Correlaçao entre reaçao de Rubino e receptores solúveis para hemácia de carneiro em pacientes com hanseníase / Correlation between Rubino's reaction and soluble sheep Erythrocyte receptors in hansen's patients

Neste trabalho correlaciona-se fatores, presentes em soros de pacientes hansenianos, que interagem com hemácias de carneiro formoladas (Reaçao de Rubino), com receptor solúvel de linfócitos T para eritrócitos de carneiro (Rs2) presente em nível elevado em soros de pacientes com hanseníase. A análise de fraçoes obtidas por cromatografia em Sephadex G-2OO demonstrou reaçao de Rubino na fraçao que corresponde ao Rs2 com peso molecular superior a 150kDA, detectado por soro anti-receptor. Adicionalmente, analisou-se o nível de receptor solúvel em soros de pacientes com hanseníase, apresentando reaçao de Rubino positiva e negativa, por imonodifusao (Mancini). Os resultados obtidos nao demonstram diferença significativa entre nível de receptor solúvel em reaçao de Rubino positiva (7.3mm) ou negativa (7.8mm) indicando que o receptor solúvel para E de alto peso molecular nao é responsável pela reaçao de Rubino

Comparative antigenic analysis of Mycobacterium avium complex (MAC) isolates from AIDS patients.

Sonicates of several Mycobacterium avium complex (MAC) strains isolated from acquired immunodeficiency syndrome (AIDS) patients were characterized in order to study the prominent antigens of these strains. Sonicates of 6-week-old cultures were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. A major 12 kDa glycoprotein antigen was observed in all the sonicates along with other proteins ranging up to 100 kDa. Western blotting, using the 12 kDa M. leprae 'specific' murine monoclonal antibody (MAb) MLO6, indicated the presence of a determinant in the 12 kDa antigen (in all the MAC isolates studied) which was immunologically cross-reactive with the 12 kDa antigen of M. leprae. The transparent variant of MAC 101 also demonstrated MLO6 reactivity while the opaque variant did not. Polyclonal antiserum raised against MAC 101 sonicate reacted with all the MAC isolates in immunodiffusion. These observations point to the cross-reactivity between these strains and suggest that they possess a M. leprae 'specific' determinant on a cross-reacting component which could be involved in virulence.

Immunoreactivity of a mammalian liver component with leprosy sera.

Sera from 77 leprosy patients in various stages of infection--tuberculoid (TT), lepromatous (LL), borderline tuberculoid and borderline lepromatous--15 contacts and 21 normal healthy individuals, were assayed in an indirect enzyme-linked immunosorbent assay and dot enzyme immunoassay using ethanol-soluble and thermostable extract of liver as the antigen. The highest incidences of reaction were found in untreated LL patients (100%) and in TT patients (91%), while the sera from borderline patients showed a comparatively lower incidence (43%). Some of the sera from contacts of leprosy patients (6/15) also showed high reactivity. Assays using lecithin as an antigen did not exhibit any reaction.

A method for the detection of M. leprae antigens in urine of leprosy patients by gel diffusion and gel electrophoretic techniques and significance in diagnosis.

A simple and novel method has been developed for the first time to detect M. leprae antigens in the urine of leprosy patients by treating the urine samples with sodium dodecyl sulphate (SDS). The antigens thus released can be demonstrated by simpler techniques like gel diffusion. By this method about 86% antigen positivity is observed in the urine of TT/BT and 83% positivity in BB patients. In BL/LL patients the antigen positivity is observed in 87% of the subjects. The high rate of M. leprae antigen positivity by the present method in the urine of patients with early leprosy may prove to be of diagnostic significance.

A simple method to quantitate circulating immune complexes in different diseases.

Immune Complexes are involved in the Pathogenesis of many diseases of varied aetiology such as autoimmune disorders, protozoal diseases, bacterial and viral infections. Quantitation of immune Complexes in these diseases can be used for diagnosis and to ascertain the prognosis. The simple method of precipitation by polyethylene glycol and quantitation by single Radial Immunodiffusion has been used in leprosy, syphilis, bacterial endocarditis and systemic lupus erythematosus (SLE). This method found significantly higher levels of circulating immune complexes (CICs) in erythema nodosum leprosum, culture positive bacterial endocarditis and SLE where CICs are known to play an important role in the pathogenesis.

Unusual precipitins in pathological human sera.

In conducting double diffusion gel precipitation tests, unexpected, strong precipitation reactions were noticed between a serum originating from a renal graft recipient and many pathological human sera. The highest frequency of positive reactions were produced by sera of patients with infectious mononucleosis (83%), chronic Epstein-Barr virus infections (72%), rheumatoid arthritis (57%), lepromatous leprosy (57%), and AIDS (44%). The precipitin in these pathological sera was identified as an antibody of IgM variety and the precipitinogen in the transplantation serum was shown to be a thermostable component with beta-globulin mobility. No explanation of the nature of reactions observed can be given at present.

Serum immune complexes in erythema nodosum leprosum reactions of leprosy.

Serum estimations of immunoglobulins, complement components and their presence in circulating immune complexes were carried out in 39 Lepromatous, 44 ENL and 22 Post ENL leprosy patients. Serum IgG, IgA, IgM, C3 and C4 levels were determined by single radial immunodiffusion. Serum immune complexes were precipitated with Polyethylene Glycol (PEG) and IgG, IgA, IgM, C3 and C4 were estimated by single radial immunodiffusion and expressed as % of precipitation of their serum level. Decreased IgG, IgM; increased IgA and C3; and no change in C4 levels are observed in ENL than Lepromatous and Post ENL patients. However, a gradual insignificant reduction of IgG, IgA, and IgM was found from Lepromatous to ENL and Post ENL patients in the PEG-precipitates. Similarly, C3 and C4 was found reduced insignificantly in ENL than Lepromatous and Post ENL patients. The significance of these estimations in relation to immune status of ENL reactions are discussed.

Immunogenic "subunit" of the ICRC antileprosy vaccine.

The administration of a vaccine containing ICRC bacilli, which is currently undergoing clinical trials in India, induces persistent lepromin conversion in lepromatous leprosy (LL) patients and lepromin-negative healthy subjects, with "upgrading" of tissue response in the former. A sonicate of ICRC bacilli, when subjected to gel-filtration chromatography using high-pressure liquid chromatography (HPLC), yields a high molecular weight glycolipoprotein (PP-I) with an apparent molecular weight of 10(6) daltons. PP-I, which brings about lepromin conversion in lepromin-negative healthy subjects, is a major immunogen of the organism, and carries epitopes for both B and T cells. A similar high molecular weight glycolipoprotein (PP-I Mycobacterium leprae) has been isolated from the sonicate of M. leprae. The two PP-I fractions exhibit a close antigenic relatedness at both B- and T-cell levels. However, they differ in their chemical composition and carry different charges. PP-I of ICRC is not only a good immunogen. Its high lipid content provides the necessary built-in adjuvant that would make it a good candidate for a "subunit" antileprosy vaccine. Also, since it carries epitopes for both B and T cells, PP-I ICRC could be used for "molecular engineering" to obtain molecules which selectively stimulate T-cell immunity which is the dominant host defense against M. leprae.

Non-organ specific thermostable tissue antigens.

Thermostable ethanol insoluble antigens (BE antigens) were identified that occur in all human tissues and human dispersed cells, but which are absent from normal human serum. In contradistinction to previously described organ-specific BE antigens, these antigens were referred to as non-organ-specific tissue antigens (NOST). Antisera obtained by immunization of rabbits with BE preparations of human organs had been selected for being devoid of any organ specificity and had been absorbed by BE preparations of pooled human serum. Such antisera could be used as reagents for detection of NOST BE antigens in pathological human sera. Inhibition of enzyme immunoassay proved to be a convenient procedure for these studies. Inhibition of 35% or more was only exceptionally noted in studying sera of normal subjects 20-40 years old, but inhibition exceeding 35% (positive results) was noted in 48% of sera from subjects at the age of 70 years or more. A high incidence of positive results was also encountered in sera of patients with end-stage renal disease (30%), renal graft recipients (18%), and in patients with lymphoma or leukemia (44%), but interestingly enough, no positive tests were noted in patients with lepromatous leprosy.

Immunologic aspects of leprosy.

Twenty seven cases of leprosy from Egypt were examined. Monocytes were found to be high in peripheral blood in lepromatous cases. The percentage of small lymphocytes in blood was increased in tuberculoid patients. The immunoglobulins were higher in all types of leprosy patients than in normal persons.

Detection of mycobacterial antigens in leprosy serum immune complex.

The antigens from immune complexes of sera from patients with mycobacterial diseases were released by sodium dodecyl sulfate. The antigenic activity of the released proteins was tested by agar gel diffusion and immunoelectrophoresis. This simple method provided direct evidence for the presence of mycobacterial antigens in the immune complexes of sera from patients with leprosy and tuberculosis.

A simplified serological test for leprosy based on a 3,6-di-O-methylglucose-containing synthetic antigen.

The recent advent of synthetic antigens containing the Mycobacterium leprae-specific epitope, 3,6-di-O-methyl-beta-D-glucopyranoside, has allowed the development of simple specific serological tests for leprosy. The incorporation of one such product, 8-carbonyloctyl O-[4-O-(3,6-di-O-methyl-beta-D-glucopyranosyl)-alpha-L- rhamnopyranoside]-BSA, into a simple "spot" test, diffusion-in-gel enzyme-linked immunosorbent assay (ELISA), allowed an over 90% detection rate of untreated lepromatous leprosy, and the results showed good concordance with conventional ELISA based on the native phenolic glycolipid I.

Alpha-1-antitrypsin in leprosy.

Estimation of Alpha-1-antitrypsin (AAT) levels was carried out in 52 patients of various types of leprosy. Fifty age and sex matched healthy individuals served as controls. The mean level of AAT in controls was 290.12 +/- 59.56 mg/dl. In patients of tuberculoid leprosy (TT), borderline tuberculoid leprosy (BT) and borderline leprosy (BB), the AAT levels were found to be 284 +/- 47.03, 314.37 +/- 31.56 and 324.44 +/- 32.05 mg/dl respectively. These were statistically insignificantly raised when compared with controls. In borderline lepromatous leprosy (BL), lepromatous leprosy without erythema nodosum leprosum (LL without ENL) and in LL with ENL there was a statistically significant rise in AAT levels. The maximum levels of AAT were observed in patients of LL with ENL (mean 500.8 +/- 93.44 mg/dl. P less than 0.001).

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.

Specificity of antibodies to Newcastle disease virus.

Specificity of antibodies to Victoria strain of Newcastle disease virus (NDV) found in infectious mononucleosis (IM) and other pathologic sera was investigated by agglutination of NDV-modified human O red blood cells, as well as by immunodiffusion and enzyme immunoassay with various preparations of the virus. These studies clearly demonstrated that the NDV antibodies are distinct from P-B or H-D antibodies. The unexpected observation that guinea pig kidney (GPK) tissues absorbed NDV antibodies allowed their classification into a group of 'GPK-positive' heterophile antibodies. The simultaneous occurrence of the NDV antibodies and H-D antibodies in IM and other diseases suggests the possibility that multiple new antigenic determinants, especially those of carbohydrate nature, may appear due to the alteration of self-antigens as a result of various pathologic processes.

Mixed agglutination with guinea pig tissue sediments for detection of heterophile antibodies.

Mixed agglutination (MA) test with sediments of guinea pig kidney (GPK) homogenates and indicator red blood cells of bovine (BRBC) or sheep (SRBC) origin was established for detection of human heterophile antibodies. By means of MA test with BRBC indicator cells, heterophile antibodies of Hanganutziu-Deicher (H-D) specificity were demonstrated in sera of patients with syphilis (20%), lepromatous leprosy (57%), infectious mononucleosis (45%), Chediak-Higashi syndrome (73%), Kawasaki disease (58%), multiple sclerosis (58%), and leukemias (13%), as well as in sera of subjects who received injections of foreign species sera (20%). Some but not all BRBC-positive sera gave positive MA tests when SRBC were employed as indicator cells. None of 13 multiple myeloma sera tested gave positive results. The incidence of positive reactions in normal human sera was 3%. Neutralization of H-D antibodies in representative pathologic sera by purified heterophile antigens showed that the antibodies under investigation were mostly directed against antigen(s) of high molecular weight glycoprotein, but not N-glycolyl-neuraminic acid (NGNA) ganglioside fraction of BRBC.