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Biomedical subjects

M Harboe

Publications and source records attributed to M Harboe.

At least 127 records · Page 7Linked to original sources
PubMed1985

Quantification of the terminal complement complex in human plasma by an enzyme-linked immunosorbent assay based on monoclonal antibodies against a neoantigen of the complex.

The fluid-phase terminal complement complex (TCC), consisting of the components C5b, C6, C7, C8, C9, and the S-protein, has recently been detected in normal human plasma by using antibodies against native terminal complement components. Increased amounts of TCC were then found in several patients with in vivo activation of complement. We now describe a sensitive, specific, and reliable enzyme-linked immunosorbent assay for quantification of the TCC, based on monoclonal antibodies against a neoantigen of the complex. The results indicate that the TCC is present in normal human plasma and in increased amounts in patients with complement activation in vivo, thus confirming previously obtained results. The assay is easy to perform and can be used for examination of large numbers of plasma samples.

Antibodies, Monoclonal↗
PubMed1985

Characterization of the immune response to an epitope on Mycobacterium leprae antigen 7 defined by a monoclonal antibody.

A mouse monoclonal antibody (038D-C6) was shown by crossed immunoelectrophoresis and radioimmunoassay to react with an epitope on the Mycobacterium leprae antigen 7. This epitope was highly crossreactive with BCG/M. tuberculosis and of a non-arabinogalactan-arabinomannan nature. A solid-phase radioimmunoassay (SPRIA) was applied, based on competitive inhibition by human sera of antigen binding by this anti-M. leprae monoclonal antibody. Inhibitory activity determined by this assay decreased markedly upon treatment in both lepromatous and tuberculoid leprosy patients. A correlation was found between the bacterial load of the patient and the inhibitory activity measured in the SPRIA assay. Serum-inhibitory activity could therefore perhaps be used as a follow-up test for patients on treatment or as a screening method to detect infective cases. A dot enzyme-linked immunosorbent assay based, like the SPRIA assay, on competitive inhibition by human sera, was explored as an inexpensive and technically simple alternative also applicable under field conditions.

Animals↗
PubMed1985

Increased plasma levels of the terminal complement complex in patients with evidence of complement activation.

The terminal C5b-9 complex of human complement has recently been described and quantified in normal human plasma by an enzyme-linked immunosorbent assay (ELISA). We collected EDTA plasma samples from 20 patients clinically suspected to have complement activation. The terminal complement complex (TCC) and C3d split products were measured. The TCC was increased in 8 patients, and 6 of these also had increased C3d values, whereas 4 patients had increased C3d and normal TCC values. Two different double-antibody assays were used to detect terminal pathway activation: the combination of anti-C6 and anti-C9 detecting only the whole complex, and the combination of anti-C6 and anti-C5 detecting intermediate complexes as well. There was a close correlation between the observations in these two assays, suggesting that in general the whole cascade including C9 is involved when the terminal pathway of complement is activated in vivo. Quantification of TCC in plasma is an important supplement to already established methods for the evaluation of complement activation in vivo.

Adult↗
PubMed1984

Immunochemical studies on human monoclonal macroglobulins with specificities for 3,4-pyruvylated D-galactose and 4,6-pyruvylated D-glucose.

Four of six human monoclonal IgM proteins were found to react best with Klebsiella polysaccharides containing 3,4py beta DGal (pyruvic acetalated D-galactopyranose), one with Klebsiella polysaccharides with 4,6pyDGlc; the sixth is uncharacterized. The combining sites of two of these (IgMWEA and IgMNAE) were essentially indistinguishable by quantitative precipitin studies at varying pH and by quantitative precipitin inhibition assays, but the other two differed in specificity of their combining sites from these and from each other. These differences were detected by precipitin inhibition assays with 3,4py beta DGal-containing oligosaccharide alditols, the R and S isomers of methyl 4,6py alpha DGal, the R isomer of methyl 4,6py beta DGal, or the R and S isomers of methyl 4,6py alpha DGlc, and -beta DGlc. In all of these except the S isomer of methyl 4,6pyDGal and R isomer of methyl 4,6pyDGlc, the carboxyl group is axial to the plane of the acetal ring. Their specificity appears to be determined by the nonreducing ends of chains and is considered to be cavity-type.

Antibody Specificity↗
PubMed1984

Anti-Mycobacterium leprae antibodies in urine from lepromatous patients examined by crossed immunoelectrophoresis and radioimmunoassay.

Precipitating anti-Mycobacterium leprae antibodies were found in concentrated urine samples from 21 out of 42 lepromatous patients. These antibodies were directed against M. leprae antigens 5, 6, and 7. In a radioimmunoassay for anti-M. leprae antibodies, 90% of these patients had higher antibody levels in their urine than control persons. There was a positive correlation between anti-M. leprae antibody levels in serum and urine. The advantages of using atraumatically collected samples like urine in epidemiological work are pointed out. The present report shows that urine can be used to measure the antibody response to a specified microorganism causing infection outside the urinary tract. The possible presence of antibodies in urine should alert researchers who look for antigens in urine to choose assays that minimize interference by such antibodies.

Antibodies, Bacterial↗
PubMed1984

Detection and quantification of the terminal C5b-9 complex of human complement by a sensitive enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay for detection and quantification of the terminal complexes (SC5b-9 and membrane attack complex) of human complement is described. We separate the complex from the native complement components, to use antibodies against the native components in a 'double-antibody sandwich' technique. It is thereby possible to detect the terminal complement complex in solution without the requirement of specific antibodies against the neoantigens. The results show that the assay is both sensitive and specific. Evidence is presented that a terminal complement complex occurs in a normal plasma pool. The terminal complement complex may be valuable for evaluating both the physiology and pathophysiology of the complement system in vivo.

Antibody Specificity↗
PubMed1984

MPB70, a unique antigen of Mycobacterium bovis BCG.

MPB70 is a constituent of Mycobacterium bovis BCG, which accounts for about 10% of the total protein content in the nutrient fluid after culture on Sauton medium. Previous studies of cell-mediated immune reactions have shown that MPB70 is a highly BCG-specific antigen occurring in markedly different concentrations in different substrains of BCG. The present studies were based on the reactivity of MPB70 with antibodies. The precipitate caused by MPB70 was identified in the crossed immunoelectrophoresis pattern of sonicates and concentrated culture fluids of BCG Tokyo. The antigen was quantified by radioimmunoassay inhibition tests on culture fluids standardized on the basis of identical total protein content. It was present in high concentrations in BCG substrains Tokyo, Moreau, Russia, and Sweden, and in M. bovis strain Ravenel. It was also detected in BCG substrains Glaxo, Copenhagen, Tice, and Pasteur, and in M. tuberculosis H37Rv, but in the latter 4 in only 1% or less of the concentration in BCG Tokyo. Six different species of slow- and rapid-growing mycobacteria showed no evidence of cross-reactivity, but a cross-reacting antigen was demonstrated in Nocardia asteroides. The antibody response after immunization of rabbits with BCG Tokyo and BCG Copenhagen was very similar in a polyvalent assay for antibodies reacting with various components of BCG. By contrast, there was a striking difference in anti-MPB70 antibodies, which were rapidly formed in large amounts after immunization with BCG Tokyo and only in very small amounts after immunization with BCG Copenhagen.

Animals↗
PubMed1983

Immunohistological studies of skin biopsies from patients with lepromatous leprosy.

Forty-six skin biopsies from lepromatous leprosy patients were examined for immunoglobulin and complement deposits as well as mycobacterial antigens. Rabbit anti-human immunoglobulin, rabbit anti-human C3, and rabbit anti-Mycobacterium bovis (BCG) were used as the primary antigen-detecting antibodies in a peroxidase antiperoxidase technique. Of the 26 biopsies from active erythema nodosum leprosum lesions, 6 were positive for immunoglobulin or complement deposits. These deposits were found in the dermoepidermal junction, within the foamy cells, and, in one patient, around a blood vessel. Five of twenty patients with lepromatous leprosy without erythema nodosum leprosum showed similar deposits in the dermoepidermal junction and within foamy cells. None of these patients had these deposits around blood vessels. Mycobacterial antigens were seen in all biopsies studied. The presence of acute inflammatory infiltrates did not correlate with the presence or absence of immunoglobulin or complement deposits. It is felt that immunoglobulin or complement deposits are not a constant feature of early erythema nodosum leprosum lesions and that these deposits may be secondary rather than primary in these lesions.

Antigen-Antibody Complex↗
PubMed1983

Demonstration of mycobacterial antigens in nerve biopsies from leprosy patients using peroxidase-antiperoxidase immunoenzyme technique.

Peripheral nerve biopsies from patients with leprosy were stained with anti-Mycobacterium bovis (BCG) in a peroxidase-antiperoxidase (PAP) system to demonstrate intraneural mycobacterial antigens. Most M. leprae antigens have been shown to cross-react with BCG. Of the 30 biopsies from borderline tuberculoid (BT) patients 18 had acid-fast bacilli while 26 of them had demonstrable mycobacterial antigens in their nerves. All borderline lepromatous (BL) and lepromatous leprosy (LL) nerve biopsies had both M. leprae and mycobacterial antigens within them. Most of the antigens in the BT patients were seen to be extracellular. In BL and LL patients antigens were seen both extracellularly and intracellularly in Schwann cells and infiltrating macrophages. Mycobacterial antigens in BT nerves were always seen to be surrounded by a mononuclear cell reaction while in the BL and LL patients antigens could be seen with minimal cellular infiltrate and the neural architecture was more or less preserved. While bacilli could not be seen in BT patients who had been released from treatment for more than 4 years, mycobacterial antigens could still be seen in some patients who had been released from treatment for up to 5 years. Patients with no skin lesions but with large, painful, or tender nerves were found to have intraneural inflammation surrounding mycobacterial antigens, while those with a similar clinical picture but without tender or painful nerves showed no marked inflammation within their nerves despite the presence of mycobacterial antigens. From these findings it was concluded that immunologically mediated inflammatory response toward intraneurally located M. leprae antigens in conjunction with other host factors may be necessary for nerve damage in the BT leprosy patients. In the BL and LL patients the mechanisms of nerve damage are still unknown with certainty but local effects and immune-complex damage secondary to abundant M. leprae antigens are worth exploring. The use of immunohistological techniques should offer a new approach in the study of the immunopathology of leprosy.

Antigens, Bacterial↗
PubMed1983

Comparison of the cytoplasmic antigens of leprosy-derived corynebacteria and some mycobacteria.

The immunological relationship between leprosy-derived corynebacteria (LDC) and reference mycobacteria was analysed by crossed immunoelectrophoresis with intermediate gel. For this purpose, three reference systems (LDC15/anti-LDC15, LDC18/anti-LDC8, and LDC8/anti-LDC8) were developed. They showed 15 to 20 distinct antigenic components in LDC cytoplasm. Extensive cross-reactivity was observed among different LDC isolates, affecting 3 to 17 components. Moreover, several components were shown to cross-react with mycobacteria when anti-bacillus Calmette-Guérin (BCG), anti-Mycobacterium leprae, other antisera and lepromatous leprosy sera were incorporated in the intermediate gel. The major cross-reactive component, antigen M, was present in all LDC isolates and cross-reacted with antigen 7 of M. leprae and antigen 60 of M. bovis BCG. The thermostability of these antigens and the specificity of the cross-reacting antigens were assessed. The data underline the degree of immunochemical homogeneity within the LDC group of micro-organisms and relatedness with M. leprae and other mycobacteria.

Corynebacterium↗
PubMed1983

Nerve damage following intraneural injection of Mycobacterium leprae into rabbits pre-sensitized to mycobacteria.

Nerve damage is a common feature of leprosy although the mechanism responsible for the damage is not clearly understood. In the tuberculoid end of the leprosy spectrum where both intraneural Mycobacterium leprae or their antigens and cell-mediated hypersensitivity to M. leprae co-exist, acute neuritis affecting major nerve trunks can occur during reversal reactions. These reactions are known to be associated with increased hypersensitivity to M. leprae antigens. The nerve involvement is therefore thought to be a direct consequence of the patient's hypersensitivity to M. leprae. So far the only indirect evidence based on in vitro studies have been produced to support such a contention. We sensitized rabbits with M. leprae and then injected M. leprae sonicate into the sciatic nerves at the peak of hypersensitivity. Seventy-two hours later, the nerves were dissected out and studied histologically. Our results show that cellular infiltration and axonal degeneration can occur as a direct consequence of hypersensitivity to intraneural M. leprae antigens. This study, therefore, offers direct evidence for the involvement of specific cell-mediated hypersensitivity to M. leprae antigens in the pathogenesis of major nerve trunk damage in the tuberculoid end of the leprosy spectrum especially during acute reversal reactions.

Animals↗
PubMed1983

Relation between anti-Mycobacterium leprae antibody activity and clinical features in borderline tuberculoid (BT) leprosy.

Antibody activity against Mycobacterium leprae antigen 7 was determined by radioimmunoassay and IgG antibodies against various antigens present in an M. leprae sonicate by a solid phase radioimmunoassay in 77 patients with borderline tuberculoid (BT) leprosy. In both assays there was a wide variation in antibody activity in individual patients although all were diagnosed as having BT leprosy. The median antibody activity was lower in newly diagnosed cases than in patients appearing with active skin lesions or new skin lesions despite dapsone (DDS) treatment of long duration. Further comparison of patients with high and low antibody activity revealed that high antibody activity was significantly correlated statistically with active skin lesions, new skin lesions and neuritis despite DDS treatment of long duration. The reason for variation in antibody activity in newly diagnosed BT leprosy remains unclear, and this patient group is of particular interest for further characterization of the basis for variation in antibody activity in tuberculoid leprosy.

Antibodies, Bacterial↗
PubMed1983

Effect of treatment on antibody activity against Mycobacterium leprae antigen 7 in tuberculoid leprosy.

Anti-Mycobacterium leprae antigen 7 antibody activity was determined by radioimmunoassay during treatment in a longtime study of individual patients with newly diagnosed borderline tuberculoid (BT) leprosy and in BT leprosy patients who were suspected from their case histories to have dapsone (DDS) resistant leprosy. There was a strong correlation between clinical and antibody activity, and clinical improvement following treatment led to a marked decrease in antibody activity in most cases. A characteristic pattern of rapid and marked increase in antibody activity shortly after the initiation of treatment was observed in patients with newly diagnosed BT leprosy. This pattern may become of practical importance in the evaluation of patients with BT leprosy as an indicator that the therapy is effective, even though this pattern was associated with a transient increase in inflammatory activity in the skin lesions. The association of inflammatory activity with increased antibody activity strongly indicates that the underlying processes are associated with the stimulation of both humoral and cellular immune responses.

Antibodies, Bacterial↗