Leila Maria Machado Vieira

Role of Tumor Necrosis Factor–α and Interleukin-10 Promoter Gene Polymorphisms in Leprosy

Single-nucleotide polymorphismswithin thegenescodingfortumornecrosisfactor(TNF)‐a and interleukin (IL)‐10 have been associated with several infectious diseases. To determine whether such polymorphisms are associated with leprosy, genotyping was performed at the 308 and 238 positions of the promoter of the TNF-a gene in 210 and 191 patients with multibacillary (MB) leprosy, respectively; 90 and 79 patients with paucibacillary (PB) leprosy; and 92 control subjects. For the 592 and 819 positions within the promoter of the IL-10 gene, 143 patients with MB leprosy, 79 patients with PB leprosy, and 62 control subjects were included in the analysis. TNF2 allele frequency was significantly higheramong controlsubjects than among all patients with leprosy or in the MB group ( and ). For the ILP ! .05 P ! .01 10 gene, the frequency of the homozygous 819TT genotype was significantly higher among patients than among control subjects. These data indicate that a relationship exists between TNF-a and IL-10 promoter polymorphisms and the development of PB leprosy. The more benign paucibacillary (PB) forms of leprosy— borderline tuberculoid (BT) and tuberculoid tuberculoid (TT) leprosy—are characterized by the predominance of a Th1-type immune response, the presence of well-formed granulomas at the site of the lesion, and control of mycobacterial replication. In contrast, in the multibacillary (MB) forms—borderline borderline (BB) and borderline lepromatous (BL) leprosy and lepromatous leprosy (LL)—no granuloma is seen, and high bacterial load and antibody levels are detected. Cytokines evidently play a critical role in triggering host-pathogen interactions. On one hand, greater tumor necrosis factor (TNF)–a production,

REACTIONAL STATES IN MULTIBACILLARY HANSEN DISEASE PATIENTS DURING MULTIDRUG THERAPY

It is well known that reactions are commonplace occurrences during the course of leprosy disease. Stigmatization may even be attributable to reactions which are also responsible for the worsening of neural lesions. A cohort of 162 newly-diagnosed baciloscopically positive patients from the Leprosy Care Outpatient Clinic of the Oswaldo Cruz Foundation (FIOCRUZ) was selected for this study. While 46% of the multibacillary (MB) patients submitted to the 24 fixed-dose multidrug therapy (MDT) regimen suffered reactions during treatment, it was found that all MBs were susceptible and that constant attention and care were required at all times. Fourteen per cent were classified as BB, 52% as BL, and 33% as LL. None of the variables under study, such as, sex, age, clinical form, length of illness, length of dermatological lesions, baciloscopic index (BI), or degree of disability proved to be associate with reaction among the patients studied. Reversal Reaction (RR) occurred in 45%, and Erythema Nodosum Leprosum (ENL) occurred in 55%. Among BB patients who developed reactions (15 patients), 93% presented RR; while among the LL patients who developed reactions (34 patients), 91% presented ENL. Likewise, ENL was very frequent among those with disseminate lesions, while RR was most often observed in patients with segmentary lesions. RR was also most likely to occur during the initial months of treatment. It was demonstrated that the recurrence rate of ENL was significantly higher than that of RR. Neither grade of disability nor BI was shown to be associated with RR and ENL reaction. However, the RR rate was significantly higher among patients showing BI < 3, while ENL predominated among those patients with BI > 3.

Edema na hanseníase: aspectos clínicos e terapêuticos

Edema, which is commonly described as a symptom of reactional states, may occur during the course of leprosy. Both diagnosis and adequate treatment measures are often difficult to achieve and failure to do so may result in permanent damage to the lower limbs. In a one-year follow-up study of leprosy patients--ten multibacillary and one paucibacillary--who had been submitted to a clinical protocol for diagnosis and pathological classification, a clinical pattern of localized and/or systemic edema was observed. Among these patients, five simultaneously presented other symptoms related to reactional states, 4 were diagnosed as Type I, and one as Type II. On the other hand, while three of the patients did not present reaction at the time when edema was diagnosed, they did develop some aspects of reactional disease later on (two had neuritis e one had Type I reaction). The edemas that preceded or were associated with reactional episodes showed clinical regression as a result of specific treatment against reactions (corticosteroids and/or pentoxifylline and/or thalidomide) in the absence of another treatment normally used for edemas. Although these data need to be confirmed by controlled studies, they strongly suggest that immunological mechanisms are involved in the physiopathology of edema in leprosy.Edema, which is commonly described as a symptom of reactional states, may occur during the course of leprosy. Both diagnosis and adequate treatment measures are often difficult to achieve and failure to do so may result in permanent damage to the lower limbs. In a one-year follow-up study of leprosy patients - ten multibacillary and one paucibacillary - who had been submitted to a clinical protocol for diagnosis and pathological classification, a clinical pattern of localized and/or systemic edema was observed. Among these patients, five simultaneously presented other symptoms related to reactional states, 4 were diagnosed as Type I, and one as Type II. On the other hand, while three of the patients did not present reaction at the time when edema was diagnosed, they did develop some aspects of reactional disease later on (two had neuritis e one had Type I reaction). The edemas that preceded or were associated with reactional episodes showed clinical regression as a result of specific treatment against reactions (corticosteroids and/or pentoxifylline and/or thalidomide) in the absence of another treatment normally used for edemas. Although these data need to be confirmed by controlled studies, they strongly suggest that immunological mechanisms are involved in the physiopathology of edema in leprosy.

Métodos histoquímicos para a identificação de leucócitos e macrófagos

The authors report techniques to detect acid-phosphatase, aplha-naphthyl esterase and peroxidase activity in lymphocytes and macrophages in cutaneous lesions of Leprosy. They recomend fixation in formol sucrose buffered followed by inclusion in gumma of Holt as the best method. This method mantain the activity and show sharply the details of cells.

Caracterização dos macrófagos presentes nas lesões cutâneas da hanseníase: estudo por monoclonais

The skin lesions from 16 leprosy patients were studied by immunofluorescence technique using monoclonal antibodies against monocytes (OKM1 and Anti-Mo) and la-like antigen. Acid phosphatase activity was evaluated using naphthol AS-BI phosphate as substrate. The macrophages seem to be a heterogeneous population in concern with the antigens here studied as well as the enzimatic activity. Ia-like antigen was expressed in a great number of cells throughout the clinical spectrum.The skin lesions from 16 leprosy patients were studied by immunofluorescence technique using monoclonal antibodies against monocytes (OKM1 and Anti-Mo) and la-like antigen. Acid phosphatase activity was evaluated using naphthol AS-BI phosphate as substrate. The macrophages seem to be a heterogeneous population in concern with the antigens here studied as well as the enzimatic activity. Ia-like antigen was expressed in a great number of cells throughout the clinical spectrum.

Immunofluorescence Studies in Paraffin-Embedded Tissue

Euzenir Nunes Sarno, Disciplina de Patologia Geral, Faculdade de Ciências Médicas, Av. 28 de Setembro, 87 4° Andar – Vila Isabel, Rio de Janeiro (Brazil) Dear Sir, The recent requirement to carry out immunofluorescence (IF) studies in renal biopsies justifies all attempts made to simplify this procedure. Many authors have reported successful comparative studies between frozen sections and paraffin-embedded tissue [2–5]. We have previously reported that paraffin-embedded tissue has given a high rate of false-negative results. Therefore, we continue to use frozen sections [6]. The fact is that the traditional IF method, using frozen sections, is an easy, quick and efficient method in spite of some inherent problems. The major difficulty seems to occur in the handling of the specimen. Renal biopsies are very thin cylinders of tissue and must be divided lengthwise into two equal parts. Frequently, it is difficult to obtain equal samples for IF and light-microscopic studies and if the biopsy is not large enough to be divided, IF studies cannot be done. The other practical problem is that the biopsy must be immediately snap-frozen. In order to simplify the traditional IF study, a procedure that avoids biopsy division should be used, i.e., the same sample might be used for both studies (IF and light microscopy). The method reported by Bolton and Mesnard↓X¦ seems to fit in with the above criteria. The same specimen can be processed for IF, histochemical, and light-microscopic analysis. Recently, we tried to further simplify this method by skipping several steps. 10 consecutive needle biopsies from patients with kidney disease (their histologic diagnoses are included in table I) were processed as follows: immediately after its removal, the specimen was divided into two parts: one part was placed in a small vial and snap-frozen in liquid nitrogen and the second part was fixed in formol-sucrose at 4 °C. 3-μm-thick frozen sections were obtained by cutting Table I. Histologic Diagnosis

Técnica de peroxidase-antiperoxidase (PAP) em biópsia renal: estudo comparativo

Immunofluorescent (IF) and immunoperoxidase (PAP) technics have a lot in commum in the detection of cellular and tissue antigens. In renal diseases they have been used in the detection of immunoglobulins and immune complexes. In the present paper the authors compare the technics of IF and PAP in paraffin sections to frozen renal biopsy sections by in 25 cases. There was significant correlation between IF of frozen renal biopsies with PAP of paraffin sections for immunoglobulins. There was on correlation for complement components and fibrinogen as well as for IF on paraffin sections for all components investigated. IF in paraffin sections was inadequate for the detection of deposits in renal glomeruli when perfomed in routine sections.