Undiagnosed purpura: a case of autoerythrocyte sensitization syndrome associated with dermatitis artefacta and pseudo-ainhum.

A 23-year-old young woman presented with recurrent episodes of painful bruising along with linear erosions on the accessible areas of the body of nine years duration with a pseudo-ainhum of her left nipple for the past three months. Her case history included repeated visits to various physicians at different centers and an extensive investigative profile. A diagnosis of autoerythrocyte sensitization was made on the basis of the clinical history, dermatological examination complemented by a positive autoerythrocyte sensitization test, psychiatric evaluation and absence of any organic cause for her ailment. She was placed on psychiatric management and has remained symptom-free after six months follow-up. The case is reported for its rarity, as well as for the association of autoerythrocyte sensitization syndrome with frank dermatitis artefacta and pseudo-ainhum, which to the best of our knowledge has not yet been reported in the literature.

Immunomodulatory assays to study structure-activity relationships of thalidomide.

Thalidomide, which has a long history of tragedy because of its ability to cause severe birth defects, is very effective in alleviating erythema nodosum leprosum in leprosy patients and aphthous ulcers in AIDS patients. The causes of these inflammatory diseases and the mechanism by which thalidomide diminishes them are unknown. It has been suggested that modulation of the immune response plays an important role. We found that thalidomide exerts immunomodulatory activity in three bioassays. It suppresses an IgM plaque forming cell response in mice injected with sheep erythrocytes: it inhibits TNF-alpha production by LPS stimulated human mononuclear cells: and it enhances IL-2 production by Con-A stimulated human mononuclear cells. We employed these bioassays to compare the activity of 15 analogs of thalidomide with thalidomide itself. Eight of the compounds were derivatives of the glutarimide moiety of thalidomide and the others were phthalimide or derivatives of the phthalimide moiety of thalidomide. N-hydroxyphthalimide, a simple derivative of phthalimide, was more effective than thalidomide and was also the most effective of the compounds assayed in suppressing the IgM plaque and TNF-alpha responses, but it did not enhance the IL-2 response, instead, it significantly suppressed it.

Efeito de Rs1 e Rs2 de soros de pacientes com hanseníase na cultura de linfócitos / The effect of seric Rs1 and Rs2 from Hansen's disease (HD) patients in lymphocytes cultures

No presente trabalho foi analisado efeito de receptores solúveis para E de pesos moleculares distintos (Rs1 e Rs2), presentes em níveis elevados em soros de pacientes com hanseníase, na cultura de linfócitos estimulados com fitohemaglutinina (PHA). Foram realizadas culturas de linfócitos de pacientes com hanseníase na forma lepromatosa (LV), na forma tuberculóide (LT) e linfócitos de indivíduos normais (LN), tratados com fraçöes de Rs1 e Rs2 adsorvidas com E (para adsorver Rs) e com hemácias de carneiro tripsinizadas (ET). Foi utilizado meio RPMI, contendo 10 porcento de soro bovino fetal, penicilina, estreptomicina, herpes a 37§C, com 5 porcento de tensäo de CO2, durante 72 horas. Os resultados obtidos demonstraram maior resposta proliferativa nas fraçöes Rs1 e Rs2 absorvidas com E, em relaçäo às fraçöes absorvidas com ET. Adicionalmente foram realizadas cultura de linfócitos com Rs1 e Rs2 previamente purificados, observando-se também inibiçäo na resposta linfoproliferativa à PHA. Esses resultados sugerem que tanto Rs1 como Rs2 apresentam atividade inibitória na resposta linfoproliferativa à PHA, tanto com linfócitos de pacientes com hanseníase na forma lepromatosa ou tuberculóide, bem como linfócitos de doadores normais

Evaluation of phagocytic function in Mycobacterium lepraemurium infection.

Infection of mice with Mycobacterium lepraemurium caused significant functional alterations of the mononuclear phagocyte system. Accelerated clearance of sheep red blood cells was consistently demonstrated throughout the infection and the infected mice showed progressive anaemia. Infected mice showed an enhanced ability to limit growth of phagocytosed Listeria monocytogenes in spleens during the early stages of infection, whereas moribund leprous mice lost this ability. Autoradiography showed that uninfected Kupffer cells and splenic macrophages of moribund mice could still phagocytose Listeria, suggesting that MLM infection did not affect the capacity of Listeria to localize to macrophages but interfered in some way with subsequent killing of such bacteria. The possible mechanisms underlying these observations are discussed.

Leprosy: altered complement receptors in disseminated disease.

We have studied the expression of the C3b receptor (CR1) on erythrocytes of 55 patients with Hansen's disease. We developed a radioimmunoassay utilizing a monoclonal antibody that recognized an epitope different from the C3b binding site, which therefore enabled us to measure total number of CR1 regardless of receptor occupancy. We observed that patients in the lepromatous pole of the disease had a mean of 310 CR1/erythrocyte, whereas the ones in the tuberculoid pole showed a mean of 577 CR1/erythrocyte; 77 normal controls had a mean of 512 CR1/erythrocyte. The number of C3b receptors on the cells of lepromatous patients was significantly decreased (p less than .001) when compared to the normal population or tuberculoid patients. The presence of receptors for the C3b fragment of complement (CR1) on the surface of human erythrocytes enables these cells to participate in a number of immune functions including the clearance of circulating immune complexes. These findings could bear importance in the ability of the host to clear immune complexes from the circulation in patients with lepromatous leprosy.

Antibodies to Newcastle disease virus in various human diseases.

Sera of patients with infectious mononucleosis (IM) and various other diseases were studied for agglutinins against Newcastle disease virus (NDV)-modified human group O red blood cells (NDVO) and antibodies to the NDV preparations. In agreement with previous studies, the NDVO antibodies are found in a wide variety of diseases in addition to IM, including Japanese IM-like syndrome (22%), syphilis (24%), lepromatous leprosy (30%), systemic lupus erythematosus (29%), multiple sclerosis (18%) and cancer (17%); these antibodies were also found in patients with renal allografts (29%). It was also noted that the Victoria (VIC), Roakin and Herts strains, but not B1 strain of NDV are active in the NDVO agglutination, and VIC and Roakin strains, but not B1 strain in the immunodiffusion. Immunodiffusion and enzyme immunoassay with various preparations of the VIC strain revealed that the major antigen(s) of the virus under study is carried by the hemagglutinin-neuraminidase (H-N) glycoproteins. The H-N molecule was also shown to be able to modify human erythrocytes for the agglutination by the pathologic sera.

The in situ characteristics of mononuclear cell infiltrates in dermal lesions of leprosy.

The in situ nature of mononuclear cell infiltrates in the dermal lesions of 38 untreated leprosy patients was studied by means of conventional surface marker techniques using erythrocytes coated with AET, anti-erythrocyte IgG antibody, hemolysin and complement for the identification of T cells, and cells bearing Fc and C3 receptors, respectively. In general, T cells were the predominant lymphocytes in the leprosy lesions. They were mostly seen to be associated with epithelioid cell granulomas and showed maximal density in tuberculoid leprosy. A graded reduction of T cells was observed in borderline leprosy with a severe reduction/absence in polar lepromatous leprosy. The cells of the mononuclear phagocyte series in the various granulomas of the leprosy spectrum showed the presence of Fc and C3 receptors. Cells bearing a higher density of these receptors had a peripheral localization; whereas cells showing diffuse staining for nonspecific esterase were located more in the central regions of the granulomas. The differences in the individual cells of the phagocytic series appeared to be related to cell maturity; whereas the quantity of T cell infiltration in the lesions showed a correlation with the leprosy spectrum.

Serology of leprosy. I. Indirect hemagglutination test with stabilized sensitized red cells.

An indirect hemagglutination (IHA) test has been described for the qualitative and quantitative detection of antibodies specific to Mycobacterium leprae. Aldehyde stabilized red cells were sensitized with a sonicate antigen prepared from M. leprae purified from armadillo liver. These cells were titrated against sera from patients with different types of leprosy, their healthy household contacts and patients with tuberculosis. Specific antibodies were demonstrated in leprous sera by IHA test after absorption of sera with M. tuberculosis and M. vaccae. All advanced forms of leprosy (LL and BL) and a variable number of other forms of leprosy (BB, BT and TT) showed a positive result with an IHA titre of 1 in 32 or above. None of the household contact sera nor sera from tuberculosis patients showed a positive IHA test. The application of the simple hemagglutination test in the immunoepidemiology of leprosy is discussed.

In-vitro modification of membrane receptors on cells of the mononuclear phagocyte system.

EA and EAC receptors have been studied on non-elicited and paraffin-induced macrophages, under a variety of culture conditions in vitro, for up to 7 days. A large decrease in the number of macrophages showing EAC receptors was found after treatment of the cells with BCG, but not "inert" particles such as latex and zymosan. This was reversible by 7 days. In the presence of a toxic material, Al(OH)3, both EA and EAC receptors were partially lost. The results obtained have been related to previous results obtained with cryostat sections of human leprosy skin lesions.

Impairment of cell-mediated immune responses by infection with Mycobacterium lepraemurium.

The effect of chronic infection with Mycobacterium lepraemurium upon cell-mediated immune responses was studied in Lewis rats. Rats infected for 40 to 175 days were completely protected from attempted induction of experimental adjuvant disease, and the severity of experimental allergic encephalomyelitis in leprous rats was markedly attenuated. Full manifestations of each autoimmune disease were expressed in littermate control groups. Skin homograft rejection by infected rats was significantly impaired (P less than 0.001) as was the delayed-type hypersensitivity response to sheep erythrocytes (P less than 0.02). It is suggested that chronic infection with M. lepraemurium exerts a nonspecific inhibitory effect on cell-mediated immunity by perturbation of normal lymphocyte recirculation and by induction of immuno-suppressor cell activity.

Evaluation of the immune response in mice infected with Mycobacterium marinum.

Foot pad infection of mice with Mycobacterium marinum carried out with a view to comparing the immune response on the humoral level of such mice, with that observed previously in mice infected with M. leprae, indicated that there was a similarity in terms of the first appearance and proliferation of immunocytes and the time at which the peak and decline in the antibody-producing cells occurred. The significant difference appeared to be in the immunoglobulin G response, which was absent in the M. leprae infected mice, but occurred simultaneously with the immunoglobulin M response at a high level, both during a primary and after a secondary challenge administered 15 days post-primary infection in the M. marinum infected mice. Further confirmation was obtained through additional studies on the specific immunoglobulin levels and determination of both immunoglobulin M and immunoglobulin G antibodies by hemagglutination. Although the growth temperature requirement of the two organisms and their ability to initiate infection in areas of the body with lower temperatures are similar, it is suggested that the type of infection induced by each one of these species in the mouse may disallow the serious consideration of the M. marinum infection model as a possible alternative experimental model for studying the role of host immunity to M. leprae infections in mice.

Immunologic aspects of leprosy with special reference to the circulating antispermatozoal antibodies.

Macroscopic sperm agglutination in gelatin, sperm immobilization and tanned red cell hemagglutination tests could detect antispermatozoal antibodies respectively in 41%, 37%, and 23% sera of 35 leprosy patients, including 5 female cases. Interestingly, all of the above tests were positive in one serum from a female patient with borderline leprosy. Sperm antibodies were detected in both lepromatous and tuberculoid forms of leprosy by the above three technics and no significant difference was observed in their incidences among the two groups of patients. A three dimensional correlation was observed in 57% of 42 tests performed with 14 sera. Head-to head type of agglutination was the predominant feature of spermagglutination observed in the sera of these patients. In the control group, only 1 of 50 normal fertile male showed a positive spermagglutination test. Not one in this group showed positive sperm immobilization and tanned red cell hemagglutination tests. Antihuman globulin consumption test, presumably a very sensitive test, was also employed to demonstrate sperm-specific antibodies in the sera of these leprosy patients. These antibodies were adsorbed on the surface of the normal donor's spermatozoa when the latter were incubated with the patients sera. Antispermatozoal antibody could be demonstrated by this sensitive technic in the sera of two female patients. Moreover, antihuman globulin was consumed more intensely by the antispermatozoal antibodies present in the sera in the lepromatous than in the tuberculoid and borderline leprosy groups.

Ability of the leprous macrophage to accept cytophilic antibodies.

The ability of leprous macrophages to accept cytophilic antibodies was not damaged. Blood macrophages derived from both types of leprosy formed rosettes with cytophilic antibody sensitized goat red cells. The rosette forming rates of tuberculoid (65.0%) and lepromatous macrophages (66.4%) were essentially the same as that of normal blood macrophages (68.8%).

Serologically active glycolipid families from Mycobacterium bovis BCG. I. Extraction, purification and immunologic studies.

Glycolipids were extracted from mycobacteria with methanol and chloroform and purified by silicic acid chromatography. These glycolipids were studied for their serologic activity by direct and indirect (Coomb's) passive hemagglutination, and by inhibition methods. Three families of serologically active glycolipids called A, B and C, plus cardiolipin, were isolated. The B and C families of glycolipids were reactive with sera from BCG immunized rabbits and also with sera from patients with tuberculosis and leprosy; the A family was reactive only with the human sera. None of the serologically active glycolipids was able to protect rabbits against tuberculosis.

Immune response to Mycobacterium leprae: plaque-forming cells in mice.

Intravenous immunization with a cell extract of Mycobacterium leprae produced a primary immune response of considerable magnitude, followed by an equally large response after secondary stimulation, as measured by assay of plaque-forming cells (PFC). Infection with M. leprae or immunization with cell extract by the footpad route produced a lower level of response than that seen in the intravenous group. Identical patterns of response, although not of the same magnitude, were observed after both primary and secondary challenges in the two footpad groups, one infected with viable M. leprae and the other immunized with M. leprae cell extract. The secondary response after a booster dose to all these groups appeared to be an enhanced immunoglobulin M response. Control studies confirmed that the immune response was a direct result of the host-parasite interaction and that the PFC observed resulted from stimulation of antibody-forming cells by antigens of M. leprae. The similarity in time of appearance of peak PFC levels in the two footpad groups may be attributed to the live challenge passing through a latent phase. Alternatively, the challenge is known to contain a large proportion on nonviable cells, and it may also contain soluble M. leprae antigens. Studies of the cross-reactivity of the antigens have extended previous observations on antigens shared between M. leprae and other mycobacterial species. Use of the two antigen-containing fractions of the M. leprae cell extract has suggested that one of the fractions contains some shared antigens, whereas the other has an antigen specific to M. leprae.