The curli regulator CsgD mediates stationary phase counter-silencing of csgBA in Salmonella Typhimurium.

Integration of horizontally acquired genes into transcriptional networks is essential for the regulated expression of virulence in bacterial pathogens. In Salmonella enterica, expression of such genes is repressed by the nucleoid-associated protein H-NS, which recognizes and binds to AT-rich DNA. H-NS-mediated silencing must be countered by other DNA-binding proteins to allow expression under appropriate conditions. Some genes that can be transcribed by RNA polymerase (RNAP) associated with the alternative sigma factor σS or the housekeeping sigma factor σ70 in vitro appear to be preferentially transcribed by σS in the presence of H-NS, suggesting that σS may act as a counter-silencer. To determine whether σS directly counters H-NS-mediated silencing and whether co-regulation by H-NS accounts for the σS selectivity of certain promoters, we examined the csgBA operon, which is required for curli fimbriae expression and is known to be regulated by both H-NS and σS . Using genetics and in vitro biochemical analyses, we found that σS is not directly required for csgBA transcription, but rather up-regulates csgBA via an indirect upstream mechanism. Instead, the biofilm master regulator CsgD directly counter-silences the csgBA promoter by altering the DNA-protein complex structure to disrupt H-NS-mediated silencing in addition to directing the binding of RNAP.

Dedicated Followers of Fashion? Bioarchaeological Perspectives on Socio-Economic Status, Inequality, and Health in Urban Children from the Industrial Revolution (18th-19th C), England.

The 18th and 19th centuries in England were characterised by a period of increasing industrialisation of its urban centres. It was also one of widening social and health inequalities between the rich and the poor. Childhood is well-documented as being a stage in the life course during which the body is particularly sensitive to adverse socio-economic environments. This study therefore aims to examine the relationship between health and wealth through a comprehensive skeletal analysis of a sample of 403 children (0-17 years), of varying socio-economic status, from four cemetery sites in London (c.1712-1854). Measurements of long bone diaphyseal length, cortical thickness, vertebral neural canal size, and the prevalence of a range of pathological indicators of health stress were recorded from the Chelsea Old Church (high status), St Benet Sherehog (middle status), Bow Baptist (middle status), and Cross Bones (low status) skeletal collections. Children from the low status Cross Bones site demonstrated deficient growth values, as expected. However, those from the high status site of Chelsea Old Church also demonstrated poor growth values during infancy. Fashionable child-care practices (e.g. the use of artificial infant feeds and keeping children indoors) may have contributed to poor infant health amongst high status groups. However, differing health risks in the lower status group revealed the existence of substantial health inequality in London at this time.

Candida albicans is phagocytosed, killed, and processed for antigen presentation by human dendritic cells.

Candida albicans is a component of the normal flora of the alimentary tract and also is found on the mucocutaneous membranes of the healthy host. Candida is the leading cause of invasive fungal disease in premature infants, diabetics, and surgical patients, and of oropharyngeal disease in AIDS patients. As the induction of cell-mediated immunity to Candida is of critical importance in host defense, we sought to determine whether human dendritic cells (DC) could phagocytose and degrade Candida and subsequently present Candida antigens to T cells. Immature DC obtained by culture of human monocytes in the presence of granulocyte-macrophage colony-stimulating factor and interleukin-4 phagocytosed unopsonized Candida in a time-dependent manner, and phagocytosis was not enhanced by opsonization of Candida in serum. Like macrophages (Mphi), DC recognized Candida by the mannose-fucose receptor. Upon ingestion, DC killed Candida as efficiently as human Mphi, and fungicidal activity was not enhanced by the presence of fresh serum. Although phagocytosis of Candida by DC stimulated the production of superoxide anion, inhibitors of the respiratory burst (or NO production) did not inhibit killing of Candida, even when phagocytosis was blocked by preincubation of DC with cytochalasin D. Further, although apparently only modest phagolysosomal fusion occurred upon DC phagocytosis of Candida, killing of Candida under anaerobic conditions was almost equivalent to killing under aerobic conditions. Finally, DC stimulated Candida-specific lymphocyte proliferation in a concentration-dependent manner after phagocytosis of both viable and heat-killed Candida cells. These data suggest that, in vivo, such interactions between DC and C. albicans may facilitate the induction of cell-mediated immunity.

Cell-mediated immunity to Histoplasma capsulatum.

Histoplasma capsulatum is a facultative intracellular pathogen, and the causative agent of the most common systemic fungal infection. Over the past several years, many new insights have been learned concerning the biology, biochemistry, and molecular genetics of this microorganism. This review focuses on the immunology of host defense against H. capsulatum yeasts with emphasis on the development of cell-mediated immunity, and the strategies used by the fungus to survive and multiply within macrophages.

Histoplasma capsulatum yeasts are phagocytosed via very late antigen-5, killed, and processed for antigen presentation by human dendritic cells.

Histoplasma capsulatum (Hc) is a facultative, intracellular parasite of world-wide importance. As the induction of cell-mediated immunity to Hc is of critical importance in host defense, we sought to determine whether dendritic cells (DC) could function as a primary APC for this pathogenic fungus. DC obtained by culture of human monocytes in the presence of GM-CSF and IL-4 phagocytosed Hc yeasts in a time-dependent manner. Upon ingestion, the intracellular growth of yeasts within DC was completely inhibited compared with rapid growth within human macrophages. Electron microscopy of DC with ingested Hc revealed that many of the yeasts were degraded as early as 2 h postingestion. In contrast to macrophages, human DC recognized Hc yeasts via the fibronectin receptor, very late Ag-5, and not via CD18 receptors. DC stimulated Hc-specific lymphocyte proliferation in a concentration-dependent manner after phagocytosis of viable and heat-killed Hc yeasts, but greater proliferation was achieved after ingestion of viable yeasts. These data demonstrate that human DC can phagocytose and degrade a fungal pathogen and subsequently process the appropriate Ags for stimulation of lymphocyte proliferation. In vivo, such interactions between DC and Hc may facilitate the induction of cell-mediated immunity.

Identification of constituents of human neutrophil azurophil granules that mediate fungistasis against Histoplasma capsulatum.

Previously we demonstrated that human neutrophils mediate potent and long-lasting fungistasis against Histoplasma capsulatum yeasts and that all of the fungistatic activity resides in the azurophil granules. In the present study, specific azurophil granule constituents with fungistatic activity were identified by incubation with H. capsulatum yeasts for 24 h and by quantifying the subsequent growth of yeasts via the incorporation of [(3)H]leucine. Human neutrophil defensins HNP-1, HNP-2, and HNP-3 inhibited the growth of H. capsulatum yeasts in a concentration-dependent manner with maximum inhibition at 8 microg/ml. At a concentration of 4 microg/ml, all possible paired combinations of defensins exhibited additive fungistatic activity against H. capsulatum yeasts. Cathepsin G and bactericidal-permeability-increasing protein (BPI) also mediated fungistasis against H. capsulatum in a concentration-dependent manner. The fungistatic activities of combinations of cathepsin G and BPI were additive, as were those of combinations of cathepsin G or BPI with HNP-1, HNP-2, and HNP-3. Lysozyme and elastase exhibited modest antifungal activity, and azurocidin and proteinase 3 exhibited no significant fungistasis against H. capsulatum yeasts. Thus, defensins, cathepsin G, and BPI are the major anti-H. capsulatum effector molecules in the azurophil granules of human neutrophils.

Regulation of the macrophage vacuolar ATPase and phagosome-lysosome fusion by Histoplasma capsulatum.

Histoplasma capsulatum (Hc) maintains a phagosomal pH of about 6.5. This strategy allows Hc to obtain iron from transferrin, and minimize the activity of macrophage (Mo) lysosomal hydrolases. To determine the mechanism of pH regulation, we evaluated the function of the vacuolar ATPase (V-ATPase) in RAW264.7 Mo infected with Hc yeast or the nonpathogenic yeast Saccharomyces cerevisae (Sc). Incubation of Hc-infected Mo with bafilomycin, an inhibitor of the V-ATPase, did not affect the intracellular growth of Hc, nor did it affect the intraphagosomal pH. In contrast, upon addition of bafilomycin, phagosomes containing Sc rapidly changed their pH from 5 to 7. Hc-containing phagosomes had 5-fold less V-ATPase than Sc-containing phagosomes as quantified by immunoelectron microscopy. Furthermore, Hc-containing phagosomes inhibited phagolysosomal fusion as quantified by the presence of acid phosphatase, accumulation of LAMP2, and fusion with rhodamine B-isothiocyanate-labeled dextran-loaded lysosomes. Finally, in Hc-containing phagosomes, uptake of ferritin was equivalent to phagosomes containing Sc, indicating that Hc-containing phagosomes have full access to the early "bulk flow" endocytic pathway. Thus, Hc yeasts inhibit phagolysosomal fusion, inhibit accumulation of the V-ATPase in the phagosome, and actively acidify the phagosomal pH to 6.5 as part of their strategy to survive in Mo phagosomes.

Macrophages in host defense against Histoplasma capsulatum.

Macrophages function in both innate and cell-mediated immunity in host defense against pathogenic fungi. They initially serve as a protected environment in which the primary fungal pathogen Histoplasma capsulatum multiplies and disseminates from the lung to other organs. Upon induction of cell-mediated immunity, cytokines activate macrophages to destroy the yeasts and thus remove them from the host.

Modulation of the effector function of human macrophages for Histoplasma capsulatum by HIV-1. Role of the envelope glycoprotein gp120.

We have demonstrated that monocyte-derived macrophages (Mphi) from HIV+ individuals are deficient in their capacity to phagocytose Histoplasma capsulatum (Hc) yeasts, and are more permissive for the intracellular growth of Hc. To determine whether these defects in Mphi function were caused by HIV infection of the Mphi and/or by pathological events associated with HIV infection, cultured normal human Mphi were infected with the HIV-1BaL strain. Virus production, quantified by reverse transcriptase activity and p24 antigen, was evident on day 8 after infection and peaked on day 16. On days 12, 16, and 20 after infection, HIV-1-infected Mphi were deficient in their capacity to recognize and bind Hc yeasts compared with control Mphi, and also were more permissive for the intracellular growth of Hc. Culture of normal Mphi with the envelope glycoprotein gp120 inhibited phagocytosis of Hc yeasts by Mphi in a concentration-dependent manner, but did not cause more rapid intracellular growth of Hc. Normal Mphi cultured in the serum of HIV+ individuals with impaired Mphi function subsequently were deficient in their capacity to phagocytose Hc yeasts, and were more permissive for the intracellular growth of yeasts compared with Mphi cultured in normal serum. Conversely, culture of normal Mphi in the serum of HIV+ patients with normal Mphi function did not affect the interaction of Hc yeasts with Mphi. Moreover, when Mphi from HIV+ individuals that were initially defective in host defense against Hc were cultured in normal HIV- serum, normal Mphi function was demonstrated. Adsorption of gp120 from the serum of two HIV+ patients removed the capacity of the serum to cause a Mphi defect in phagocytosis of Hc, but had no effect on the capacity of the serum to cause accelerated intracellular growth. These data demonstrate that observed defects in Mphi interaction with Hc yeasts may be caused by gp120 and other, as yet unknown serum component(s) probably released into serum by HIV-infected cells.

Activation of human macrophage fungistatic activity against Histoplasma capsulatum upon adherence to type 1 collagen matrices.

Human monocyte/macrophages (Mphi) were adhered to extracellular matrix proteins, and the intracellular growth of Histoplasma capsulatum (Hc) yeasts were quantified and compared with their growth in Mphi adhered to plastic. Freshly isolated monocytes and cultured monocyte/derived Mphi adhered to type 1 collagen gels, but not to nongelled collagen-, fibronectin-, laminin-, or vitronectin-coated surfaces, demonstrated significant fungistatic activity against Hc yeasts. Activation of Mphi developed immediately upon adherence to the collagen matrices (1 h) and did not require additional time in culture. In addition, many of the yeasts were digested by 24 h postinfection. Mphi adhered to collagen maintained their fungistatic activity for up to 4 days, during which time monolayers cultured on plastic were destroyed. Culture of Mphi in the presence of IFN-gamma or TNF-alpha for 24 h before infection did not augment the fungistatic activity of collagen-adherent Mphi. Likewise, culture of monocytes on collagen gels with IL-3, granulocyte-Mphi CSF (GM-CSF) or Mphi CSF (M-CSF) for 7 days did not enhance Mphi fungistatic activity above that obtained by monocytes cultured on collagen alone. The mechanism(s) of Mphi-mediated fungistasis was not associated with production of toxic oxygen radicals, nitric oxide, or the restriction of intracellular iron. However, experiments with horseradish peroxidase-labeled gold colloids and immunoelectron microscopy demonstrated that phagolysosomal fusion, which is minimal in Hc-infected Mphi adhered to plastic, is enhanced significantly at both 1 h and 24 h postinfection in Mphi adhered to collagen matrices. These data suggest that in vivo, matrix-bound Mphi may express a previously unrecognized antifungal activity that proceeds in the absence of exogenous cytokines and is mediated, in part, by overcoming the capacity of Hc yeasts to inhibit Mphi phagolysosomal fusion.

Role of cell-surface molecules of Blastomyces dermatitidis in host-pathogen interactions.

The fungal pathogen Blastomyces dermatitidis produces an adhesin (WI-1) in yeast stages, which contains repetitive regions that bind host-cell receptors. Adhesin and glucan may modulate fungal interactions with macrophages; their level of expression is altered in hypovirulent mutants. Adhesin is also involved in immune responses, and may be important in eliciting the clearance of the fungus.

Oxidative killing of Cryptococcus neoformans by human neutrophils. Evidence that fungal mannitol protects by scavenging reactive oxygen intermediates.

Polymorphonuclear neutrophils (PMN) kill Cryptococcus neoformans (Cn) by oxidative mechanisms, but the roles of various reactive oxygen intermediates (ROIs) are not known. We used a mannitol low-producing Cn mutant (Cn MLP) and its wild-type parent (Cn H99) to examine the role of ROIs distal to H2O2 in PMN killing and to determine whether mannitol produced by Cn protects the fungus against ROIs. At PMN:Cn cell ratios of 1:1, 10:1, and 100:1, PMN killed significantly more Cn MLP than Cn H99 cells after 2 and 4 h (p less than 0.05). Superoxide dismutase and the hydroxyl radical (OH.) scavengers mannitol and DMSO inhibited killing of both strains (p less than 0.05), but catalase did not. Cn H99 and Cn MLP stimulated PMN to produce similar amounts of O2- and H2O2. In contrast, Cn MLP stimulated greater luminol-dependent chemiluminescence than did Cn H99 (p less than 0.05). Finally, H2O2 alone killed similar numbers of Cn H99 and Cn MLP cells, but oxidants generated by FeSO4 (1 microM), H2O2 (10 microM), and iodide (1 to 3 microM) killed significantly more Cn MLP than Cn H99 cells in 1 h (p less than 0.05). Mannitol, DMSO, and catalase completely inhibited killing of both Cn strains by this cellfree system, but superoxide dismutase did not. These results suggest that 1) distal ROIs such as OH. and HOCI are key effector molecules against Cn, and 2) mannitol produced by Cn may protect against oxidative killing by scavenging distal ROIs.

Inhibition of growth of Histoplasma capsulatum yeast cells in human macrophages by the iron chelator VUF 8514 and comparison of VUF 8514 with deferoxamine.

Histoplasma capsulatum requires intracellular iron to survive and multiply within human and murine macrophages (M phi). Thus, iron chelators may be useful compounds in the treatment of histoplasmosis. In the present study we compared the efficacies of five different iron chelators with deferoxamine (DEF) for their capacity to inhibit the growth of H. capsulatum yeast cells in culture medium and within human M phi. Of the agents tested, only one, VUF 8514, a 2,2'-bipyridyl analog, was found to be effective. VUF 8514 inhibited the growth of yeast cells in tissue culture medium and within M phi in a dose-response fashion. In tissue culture medium, the 50% effective dose (ED50) of VUF 8514 was 30 nM and the ED50 of DEF was 1 mM. In human M phi, the ED50 of VUF 8514 was 520 nM and the ED50 of DEF was 4 mM. Thus, VUF 8514 was effective at a concentration 7.7 x 10(3)-fold lower than DEF in inhibiting the growth of yeast cells in M phi. Inhibition of the intracellular growth of yeast cells by VUF 8514 was reversed by holotransferrin and iron nitriloacetate, an iron compound that is soluble at neutral to alkaline pH. Thus, VUF 8514 inhibits the intracellular growth of yeast cells by acting as an iron chelator rather than through its capacity as a weak base. These data suggest that the hydroxamic acid siderophore of H. capsulatum yeast cells competes successfully for iron against some iron chelators but not others and that VUF 8514 may be a potential therapeutic agent for the treatment of histoplasmosis.

Myelinogenic potential of an immortalized oligodendrocyte cell line.

The myelinogenic potential of an oligodendrocyte cell line (N20.1) immortalized by transformation with a temperature-sensitive retrovirus (Verity et al., J Neurochem 60:577-587, 1993) has been evaluated in a co-culture system utilizing dorsal root ganglion neurons. When N20.1 cells were placed in co-culture with dorsal root ganglion neurons at 39 degrees C, the temperature at which TAg expression is decreased relative to that in cells maintained at 34 degrees C, there was a dramatic decrease in the N20.1 proliferation rate compared to cells maintained in the absence of neurons at either temperature. This decrease in proliferation was observed within 3 days of co-culture and appeared to precede a further decrease in TAg expression that occurred with time in response to the neurons. In co-cultures the immunoreactivity of N20.1 cells for galactocerebroside increased with time, and the cells appeared to establish contact with neurites and initiate formation of membranous sheets. When the duration of co-culture was extended to 52 days, myelin-like figures were noted by electron microscopy. Thus, the extent of N20.1 differentiation is dependent on the presence of neurons and the duration of co-culture. This culture system represents a potentially powerful tool for the study of neuronal-glial interactions influencing myelinogenesis and remyelination.

Macrophages from human immunodeficiency virus-positive persons are defective in host defense against Histoplasma capsulatum.

The phagocytic and fungistatic activity of monocyte-derived macrophages from human immunodeficiency virus (HIV)-positive persons against Histoplasma capsulatum yeasts was determined. Macrophages from HIV-positive patients were profoundly deficient in their capacity to recognize and bind H. capsulatum, but ingestion of bound yeasts was normal. The binding of H. capsulatum by patient macrophages tended to decrease with a decrease in CD4+ T lymphocyte counts. Another major defect was that patient macrophages were more permissive for the intracellular growth of H. capsulatum. Macrophages from 22 of 58 patients showed a > or = 2-fold increase in intracellular growth compared with control macrophages. Thus, in addition to defects in cell-mediated immunity caused by a loss of CD4+ T cells, macrophages from HIV-positive patients exhibit intrinsic defects in macrophage function against H. capsulatum that may contribute to the increased susceptibility of HIV-positive patients to disseminated histoplasmosis.

The WI-1 antigen of Blastomyces dermatitidis yeasts mediates binding to human macrophage CD11b/CD18 (CR3) and CD14.

Three genetically related strains of Blastomyces dermatitidis (Bd) yeasts that differ in their expression of WI-1, an immunodominant cell wall Ag, were tested for their capacity to bind to human macrophages (M phi) in the absence of serum. These strains included American Type Culture Collection (ATCC, Rockville, MD) ATCC 26199, which is virulent for mice; an attenuated mutant strain ATCC 60915, which expresses 2.5-fold more WI-1 than strain 26199; and an avirulent mutant strain, ATCC 60916, which has sixfold more WI-1 than strain 26199. Attachment of both mutant strains to M phi was rapid and was maximum after 10 min at 37 degrees C. Attachment of strain 26199 to M phi was approximately 40% of that obtained with the mutants. Binding of Bd to M phi was temperature- and Mg(2+)-dependent, and heat-killed yeasts bound to M phi as well as viable yeasts. Experiments with receptor-specific mAbs demonstrated that 26199 yeasts bound predominantly to the LPS binding site on CD11b/CD18 (CR3). However, the mutants bound to M phi CD14 as well as CR3. Fab anti-WI-1 inhibited the binding of all strains to M phi by 69 to 78%. Latex microspheres coated with purified WI-1 or a 25-amino acid tandem repeat located within WI-1 also bound to M phi CR3 and CD14. These data demonstrate that: 1) WI-1 is a major ligand on Bd that mediates attachment of yeasts to human M phi; 2) the binding activity of WI-1 is located within the 25-amino acid tandem repeat; and 3) binding of Bd yeasts to M phi is mediated through the LPS binding site on CR3 and CD14.

Clinically significant mucosal candidiasis resistant to fluconazole treatment in patients with AIDS.

Eight cases of severe mucosal candidiasis in patients with AIDS who were taking fluconazole at a dosage of 400-800 mg/d are described. Candida albicans alone or in conjunction with Torulopsis glabrata or Candida stellatoidea was isolated from each patient. In vitro susceptibility testing demonstrated resistance to fluconazole in all eight cases. All tested isolates were susceptible to amphotericin B, and six of eight isolates tested were susceptible to itraconazole. All individuals were severely immunocompromised (CD4 lymphocyte counts: mean, 15/mm3; range, 6-39/mm3) and had been receiving prophylaxis with fluconazole for a mean of only 3 months (range, 1-7 months). The occurrence of candidal mucositis in patients receiving high doses of fluconazole is a matter of concern that requires further study in regard to the causes, prevention, and treatment of the disease.

Altered expression of surface protein WI-1 in genetically related strains of Blastomyces dermatitidis that differ in virulence regulates recognition of yeasts by human macrophages.

The molecular basis for pathogenicity and virulence of the dimorphic fungus Blastomyces dermatitidis remains unknown. WI-1 is a major cell wall protein of B. dermatitidis yeasts and is a recognition target of both humoral and cell-mediated immunity. As an initial study to determine if WI-1 might be linked to virulence of B. dermatitidis, we quantified WI-1 expression on three genetically related strains that differ in their virulence for mice: wild-type virulent ATCC strain 26199, mutant ATCC strain 60915 (which is 10,000-fold reduced in virulence), and mutant ATCC strain 60916 (which is avirulent). Two principal alterations in WI-1 expression were observed in the mutants. First, the mutants express more WI-1 on their surface, as quantified by flow cytometry with monoclonal antibody to WI-1 and by radioimmunoassay, but the WI-1 on their cell wall is less extractable than that on the wild-type strain. Second, the mutants shed less WI-1 during culture and demonstrate impaired processing of shed WI-1. Surface alterations in WI-1 were accompanied by significant differences in the binding of the virulent and mutant strains to human monocyte-derived macrophages. Attachment of yeasts to macrophages paralleled and was proportional to the expression of WI-1. Compared with wild-type yeasts, both mutants bound to macrophages more rapidly and in two- to threefold-greater magnitude. Furthermore, about 75% of yeast binding to macrophages was inhibited by a Fab anti-WI-1 monoclonal antibody. These results suggest that altered WI-1 expression on attenuated and avirulent mutant B. dermatitidis yeasts greatly facilitates macrophage recognition and binding of yeasts and, in turn, may contribute to more rapid ingestion and killing in the host.

Chloroquine induces human macrophage killing of Histoplasma capsulatum by limiting the availability of intracellular iron and is therapeutic in a murine model of histoplasmosis.

We investigated the role of intracellular iron on the capacity of Histoplasma capsulatum (Hc) yeasts to multiply within human macrophages (Mphi). Coculture of Hc-infected Mphi with the iron chelator deferoxamine suppressed the growth of yeasts in a concentration-dependent manner. The effect of deferoxamine was reversed by iron-saturated transferrin (holotransferrin) but not by iron-free transferrin (apotransferrin). Chloroquine, which prevents release of iron from transferrin by raising endocytic and lysosomal pH, induced human Mphi to kill Hc. The effect of chloroquine was reversed by iron nitriloacetate, an iron compound that is soluble at neutral to alkaline pH, but not by holotransferrin, which releases iron only in an acidic environment. Chloroquine (40-120 mg/kg) given intraperitoneally for 6 d to Hc-infected C57BL/6 mice significantly reduced the growth of Hc in a dose-dependent manner. At 120 mg/kg there was a 17- and 15-fold reduction (P < 0.01) in CFU in spleens and livers, respectively. The therapeutic effect of chloroquine also correlated with the length of treatment. As little as 2 d of chloroquine therapy (120 mg/kg), when started at day 5 after infection, reduced CFU in the spleen by 50%. Treatment with chloroquine for 10 d after a lethal inoculum of Hc protected six of nine mice; all control mice were dead by day 11 (P = 0.009). This study demonstrates that: (a) iron is of critical importance to the survival and multiplication of Hc yeasts in human Mphi; (b) in vitro, chloroquine induces Mphi killing of Hc yeasts by restricting the availability of intracellular iron; and (c) in vivo, chloroquine significantly reduces the number of organisms in the spleens and livers of Hc-infected mice and can protect mice from a lethal inoculum of Hc yeasts. Thus, chloroquine may be effective in the treatment of active histoplasmosis and also may be useful in preventing relapse of histoplasmosis in patients with acquired immunodeficiency syndromes.