The defence mechanism in sporotrichosis and the influence of corticosteroid (Cs) application were studied.
A healthy adult man was intracutaneously injected 0.2ml Sporothrix schenckii spore suspension (1.8×106 spores) at 12 sites on his forearm. A half of the sites were applied Cs ointment. The injected sites were successively biopsied on day 3, 7, 14, 14, 21 and 28. The specimens were examined histopathologically and immunohistochemically using PAP method (antibodies used were; anti-MT-1, IgG, IgA, IgM, IgE, C3, lysozyme, α-antitrypsin and α1-antichymotrypsin, S-100 protein), The specimen taken on day 28 was also examined using ABC method (antibodies used were; anti-leu 1, 2a, 3a, 4, 6, 7, 14, M1 and anti-OKIA1).
Histopathologically, control sites showed in its course abscess formation, transepidermal elimination of the abscess, maturation of Mo-Mφ around the abscess, and increase of the lymphocytes surrounding the granuloma. Fungal elements were seen mostly in the granuloma, but gradually decreased during the course. Cs inhibited these processes, and in Cs-applied group the number of fungal elements increased during the course to form a mass of innumerable spores seen in the specimen taken on day 28. Immunohistochemically, PMN, T cells (h/iT, s/cT), B cells, plasma cells (IgG-, IgA- and IgM-positive), histiocytes and giant cells were identified. IgE-, S-100 protein positive cells, NK cells, Langerhans cells, and C3-positive substances were not detected. The subpopulations in both groups were the same in essential.
From these results it is assumed that PMN, Mφ T and B cells contribute and interact in the defence mechanism of sporotrichosis. It is also shown that Cs application facilitated the fungal proliferation. The inhibition of PMN aggregation may be a cause of this state.

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In order to clarify the defense mechanism against Sporothrix schenckii infection, we examined the tissue reactions of mice against the organism with time by both light and electron microscopy. The histological feature at an early stage was a mixed cell granuloma consisting of polymorphonuclear leukocytes (PMNs) and macrophages and later on the macrophages at the periphery had become enlarged and vacuolated. Under electron microscopy, blastopores were seen to have been phagocytized by the PMNs and macrophages, and extracellular blastopores were not seen. PMNs taking up blastopores were phagocytized by other PMNs or macrophages. At the third and fourth months, the phagosomes of the macrophage had grown in size, and contained a number of blastopores. During the experiment, the ultrastructure of the blastopores was well preserved, and their viability was considered high. In the defense mechanism against S. schenckii infection, PMN phagocytosis is of great importance, but the dynamics of the macrophages may be considered the predominant factor determining the course of the disease.

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Tissue responses to the causative organisms and their parasitic forms were re-examined in cases of chromomycosis and phaeomycotic cyst caused by Fonsecaea pedrosoi, Phialophora verrucosa and Exophiala jeanselmei. Additionally, tissue responses developed in the brain of congenitally athymic nude (nu/nu) mice and their heterozygous littermates (nu/+) which were experimentally infected with F. pedrosoi or Exophiala dermatitidis were investigated.
The results are as follows :
1) The tissue responses to the dematiaceous fungus infection depend on the strength of host defense rather than the species of the fungi. Among various defense mechanisms, the role which cell-mediated immunity plays, in particular, is important. For example, in nu/nu mice inoculated with F. pedrosoi remarkable pyogenic response occurred to the causative organisms instead of the granulomatous appearing response usually in nu/+ mice.
2) Parasitic forms of the dematiaceous fungi distributed continuously from a hyphal form to sclerotic cells, and the parasitic forms were fairly characterized by the species. On the other hand, the parasitic forms also depend strongly on the strength of host defense. For example, in compromised host the parasitic forms become almost hyphae, while in non-compromised host sclerotic cells are easily formed.
3) Due to the strength of host defense or the species of the causative organisms, clinical types of dematiaceous fungus infection are continuously changing between chromomycosis and phaeomycotic cyst. It is difficult to draw a line between the two diseases. We need on overall understanding of dematiaceous fungus infection.

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Phaeohyphomycosis and hyalohyphomycosis, relatively new terms introduced by Ajello and co-workers, encompass infections caused by various opportunistic fungi with dematiaceous and non-dematiaceous filamentous tissue forms, respectively. Clinical, histopathologic, and mycologic characteristics of these clinical entities are critically discussed. We emphasize that they serve as complements to each other for the sake of utilizing a rational and meaningful terminology for mycotic diseases.
In addition, we clarify the disease entity of “mycetoma, ” and point out criticisms on the erroneous use of this term to denote pulmonary “fungus balls.”

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This study was carried out to examine a non-specific defense reaction in experimental acute and chronic dermatophytosis which was induced by puncture inoculation of Trichophyton mentagrophytes to the human skin grafted in the nude mouse. In the acute stage, two weeks after inoculation, dense cell infiltration and microabscess consisting primarily of polymorphonuclear leukocytes was observed in the area around the depressed horny material and under the stratum corneum which contained fungal elements. This histological profile was suggestive of non-specific defense reaction to the fungus that invades through the stratum corneum into the stratum granulosum and stratum spinosum. In the chronic stage, 5 and 11 weeks after inoculation, a large number of fungal elements was found to be localized in the entire stratum corneum and even in the parakeratosis of the deep horny layer. The epidermis showed marked acanthosis accompanied by irregular elongation of rate ridges, and no intraepidermal invasion of inflammatory cells. These proliferative changes of the epidermis were also suggestive of one of the non-specific defense reactions to the fungus invasion. Electron microscopic findings of the parasites in the stratum corneum 11 weeks after inoculation were similar to those in clinical dermatophytosis. This experimental system is considered to be useful as a model for histopathological and immunological study of dermatophytosis in the human skin.

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Tinea Profunda are divided into three types according to the parasitic forms of the fungus in the dermis : acute, subacute and chronic. Kerion and tinea barbae are known to be the acute type, granuloma trichophyticum belongs to the chronic type. Recently an unusual type of granuloma trichophyticum has been reported in immune compromised host. Histologically, subcutaneous nodules consist of three layers. The inner one is neutrophilic abscesses, the intermediate one is granulomatous, and the outer one consists of a thin connective tissue. Vivid slender hyphae are found in the inner and granulomatous layers.
Next, we studied the conversion of trichophyton rubrum from hyphae to spherical cells using the agarimplantation method. Because T. rubrum affects only keratinized tissue and hardly invades the deep tissue. However, in patients with granuloma trichophyticum, hyphae grow in granulomatous tissue. Therefore, it is important to clarify whether cultures of T. rubrum isolated from the disease differ from those from trichophytia superficialis. Morphologically there are no differences between them. Thirteen cultures of T. rubrum were isolated from tinea superficialis and agranuloma trichophyticum and were implanted into the peritoneal cavities of ddY male mice. In conclusion, the parasitic forms of the cultures isolated from granuloma trichophyticum were not spherical but filamentous and were different from the cultures isolated from tinea superficialis.

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In order to clarify the tissue responses to Aspergillus in the lung and its morphologic changes in the lesions, 43 autopsy cases (invasive type 39, aspergilloma 2, and granuloma type 2) and 10 surgical cases (aspergilloma 4, allergic type 5 and other 1) were examined histopathologically, and compared with the results of experimental studies.
In the autopsy cases, the lesions were purulent or granulomatous, necrotizing, non-inflammatory, in order of decreased defense mechanisms of hosts. However, the principal lesion was a chronic necrotizing one. These lesions were associated with thrombosis, hemorrhage, infarction and histolysis. The fungal elements in the noninflammatory, necrotizing and purulent lesions revealed a radiating pattern of hyphal growth with typical dichotomous branches and septation, while those in the granulomatous lesion and a part of the purulent were a small number of large-sized spores and irregular shaped hyphae.
In the surgical specimens, the tissue lesion of primary as well as secondary aspergilloma was non-specific chronic inflammatory and the fungal elements in the tissue were occasionally phagocytized in macrophages. The granulomatous lesions were remarkable in allergic aspergillosis, but these lesions were considered to be formed by allergic mechanism to fungus rather than by its invasive growth in tissues.
It was suggested in experimental studies that the spores reached in the lung were easily eliminated by polymorphonuclear leukocytes in the healthy hosts, however the spores in compromised hosts developed variable lesions similar to human autopsy cases.

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Studies on experimental pulmonary aspergillosis in mice were carried out using a newly devised aerosol exposure apparatus.
SPF, ddy mice were pretreated with cyclophosphamide and prednisolone, and then exposed to aerosolized spores of Asp. fumigatus or Asp. ochraceus for 5hrs. Experimental mice were observed for 3 weeks. In some experiments designed to study the sequence of the pathologic lesions and fate of the inhaled spores, mice were sacrificed at intervals following exposure.
The results obtained were as follows : 1. A linear relationship was observed between times of spore-inhalation and number of fungus cells deposited in the lungs. 2. An accurate reproducibility of experimental Aspergillus infection was confirmed. 3. The inhaled fungus cells in the lung were rapidly decreased in number in healthy mice, but a number of the infected foci with abundant fungal growth were recognized 72hrs after exposure in the compromised mice. 4. By exposure to Asp. ochraceus, mild infected foci were recognized in compromised mice 72hrs after exposure. 5. It was confirmed that an inhalation of Amphotericin B aerosol was effective for experimental pulmonary Aspergillus infection in mice.

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The histopathologic study of fungal lesions in autopsy cases with leukemia was performed and compared with an animal experiment.
Necrotic lesion was predominant in the autopsy cases of candidiasis and aspergillosis. However, mucormycosis and cryptococcosis showed their own characteristic histopathologic findings, namely, thrombosis and hemorrhagic infarction due to intravascular growth of hyphae in mucormycosis and cystoid or granulomatous lesion in cryptococcosis. Neutropenia less than 1, 200/mm³ which was considered to be the lowest level of the normal range of the number of mature neutrophils in the peripheral blood was demonstrated in approximately 70% of the cases with fungal infection and this result may suggest the compromised state of the hosts. However, neutropenia was not present in about 30% of the cases with fungal infection.
Also, necrotic lesion was predominant in experimental candidiasis and aspergillosis using leukemic mice. The infiltration of neutrophils in fungal lesion was rather more marked in the animals without treatment of steroid hormone than in those with it, although the number of peripheral neutrophils increased by administration of steroid hormone in this experiment. This result suggested that an increased susceptibility to fungi was related also to impaired function of neutrophils and showed a possibility that neutrophil function was impaired in the autopsy cases without neutropenia.
According to these results, an increased susceptibility to fungi in leukemic patients is suggested to be due to neutropenia and/or impaired function of neutrophils.

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Opportunistic fungal infections occurring frequently in patients with acute leukemia are candidiasis, aspergillosis and cryptococcosis in that order.
When we try to clarify the relationship between acute leukemia and the fungal infections, it is useful to know about fungal infections to renal transplant recipients. Even though the fungal diseases occurring frequently in renal transplant recipients are the same as those in patients with acute leukemia, the frequency of each fungal disease is definitely different namely, in the renal transplant recipients, cryptococcosis is the fungal disease observed most frequently, followed by candidiasis and aspergillosis in that order.
An explanation for the difference is thought to be as follows :
1. The defense mechanisms of host against Candida albicans, Aspergillus fumigatus or Cryptococcus neoformans are different from each other.
2. The defense mechanisms in patients with acute leukemia are different from those in renal transplant recipients.
3. Ca. albicans is one of the members of normal human flora and A. fumigatus is frequently isolated from air. We contact them frequently in daily life. However, it is usually difficult to isolate Cr, neoformans from living circumstances or nature.
Due to the difference in combinations of these factors, the frequency of each fungal disease occurring in patients with acute leukemia is different from that in renal transplant recipients.

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A 48-year-old man noted itchy erythematous lesions on his face, trunk and extremities in August 1984. He was treated with topical and systemic steroid by a local dermatologist for about a month; however, the lesions spread further and in addition nodules with a thick crust developed on his face. Upon his initial visit to our hospital in October 1984, physical examination revealed many walnut sized nodules, papules and erythema with scales and crusts, especially on his face. His oral mucous membrane was covered with white plaques. There were many annular erythemas with pityroid scales on his trunk and extremities. All his toenails were thickened and discolored. His soles were also thickened with fissuring lesions. Candida albicans was cultured from the affected skin and mucosal lesions. Trichophyton mentagrophytes was cultured from his soles and toenails. Biopsy of the nodule on his forehead revealed hyperkeratotic granulomatous lesions with numerous fungal elements. A chest x-ray showed a tumor shadow on his left lung. On detailed examination, it was confirmed as a malignant thymoma. After an oral dosage of 200mg of ketoconazole for six weeks, the cutaneous lesions almost disappeared. However, the thymoma developed further despite medication. He died of dyspnea in October 1985. This case corresponds to the CMCC-Thymoma Syndrome, which Kirkpatrick described in 1979.

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Immunoelectrophoresis (IEP) and radioimmunoassay (RIA) were performed to investigate the activation pathway of complement components, C3 and C5, of normal human serum by Sporothrix schenckii. C3 conversion in normal serum was observed immuno electrophoretically, when serum was incubated either with S. schenckii cells (both yeast and mycelial forms) or with cell-wall polysaccharides. C3a and C5a concentrations as measured by RIA were 17, 800-57, 800 ng/ml and 82-925 ng/ml, respectively, and were correlated with antigen doses. C3 conversion did not occur in serum heated at 50°C or 56°C for 30min, nor chelated with EDTA, but occurred only in serum chelated with Mg#-EGTA. It was suggested that both mycelial and yeast form cells and cell-wall polysaccharides of S. schenckii activate the complement system via an alternative pathway.

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It was found in the previous report that Sporothrix schenckii cell wall polysaccharides activate the alternative pathway of a complement system. In this report, C3 conversion of 2 sera from patients with sporotrichosis was evaluated by immunoelectrophoresis. Immunoelectrophoretic C3 conversion did not occur in untreated sera, but occurred in the sera activated by S. schenckii cells. Radioimmunoassay was performed to measure C3a and C4a concentrations in sera of 7 patients with sporotrichosis which were untreated or activated in vitro by S. schenckii yeast form cells. Almost all cases showed high levels of C3a and C4a. From these data, it was possible to speculate the involvement of either classical or alternative pathway or both pathways in each patient with sporotrichosis.

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The study was performed in an attempt to clarify the present state of deep-seated mycoses. All autopsy cases during the ten-year period from 1975 to 1984 in the 2nd Department of Pathology of Shinshu University School of Medicine were used for this study. Ninety three cases of mycoses were present, which was 13.0 percent of the 714 total autopsy cases. The principal mycoses were candidiasis, aspergillosis, cryptococcosis and mucormycosis in this order. Frequent antecedent disorders were leukemia, carcinoma, malignant lymphoma and hemodyscrasia other than leukemia in this order. The rate of incidence of mycoses was the highest in malignant lymphoma (80.0%), followed by leukemia, hemodyscrasia other than leukemia and carcinoma. In candidiasis and aspergillosis, necrotic lesion was predominant. This result precisely reflected the compromised state of the hosts. In cryptococcosis and mucormycosis, initial tissue lesions were the ones common in each mycosis. In the 82.8% of mycoses associated with leukemia, the count of neutrophil leukocytes decreased less than 2, 500/mm³, which showed the immunocompromised state of the hosts. The count of lymphocytes had the same tendency to decrease when the count of neutrophil leukocytes decreased less than 2, 500/mm³.

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We compared the mycelial and granular forms of E. jeanselmei isolated from a 23-year-old male patient with chromoblastomycosis with 8 preserved strains of E. jeanselmei and 7 of E. dermatitidis morphologically, physiologically and serologically. The mycelial form of the isolated strain showed the characteristics of E. jeanselmei. The granular form of this strain differed from that of E. dermatitidis in colonial appearance, microscopic morphology and serologic reactions.
The comparison between E. jeanselmei and E. dermatitidis revealed that these 2 species had annellations but differed from each other in colonial appearance and growth temperature. It is concluded that the 2 species can be differentiated and should be considered as a different species.

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D-Arabinitol/creatinine ratios were determined in urine and serum specimens simultaneously obtained from control subjects and patients with invasive candidiasis. The mean (±SD) of urinary D-arabinitol/creatinine ratios of male (ages 0-15, 16-65, 66 years old) and female (ages 0-15, 16-65, 66 years old) were 0.4 (±0.4), 0.5 (±0.5), 0.9 (±0.8) and 0.5 (±0.3), 0.6 (±0.9), 0.6 (±0.4) μmol/mg, respectively. The higher incidence of high urinary values was recognized in the older control subjects. These results were similar to the data obtained from serum D-arabinitol values in control subjects. In the patients with invasive candidiasis there was a disagreement between the urinary D-arabinitol/creatinine ratios and the serum ratios. When serum D-arabinitol was increasing, the serum D-arabinitol/creatinine ratio was higher than the urinary D-arabinitol/creatinine ratio. After the effective treatment, the urinary ratio was higher than the serum ratio. Although the determination of serum D-arabinitol is more helpful than the urinary D-arabinitol/creatinine ratio for early diagnostic purposes, the comparison between the urinary ratio and the serum ratio also may be useful to know whether the production of D-arabinitol by Candida is increasing or decreasing in the body.

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The bacterial vaccine, heat-killed Clostridium butyricum exhibits various immunomodulating activities including a strong protection activity against Candida albicans infection in mice. The analyses of immunomodulating activities induced by the vaccine showed that the vaccine stimulated macrophage and natural killer cell activity. Stimulation of delayed type hypersensitivity, IgM antibody formation and induction of γ-type interferon were observed. The vaccine was also mitogenic for B-cell lymphocytes. Among these immunomodulating activities, interferon was considered to be one of the important factors in the manifestation of in vivo anti-candida activity. Futrher studies using recombinant interferon (IFNa-A/D type interferon) confirmed the active role of interferon in the manifestation of the protection. The interferon production by the vaccine was also found to be dependent on the mouse strain used and was most active in DDY mice.

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