anti-Candida Antibodies Research Articles

Value of detection of antibodies toCandida albicans germ tube in the diagnosis of systemic candidosis

To test the value of detection of anti-Candida albicans germ tube antibodies by indirect immunofluorescence assay in the diagnosis of systemic candidosis, a retrospective study was done using 126 sera from 27 patients with presumptive systemic candidosis (13 immunocompromised), 165 sera from 45 patients with aspergillosis (29 immunocompromised), 35 sera from eight patients with cryptococcosis (6 immunocompromised), and 101 sera from 101 blood donors. While 21 of 27 patients with systemic candidosis (77.8%) had anti-germ tube antibodies, these antibodies were absent in all patients with cryptococcosis and in all blood donors. They were however detected in 5 of 45 patients with aspergillosis (11.1%). Ten of 13 (76.9%) immunocompromised patients with candidosis had anti-germ tube antibodies; similar results were obtained in immunocompetent patients with candidosis (78.6%). The specificity was 96.8%, indicating a high degree of discrimination was possible between systemic candidosis and other invasive mycoses in the patients studied. Anti-germ tube responses did not appear to be significantly reduced in immunocompromised patients.

Immunoglobulin G (IgG) antibodies in serum of autoimmune infertile men and their wives react against special sperm antigens of the husbands

A rapid, specific, and sensitive assay using passive hemagglutination and Coombs' tests has been established for detecting male autoimmunity and female isoimmunity to sperm antigens. Antisperm antibody titers evaluated by this method were similar in 156 normal females, 400 pregnant females, 25 gonadal dysgenetics, 43 users of Oracon (an oral contraceptive), and 25 normal males. In contrast, in the evaluation of 50 couples with unexplained infertility, 16 males were found to have significantly elevated Coombs' autoantibody titers (primarily IgG) and correspondingly higher antisperm antibody titers in their serum and seminal plasma; another 28 had low autoantibody titers and low serum and seminal plasma antibody titers; and 6 had intermediate titers. The wives of 63% of the males with high autoantibody titers were found to be ‘isoimmune’, as compared with 7% of those with low titers. The isoimmune females had significantly higher antisperm antibody titers than did those with normal titers. In general, comparable titer values were obtained whether sperm extract or seminal plasma was used as the erythrocyte coat for passive hemagglutination. The titers were consistently higher in the seminal plasma than in the serum of males. Very little crossreactivity was apparent between anti-Candida and antisperm antibodies, and antibodies to blood group antigens did not appear to influence the results. When the results obtained by passive hemagglutination were compared with those obtained by immunofluorescence, Franklin-Dukes, and hanging drop interface methods, only those of the passive hemagglutination and Coombs' test correlated well with the results of postcoital tests and seminal analysis.

Coexistence of free antigens, free antibodies and immune complexes in sera from patients with suspected deep-seated candidosis.

Two commercially available latex agglutination tests: Cand-TEC(TM) from RAMCO Laboratories, Inc., Houston, Texas, USA, for the detection of a heat labile Candida antigen, and LA-Candida Antigen Detection System from Immuno-Mycologics (IMMY), Inc., Norman, Oklahoma, USA, for the detection of Candida mannan antigen, and Own LAT, a self-prepared latex agglutination test for the detection of Candida mannan antigen were tested on 467 sera of patients at risk for deep-seated Candidosis. Separate, parallel investigations were made to demonstrate anti-Candida antibodies (3 methods), circulating immune complexes (not antigen-specific, 2 methods), and rheumatoid factor (2 methods). Specificity and sensitivity of the mannan antigen detection systems were studied. Conditions of specific and non-specific mannan binding to antisera were studied in a defined mannan-antiserum system. The potential and the limitations of the Candida mannan antigen and protein antigen detection systems using latex particle agglutination are evaluated.

Immunological analysis of the aqueous humour in candida endophthalmitis. I: Experimental study.

An immunological study of aqueous humour and serum was carried out in experimental ocular candidosis in rabbits. Anti-candida antibody titres, immunoglobulin G, and albumin levels were evaluated in the aqueous humour and in the serum. A local synthesis of anti-candida antibodies was detected in numerous rabbits at the height of clinical activity of the lesions.

Detection of antibodies to Candida albicans germ tube in the diagnosis of systemic candidiasis.

Sera from 109 subjects were tested for the presence of anti-Candida albicans antibodies by an indirect immunofluorescence assay. Aliquots of the sera were adsorbed with heat-killed blastospores to remove the antibodies against the surface of the yeast-phase cell wall and tested for anti-germ tube cell wall antibodies. Unadsorbed sera stained the entire cell wall of yeast and germ-tubes. Immunoglobulin G (IgG) antibodies were found in all patients with systemic candidiasis and in 81.2% of patients with Candida albicans isolated from skin and mucous membranes. IgA and IgG were found in 67.4 and 57.1%, respectively, of controls without evidence of candidiasis. After the adsorption only sera from patients with systemic candidiasis showed antibodies, predominantly IgA, against germ tube cell wall. Adsorption of the sera thus increased the specificity, efficiency, and positive and negative predictive values of the test. The test achieved the highest sensitivity in adsorbed sera for the combination of IgA and IgG.

Idiotypy of human anti-Candida albicans antibodies: recurrence, presence of a cross-reactive autoanti-idiotypic-like activity, and role in the induction of specific in vitro antibody response.

Rabbit anti-idiotypic antibodies (L12) were raised against human anti-mannan of Candida albicans (CA) antibodies isolated from the serum of a normal donor. The absorbed anti-idiotypic antiserum bound to donor anti-CA mannan antibodies but not to control immunoglobulins. Binding was inhibited by CA mannan but not by other polysaccharide antigens. L12 was shown to cross-react with anti-CA mannan-isolated antibodies or with anti-CA antibody-containing sera from individuals unrelated to the donor. IgG fraction isolated from the donor serum was repeatedly absorbed on CA mannan Sepharose to remove anti-mannan antibodies. This IgG fraction (named autoanti-idiotypic fraction) blocked, in a dose-dependent fashion, the binding of rabbit anti-idiotype to donor anti-CA mannan antibodies. Moreover, this CP-depleted IgG fraction cross-reacted with public idiotypic determinants of unrelated anti-CA mannan antibodies. Finally, L12 induced sensitized lymphocytes to produce anti-CA mannan antibodies in vitro in the absence of antigen.

Proliferation of human peripheral blood mononuclear cells induced by Candida albicans and its cell wall fractions.

Glutaraldehyde-inactivated cells and cell-wall fractions of Candida albicans were studied for their capacity to induce or inhibit the in-vitro proliferation of human peripheral blood mononuclear cells (PBMC), as measured by 3H-thymidine incorporation. Both the intact cells (CA) and a phosphorylated gluco-mannan-protein complex of the cell wall (GMP), in microgram doses, were strong inducers of PBMC proliferation, with a peak of activity at 6-9 days of culture and varying with the PBMC donor. A significant but much lower proliferation was observed on exposure of PBMC to a low-protein (less than 3% by weight) mannan component (M) of the cell wall. Both a hot-alkali extracted mannan-protein complex (M-alk), comparable to GMP in crude chemical composition, and an alkali-insoluble cell-wall glucan (GG) were inactive. None of the Candida fractions induced a lymphoproliferation of umbilical cord blood cells and all fractions, except GG, were equally effective in binding human anti-Candida antibodies as shown by a sensitive ELISA-inhibition assay. Moreover, a monoclonal antibody against the class II determinant of the HLA complex inhibited PBMC proliferation irrespective of the Candida antigen used. Taken together, the data shows that in inducing lymphoproliferation, Candida fractions act as specific antigens rather than as non-specific mitogens. Use of intact Candida cells and chemically-defined cell-wall components appears preferable to use of undefined antigenic mixtures as stimulators of PBMC proliferation.

Candida endophthalmitis after heroin abuse

Three cases of ocular candidosis involving heroin abusers have been observed in 1983 in Toulouse department of ophthalmology. These three patients had used iranian brown heroin. Twenty similar cases have been published in these last years. This new pathology can be explained on two reasons. The first is that the drug abusers have some immunity pertubation; however, immunity exploration in these patients does not reveal any immunodeficiency. The second reason, certainly more important, is the method of using heroin. The diagnosis of Candida endophthalmitis of course based on clinical context must be proved by biological tests. Candida albicans is never identified in aqueous humor. For this reason, it seems very interesting to detect anti-candida antibodies in aqueous humor. It has been used as methods of dosage laser Nephelemetry for IgG and immunofluorescence for candidosis antibodies. The criterion used is similar to the toxoplasmosis coefficient established by Desmonts (3). In two cases, this test was the only way that permits us to have certitude of candidosis ocular diagnosis. Otherwise the observations show that anterior chamber punction is more significant when there is an anterior uveitis.

Effect of antigen used in the detection of anti -Candida albicans antibodies

Antigens of different origins were used in the investigation of anti-Candida albicans antibodies. This can influence the results obtained. We have assayed three different antigenic preparations weekly for 8 weeks in the study of anti-C. albicans antibodies induced by cutaneous, digestive, and systemic inoculations with C. Albicans ATCC 26555 in rabbits free of specific antibodies, and using indirect immunofluorescence (IIF), direct agglutination (DA) and counterimmunoelectrophoresis (CIE) as serological methods. In IIF and DA, two antigens were used (C. albicans ATCC26555 and C. albicans NCPF3153). In CIE we also used a third commercial antigen. All three somatic antigens were used at three different concentrations. Using IIF and DA the titres obtained with both antigens were similar in different inoculations. The IIF was somewhat earlier in the detection of antibodies, and the titre reached was higher when the antigen used was obtained from the inoculated strain. The detection of precipitins by CIE was in most cases only positive with the antigen obtained from the homologous strain, the highest level being reached in the systemic inoculation.

Langerhans cells in candidal leukoplakia.

Using histochemical and immunohistochemical methods, we examined specimens of candidal leukoplakia from the oral mucosa of 5 smokers to determine the morphological relationships between Candida and Langerhans cells (LC) in tissue sections. LC were fairly evenly distributed in control sections, but had a patchy distribution in lesions. Fewer LC were found in lesions than in control tissue, but the difference was not statistically significant. Candidal antigens were not detected on LC by the methods used, but we found ATPase-positive LC among, or at least near, intraepithelial candidal hyphae. However, sections double-reacted with anti-Candida and T6 antibodies to label candidal antigens and LC, respectively, showed a clear zone of epithelium between the T6-positive LC and the candidal hyphae. The difference in distribution of ATPase-positive and T6-positive LC may indicate locations of 2 subtypes of LC, or a change in T6 antigen expression by the LC closest to the candidal hyphae.

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.

Effect of cutaneous and digestive colonization in the induction of anti-Candida albicans antibodies: experimental study.

Candida albicans colonization induces antibodies, which must be taken into account in the serological diagnosis of candidiasis. In order to determine the degree of this effect, an experimental study in rabbits free of specific anti-Candida antibodies by cutaneous and digestive inoculation has been carried out. The evolution of humoral response was studied over 8 weeks by indirect immunofluorescence (IIF), direct agglutination (DA), counterimmunoelectrophoresis (CIE) and double diffusion (DD). The cutaneous colonization detectable by culture was maintained until the second week in 70% of the animals and the presence of antibodies detectable by IIF and DA was observed after the 2nd week. The highest antibody titre by IFF and DA was 1/64, and was reached in the 5th week, with a tendency to drop in the following weeks. Precipitins were only detected by CIE in 15% of the animals in the 7th week. Elimination of yeast in stools continued only in 20% of the animals in the 2nd week of the experiment. Antibodies were detected by IIF and DA after the 2nd week, with the highest titres detectable by IFF in the 5th week. Precipitant antibodies detectable by CIE appeared in 15% of the animals in the 8th week.

A purified polysaccharide isolated from Candida albicans induces antibody response in vitro by human peripheral blood lymphocytes and discriminates between sera from normal and Candida albicans-infected individuals

A purified polysaccharide extracted from Candida albicans (MPPS), stimulates in vitro synthesis of specific antibodies by human peripheral blood lymphocytes. These antibodies can be detected by a sensitive enzyme-linked immunoassay. The same assay can be applied to the quantitation of anti-candida antibodies in serum. Statistically significant differences were found between sera of normal and candida-infected individuals.

In vitro and in vivo adherence of Candida albicans to mucosal surfaces

In continuation of a previous study, chitin soluble extract (CSE) and anti-Candida albicans antibodies were found to inhibit the in vitro adherence of C. albicans to human vaginal epithelial cells by 60-70%. Pretreatment of epithelial cells with chitin, CSE or N-acetyl-glucosamine (NAG) decreased the percentage of adherence, while pretreatment of yeasts had no such effect, indicating that chitin or its derivatives may be involved in mediating the adherence of C. albicans. In vivo attachment of C. albicans to epithelial cells was studied in an experimental murine vaginitis created by inoculating mice intravaginally with yeasts at the oestrus stage. Attachment of the yeasts to exfoliated vaginal epithelial cells was followed microscopically, and the course of infection was assessed histopathologically in tissue sections at various times post-yeast inoculation. The infection rate at 24-h post-yeast inoculation was approximately 50%. Attempts were made to block in vivo attachment and prevent infection by pretreating 208 mice with either CSE, NAG, mannan or phosphate-buffered saline (PBS) prior to inoculation with yeasts. The infection rate among mice pretreated with mannan or PBS was 41.6% and 43.8%, respectively. Among CSE- or NAG-pretreated animals, the rate ranged from 0-9%. Treatment of mice with CSE or NAG after inoculation of the yeasts did not prevent infection. The data from the in vivo experiments reveal that pretreatment of animals with chitin derivatives blocks attachment of yeasts to the vaginal mucosal surfaces and leads to the prevention of vaginal infection in an experimental model.

Effects of specific antibodies on the interaction between the fungus Candida albicans and human oral mucosa

Host-parasite interactions were studied in four groups: non-infected controls; infected carriers of Candida albicans without evidence of candidiasis; subjects with acute candidiasis; and subjects with chronic candidiasis. Specific anti-candida antibodies were demonstrated in saliva from subjects of all four groups; the titres reflected the degree of antigenic stimulation, being significantly higher in candidiasis than in controls or infected carriers. The adherence of C. albicans to buccal epithelial cells was not significantly different in a given saliva, regardless of whether the assay was carried out with autologous C. albicans and epithelial cells or with a stock strain in a standardized assay. Therefore, the standard assay was used to study the effects of specific salivary antibodies on adherence. A significant inverse correlation was found between salivary IgA anti-candida antibodies and the adherence of C. albicans to buccal epithelial cells, suggesting that IgA antibodies can inhibit adherence of Candida to the oral mucosa. In some instances, removal of antibodies led to a significant increase in adherence; however, often this was not the case, indicating that some but not all of the antibodies present were capable of inhibiting the adherence of C. albicans to epithelial cells.

Specific inhibition of in vitro Candida-induced lymphocyte proliferation by polysaccharidic antigens present in the serum of patients with chronic mucocutaneous candidiasis.

A specific inhibitory activity of in vitro proliferative responses of normal human lymphocytes to Candida metabolic antigen was found in the serum of 6 out of 23 children with chronic mucocutaneous candidiasis. In each of the six patients, the presence of an inhibitory activity was associated with Candida-specific cellular defects, characterized by a negative-skin test and a lack of in vitro lymphocyte proliferation. The presence of a circulating inhibitor was detected during relapses of the disease and disappeared under antifungal therapy. This inhibitory effect was not associated with any toxicity on tested lymphocytes. The factor was shown to be nondialysable, thermostable, nonprecipitable with ammonium sulfate and absorbable on anti-Candida antibodies or concanavalin A-coupled agarose columns. Altogether, these results suggest that the inhibitory factor is not an immunoglobulin, but rather a polysaccharidic antigen of Candida albicans. An inhibition of Candida-induced proliferative response of normal human lymphocytes was also obtained by addition of polysacharide antigens or purified mannans from C. albicans to cultures. Candida polysaccharidic antigens appeared, therefore, to be involved in specific depression of cellular functions observed in chronic candidiasis.

Experimental mycosis in immunosuppressed rabbits

Chronic candidosis was established in rabbits by the injection I.V. of 2 X 10(6) cells of C. albicans. The rabbits were assayed every week for 14 weeks for the appearance of candida antigen and anticandida antibodies in serum and other body fluids. Tests were carried out in double diffusion plates; antigen against hyperimmune rabbit sera and antibody against Candida cell sap antigen preparation. A sensitive specific passive hemagglutination procedure was also developed which used chromate treated cells. In rabbits with chronic candidosis not treated with cyclophosphamide antigen was detected in 4x concentrated serum between the fifth and sixth week. At about the same time antibodies were demonstrable and thereafter antigen was no longer detected. Maximum antibody titer occurred between the eight to 10th week and disappeared thereafter. If cyclophosphamide 30 mg/kg was given at this point, anticandida antibodies reappeared in high titers, persisted from three to four weeks and then disappeared. At autopsy no evidence of candidosis was present. If rabbits were pretreated with cyclophosphamide 60 mg/kg for one week before inoculation and given the drug weekly thereafter no antibody was detectable but antigen and antibody were present in body fluids (not serum) at post mortem.

Immunoglobulin E anti-Candida antibodies and candidiasis.

Elevated levels of immunoglobulin E anti-Candida antibodies were observed in the sera of patients with systemic and vaginal candidiasis and in cervicovaginal washings of the latter.

Anti-Candida antibodies in patients with acute leukemia: A prospective study

Serial determinations of anti-Candida agglutinating and precipitating antibodies were carried out prospectively over a period of eighteen months on 154 patients with acute leukemia. During this study visceral candidiasis developed in fourteen patients. A significant rise in agglutinating antibody titer was noted in seven of these patients, and in three additional patients blood cultures grew Candida species. Thus an antemortem diagnosis of candidiasis was made in ten of fourteen patients with this infection. A “false” rise in antibody titer occurred in patients with other disseminated fungal infections, in patients with fever of unknown origin which resolved spontaneously with regression in the patient's leukemia and in one patient with pulmonary tuberculosis. We recommend serial determinations of anti-Candida agglutinating antibody titers in groups of patients known to have increased susceptibility to fungal infections, as this is the most reliable means for diagnosing visceral candidiasis.