mice which received injection ofCandida albicans to the spleen, provides circumstantial support that the observed increase of carbohydrate level as reported here in the spleen and thymus represents the pathological reaction towards metabolic products ofCandida albicans and forms the basis of a collapse of the defense system of the experimental animals.Thymus and spleen appear to act synergically in the reported experimental conditions and as a part of enterodefense system. The possibility of relevance of observed pathology to humans is discussed.This work was supported partially byDora Kaplan, Joan Sloan, Cathy Cooper Memorial Funds and the Roon Foundation.     

The serological grouping of Candida albicans strains isolated in the Kraków province in the years 1963–1965 by means of agar gel diffusion test

Summary One hundred forty sevenCandida albicans strains isolated from different clinical specimens were serologically differentiated. Agar gel diffusion test inBjörklund modification was applied inCandida albicans grouping. Autolysates prepared from the strains under study and from the group A and B standard strains ofHasenclever were used as antigens against anti-Candida albicans group A rabbit immune sera. Among 147 examined strains isolated from 107 patients, 137 strains were classified as belonging to group A and only 10 (9.3 %) strains as belonging to group B.     

Amylolytic activity of yeasts

The total amylolytic activity of 14 selected fermentation type I members of the generaSaccharomyces, Zygosaccharomyces, Candida andTorulopsis was studied. Fractions with α-glucosidase activity, the specificty of which was tested for maltose and sucrose, were isolated on carboxymethyl-cellulose from the intracellular contents of two strains ofCandida albicans and one ofCandida stellatoidea. The fraction from the strainCandida albicans 29-3-109, which is more virulent for mice, displayed the greatest α-glucosidase activity, moderate activity was present in the strainCandida stellatoidea 29-64-1, while the lowest activity was found in the less virulent strain ofCandida albicans, 29-3-19.     

Antigenic relationships between Aracoccidioides loboi and other pathogenic fungi determined by immunofluorescence

Summary The fluorescent antibody technique was used to study antigenic relationships betweenParacoccidioides loboi and other pathogenic fungi. The findings suggest thatP. loboi is more closely related antigenically to certainP. brasiliensis strains than to others and that it has antigens in common with the yeast form ofHistoplasma capsulatum, H. duboisii, Blastomyces dermatitidis, Candida albicans and also the mycelial form ofCoccidioides immitis. Serum globulins from 3 cases of keloidal blastomycosis were labelled with fluorescein isothiocyanate. These conjugates showed slight or no reactivity withP. loboi, the yeast forms ofP. brasiliensis, H. capsulatum, H. duboisii andB. dermatitidis, However, they stained brightlyC. albicans, serotypes A and B, the tissue form ofC. immitis and the yeast form ofSporotrichum schenckii. Adsorption of these reagents withC. albicans eliminated all staining except that forS. schenckii. These patients had no history of clinical sporotrichosis.Deceased. Last address: Fundacão Gonçalo Moniz, Salvador, Bahia, Brazil. Requests for reprints should be sent to Dr.William Kaplan.Dr.Miranda is in private practice in Rio de Janeiro, Brazil.     

Immunochemical studies on mannans of the genusSaccharomyces

Antigenic mannans isolated from the cells ofSaccharomyces fermentati, Saccharomyces rosei,Saccharomyces delbrueckii, Torulopsis colliculosa, Candida albicans andSaccharomyces cerevisiae were examined for their reactivity withSaccharomyces fermentati andCandida albicans antisera. Mannans ofTorulaspora as well asCandida albicans showed high cross-reactivity with the investigated antisera, which could be due to the presence of long side chains established by the partial acetolysis method. The low specific rotations ofSaccharomyces fermentati, Saccharomyces rosei andTorulopsis colliculosa mannans indicate a predominance of β-glycosidio linkages, whereasSaccharomyces delbrueckii andCandida albicans mannans possess predominantly α-linkages.Saccharomyces cerevisiae mannan showed different structural and immunological properties.     

Occurrence of lymphomas and lupus erythematosus-like syndrome in the course of experimental Candida albicans infection in mice

Occurrence of four lymphomas was seen following injection of 1.4×10³ Candida albicans cells to the spleen of eighty non - inbred albine mice. These neoplastic changes occurred in three to seven months of the duration of the experiment. Extensive invasion of the subcutaneous tissue by the tumor was found in one animal. Two of the animals with lymphoma also had changes in the kidneys compatible with lupus erythematosus and presence of LE cells in the blood. In other animals most frequent and extensive pathological changes were found in the kidneys and in order of frequency were as follows: thickening of the basement membrane, fibrinoid degeneration, presence of hematoxylin bodies, wireloop formation. In total, the presence of positive LE cells in blood was found in seven instances and suggestive LE cells was observed in eight animals. Finding of perisplenitis, exudative pleuritis and pericarditis gave additional support to the resemblance of observed pathological changes with human lupus erythematosus. The fact that occurrence of lymphomas and lupus erythematosus together has been reported in humans provides additional interest to the results reported.Results presented here and previously suggest that there may be more extensive interrelationship among variety of pathological changes observed than is accepted until now. Better understanding of host — parasite relationship ofCandida albicans, and possible other fungi, could advance out knowledge of pathogenesis of these diseases.This work was supported partially byDora Kaplan, Joan Sloan, Memorial Funds and the Roon Foundation.     

Interactions ofCandida albicans with bacteria and salivary molecules in oral biofilms

The yeastCandida albicans coaggregates with a variety of streptococcal species, an interaction that may promote oral colonization by yeast cells.C. albicans andCandida tropicalis are the yeasts most frequently isolated from the human oral cavity and our data demonstrate that both these species bind toStreptococcus gordonii NCTC 7869 while two otherCandida species (Candida krusei andCandida kefyr) do not. Adherence ofC. albicans was greatest when the yeast had been grown at 30° C to mid-exponential growth phase. For 21 strains ofC. albicans there was a positive correlation between the ability to adhere toS. gordonii and adherence to experimental salivary pellicle. Whole saliva either stimulated or slightly inhibited adherence ofC. albicans toS. gordonii depending on the streptococcal growth conditions. The results suggest that the major salivary adhesins and coaggregation adhesins ofC. albicans are co-expressed.     

Epidemiological investigations of oral Candida Albicans

Using a gargle-rinse technique, the oral cavities of 103 volunteers were sampled and cultured for the presence ofCandida albicans. Thirty-six (33.95 %) were positive forC. albicans, including 14 females and 22 males. Sixty-four subjects, including negative controls, were placed on treatment regimes of a pre-sleep gargle-rinse with either sterile distilled water (W) or Cepacol® Mouthwash/Gargle (C). The possible effects of ambient temperature, diet, age, sex, and mouthwash use on oralC. albicans levels are illustrated and discussed, including some evidence for familial endemicity. On simulated sporadic or continuous mouthwash use, some individuals showed statistically significant reductions in oralC. albicans flora, whereas others had biologically significant reductions that were not confirmed statistically. A few originally negative individuals developed non-persistent lowC. albicans counts on one or two days. Total bacterial counts were made for 32 subjects, for most of whom biologically significant reductions were obtained, although the counts were highly variable and erratic. The data support the concept that a reduction in oralC. albicans does not lead to an increase in total bacterial flora, and vice versa.with the technical assistance ofAlyce R. Schmitt Paper 741, Department of Botany, The Ohio State University. This investigation was supported by a research grant form the Wm. S. Merrell Co., Cinninnati, O.     

Effect of Candida albicans dsDNA in Gastrointestinal Candida Infection

Neonates are highly sensitive to infections because they are biased to develop Th2 immune responses. When exposed to certain agents, such as DNA vaccines or CpG DNA motifs, neonates are capable to mount adult-like Th1 protective responses. This study investigates the capacity of Candida albicans (C. albicans) dsDNA to induce host resistance in newborn mice against gastrointestinal C. albicans infection. The protective properties of dsDNA are related to an increased number of spleen CD4+ T cells secreting IFN-γ. In infected DNA-treated mice, an enhanced production of IFN-γ by Peyer’s patch cells was observed together with reduced colonization and histopathological changes in the stomach. Our results indicated that C. albicans dsDNA administration in neonates elicited the protective immune response against gastrointestinal Candida infection.     

A possible function of the chilamydospores of Candida albicans

Strains ofCandida albicans produce large numbers of chlamydospores in a liquid medium composed of 1% sodium taurocholate in distilled water. Studies made of these chlamydospores revealed that they do not contain endospores or ascospores but they do contain globules which are lipoid in character. These globules are extruded when chlamydospores split in the liquid medium or under slight pressure. Evidence was not obtained which could support van der Walt's (1967, 1969) statements to the effect that chlamydospores bud, germinate or change into other types of cells. Furthermore, no morphological or cytological evidence was found to substantiate the proposed life cycle ofC. albicans as outlined by van der Walt (1969). It is suggested that a possible function of the chlamydospores ofC. albicans is that of a storage cell.     

Protective effect of fungal extracellular vesicles against murine candidiasis

Extracellular vesicles (EVs) are lipid bilayered compartments released by virtually all living cells, including fungi. Among the diverse molecules carried by fungal EVs, a number of immunogens, virulence factors and regulators have been characterised. Within EVs, these components could potentially impact disease outcomes by interacting with the host. From this perspective, we previously demonstrated that EVs from Candida albicans could be taken up by and activate macrophages and dendritic cells to produce cytokines and express costimulatory molecules. Moreover, pre‐treatment of Galleria mellonella larvae with fungal EVs protected the insects against a subsequent lethal infection with C. albicans yeasts. These data indicate that C. albicans EVs are multi‐antigenic compartments that activate the innate immune system and could be exploited as vaccine formulations. Here, we investigated whether immunisation with C. albicans EVs induces a protective effect against murine candidiasis in immunosuppressed mice. Total and fungal antigen‐specific serum IgG antibodies increased by 21 days after immunisation, confirming the efficacy of the protocol. Vaccination decreased fungal burden in the liver, spleen and kidney of mice challenged with C. albicans. Splenic levels of cytokines indicated a lower inflammatory response in mice immunised with EVs when compared with EVs + Freund's adjuvant (ADJ). Higher levels of IL‐12p70, TNFα and IFNγ were detected in mice vaccinated with EVs + ADJ, while IL‐12p70, TGFβ, IL‐4 and IL‐10 were increased when no adjuvants were added. Full protection of lethally challenged mice was observed when EVs were administered, regardless the presence of adjuvant. Physical properties of the EVs were also investigated and EVs produced by C. albicans were relatively stable after storage at 4, ?20 or ?80°C, keeping their ability to activate dendritic cells and to protect G. mellonella against a lethal candidiasis. Our data suggest that fungal EVs could be a safe source of antigens to be exploited in vaccine formulations.     

The effect of the supernatants obtained fromSporothrix schenkii andCandida albicans culture on the generation of reactive oxygen species by polymorphonuclear leukocytes

The effect of the supernatants obtained from the liquid culture medium ofSporothrix schenkii andCandida albicans on the generation of superoxide anion (O ₂ ^– and hydroxyl radicals OH_., the elements of reactive oxygen species (ROS), and chemilimunescence (CL), a measure of several ROS, by polymorphonuclear leukocytes (PMNs) was examined. In our study, it was shown that the supernatant ofS. schenkii increased all types of ROS generation examined and CL, while that ofC. albicans increased OH_. generation and CL. The effect of the supernatants ofS. schenkii on OH_. generation and CL and that ofC. albicans on CL were most remarkable when the supernatant obtained 8 weeks after the inoculation was used. The supernatant ofS. schenkii was shown to be a much more potent stimulant than the supernatant ofC. albicans. This ROS-stimulating effect of the supernatant ofS. schenkii was heat stable but not dialyzable. These findings suggest the possible role of ROS produced by infiltrated PMNs in the inflammatory skin lesions induced byS. schenkii.     

Pathogenicity in mice of strains of Candida albicans (Robin) Berk. Isolated from burn patients

Three strains ofCandida albicans, isolated from burn patients who succumbed to systemic candidiasis, were studied in random-bred Swiss white mice to determine their relative pathogenicity and their histopathology. The most virulent strain caused 89.5 % mortality, compared to 70 % and 65 % at 21 days for the other two strains. Histopathological studies showed extensive involvement of the kidneys, with heavy colonization of the renal pelvis; pseudomycelia, budding cells, and blastospores were observed in all animals that died. Animals surviving after 21 days were sacrificed; no extreme pathological reaction nor cells ofC. albicans could be detected.Paper No. 673, Dept. of Botany, Ohio State Univ., 1735 Neil Ave., Columbus, 43210; based, in part, on a thesis presented in partial fulfillment of the M.Sc. degree.Reprint requests should be sent to the junior author at the address above.     

A combination rapid and standard method for identification of Candida albicans

A medium consisting of an aqueous extract of zein, lactose, and Tween 80 is used together with an overlay of 1 % Tween 80 and coverslipping to provide a combined rapid (germ tube) and standard (chlamydospore) method for diagnosis ofCandida albicans. The method is exquisitely sensitive for diagnosis ofC. albicans but lumps chlamydospore-producing strains ofC. tropicalis withC. albicans.     

Comparative media studies in the isolation ofCandida albicans from pregnant women

Summary Rice agar and corn meal agar, with and without Tween 80, were evaluated clinically as directly inoculable selective and differential media for the isolation ofC. albicans from vulvovaginal specimens taken from pregnant women. Chlamydospore formation on these media was investigated as a criterion for the identification ofC. albicans.Of 301 patients cultured, 118 (39.2 %) gave positive cultures for yeast-like fungi of the genusCandida. Of 118 strains for which fermentation patterns were determined, 69 (58.5 %) gave the pattern forC. albicans. Of these, 56 (81.1 %) formed chlamydospores.Tween 80 was found to exert a very stimulating influence on chlamydospore production. Rice agar with Tween 80 appeared to be the most efficient medium for eliciting chlamydospores. However, since strains of 4 out of 6 species ofCandida isolated were found to sporulate it was concluded that chlamydospore formation is not a reliable criterion for the speciation ofC. albicans.Each of the 4 media served satisfactorily as a directly inoculable selective medium for the isolation of yeast-like fungi of the genusCandida from vulvovaginal specimens. None of the media appeared to preferentially stimulate chlamydospore production inC. albicans.Dr.Smith is Associate Professor in the Department of Microbiology; Dr.Taubert is a Fellow in the Department of Obstetrics and Gynecology; Mr.Towns is Laboratory Assistant in the Department of Microbiology.Supported in part by a grant from the Lederle Medical Faculty Awards Committee and in part by United States Public Health Service Grant E-3068.