A direct enzyme-linked immunosorbent assay (ELISA) for antibodies to enterobacterial Re core glycolipid and lipid A

Nys, Monique; Damas, Pierre; Damas, François; Joassin, L.; Demonty, J

doi:10.1007/bf00190532

Cited by 11 publications

(11 citation statements)

References 19 publications

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“…In this study, we used different ELISAs and an immunoprecipitation procedure to detect the LPS-precipitable antibodies in a sample of the Belgian blood donor population, with the aim of preparing effective immune globulin with high and consistent levels of functional antibodies. In agreement with other reports [35][36][37][38][39][40], we found a natural occurrence of antibodies to LPS, core glycolipid and lipid A in normal subjects. The presence of antibodies to antigenic determinants of various LPSs was expected.…”

Section: Discussionsupporting

confidence: 93%

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Screening and characterization of specific anti‐lipopolysaccharide antibodies in Belgian blood donors by enzyme‐linked immunosorbent assays

Nys¹,

Laub²,

Damas³

et al. 1996

Eur J Clin Investigation

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The goal of this project was to find and collect high concentrations of endotoxin-specific antibodies for therapeutic IgG- or IgM-enriched preparations. Various enzyme-linked immunosorbent assays (ELISAs) were developed to perform longitudinal studies of the serological response to a large panel of smooth and rough purified lipopolysaccharide (LPS) extracts in a population of healthy blood donors. To accomplish this, 1612 human serum samples from volunteer blood donors collected by seven different blood banks in Belgium were screened and specific IgM and IgG activities were measured. Approximately 17% of the donors had anti-LPS concentrations higher than 40 mg L-1. Of these, 10.9% had anti-smooth LPS antibodies, 3.7% had anti-rough LPS antibodies and 2.8% were found to be positive towards both types of LPS. The mean anti-LPS antibody concentration was 8 mg L-1 for rough LPS and 14 mg L-1 for smooth LPS. Age- and sex-related distributions of the activities indicated that the greatest prevalence of high anti-LPS concentration was in women aged 40-49 years and in men older than 60 years. Differential absorption experiments showed that the pooled serum of selected blood donors contained a mixture of specific and cross-reacting antibodies. We detected predominantly anti-LPS activities due to the IgG1 and IgG2 subclasses. The range of specificities to different LPS was increased by the pooling of selected sera. It was concluded that pools of naturally occurring specific anti-LPS immunoglobulin antibodies may be obtained in Belgium by screening blood donors using ELISAs that we have developed.

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Section: Discussionsupporting

confidence: 93%

“…Other authors have observed that naturally occurring antibody to LPS is present in most individuals to a variable degree [38][39][40]. A response may be boosted by infections but also by vaccination [39,44,45].…”

Section: Discussionmentioning

confidence: 99%

Screening and characterization of specific anti‐lipopolysaccharide antibodies in Belgian blood donors by enzyme‐linked immunosorbent assays

Nys¹,

Laub²,

Damas³

et al. 1996

Eur J Clin Investigation

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“…The presence of endogenous anti-LPS antibodies (IgG and/or IgM) to the panel of smooth LPS, rough LPS, and lipid A antigens was evaluated by a direct enzyme-linked immunosorbent assay (ELISA) as previously described with minor modi®cations (Nys et al 1987;Camus et al 1997). All samples were assayed, and were screened using twofold dilutions of individual preparations from 1:100 to 1:51200 in contact buer and negative controls (uncoated wells).…”

Section: General Proceduresmentioning

confidence: 99%

Endotoxaemia, production of tumour necrosis factor ? and polymorphonuclear neutrophil activation following strenuous exercise in humans

Camus

Nys

Poortmans

et al. 1998

European Journal of Applied Physiology

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To examine whether endotoxaemia accompanying long-term, strenuous physical exercise is involved in exercise-induced increase in plasma tumour necrosis factor alpha (TNF-alpha) concentration and polymorphonuclear neutrophil (PMN) activation, 14 male recreational athletes [mean age 28 (SEM 1) years] were studied. Exercise consisted of a 1.5-km river swim, a 40-km bicycle race, and a 10-km road race. Mean time to complete the race was 149.8 (SEM 4.8) min. The plasma concentrations of granulocyte myeloperoxidase (MPO) and TNF-alpha were significantly higher than baseline values immediately and 1 h after exercise (P<0.001). Both variables returned to pre-race levels the day after exercise. Marked, transient decreases in plasma concentrations of anti-lipopolysaccharide (LPS) immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies directed against a panel of selected smooth gram-negative LPS were observed after the race, reaching in most cases minimal values in the blood sample drawn immediately following the completion of the triathlon. There was no significant correlation between the magnitude of PMN activation, as assessed by the increase in plasma concentrations of MPO, and the humoral markers of endotoxaemia and TNF-alpha. An inverse, highly significant relationship between the increase in plasma TNF-alpha concentrations and the changes in circulating anti-LPS IgM antibodies concentrations was observed (r = -0.7; P<0.01). These findings suggest that exercise-induced endotoxaemia was involved in the release of TNF-alpha, that the magnitude of the TNF-alpha response to exercise was down-regulated by anti-LPS antibodies of the IgM class, and that the production of TNF-alpha and endotoxaemia did not seem to play a role in the activation of circulating PMN in the exercising subjects.

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“…The presence of endogenous anti-LPS antibodies (IgG) to the panel of smooth LPSs, rough LPSs, and lipid A antigens was measured by a direct ELISA, as previously described with minor modifications [25]. Briefly, the plates (Nunc) were coated either with a mixture of the 13 smooth (S)-form LPSs (10 &ml of each), with a mixture of the nine rough (R)-form LPSs (10 pg/ml of each), or with individual purified smooth or rough LPSs at a concentration of 10 pg/ml and lipid A at a concentration of 5 pg/ml.…”

Section: Anti-lps Antibody Assaymentioning

confidence: 99%

Mild Endotoxaemia and the Inflammatory Response Induced by a Marathon Race

Camus¹,

et al. 1997

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1. To address the question of whether endotoxaemia could be involved in the inflammatory response induced by long-term strenuous exercise, 18 male marathon runners [mean age 41 +/- 2 (SEM) years] were studied. Their performance in the marathon ranged from 2 h 46 min to 4 h 42 min. 2. Four venous blood samples were drawn: at rest, just before the race (baseline); within 15 min following the completion of the marathon; after 1 h of recovery; and the morning after the race. 3. The following humoral markers of the inflammatory response to exercise were measured: polymorphonuclear myeloperoxidase (MPO), anaphylatoxin C5a (C5a), tumour necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6). Plasma endotoxin was measured by a sensitive and rapid chromogenic Limulus assay. All inflammatory markers were significantly increased (P < 0.001) after the race, reaching in most cases peak values in the first blood sample drawn following the completion of the marathon [MPO, 298 +/- 19 ng/ml (SEM); C5a, 1.45 +/- 0.32 ng/ml; TNF-alpha, 20 +/- 3 pg/ml; IL-6, 88 +/- 13 pg/ml] when compared with baseline [MPO, 146 +/- 16 ng/ml (SEM); C5a, 0.27 +/- 0.2 ng/ml; TNF-alpha, 12 +/- 1.5 pg/ml: IL-6, 1.0 +/- 0.5 pg/ml]. Traces of plasma endotoxin (ranging from 5 to 13 pg/ml, with one exceptionally high value of 72 pg/ml measured in one runner) were detected in seven subjects within the first hour of recovery. An ELISA method was used to determine the endogenous IgG antibodies toward a range of Gram-negative bacterial lipopolysaccharides (LPSs) of different sizes and structures. A transient decrease in certain anti-LPS activities, mainly against rough LPS, occurred in most cases in the first blood sample drawn after the race. There was no correlation between the magnitude of the inflammatory response to exercise, as assessed by the increase in blood levels of humoral markers of inflammation, and the changes in circulating endotoxin levels of anti-LPS IgG activity following the race. 4. From these results, we conclude that the mild, transient endotoxaemia detected in some of our subjects does not play a major role in the observed inflammatory response to a marathon competition.

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A direct enzyme-linked immunosorbent assay (ELISA) for antibodies to enterobacterial Re core glycolipid and lipid A

Cited by 11 publications

References 19 publications

Screening and characterization of specific anti‐lipopolysaccharide antibodies in Belgian blood donors by enzyme‐linked immunosorbent assays

Screening and characterization of specific anti‐lipopolysaccharide antibodies in Belgian blood donors by enzyme‐linked immunosorbent assays

Endotoxaemia, production of tumour necrosis factor ? and polymorphonuclear neutrophil activation following strenuous exercise in humans

Mild Endotoxaemia and the Inflammatory Response Induced by a Marathon Race

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