IgM anti-phenolic glycolipid I and IgG anti-10-kDa heat shock protein antibodies in sera and immune complexes isolated from leprosy patients with or without erythema nodosum leprosum and contacts.

The aim of the present work was to evaluate the levels of anti-PGL-I and anti-10-kDa heat shock protein antibodies in serum and immune complexes isolated from leprosy patients, convivients and controls. Leprosy patients with erythema nodosum leprosum or without it were included and a comparative study was done to investigate intergroup differences. Immune complexes were precipitated from serum by polyethylene glycol 3.5%; antibody levels were measured in sera and in dissociated immune complexes by ELISA. Serum antibody levels were then correlated with immune complex-associated antibody levels. The results showed that the erythema nodosum leprosum group differed from controls, contacts and non-erythema patients in their immune complex levels. IgM anti-PGL-I and IgG anti-10-kDa heat shock protein antibodies were constituents of the immune complexes in patients with erythema nodosum leprosum, who exhibited a significant difference in their immune complex composition compared with controls, contacts and non-erythema patients; while free antibody levels (anti-PGL-I and anti-10-kDa) did not differentiate between erythema and non-erythema patients, the measurement of immune complex-associated antibodies demonstrated a significant difference between the two clinical conditions. Furthermore, the measurement of immune complex-associated anti-PGL-I IgM made it possible to differentiate between contacts and controls. The significance of these results is discussed.

[Autoantibodies in lepromatous leprosy]. / Autoantikörper bei lepromatöser Lepra.

84 Turkish patients with lepromatous and borderline lepromatous leprosy were investigated with regard to the presence of antibodies to a variety of tissue constituents and immunoglobulins. Thirty Turkish-born volunteers and two groups of patients with systemic lupus erythematosus and primary biliary cirrhosis respectively served as controls. No increased incidence of autoantibodies to smooth muscle cells, nuclei, mitochondria, striated muscle, thyroid microsomes and thyroglobulin was detected. A high incidence of rheumatoid factor and circulating immune complexes was found.

[Isolation and characterization of immune complexes associated with malignant tumors and leprosy]. / Aislamiento y caracterización de complejos inmunes asociados a tumores malignos y a lepra.

We are dedicated to the study of circulating immune complexes (CIC) associated with different diseases: malignant tumors, leprosy and rheumatoid arthritis. Immune complexes were evaluated by various methods: 125I-Clq binding assay, 125I-IgG binding test, 125I-bovine conglutinin binding assay and polyethylene glycol precipitation test (3.5% and 2.5%). Techniques for the isolation and splitting of CIC in their components were performed in sera from patients with tumors and with leprosy. These methods consisted in the combination of CIC with protein A followed by elution with different buffers. CIC splitting techniques were first applied on immune complexes formed in vitro (BSA-aBSA, OVA-aOVA). The analysis of CIC fractions was done by SDS-PAGE, immunoelectrophoresis and immunoblotting techniques. Results were as follows: CIC levels correlated with active stages of disease, decreasing during remission so that CIC detection can be useful to evaluate response to treatment. The isolation and splitting of immune complexes into their components resulted in the obtention of immunologically active fractions, especially in sera from patients with gastrointestinal and breast cancer and with leprosy.

Aislamiento y caracterización de complejos inmunes asociados a tumores malignos y a lepra / Isolation and characterization of immune complexes associated with malignant tumors and leprosy

Hemos estudiado los complejos inmunes circulantes (CIC) asociados a diversas enfermedades tales como neoplasias malignas, lepra y artritis reumatoidea. Se realizó la detección de CIC empleando diferentes técnicas: test de combinación a 125I-Clq, a IgG radioactiva, a conglutinina bovina marcada, test de aglutinación de plaquetas y precipitado en polietilen glicol 3,5% y 2,5%. Posteriormente, se desarrollaron técnicas de asislamiento y separación de los CIC para la obtención de sus fracciones constituyentes en ciertos tumores malignos y en lepra, basadas en la afinidad de los CIC por proteína A y en la acción de diferentes buffers. Estos métodos se aplicaron previamente a complejos inmunes preparados in vitro (BSA-aBSA, OVA-aOVA, etc.). El análisis de las fracciones se realizó por SDS-PAGE, inmunoelectroforesis, inmunoblotting, etc. Los resultados fueron los siguientes: se observó una elevada incidencia de niveles de CIC positivos coincidentes con los estadios de actividad clínica, disminuyendo en la remisión. Así mismo en algunos grupos de pacientes los hallazgos de CIC permitieron evaluar la eficacia terapéutica. Con el desarrollo de técnicas de separación de CIC se obtuvieron fracciones inmunológicamente activas provenientes de los CIC aislados del suero de pacientes portadores de tumores malignos, especialmente del tubo digestivo y de mama y posteriormente en el suero de individuos con lepra. Los resultados obtenidos hasta la fecha han demostrado la utilidad clínica de los estudios...

Aislamiento y caracterización de complejos inmunes asociados a tumores malignos y a lepra / Isolation and characterization of immune complexes associated with malignant tumors and leprosy

Hemos estudiado los complejos inmunes circulantes (CIC) asociados a diversas enfermedades tales como neoplasias malignas, lepra y artritis reumatoidea. Se realizó la detección de CIC empleando diferentes técnicas: test de combinación a 125I-Clq, a IgG radioactiva, a conglutinina bovina marcada, test de aglutinación de plaquetas y precipitado en polietilen glicol 3,5% y 2,5%. Posteriormente, se desarrollaron técnicas de asislamiento y separación de los CIC para la obtención de sus fracciones constituyentes en ciertos tumores malignos y en lepra, basadas en la afinidad de los CIC por proteína A y en la acción de diferentes buffers. Estos métodos se aplicaron previamente a complejos inmunes preparados in vitro (BSA-aBSA, OVA-aOVA, etc.). El análisis de las fracciones se realizó por SDS-PAGE, inmunoelectroforesis, inmunoblotting, etc. Los resultados fueron los siguientes: se observó una elevada incidencia de niveles de CIC positivos coincidentes con los estadios de actividad clínica, disminuyendo en la remisión. Así mismo en algunos grupos de pacientes los hallazgos de CIC permitieron evaluar la eficacia terapéutica. Con el desarrollo de técnicas de separación de CIC se obtuvieron fracciones inmunológicamente activas provenientes de los CIC aislados del suero de pacientes portadores de tumores malignos, especialmente del tubo digestivo y de mama y posteriormente en el suero de individuos con lepra. Los resultados obtenidos hasta la fecha han demostrado la utilidad clínica de los estudios... (AU)

Isolation and analysis of circulating immune complexes in leprosy.

Circulating immune complexes (CIC) were isolated by two antigen nonspecific methods from 60 leprosy patients belonging to borderline tuberculoid (BT) and lepromatous (LL) types with and without reactions. CIC were elevated in both BT and LL reactions. CIC from BT in reaction (BTR) were found to consist largely of IgG and C3, whereas, C-reactive protein could be found in CIC from LL reactions (LR). In addition, IgM and rheumatoid factor were demonstrated in the complexes of LR patients who had mainly arthritis. Antimycobacterial antibody was seen in the complexes of two-thirds of LR patients who had predominantly skin manifestations as part of their reaction. The relevance of these findings to the clinical manifestations of different types of reactions is discussed.

Identification of components of IC purified from human sera. II. Demonstration of mycobacterial antigens in immune complexes isolated from sera of lepromatous patients.

Immune complexes have been purified from sera of patients with lepromatous leprosy, using solid phase conglutinin and analysed by SDS-PAGE. Some of their components have been immunologically identified after electrophoretic blotting on nitrocellulose. First, immunoglobulins, complement components (C1q, C1s, C3) and CRP were found in IC. Secondly, one mycobacterial antigen of 67 kD was directly identified in IC while two other components (20 kD and 14.4 kD) of possible M. leprae origin were also found in IC. This study suggests that lepromatous patients develop a good antibody response against some M. leprae antigens (33 kD and 12 kD), which are rapidly eliminated from circulation, while other M. leprae antigens (e.g. 67 kD) can persist in relative antigen excess, within circulating IC.