Suppressive action of cytoplasmic and metabolite extracts of Candida albicans on the immune response in guinea pigs

This study investigated the possibility that Candida albicans components exert a suppressive effect on the immune response of guinea pigs (GP), similar to that of live C. albicans organisms as was previously shown. Hartley GP were inoculated with C. albicans crude cytoplasmic or metabolite (culture filtrate) extracts (containing most of the organism's cell contents or its metabolite and degradation products, respectively). Their immunological responses towards sheep red blood cells (SRBC) were compared with those of GP inoculated with SRBC alone or with SRBC together with live C. albicans organisms. The immunological responses were measured by: 1) rosette formation (RF) of SRBC with peritoneal macrophages, 2) haemolytic plaque formation (PFC) with lymph node-cells, 3) haemagglutination and 4) haemolysis tests. According to the RF tests, inoculation of GP with either cytoplasmic or metabolite extracts resulted in descreased RF as compared to GP inoculated with SRBC only; the decrease was correlated with the protein concentration of the extracts. Inoculation with metabolite extract led to a more diminished RF than with cytoplasmic extract, but less than with live C. albicans organisms. Inoculation of cytoplasmic extracts did not affect the haemagglutinin and haemolysin titers, while that of metabolite extracts resulted in a slight decrease of these titers. The assays for PFC were not conclusive enough to point to a suppressive effect of C. albicans extracts. In summary, it appears that both the cytoplasmic and metabolite extracts of C. albicans exert a partial suppressive effect on the immune response in GP, as judged primarily on the basis of the RF results.     

Induction of the immune response suppression in mice inoculated with Candida albicans

There is a controversy in respect to the immunological response (humoral or cellular) concerning the defense against Candida albicans. Candidosis would induce sub-populations of suppressor cells in the host cell-immune response. This report tries to show the effect of different doses of C. albicans (alive or heat-killed) on the expression of cell-mediated and humoral immunity. The effect upon cell immunity was determined by inoculating different lots of singeneic mice, doses of varied concentration of C. albicans and checking for delayed-type hipersensitivity (D.T.H.). D.T.H. was also controlled in syngeneic normal mice which had previously been injected with inoculated mice spleen cells. Humoral immunity was assayed by measuring the induced blastogenesis by Pokeweed Mitogen on spleen mononuclear cells with different doses of C. albicans. Results obtained show that the different doses gave origin to: Suppression of humoral and cell response (10(8) alive); Suppression of only humoral response (10(6) alive); Suppression of cell response and increase of humoral response (10(9) dead); Increase of both responses (10(8) dead).     

Immune response in mice infected with Candida albicans in the mycelial form

The effect of the infection with the mycelial form of a Candida albicans strain (Mycology Dept.) upon the immune system in mice was studied.BALB/c mice were infected intraperitoneally in a single dose of a 3×10⁶, 6×10⁶ and 12×10⁶ cell suspension of the strain.Macrophages's activity was studied the days 7,14,21,28, 35, and 42 after inoculation, by the following assays: phagocytosis in vitro, mononucleated phagocytic system by the colloidal carbon clearance technique, the lymphocyte's activity by the direct plaque forming cells technique (PFC) and delayed hypersensitivity (DTH).Infection with the mycelial form did not affect the peritoneal macrophage's phagocytic ability, neither modified the delayed hypersensitivity to sheep red blood cells (SRBC). However, a slight and transient depression of the lymphocyte stimulation was found. Suppression of PFC to SRBC was high when a 12×10⁶ cell suspension was used in contrast to the infection with blastospores.These results suggest that systemic infection by Candida albicans in its mycelial form do not induce a non specific immunosuppression.     

Low dose of Concanavalin-A enhances innate immune response and prevents liver injury in mice infected with Candida albicans

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.     

Cellular and humoral immune responses to Candida albicans in subcutaneously infected mice

Pure mycelial and yeast cultures of Candida albicans were produced in a low sulphate medium. Groups of mice were injected subcutaneously with increasing doses of viable or heat-killed mycelial or yeast cells and the kinetics of delayed-type hypersensitivity (DTH), anti-mycelial and anti-yeast antibodies were studied. Both the dose and the morphological phase of C. albicans showed an influence on the development of the DTH, but the viability is the factor which showed the highest influence on this reaction, since on the one hand mice infected with viable yeast or mycelial cells developed higher DTH levels than mice injected with heat killed cells, and on the other hand this factor seems to play an important role in the kinetics of DTH response. The enzyme-linked immunosorbent assay has been adapted to detect antibodies to yeast and mycelial phase cytoplasmic antigens of C. albicans. In contrast with the DTH reactions, neither dose, morphological phase nor viability played an important role on the antibody titer developed. However, the use of mycelial cytoplasmic antigens seems to be better than the yeasts to detect anti -Candida antibodies over the last days studied.     

Suppression of humoral response during the course of Candida albicans infection in mice

This paper aims at demonstrating the non-specific immunosuppression as regards thyme-dependent antigens sheep erythrocytes (SRBC) during the course of Candida albicans systemic infection.Three lots of syngeneic /BALB/c mice, 8–12 weeks of age, were used. The first normal lot was inoculated via the intraperitoneal route with a (SRBC) suspension (4×10⁸ cells ml) in a Hank's balanced saline solution. The primary response of antibodies formed by splenic cells was measured from 4 to 8 days after inoculation using the direct plaque forming cells technique. The second lot was infected by the same route with a suspension of Candida albicans (1×10⁷ cells). Positive retrocultures from the blood and kidneys of these infected mice were obtained. These yeasts cultivated in a Sabouraud medium were harvested after 20 h at 37 °C. Following the same methodology the immune response to SRBC was determined. The serum obtained from infected mice was transferred to a third lot of mice at different intervals during the course of the infection. The immune response to SRBC was done by the direct plaque-forming cells technique. Controls were carried out using normal donors and recipients.A suppression of the immune response was obtained as from the 2nd day of inoculation up to the 28th day. It was not possible to transfer such suppression passively by means of the serum.These results suggest that the systemic infection by Candida albicans induce a non-specific immunosuppression in the organism, already demonstrated in viral infections, bacteria, protozoaria and metazoaria in mammals.In some way, this will contribute to explain the mechanisms of immune response to Candida albicans.     

Necrostatin-1抑制白色念珠菌诱导的巨噬细胞自噬而影响免疫应答

随着现代医疗技术的发展,人口老龄化加剧,抗生素滥用等降低人体免疫力的因素越来越多,使得由白色念珠菌Candida albicans感染引起的疾病发生率越来越高。本研究通过使用程序性坏死的选择性抑制剂Necrostatin-1,初步探索了细胞自噬和程序性死亡在巨噬细胞抗白色念珠菌感染中的作用。研究表明:细胞自噬参与了白色念珠菌侵染巨噬细胞的过程,而Necrostatin-1可能通过抑制白色念珠菌感染引起的自噬促进IL-6的表达,并抑制TNF-α(肿瘤坏死因子α)的表达,进而对白色念珠菌引起的天然免疫反应产生影响。细胞自噬在抗真菌感染方面的研究已有报道,而程序性坏死在抗真菌感染方面的研究鲜有报道,同时Necrostatin-1在巨噬细胞抗白色念珠菌感染中的作用尚未见报道。本研究对白色念珠菌侵染机理的探索具有一定的意义。     

Thermotolerance and the heat-shock response in Candida albicans

At elevated temperatures, yeast cells of Candida albicans synthesized nine heat-shock proteins (HSPs) with apparent molecular masses of 98, 85, 81, 76, 72, 54, 34, 26 and 18 kDa. The optimum temperature for the heat-shock response was 45 degrees C although HSPs were detected throughout the range 41-46 degrees C. Protein synthesis was not observed in cells kept at 48 degrees C. Yeast cells survived exposure to an otherwise lethal temperature of 55 degrees C when they had previously been exposed to 45 degrees C. The thermotolerance induced during incubation at 45 degrees C required protein synthesis, since protection was markedly reduced by trichodermin. Mercury ions induced a set of three stress proteins, one of which corresponded in size to an HSP, and cadmium ions evoked one stress protein seemingly unrelated to the HSPs observed after temperature shift.     

Physical stress primes the immune response of Galleria mellonella larvae to infection by Candida albicans

Larvae of the greater wax moth (Galleria mellonella) that had been subjected to physical stress by shaking in cupped hands for 2 min showed reduced susceptibility to infection by Candida albicans when infected 24 h after the stress event. Physically stressed larvae demonstrated an increase in haemocyte density and elevated mRNA levels of galiomicin and an inducible metalloproteinase inhibitor (IMPI) but not transferrin or gallerimycin. In contrast, previous work has demonstrated that microbial priming of larvae resulted in the induction of all four genes. Examination of the expression of proteins in the insect haemolymph using 2D electrophoresis and MALDI TOF analysis revealed an increase in the intensity of a number of peptides showing some similarities with proteins associated with the insect immune response to infection. This study demonstrates that non-lethal physical stress primes the immune response of G. mellonella and this is mediated by elevated haemocyte numbers, increased mRNA levels of genes coding for two antimicrobial peptides and the appearance of novel peptides in the haemolymph. This work demonstrates that physical priming increases the insect immune response but the mechanism of this priming is different to that induced by low level exposure to microbial pathogens.     

The distribution in mice of intravenously administered labelled Candida albicans.

The time dependence of the distribution of intravenously injected radiolabelled Candida albicans in the body of mice was studied. The Candida cells were labelled by cultivating them 7 days at 28 degrees C in a medium containing one of the following radionuclides: 46Sc, 95Nb, 59Fe, 144ce, 89Sr, 60Co, 65Zn, 54Mn, 45Ca, 51Cr and 91Y, which are listed in decreasing order in respect to amount bound. The labelled cells were killed by heating them 120 min at 60 degrees C, without loss of immunologic properties. Owing to the amount and strength of binding, 144Ce labelled Candida, together with 14C labelled cells was used in animal kinetic study. A rapid disappearance of the labelled cells from blood, interrupted by a small peak, was paralleled by a transient uptake in lungs and followed by a long lasting accumulation in the liver. The kidneys and spleen revealed only small uptakes of the labelled material.     

The role of metabolic energy in the lethal action of basic proteins on Candida albicans.

Comparative studies were made on the destructive effects of certain basic proteins on a strain of Candida albicans and two of its respiration-impaired mutants. Both by direct plate counts of survivors and by quantitative ultraviolet spectrophotometric analyses of released cellular constituents, the respiration-impaired mutants were less vulnerable to the destructive actions of the basic proteins than were ordinary wild-type cells. The lethal incidence and the ultraviolet absorbing cellular substances released from wild-type cells by the proteins were markedly decreased in the presence of the oxidative phosphorylation uncouplers sodium azide, 2,4-dinitrophenol, and salicylanide and approximately equal to the effects produced on an oxidative phosphorylation mutant not treated with the uncouplers. The heightened resistance of a culture through mutational or chemical impairment of its respiratory system suggests a role of metabolic energy in the destructive action of various basic proteins on yeast cells.     

Combined action of amphotericin B and methacyclin on Candida albicans

The combined effect of amphotericin B, a polyene antibiotic, and metacycline, a tetracycline antibiotic, on the cells of C. albicans was studied. The method of square titration followed by quantitative plating of the samples was used for estimation of the combination efficiency. An attempt was also made to investigate the characteristic features of metacycline penetration into the yeast cells under the effect of various doses of the polyene antibiotic. The capacity of metacycline for fluorescence in the yellow-green pectral region was employed for this purpose. It was shown that the drugs had a synergistic effect on C. albicans. The fluorescence research methods allowed one to demonstrate that even low subinhibitory doses of amphotericin B increased the permeability level of the cytoplasmic membrane and provided penetration of metacycline into the cytoplasm almost during the first hours of the contact. The time course of metacycline cumulation in the cells was followed up and the characteristic features of the antibiotic localization were analysed.