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  • 1
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract If the determinants of shape and cell wall topography are independently regulated and induced in germ tube formation in Candida albicans, these processes may be separable in a non-germ tube forming strain. The expression of several preferentially expressed hyphal surface components in a parental, non-germ tube forming variant, and a germ tube forming revertant strain were examined by indirect immunofluorescence. The proportion of germ tubes expressing the determinants and the morphological localization of expression was similar. Few yeast cells in germ tube cultures bound probes and there was no increase in binding by yeast cells of the variant strain. Extraction with β-mercaptoethanol prior to analysis had little effect on probe binding and the shape of yeast cells were similar. These observations suggest the ability to promote apical expansion in germ tube formation and surface expression of certain markers were coordinately regulated.
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  • 2
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract We have previously reported a 37 kDa laminin-binding protein (p37) and a 58 kDa fibrinogen-binding mannoprotein (mp58) on the surface of Candida albicans. A few yeast cells expressed both functional receptors at the surface while germ tubes expressed a functional mp58 fibrinogen but not a functional p37 laminin receptor. These receptors were heterogeneously dispersed at the surface as shown by binding of rabbit antiserum to mp58 (PAb anti-mp58) and antiserum to the human high affinity laminin receptor. In this report we have used a dual fluorescence technique to determine if the two receptors colocalize, perhaps as part of a receptor complex. Fibrinogen was used as a probe for mp58 and polyclonal antiserum generated to the p37 (PAb anti-p37) was used as a probe for the 37 kDa laminin-binding protein. Both receptors were heterogeneously distributed, but the receptors were not colocalized as the areas of concentration of each receptor were different. Immunohistochemical analysis of tissue sections from patients with disseminated and superficial candidiasis with PAb anti-p37 and PAb anti-mp58 revealed that both receptors were also expressed in infected tissues. The patterns of morphological expression were similar to the in vitro patterns detected by immunofluorescence.
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  • 3
    ISSN: 1573-0832
    Keywords: Candida albicans ; Cell wall ; Germ tube ; Monoclonal antibody ; Surface antigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Monoclonal antibodies against germ-tube specific antigens ofCandida albicans were produced by a selective immunization protocol. The monoclonal antibody (MAb) DC3:H10 recognized a determinant preferentially expressed by germ tubes of the fungus compared to yeast cells. The MAb DC3:H10 antigen was expressed by nearly all germ tubes of the strains 3153A and B311. It was also expressed by germ tubes of the white phenotype of strain WO-1, and by many exponentially growing yeast cells of the opaque phenotype of this strain. The determinant appeared very rapidly after induction of germ tubes of strain 3153A. The MAb DC3:H10 epitope was sensitive to proteolytic treatment but not to periodate treatment, indicating its protein nature. The reactive material could be extracted from intact germ tubes by treatment with β-mercaptoethanol. On elution from a G-200 Sephadex column it yielded an apparent molecular weight of 519 kDa. This fraction appeared heterogeneous since at least two major bands of lower molecular weight were detected by silver staining following electrophoretic separation under denaturing conditions.
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  • 4
    ISSN: 1573-0832
    Keywords: Candida albicans ; Cell wall ; Complexes ; Mannoproteins ; Receptors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have previously described a monoclonal antibody, MAb DC3:H10, which recognized an epitope preferentially expressed on the surface ofCandida albicans germ tubes. In the present study we examined the MAb-reactive material further. Immunoblot analysis of the material purified partially by Sephadex G-200 and DEAE-Sephacel chromatography reacted with antibodies to theC. albicans C3d receptor (CR2). In an ELISA, MAb DC3:H10 captured antigen that was recognized by both anti-CR2 and anti-mp58 fibrinogen binding mannoprotein polyclonal antibodies. The MAb DC3:H10 failed to compete with either of these antisera in an ELISA. Indirect immunofluo-rescence (IIF) analysis showed differences in surface distribution for the MAb DC3:H10, the CR2, and the mp 58 epitopes. Dual labeling IIF experiments showed MAb DC3:H10 binding to be unaffected by binding of fibrinogen or anti-mp58 antibody. However, the binding patterns of MAb DC3:H10 were modified in the presence of anti-CR2 antibody, suggesting a complex interaction between these cell wall components.
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  • 5
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Lipid synthesis has been studied in the dimorphic fungus Candida albicans. 14C-acetate incorporation into lipid material was used to measure new lipid synthesis in two cultures in which either yeast or mycelial growth was initiated from stationary phase yeast cells. When resuspended in fresh medium at 37 °C, cells resume growth and change morphology while at 30 °C cells resume budding growth. When resuspended at the appropriate temperature, both yeast and germ tube cultures immediately incorporated 14C-acetate into lipid material. The labeled lipid was more or less evenly divided between neutral and phospholipid. Phosphatidyl choline was the major phospholipid fraction and along with phosphatidyl ethanolamine accounted for 60–65 % of the total phospholipid. Lipid synthesis during growth initiation of either morphology showed a similar pattern, with no significant differences observed in neutral or phospholipid or phospholipid components between yeast and mycelial forms.
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  • 6
    ISSN: 1573-0832
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract When grown and induced to form germ tubes in liquid defined media, yeast cells of Candida albicans must reach stationary phase before acquiring ability to carry out the yeast-mycelial transition. This study examined the effect of the carbon source utilized for yeast growth on the inducibility of stationary phase yeast. When grown to the same stationary phase cell density as glucose cultures, cultures grown on citrate were fully inducible while cultures grown on galactose and mannose showed a small reduction. Cultures grown on ethanol were reduced 80% in morphological conversion. When glucose grown cells were induced in the presence of these carbon sources, hexoses supported full induction while ethanol reduced induction 80%. Induction in the presence of carboxylic acids was similar to induction in the absence of added carbon source. When induced on the same source used in yeast growth, germ tube formation was reduced for all carbon sources except hexoses. When induced in the absence of added carbon source, yeasts grown on citrate and ethanol were inhibited 80–100%. Cultures starved for glucose were more inhibited than cultures starved for NH4Cl when induced without added carbon source. These observations suggest that the metabolic state of the stationary phase cell is an important factor in the ability to respond to conditions inducing germ tube formation.
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