Establishment of an ELISA system for determining soluble LAIR-1 levels in sera of patients with HFRS and kidney transplant

Ouyang, Wei; Xue, Jian; Liu, Jingmei; Jia, Wei Hua; Li, Zhouli; Xie, Xin; Liu, Xuesong; Jian, Jinlong; Li, Qi; Zhu, Yi‐Chun; Yang, An‐Gang; Jin, Boquan

doi:10.1016/j.jim.2004.06.005

Cited by 28 publications

(23 citation statements)

References 27 publications

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“…Thus, the lower expression of LAIR-1 on stimulated Tcells is not caused by decreased transcription of LAIR-1. Next, we performed an ELISA to detect soluble LAIR-1 [17]; no LAIR-1 protein was detected in the supernatant of stimulated T cells (data not shown). Thus, if LAIR-1 is shed upon T cell stimulation, it reaches concentrations less than 4 ng/mL, the detection limit of our assay.…”

Section: Lair-1 Is Downregulated Upon Polyclonal Stimulation In Vitromentioning

confidence: 99%

Regulated expression of the inhibitory receptor LAIR‐1 on human peripheral T cells during T cell activation and differentiation

et al. 2007

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The leukocyte-associated Ig-like receptor-1 (LAIR-1) is capable of inhibiting immune cell function through interaction with collagens. LAIR is expressed on the majority of peripheral blood mononuclear cells. The abundant expression of both receptor and ligand calls for regulatory mechanisms to relieve the continuous interaction between collagens and LAIR-1. This regulation may occur at the expression level of the receptor. Here, we report that LAIR-1 is indeed differentially expressed during human T cell differentiation. Naive CD4 + and CD8 + T cells as well as CD8 + T cells of the effector phenotype express higher levels of LAIR-1 compared to memory T cells. In vitro stimulation revealed a decrease in LAIR-1 expression upon activation, and the lower LAIR-1 expression on CD127 -T cells suggests that activation-induced down-modulation of LAIR-1 may also occur in vivo. Furthermore, crosslinking of LAIR-1 on primary T cells results in an inhibition of T cell function. Our data suggest that regulated expression of LAIR-1 and the subsequent change in the threshold for activation may be a mechanism to modulate inhibition of the immune system.

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Section: Lair-1 Is Downregulated Upon Polyclonal Stimulation In Vitromentioning

confidence: 99%

Regulated expression of the inhibitory receptor LAIR‐1 on human peripheral T cells during T cell activation and differentiation

et al. 2007

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“…One level of regulation occurs by modulating hLAIR-1 expression at different stages of differentiation/activation of immune cells, as was demonstrated for B cells (12), T cells (4,5), neutrophils (3), and dendritic cells (our unpublished observations). Furthermore, hLAIR-1 can be shed from immune cells upon cellular activation (18). Another system to regulate the interaction between collagens and hLAIR-1 may involve the putative secreted homolog LAIR-2 (CD306).…”

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confidence: 99%

The Soluble Leukocyte-Associated Ig-Like Receptor (LAIR)-2 Antagonizes the Collagen/LAIR-1 Inhibitory Immune Interaction

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et al. 2008

The Journal of Immunology

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Leukocyte-associated Ig-like receptor (LAIR)-1 is a collagen-receptor that inhibits immune cell function upon collagen binding. Next to LAIR-1, the human genome encodes LAIR-2, a putative soluble homolog. In this study we show, for the first time, that the LAIR-2 gene is broadly transcribed in human PBMC, mirroring the expression profile of LAIR-1. LAIR-2 protein is expressed as a soluble receptor exhibiting high affinity for various collagen molecules to which it binds in a hydroxyproline-dependent manner. In vitro stimulation of PBMC induces secretion of LAIR-2. We detect high amounts of LAIR-2 in urine of pregnant women, indicating that the soluble receptor is indeed produced in vivo and can be cleared from the body via urine. Furthermore, LAIR-2 levels are increased in synovial fluid of patients with rheumatoid arthritis as compared with osteoarthritis patients. We hypothesize that soluble LAIR-2 may function as a natural competitor for LAIR-1, thereby regulating its inhibitory potential. Indeed, LAIR-2 prevents binding of human LAIR-1 to collagens and LAIR-1 cross-linking in vitro, suggesting that the protein has an immunoregulatory function in vivo. Hence, we reveal a novel mechanism of immune regulation by a soluble LAIR receptor regulating the inhibitory potential of the membrane-bound LAIR-1 via competition for ligands.

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“…Therefore, in this research, the weight was expressed in terms of protein concentration which was determined spectrophotometrically. Due to low absorbance (purified factor VIII) obtained by many investigators at k 280 nm, this could be due to low aromatic amino acid content of factor VIII [25], thereby protein was also measured by Bradford method. The amount of factor VIII in patients is determined by PTT method, in these patients the amount of PT is usually normal and PTT will be high hence, by adding plasma of normal person, the amount of PTT will be normal.…”

Section: Discussionmentioning

confidence: 99%

Production and Purification of Rabbit’s Polyclonal Antibody Against Factor VIII

et al. 2011

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The attempt is made to produce recombinant factor VIII but the first step in producing such product is production and purification of rabbit's polyclonal antibody against factor VIII. The second and third steps involve monoclonal antibody and recombinant factor VIII production. Factor VIII is one of the most important coagulating factor where its deficiency leads to diseases like hemophilia type A or classic. It is an inherited disease. Previously, it was obtained through fractionation of blood plasma of blood donors. After processing, factor VIII could be used to manage such patients. Due to transfer of viral disease like hepatitis and HIV through factor VIII obtained by fractionation, high cost of production, insufficiency of the donors and the process of virus removal, thus production of factor VIII through recombinant technology can be useful and helpful. The reaction between antibody and antigen is one the most specific reaction; therefore, such reaction can be employed to identify factor VIII. Thereby, rabbits were injected several times with adjuvant-linked antigen to produce antibody. The antibody was separated from the blood sample, purified and used to identify factor VIII in the research.

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Establishment of an ELISA system for determining soluble LAIR-1 levels in sera of patients with HFRS and kidney transplant

Cited by 28 publications

References 27 publications

Regulated expression of the inhibitory receptor LAIR‐1 on human peripheral T cells during T cell activation and differentiation

Regulated expression of the inhibitory receptor LAIR‐1 on human peripheral T cells during T cell activation and differentiation

The Soluble Leukocyte-Associated Ig-Like Receptor (LAIR)-2 Antagonizes the Collagen/LAIR-1 Inhibitory Immune Interaction

Production and Purification of Rabbit’s Polyclonal Antibody Against Factor VIII

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