Hepatitis B and hepatitis C virus infections in dermatological patients in west Sicily: a seroepidemiological study

OBJECTIVES: To evaluate the relative frequencies and molecular epidemiological features of viral hepatitis types B and C in dermatological patients in our geographical area. METHODS: We determined the hepatitis B virus (HBV) and hepatitis C virus (HCV) antibodies and the hepatitis B virus surface antigen (HBsAg) in a cohort of 677 dermatological patients admitted to the Department of Dermatology of Palermo. An 8-mL blood sample was taken from all subjects. The following assays were used: HBsAg, anti-HB core (antigen) (anti-HBc), anti-HB surface (antigen) (anti-HBs), anti-HB early (antigen) (anti-Hbe) and anti-HCV antibodies using enzyme-linked immunosorbent assay. RESULTS: One hundred and eighty-nine (27.91%) of the 677 dermatological patients were positive for anti-HBc, anti-HBs, anti-HBe and/or anti-HCV antibodies. In particular 22% (149 patients) were anti-HBc, anti-HBs or anti-HBe positive, reflecting exposure to HBV, and six patients (0.88%) were chronic carriers of HBsAg; 2.36% of the dermatological patients (16 persons) were anti-HCV positive. Tests showed that 24 subjects (3.52%) were infected with hepatitis B or C. The peaks in the age bands were in the 55-80-year-old age groups. CONCLUSIONS: This study confirms a high rate of HBV and HCV exposure with chronic carriers in our dermatological patients. We assume that the high prevalence of HCV and HBV in dermatological patients is more likely to be age related than to represent a true and direct association with dermatological diseases in general. Definite conclusions will only be available after large epidemiological studies that can establish or refute an aetiological and pathogenetic role of HBV and HCV in certain skin diseases associated with liver infection.     

An appraisal of enzyme linked immunosorbent assay (ELISA) and serum antibody competition test (SACT) in leprosy.

Seventy-eight untreated leprosy patients, 104 treated patients and 105 healthy contacts were tested using two serological tests, SACT (serum antibody competition test based on competitive inhibition of monoclonal antibody binding to the MY2a determinant of M. leprae) and ELISA (measurement of IgM antibodies to the neoglycoproteins D-BSA and ND-BSA representing the phenolic-glycolipid antigen of M. leprae). The controls included normal healthy individuals, patients with sputum positive pulmonary tuberculosis, and active cases of rheumatoid arthritis from the department of rheumatology. The specificity of SACT was found to be very high. ELISA was found to be positive in two patients with rheumatoid arthritis, one each for D-BSA and ND-BSA ELISA. Both tests had a high sensitivity in BL and lepromatous patients. The sensitivity to both tests was considerably lower in tuberculoid and BT patients i.e., below 40%. Therefore the diagnostic value of a negative test in suspected cases of leprosy was very low employing either of the two tests. A proportion of patients with paucibacillary tuberculoid and BT leprosy were positive after six months or longer after therapy. Similarly a large number of BL and lepromatous patients were positive after considerably longer periods of treatment. The use of either tests for determining the duration of therapy is therefore limited. SACT appears to be more sensitive than ELISA with ND-BSA in detecting subclinical infection. The cumulative positivity of the two tests may be used as a measure of the infectivity of the disease in the community and for evaluating disease control methods.     

Prevalence and incidence of hepatitis B virus infection in STD clinic attendees in Pune,India

OBJECTIVES: To estimate the prevalence and incidence of hepatitis B virus (HBV) infection among patients attending three STD clinics in Pune, India, and to identify associated risk factors. METHODS: Of the 2098 patients screened at STD clinics in Pune during 1996, 497, who returned for at least one follow up visit, were screened for various markers of HBV infection (HBsAg, anti-HBs, anti-HBc), HIV antibody, and VDRL. RESULTS: Of the 497 participants 3.6%, 26.5%, and 43.2% were positive for HBsAg, anti-HBs, and anti-HBc respectively. Tattooing (AOR 1.64, 95% CI 1.03 to 2.64) was found to be independently associated with presence of core antibody. Additionally, history of being in commercial sex work and history of a genital ulcer were independently associated with a positive anti-HBc antibody test (AOR 12.45, 95% CI 5.58 to 27.82 and AOR 1.70, 95% CI 1.09 to 2.66, respectively). 72 out of 497 (14.5%) participants were HIV positive at baseline. HIV-1 antibody positive patients were more likely to have a positive anti-HBc test (69.4% v 39.0%, p<0.001). 30 out of 282 participants, negative for anti-HBc antibody at enrolment, seroconverted subsequently, resulting in an incidence of 10.86 per 100 person years (95% CI 7.2%, 14.5%) (mean and accumulated follow up of 11.7 months and 276.17 person years, respectively). CONCLUSIONS: A high prevalence and incidence of HBV infection, seen in STD clinic attendees underscore the need to provide HBV vaccine to commercial sex workers and their clients in India.     

High Incidence of IgG Antibodies to Phenolic Glycolipid in Non-Leprosy Patients in India

Purified phenolic glycolipid (PGL-1) from Mycobacterium leprae was used to detect IgG antibodies against PGL-1 in leprosy patients in an enzyme-linked immunosorbent assay (ELISA). A total of 698 sera were screened; they came from patients suffering from leprosy, autoimmune disease, myeloma, tuberculosis and sexually transmitted diseases (STDs). Cases with miscellaneous diseases and persons undergoing AIDS screening were also included. Sera from lepromatous and tuberculoid leprosy patients gave positivity rates of 60.5% and 41.7%, respectively. In non-leprosy cases, the PGL-1 ELISA showed an overall positivity rate of 6.9%; this was greatest in patients with tuberculosis (43.8%) followed by autoimmune diseases (40.9%) and miscellaneous cases including liver diseases (37.9%). This study emphasizes that PGL-1 ELISA has a low predictive value for diagnosis of active infection by Mycobacterium leprae. Positive reactions in a significant percentage of patients with autoimmune disease are intriguing and need indepth study.     

Evaluation of anti-cardiolipin antibody and its cross-reactivity in sera of patients with lepromatous leprosy

Summary Using a sensitive and modified solid-phase radioimmunoassay for detecting anti-cardiolipin antibodies, sera of 45 patients with lepromatous leprosy were examined. Nine of the 45 (20%) showed positive levels of anti-cardiolipin antibodies. Inhibition tests revealed that these antibodies significantly cross-reacted with double-stranded (ds) DNA, but not with single-stranded (ss) DNA or extractable nuclear antigens (ENA). We describe the unique pattern of antibody cross-reactivity with cardiolipin and dsDNA in sera of patients with lepromatous leprosy.     

Hepatic involvement and hepatitis B surface antigen (Hbs Ag) in leprosy

Hepatic involvement and hepatitis B surface antigenemia was studied in 80 leprosy patients and results were compared with 50 normal healthy controls. HbsAg was detected in 7.54% of lepromatous leprosy patients as compared to 2% of the normal healthy controls. There was a decrease in albumin and increase in globulin levels with significant decrease in A: G ratio. SGPT levels were significantly raised in lepromatous leprosy patients. Histopathological changes were present in 57.1% of lepromatous leprosy and 23.8% of tuberculoid leprosy patients.     

Comparative evaluation of enzyme immunoassays based on synthetic glycoconjugates and phenolic glycolipid-I for immunodiagnosis of leprosy

Enzyme immunoassays (EIAs) based on synthetic glycoconjugates containing the terminal monosaccharide (M-BGG) or disaccharide (ND-BSA) residue of the trisaccharide component of phenolic glycolipid-I (PGL-I), for immunodiagnosis of leprosy are described. The results of the assays were compared with that of the EIA using PGL-I. All the three assays were highly specific for leprosy. The per cent positivity of active lepromatous leprosy (LL) patients with M-BGG was 78.05 in comparison to 85.36 with ND-BSA and 82.11 with PGL-I. Similarly, the positivity of tuberculoid (TT) leprosy patients in M-BGG assay was lower than that in EIAs using ND-BSA or PGL-I. However, the difference in the positivity of individual category of leprosy patients in the three EIAs was not statistically significant. The correlation between absorbance values of leprosy sera in EIAs based on M-BGG and PGL-I, as well as that in assays using ND-BSA and PGL-I was statistically significant.     

Prevalence of colour blindness among patients with leprosy.

Using Ishihara test plates the prevalence of colour blindness was studied on six hundred and ninety-seven leprosy patients and two hundred and ninety-two normal healthy controls. 7.88% of male patients with tuberculoid leprosy, 12.18% of male patients with lepromatous leprosy, and 0.67% of male controls were detected to be colour blind (red-green deficiency or total colour weakness). The differences between the different groups are significant. Among female patients and controls, only one lepromatous leprosy patient was detected to have red-green deficiency. This suggests the possibility of a genetic predisposition to Mycobacterium leprae infection in patients with leprosy.     

IMMUNOLOGICAL ASPECTS OF LEPROSY

Leprosy has been regarded as a host dependent disease that presents an immunological spectrum. Various parameters of in vivo and in vitro cell-mediated and humoral responses in different forms of leprosy were studied. Lepromatous leprosy patients showed hyper gammaglobulinemia with significantly elevated levels of IgG and IgA while tuberculoid leprosy patients showed raised IgG levels. Cutaneous reactions to lepromin antigen were significantly depressed in patients with lepromatous leprosy while those to PPD were normal. Mitogenic response of peripheral leucocytes to phytoheamagglutinin was considerably depressed in untreated lepromatous leprosy and normal in tuberculoid leprosy. Dapsone-treated patients with lepromatous leprosy showed a normal or enhanced response. Contributory factors were also found in plasma. T-lymphocytes were significantly reduced in lepromatous leprosy with high bacillary load. B-cells were increased in lepromatous patients. Tuberculoid leprosy patients had normal values. Phagocytic and bacteriolytic activity of blood derived macrophages was not altered in the lepromatous or tuberculoid patients.     

Antispermatozoal antibodies in leprosy with special reference to their morphological patterns

Sixty two male patients with polar leprosy--38 lepromatous and 24 tuberculoid types were investigated for the presence of antispermatozoal antibodies with special reference to their morphological patterns. Antibodies were detected by three different immunological techniques. Sperm agglutination was found to be the most sensitive. The incidence of antibodies was higher in patients with lepromatous leprosy and was directly proportional to the duration of the disease in both types of leprosy. Morphologically, head-to-head type of agglutination was observed in 50 percent of the patients, mixed in 41.7 percent and tail-to-tail type in 8.3 percent. There was no correlation between the number of ENL attacks and the incidence of anti-bodies. In polar tuberculoid leprosy patients the histological findings of testicular biopsy indicated cell mediated tissue damage occurring in a non-infective form.     

IMMUNOHISTOCHEMISTRY IN HISTOID LEPROSY

Background. Histoid leprosy is a rare form of multibacillary leprosy as the result of secondary or even primary resistance to dapsone. The etiopathogenesis has not been clarified up to now. Methods. An immunohistochemical study was carried out for the expression of various markers on epidermal and dermal cell populations using sections of frozen skin specimens from 5 patients with histoid leprosy as compared to specimens from 7 tuberculoid and 7 lepromatous patients. Results. Dendritic epidermal cells, identified by monoclonal antibodies against CD1, hla-dr , CD45, and CD36, were found reduced in histoid leprosy as compared to both tuberculoid and lepromatous groups. A gradual reduction of keratinocytic hla-dr expression from tuberculoid to lepromatous to histoid leprosy was observed. The pattern of CD36, CD4, and CD8 expression of lymphomonocytic cells in the dermis of histoid lesions was similar to that of tuberculoid leprosy, but without the formation of an organized granuloma. CD45+ cells as well as activated lymphocytic cells, expressed by the activation immunophenotype (CD1, hla-dr , CD25, CD71, egf-r ) were found frequently in all groups. Conclusions. The in situ immunohistochemical findings support a modified hypersensitivity reaction of the cellular type that results in an inhibition of the lesional expansion, but not in the destruction of the bacilli within the histoid lesion.     

Immunotherapy in leprosy: a new approach

An immunotherapeutic agent prepared from patient's own affected skin was tried in 30 leprosy cases. 53.6% cases of lepromatous and borderline lepromatous group showed lepromin conversion from lepromin negativity to positivity after 12 weeks of immunotherapy. The clinical and bacteriological improvement was also good. 88.1% cases of borderline tuberculoid also showed fair to good clinical recovery following 12 weeks of immunotherapy.     

Lesional T-cell subset in leprosy and leprosy reaction

BACKGROUND: The T-cell-mediated immune response plays an important role in leprosy. The in situ proportion and pattern of distribution of T-cell subsets in leprosy skin lesions have been studied, but no conclusion could be drawn. METHODS: We used monoclonal antibodies for T-helper and T-suppressor surface antigen to define the nature of dermal infiltration in 17 cases of nonreactional leprosy and 20 cases of reactional leprosy. RESULTS: We found T helper admixed with T suppressor in an aggregated pattern in the granulomas of most cases of nonreactional leprosy and in type I reactional leprosy, but a diffuse infiltrate throughout the dermis of type II reactional leprosy. The T-helper/suppressor ratio was 1.68 in tuberculoid and 1.5 in lepromatous cases. The T-helper/ suppressor ratios of borderline tuberculoid (3.11) and type I reactional leprosy (2.54) were not statistically different. The T-helper/suppressor ratio of type II reactional leprosy (5.83) was statistically higher than nonreactional lepromatous cases. CONCLUSIONS: The alteration of the T-helper/suppressor ratio in our study is mainly due to the reduction of T-suppressor cells in the dermal infiltrates, especially in type II reactional leprosy. Further studies of T-suppressor functions may be important in the pathogenesis of leprosy.     

小鼠神经显示麻风病抗神经抗体的初步研究

本文检测28例各型麻风患者的抗神经抗体。28例各型患者中抗神经抗体阳性为25/28。这一结果揭示在多菌型患者抗体阳性频度和滴度水平有高於少菌型患者的趋势。对出现ENL反应的11例多菌型患者则未显示出其反应类型与抗神经抗体之间具有关系。有关麻风患者的型别、病期、及神经炎等与抗神经抗体频度和滴度之间的相互关系,有待进一步深入的研究才有可能加以阐明。     

Lymphocyte adenosine deaminase activity (L-ADA) in leprosy, during and after treatment of reactions

Twenty-five patients with type 1 (lepra) and type 2 (E.N.L.) leprosy reactions were studied for lymphocyte adenosine deaminase activity (L-ADA), during and after treatment of the reactions, using a standard technique, in order to establish its pattern and if possible, its value in assessing the course of reactions. The results were compared with those from 30 control subjects, comprising 10 normal healthy adults, 10 patients with borderline tuberculoid (BT) leprosy, four patients with borderline lepromatous (BL) leprosy and six patients with lepromatous (LL) leprosy. The level of L-ADA in the leprosy controls was higher than that of normal healthy subjects. The L-ADA values in patients with different types of reactions were about 10-fold higher than those obtained from leprosy controls, emphasizing a possible role in assessing reactions in leprosy. However, there was no significant variation in L-ADA levels, either between the various leprosy controls or reaction groups, before and after treatment.     

LDH isozymes with anomalous bands in semen of leprosy patients

Activity of LDH isozymes was evaluated electrophoretically on 7% acrylamide gel in semen of 37 leprosy patients (15 with borderline, 12 with borderline tuberculoid and ten with lepromatous leprosy) and ten fertile men of 30-45 years of age. Significantly lower activities were recorded of LDH1 in all categories of leprosy patients. Similarly, lowering of LDH2 activity was noticed in borderline and lepromatous cases only, lowering of LDH4 activity in lepromatous cases only and LDH5 activity was lowered in borderline leprosy patients. Lowest activity of LDH3 and absence of LDHx were found in lepromatous leprosy. However, in borderline tuberculoid patients, LDH3 and LDHx were significantly higher. This exceptional increase in activity was found to be due to presence of additional (anomalous) isozymes bands of LDH3, LDHx and LDH4 in 25% of borderline tuberculoid patients. Additional bands of LDH3 have also been located in 40% of the borderline leprosy patients.     

Penile tuberculoid leprosy in a five year old boy

A 5-year-old contact of a lepromatous leprosy patient with a tuberculoid lesion on the anterior aspect of the shaft of the penis is reported. The child was clinically suspected to have borderline tuberculoid leprosy during a survey of contacts of leprosy patients, which on histopathology revealed features of subpolar tuberculoid leprosy. The father of the child was recently detected as a case of lepromatous leprosy and was started on multibacillary regime of WHO multidrug therapy. The reason for the localization of the lesion to the shaft of the penis is also suggested. Skin as a route of transmission of tuberculoid leprosy is also emphasized.     

Specificity of lymphocytotoxic autoantibodies (LCAbs) found in the serum of leprosy patients: class I MHC antigens

Lymphocytotoxic autoantibodies (LCAbs) of the IgM class have been identified in patients with borderline tuberculoid (BT) and borderline lepromatous (BL) leprosy with Type I reactions (I) as well as lepromatous leprosy (LL) patients with erythema nodosum leprosum reactions (ENL). The observation that lymphocytotoxic activity (LCA) was reduced in the presence of platelets led us to determine whether LCAbs had specificities for Class I Major Histocompatibility Complex (MHC) determinants. Absorption of LCA positive sera with platelets, classically used to deplete Class I specific lymphocytotoxic antibodies, reduced LCA towards autologous as well as allogeneic target cells. This was true for LCA positive sera from all patient classifications (group BT in the autologous system, p less than 0.01; in all other patient groups, p less than 0.001). Introducing B-2m to cytotoxicity assays only marginally reduced LCA when added at high concentrations (5 mg/ml). An anti-Class I MHC antiserum which blocked the lytic activity. The data indicate that LCAbs while absorbed by platelets, are not specific for the Class I MHC antigens. The autoantigen recognized by these autoantibodies therefore remains to be identified.     

Multibacillary leprosy: lesions with macrophages positive for S100 protein and dendritic cells positive for Factor 13a

In the defense against Mycobacterium leprae , macrophages play an essential part in the mechanism of bacterial lysis but require the presence of cytokines such as interleukin 2 and gamma interferon from lymphocytes in order to effectively kill the organisms in any number. While there have been many studies of the lymphocytes in lesions of leprosy, less attention has been given to the immunohistochemical characterization of the macrophage populations. In this study, the cutaneous lesions of 69 patients with leprosy (42 lepromatous, 5 mid-borderline, and 22 tuberculoid) were evaluated by immunohistochemistry for the expression of S100 protein, CDla, CD68, muramidase, HLA-DR, and Factor 13a. The macrophages from lesions of polar, subpolar, and borderline Iepromatous leprosy patients expressed S100 protein intensely and constantly. In contrast, the lesions of polar and subpolar tuberculoid leprosy had very few cells that were immunoreactive for S100 protein ('S1000+') in the granulomas in the dermis. The macrophages in all lesions were reactive for CD68 and muramidase. In paraffin sections, macrophages of lepromatous lesions failed to stain for HLA-DR, whereas in tuberculoid lesions, they were strongly positive for HLA-DR. Three patients with histoid leprosy (relapse lesions) had lesions that were strongly positive for Factor 13a and were negative for S100 protein ('S100−'). Given the possible chemotactic and migration inhibition effects of the calcium-binding proteins of the S100 family, these data suggest a possibly important role for S100 protein in the accumulation of macrophages in lepromatous leprosy, and also reveal infection of Factor 13a+ dermal dendritic cells in histoid leprosy.