[Role of erythrocytes in mechanisms of nonspecific protection of blood in infection caused by the fungus of genus Paecilomyces.]

Paecilomyces variotii is a commonly occurring species in air and food, and it is also associated with many types of human infections. Tissue forms of the fungus Paecilomyces variotii or their cytoskeletons were revealed in the cytoplasm of erythrocytes in patients with allergy and bronchial asthma in paecilomycosis. Our study was aimed at investigating the role of red blood cells in the mechanisms of the nonspecific protection of the host in conditions of chronic persistent infection of the blood with the fungus of the genus Paecilomyces. We examined a total of eighty-four 16-to-72-year-old patients (39 men and 45 women) presenting with activation of paecilomyces infection in blood. We used laboratory, biochemical, allergic-and-immunological and microbiological methods of study. Fungal cultures were identified phenotypically and by means of phylogenetic analysis.Our findings are suggestive of a new type of the oxygen-dependent mechanism of cytotoxicity of erythrocytes, which is caused by permanent formation of reactive oxygen species as a result of non-enzymatic oxidation of haemoglobin to methaemoglobin. The resulting superoxide anion radical (O2-), hydrogen peroxide (H2O2), and hydroxyl radical (OH-) exhibit a powerful bactericidal action which is, probably, activated when the fungal cells are captured and immersed in the erythrocyte cytoplasm or in a closed cavity formed by RBCs around large fungal cells. In conditions of chronic blood infection with tissue forms of fungi of the genus Paecilomyces oxygen-dependent cytotoxicity of erythrocytes is the main mechanism of readjustment of blood from the infectious agent of Paecilomycosis.

[The clinical immunologic characteristic and laboratory diagnostic of bronchial asthma under pecilomycosis.]

The actuality of the problem is is conditioned by detection of infection of patients' blood with fungi genus paecilomyces in the form of carriage or activation of pecilomycosis infection with clinical manifestation of allergic diseases and bronchial asthma. The purpose of study is to analyze clinical and immunologic characteristics of bronchial asthma under pecilomycosis and to develop new approaches to its laboratory diagnostic in conditions of the primary health care unit. The complex examination of patients that included collection of allergy anamnesis, physical examination, application of laboratory, cytology, immunology and microbiology analysis was carried out. The diagnosis of pecilomycosis was established at the basis of express-diagnostic using direct count of mature spherules and yeast-like cells of fungi genus Paecilomyces in blood sample, detection of specific IgE- and IgG antibodies to allergen of fungus Paecilomyces spp. in blood of patients and cultural analysis. The clinical immunological characteristics of atopic and endogenous (non-atopic) forms of bronchial asthma in 143 patients were examined. The activation of pecilomycosis was established in 120 (83.9%) patients: the content of mature spherules of fungus genus Paecilomyces in blood varied from 8 000 to 55 000 (in average 17 500±755) in 1 mkl at the standard 1000-6000 (in average 3417±433) in 1 mkl. The specific IgE- and IgG antibodies to allergen of fungus Paecilomyces spр. were detected (57% and 81% correspondingly). The development of bronchial asthma is induced by activation of pecilomycosis in blood: it occurred against the background of acute respiratory viral infection in 74 patients (62%), seasonal and age fluctuations of immune status in 34 patients (28%), under stress impact in 12 patients (10%). The immune response to infection with fungus genus Paecilomyces is conditioned by mechanisms of non-specific resistance and specific immunity. The eosinophils and natural killers are involved into sanation of blood from fungal cells of non-phagocyting sizes using antibodies-mediators of specific IgE and IgG antibodies to allergen of fungus Paecilomyces as effector cells in reaction of antibody-dpendent and cell mediated cytotoxicity. The phagocytosis of small fungal forms was detected: endospores by monocytes, neutrophils, eosinophils and cytolysis of fungal cells by erythrocytes. The obtained data permits to differentiate carriage or activation of pecilomycosis in blood of patients with bronchial asthma, to mark out individuals with atopic or non-atopic profile of immune reaction determining specificity of their immune defense from agent of fungal infection and clinical picture of disease. The algorithm is proposed concerning diagnostic of bronchial asthma of pecilomycosis etiology in conditions of primary health care unit.

[The cultural features and parasitic characteristics of fungus genus Paecilomyces species Paecilomyces Variotii Bainier (1907).]

The fungi genus Paecilomyces are spread everywhere in nature. They attract attention from the middle of 1960s as a cause of development of septic endocarditis, sepsis, pneumonia and other types of pathology developed against the background of immunodeficiency. They induce development of infectious allergic alveolitis and bronchial asthma. The cultural features and pathogenic characteristics of clinical isolates (n=6) of fungus Paecilomyces variotii Bainier (1907) are studied. All isolates are separated from blood of patients with bronchial asthma visited allergologist of Moscow polyclinic. The culture of cells of embryonic kidney of bull-calf morphology and biological characteristics of tissue parasitic forms of fungus were analyzed. The typical sign of parasitic form of fungus P. variotii Bainier (1907) was a development of chlamydospores containing globe-shaped formations - endospores. The analysis of transformation of mycelial saprophyte form of fungus into tissue parasitic form established involvement of chlamydospores and apparently others organs of fruiting of vegetative body of fungus into transition to parasitic way of life in host organism with development of spherules and yeast-like cells. The cycle of development of tissue parasitic form from endospore to mature tissue form comprised 6 hours.

[Pathogenesis of myocarditis in patients with bronchial asthma and pecilomycosis].

We studied the influence of activation ofpecilomycotic infection in blood on the morphofunctional state of myocardium based on the clinical and pathomorphological data. The study included 23 patients with bronchial asthma (BA) concomitant with pecilomycosis after acute viral respiratory infection; protocols of autopsies and pathological sections of the heart muscle of the patients who died from asphixy and acute cardiac insufficiency were analyzed. Histological sections showed up vascular dystonia of the microcirculation bed, arteriole spasms, wall destruction in small and medium-size vessels and the adjacent muscular tissue, stromal oedema, valvular swelling, andperiwall endocarditis. Cardiac cavities and coronary vessels contained erythrocyte-rich thrombi with ferulas of Paecilomyces at different stages of development. Clinical manifestations of destructive changes in the heart muscle developing under effect of Paecilomyces infection had the form of rhythm and conductivity disorders. The local protective inflammatory reaction of productive type had the form of intermediate and vascular-type myocarditis passing to postmyocarditic cardiosclerosis. Periodic bursts of activation ofpecilomycotic infection in blood with the accumulation of fungal phospholipase A2 and lipid metabolites produced membranotoxic effect and can play the key role in the development of atherosclerosis and hypertensive disease in patients with BA and pecilomycosis.

[HYPEREOSINOPHILIC SYNDROME ASSOCIATED WITH SEPSIS DUE TO PAECILOMYCES FUNGI DISSEMINATED INTO THE LIVER].

A 49 year old woman with signs of chronic sepsis, hepatomegaly, and high eosinophil count was under long-term examination including consultations with an oncologist, parasitologist, and hematologist, diagnostic laparotomy, and studies of liver biopsies. Seeding blood samples onto Saburo's medium resulted in the growth of Paecilomyces variotii Bainier colonies. Counting mature spherules of the fungus revealed 59000 spherules per 1 mcl compared with the normal value of 1000-6000 which suggested paecilomycotic etiology of sepsis. The histological study of liver biopsies demonstrated hemorrhagic foci and eosinophilic infiltrates around fungal spherules. The clinical recovery of the patient was achieved after 3 courses of pulsed terbinafine therapy (500 mg/d every other day for 14 days during a month) in combination with vitamins and i/v infusion of 100 ml of a fluconazole solution (2 mg/ml) every third day (10 procedures during a course of therapy).