Indirect double sandwich ELISA for the specific and quantitative measurement of mouse IgM, IgA and IgG subclasses

Klein-Schneegans, Anne-Sophie; Gavériaux, Claire; Fonteneau, Paul; Loor, Francis

doi:10.1016/0022-1759(89)90388-8

Cited by 46 publications

(19 citation statements)

References 11 publications

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“…In C57BL/6 mice, IgG in milk is predominantly IgG2b due to its abundance in serum, although IgG1 is also present in disproportionately high concentrations compared to those of serum (30,42). The differential loading of immunoglobulin subclasses into milk may have been responsible for the discrepancy in urease-specific IgG levels in the sera from dams compared to those of their pups, but we did not confirm this by determining the subclasses of the antibodies we measured.…”

Section: Discussionmentioning

confidence: 73%

Antibody-Mediated Protection against Infection withHelicobacter pyloriin a Suckling Mouse Model of Passive Immunity

Gorrell

Robins-Browne

2009

Infect Immun

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Studies of active immunization againstHelicobacter pylori indicate that antibodies play a minor role in immunity. There is also evidence, however, that the translocation of antibodies in the stomach may be insufficient to achieve functional antibody levels in the gastric lumen. We have used a suckling mouse model of passive immunity to determine if perorally delivered antibodies can protect against infection with H. pylori. Female C57BL/6 mice were immunized parenterally with formalin-fixed cells of three clinical isolates of H. pylori (3HP) or the mouse-adapted H. pylori strain SS1 before mating. Their pups were challenged with the SS1 strain at 4 days of age and left to suckle before determination of bacterial loads 14 days later. Compared to age-matched controls, pups suckled by 3HP-vaccinated dams were significantly protected against infection (>95% reduction in median bacterial load; P < 0.0001). Pups suckled by SS1-vaccinated dams were also significantly protected in terms of both median bacterial load (>99.5% reduction; P < 0.0001) and the number of culture-negative pups (28% versus 2% for immune and nonimmune cohorts, respectively; P < 0.0001). Similar results were obtained with pups suckled by dams immunized with a urease-deficient mutant of SS1. Fostering experiments demonstrated that protection was entirely attributable to suckling from an immunized dam, and antibody isotype analysis suggested that protection was mediated by the immunoglobulin G fraction of immune milk. Analysis of the bacterial loads in pups sampled before and after weaning confirmed that infection had been prevented in culture-negative animals. These data indicate that antibodies can prevent colonization by H. pylori and suppress the bacterial loads in animals that are colonized.

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Section: Discussionmentioning

confidence: 73%

Antibody-Mediated Protection against Infection withHelicobacter pyloriin a Suckling Mouse Model of Passive Immunity

Gorrell

Robins-Browne

2009

Infect Immun

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“…Blood and urinary urea nitrogens were measured by an autoanalyzer (Hitachi model 736, Tokyo, Japan), and the clearance of urea nitrogen for 24 h (Cun) was calculated. Serum IgA, IgG, and IgM were measured by sandwich ELISA described previously [12]. Briefly, microtitration plates (Corning, Cambridge, Mass., USA) were coated with goat anti-mouse IgA (Cappel Laboratories, Cochranville, Pa., USA), IgG, or IgM antibodies (Cappel Laboratories), and nonspecific protein bindings were blocked with BSA.…”

Section: Methodsmentioning

confidence: 99%

Heminephrectomy Causes the Progression of Glomerulosclerosis and Apoptosis in High IgA Strain ddY Mice

Kusano

Muso

Ono

et al. 2002

Nephron

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Background: The reduction in nephrons in IgA nephropathy is critical to the prognosis of this disease. However, the immunopathological mechanism of the modifications seen in glomerular lesions is not clear. We thus investigated the influence of nephron reduction by heminephrectomy on renal lesions in a high immunoglobulin A inbred strain of ddY mouse (HIGA mouse), which shows progressive mesangial sclerosis with elevated renal expression of transforming growth factor (TGF)-β. Methods: Five-week-old HIGA mice were heminephrectomized (Nx), and were evaluated in comparison with a sham-operated group (S) at 40 weeks old. Histological findings, immunoglobulin depositions (IgG, IgA, and IgM), and expressions of cytokine and extracellular matrix proteins (TGF-β, fibronectin, collagen type I and IV) were analysed. PCNA and TUNEL stainings were performed with electron microscopic detection of apoptosis. Tissue renin-angiotensin systems (RAS) were also investigated by real-time quantitative RT-PCR. Results: In the Nx group, the glomerular tuft area and ratio of mesangial matrix area per tuft were significantly increased, and the glomerular cell count per tuft area was significantly decreased. Glomerular immunoglobulin deposits of IgG, IgA, and IgM in Nx were all significantly expanded in the paramesangium. The glomerular expressions of TGF-β and the extracellular matrix proteins were significantly increased in Nx mice. In contrast to the significant decrease of PCNA-positive cells, TUNEL-positive cells were significantly increased in Nx. Angiotensin-converting enzyme (ACE) was significantly increased in the renal cortex of Nx. Conclusion: Simple heminephrectomy, other than 5/6 renal ablation, of HIGA mice may be a potential model for research into the progressive glomerulosclerosis of human IgA nephropathy. The pathological role of apoptosis is apparently involved in these disease processes, possibly through upregulated RAS.

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“…We found a direct correlation between the levels of Th1 cytokines in serum of tumour-bearing animals and rejection of the tumour. Based on the day of onset of tumour rejection we classified the animals as early regressors (days 9-12) and late regressors (days [16][17][18][19][20][21][22][23][24][25][26][27][28]. Figure 1 shows the tumour growth pattern in the early and late regressors.…”

Section: Resultsmentioning

confidence: 99%

“…Cytokine levels (IL-2, IL-4, IL-10, IFN-g and TNF-a) in the serum and tumour extracts were estimated by ELISA according to Schneegans et al [20], with some modifications. Briefly, 50-ml (1:100 in PBS) aliquots were added to 96-well microtitre plates (Flow Labs, VA) and incubated at 4ЊC overnight.…”

Section: Elisamentioning

confidence: 99%

High intratumoural level of cytokines mediates efficient regression of a rat histiocytoma

Khar

Varalakshmi

1997

Clin Exp Immunol

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SUMMARYWe have studied the role of cytokines in the spontaneous regression of AK-5 histiocytoma in syngeneic rats. Animals in which the tumour regresses show high levels of cytokines in the serum compared with animals which succumb to the tumour, and levels of IL-2, IL-4, IL-12, interferon-gamma (IFN-g) and tumour necrosis factor-alpha (TNF-a) are significantly higher in tumour tissue of the former. Thus there is an association between rejection of the tumour and the levels of cytokines present in the tumour mass. Our results also suggest a predominant Th1-type of response in those rats that display early tumour rejection.

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Indirect double sandwich ELISA for the specific and quantitative measurement of mouse IgM, IgA and IgG subclasses

Cited by 46 publications

References 11 publications

Antibody-Mediated Protection against Infection withHelicobacter pyloriin a Suckling Mouse Model of Passive Immunity

Antibody-Mediated Protection against Infection withHelicobacter pyloriin a Suckling Mouse Model of Passive Immunity

Heminephrectomy Causes the Progression of Glomerulosclerosis and Apoptosis in High IgA Strain ddY Mice

High intratumoural level of cytokines mediates efficient regression of a rat histiocytoma

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