Differential development of CD4 and CD8 cytotoxic T cells (CTL) in PBMC across the leprosy spectrum; IL-6 with IFN-gamma or IL-2 generate CTL in multibacillary patients.

In the present study we evaluated the contribution of CD4 and CD8 T cells on the antigen-specific cytotoxic activity induced by whole Mycobacterium leprae in leprosy patients and normal controls (N) as well as the modulation of this activity by some cytokines. Peripheral blood mononuclear cells (PBMC) from N or from leprosy patients were stimulated with antigen in the presence or absence of cytokines for 7 days. M. leprae-stimulated PBMC were depleted of CD4 or CD8 antigen-bearing cells and employed as effector cells in a 4-hr [31Cr]-release assay against autologous M. leprae-pulsed macrophages. Our results demonstrate that both CD4 and CD8 T cells contribute to M. leprae-induced cytotoxic activity, with differences observed in paucibacillary (PB) and multibacillary (MB) patients. CD8-mediated cytotoxic activity is higher than that of CD4 cells in PB patients, while in MB patients CD4 cytotoxicity is predominant. Our data also demonstrate that the generation of CD4 and CD8 cytotoxic T lymphocytes (CTL) can be modulated differentially by interleukin-4 (IL-4), IL-6, gamma interferon (IFN-gamma), or IL-2. Although MB patients developed the lowest CTL response, cytokines such as IL-6 plus IL-2 or IFN-gamma were able to generate both CD4 and CD8 cytotoxic T cells from MB patients. In PB patients, IL-6 plus IFN-gamma displayed the highest stimulation on CD8 effector cells. Thus, an important role may be assigned to IL-6, together with IL-2 or IFN-gamma, in the differentiation of M. leprae-specific CTL effector cells.

Lack of cytotoxic activity against Mycobacterium leprae 65-kD heat shock protein (hsp) in multibacillary leprosy patients.

Cytotoxic T cells play an important role in host defence mechanisms, as well as in the immunopathology of leprosy. In this study, we evaluated whether Mycobacterium leprae hsp18, hsp65 and Myco. tuberculosis hsp71 could induce cytotoxic T cell activity against autologous macrophages pulsed with these hsp. Paucibacillary (PB) patients and normal controls generated more effector cells than multibacillary (MB) patients with all three hsp tested. There was no cross-reactivity between any of the hsp tested. Mycobacterium leprae hsp65 induced cytotoxic responses only in those MB patients undergoing an erythema nodosum leprosum (ENL) episode. Although hsp65 and hsp18 induced similar proliferation in MB patients, a high proportion of these patients did not generate cytotoxic effector cells in response to hsp65. Hence, those T cells reacting to hsp65 may play an important role in the control of Myco. leprae infection.

IFN-gamma, IL-6 and IL-4 modulate M. leprae- or PPD-specific cytotoxic T cells in leprosy patients.

Specific cytotoxic T cells against intracellular pathogens may be generated in vitro. On the other hand it is well known that cytokines can regulate almost every aspect of immune function. The aim of this study was to evaluate the effect of some cytokines on the generation of cytotoxic T cells with specificity for Mycobacterium leprae- or PPD-pulsed autologous macrophages from leprosy patients and normal controls. Peripheral blood mononuclear cells from M. bovis BCG-immunized controls or from leprosy patients were stimulated with antigen, in the presence or absence of cytokines, for 7 days. These were used as effector cells in a 4-h [51Cr]-release assay. Our results show that development of cytotoxic T cells may be enhanced by gamma-IFN, IL-6 or the combination of IL-6 and IL-2. Addition of IL-2 or TNF-alpha alone did not modify the generation of cytotoxic activity. IL-4 down-regulated the cytotoxic response and gamma-IFN was able to counteract this effect. Hence, the generation of specific cytotoxic T cells can be modulated by cytokines. Whether this cytotoxic mechanism contributes to protection or tissue damage in M. leprae infection remains to be determined.

T-cell-mediated cytotoxicity against Mycobacterium antigen-pulsed autologous macrophages in leprosy patients.

The involvement of CD4+ T lymphocytes in the defense mechanisms against intracellular pathogens is widely recognized. Little information is available on the generation and specificity of the cytotoxic cells that eliminate human monocytes/macrophages infected with mycobacteria. In this work, we tested whether mononuclear cells from leprosy patients could generate cytotoxic T-cell activity against autologous macrophages pulsed with Mycobacterium leprae or purified protein derivative (PPD) in a 4-h 51Cr release assay. Peripheral blood mononuclear cells from normal Mycobacterium bovis BCG-immunized controls or from leprosy patients stimulated with antigen for 7 days were used as effector cells. Paucibacillary (PB) patients and normal controls yielded more active effector cells in this system than multibacillary (MB) patients. MB patients were able to develop cytotoxicity against M. leprae, BCG, or PPD, in contrast with the immunological anergy widely described. We did not find cytotoxicity against unpulsed macrophages. Cross-reactivity was observed between PPD, BCG, and M. leprae. Only antigen-pulsed autologous macrophages were suitable as target cells. M. leprae-induced cytotoxic cells were found in both CD4+ CD8- and CD4- CD8+ T-cell subsets, whereas CD4+ cells were the main component of PPD-induced cytotoxicity. In MB patients, BCG-induced cytotoxic cells were better killers of M. leprae-pulsed macrophages than cells induced by M. leprae. This is an interesting finding in view of the ongoing vaccination trials. The involvement of CD4- or CD8-mediated cytotoxicity may be important in the balance between protection and tissue or nerve damage.

The treatment of melasma. 20% azelaic acid versus 4% hydroquinone cream.

The efficacy of 20% azelaic acid cream and 4% hydroquinone cream, both used in conjunction with a broad-spectrum sunscreen, against melasma was investigated in a 24-week, double-blind study with 329 women. Over the treatment period the azelaic acid cream yielded 65% good or excellent results; no significant treatment differences were observed with regard to overall rating, reduction in lesion size, and pigmentary intensity. Severe side effects such as allergic sensitization or exogenous ochronosis were not observed with azelaic acid.

Reduced suppressor cell response to Mycobacterium leprae in lepromatous leprosy.

We have previously shown that concanavalin A (ConA) induction of suppressor cell activity is impaired in patients with lepromatous leprosy (LL). In this study, we demonstrated that the proportion of cells bearing the Leu8 antigen (associated with suppressor-inducer cells) is low in LL patients and tends to normalize during the erythema nodosum leprosum (ENL) episode. Antigen-induced suppressor cell function was evaluated by a two-stage assay. In the first stage, peripheral blood mononuclear cells (PBMC) were cultured for 5 days either in the presence of gamma-irradiated Mycobacterium leprae or in tissue culture medium as a control. In the second stage, mitomycin C-treated suppressor or control cells were added to phytohemagglutinin (PHA)- or ConA-stimulated autologous PBMC. The results indicate that the ability of M. leprae to induce suppressor activity was lower in LL patients than in patients with tuberculoid (TT) and intermediate clinical (BB, BL, BT) forms and Mycobacterium bovis BCG-immunized normal controls. In ENL patients, the percent suppression was between that of TT and normal individuals. M. leprae-induced suppression was more effective on ConA- than on PHA-triggered T-cell proliferation in all groups. In contrast, normal PBMC cultured for 5 days in RPMI 1640 medium (N-C) and cells from patients with leprosy (TT-C and LL-C) had effects of their own on PHA- or ConA-induced proliferation. LL-C depressed the response to ConA and enhanced PHA-induced proliferation of autologous cells. Conversely, TT-C reduced PHA-induced proliferation and increased the ConA response. Suppression of proliferation could not be overcome with exogenous interleukin-2 and was not related to the induction of the Tac antigen. The abilities of LL, TT, ENL, and normal cells to proliferate upon PHA or ConA stimulus were similar, indicating that the defect in the generation of in vitro suppression by M. leprae in LL patients occurred during the induction period (step 1 of assay).

Experimental reproduction of leprosy in seven-banded armadillos (Dasypus hybridus).

Four autochthonous seven-banded armadillos (Dasypus hybridus) from Argentina which developed disseminated leprosy are reported. The average time from inoculation to death was 14.6 months. Acid-fast bacilli (AFB) were found in the skin, nerves, spleen, lymph nodes, lungs, meninges, and striated muscle. Several tests were used to prove that the AFB found in the infected animals were Mycobacterium leprae: a) failure to grow in standard culture media for mycobacteria, b) extraction of acid-fastness with the pyridine test, c) patterns of growth in the mouse foot pad, and d) histopathologic features.

ConA-induced suppressor cells in lepromatous leprosy patients during and after erythema nodosum leprosum.

ConA-induced suppressor activity in patients with lepromatous leprosy (LL) was studied. Patients were studied during and after erythema nodosum leprosum (ENL) reactions. The study included 16 patients with ENL, nine of whom returned once the ENL episode was over. Patients were compared to 12 normal controls. Suppressor activity was evaluated in vitro by cultivating peripheral blood lymphocytes (PBL) with an inducer of T suppressor cells, concanavalin A (ConA), and with two different mitogens, phytohemagglutinin (PHA) and ConA, in order to measure the inhibition of the proliferative responses in all cases. In contrast, in LL patients during ENL the ConA-induced suppressor response was markedly reduced. The reduction in suppressor responses was even more marked in the LL patients after the ENL episode. Reduced levels of suppressor activity in LL patients reveal a defect in central mechanisms of control in the immune response.

Arsénico y sus complicaciones

Referente a la intoxicación crónica por arsénico en Argentina, cuya fuente principal son las aguas subterráneas ya que ciertas áreas endémicas sobrepasan las concentraciones máximas aceptadas. Presenta la fisiopatología de la intóxicación y cuatro casos clínicos

Arsénico y sus complicaciones

Referente a la intoxicación crónica por arsénico en Argentina, cuya fuente principal son las aguas subterráneas ya que ciertas áreas endémicas sobrepasan las concentraciones máximas aceptadas. Presenta la fisiopatología de la intóxicación y cuatro casos clínicos

Defective blood mononuclear phagocyte function in patients with leprosy.

Patients with lepromatous leprosy possess a defective lymphocyte function in vivo and in vitro that is less evident in the tuberculoid form. Data concerning their macrophage ability to digest Mycobacterium leprae are controversial. The purpose of this study was to determine whether monocytes from patients with either tuberculoid or lepromatous leprosy were altered in their enzyme systems, that is myeloperoxidase-dependent and myeloperoxidase-independent systems. The ability of adherent blood monocytes to ingest and kill Candida pseudotropicalis after 30 and 60 min of incubation with yeast cells was tested. Mononuclear phagocytic cells from patients with either principal form of leprosy functioned similarly to normal monocytes in phagocytosis while their fungicidal activity for C. pseudotropicalis was statistically significantly altered and was more evident in the lepromatous than in the tuberculoid type. The results indicate that peripheral blood monocytes from patients with leprosy possess an impaired enzymatic candidacidal activity.

Reacción leprosa y lavamisole (R. 12.564)

Se trataron veinticinco pacientes de lepra lepromatosa con brote subintrante de eritema nudoso con Levamisole, más la medicación específica y Talidomida que recebían observándose que el agregado de Levamisole, permite reducir la dosis necesaria de Talidomida para controlar el brote.