Establishment of Sandwich Enzyme-Linked ImmunoSorbent Assay (ELISA) System for Quantitation of CD81

Hatanaka, Shiho; Mihara, Hiroki; Kohno, Sayuki; Fujimoto, Erika; Tanaka, Miyuki; Takehara, Motoharu; Yamasaki, Tsutomu; Nakanishi, Tohru

doi:10.2485/jhtb.24.401

Cited by 3 publications

(4 citation statements)

References 14 publications

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“…It was reported that TNF- and IL-1 induce synoviolin expression through activation of Erk1/2-ETS1 signalregulated kinase in synovial fibroblasts 18) . We have previously shown that CD81 mediates TNF-á-induced synoviolin expression in SW982 cells 12) . Thus, a similar mechanism acting through activation of the Erk1/2-ETS1 signal-regulated kinase may act in the CD81-synoviolin pathway and lead to CD81-dependent proliferation of synoviocytes in RA.…”

Section: Discussionmentioning

confidence: 95%

“…This eluted fraction was concentrated and the titer of IgG1 MAb was analyzed by ELISA. These IgG1 MAbs showed a higher titer to the antigen compared with the titer of antiserum 11,12) .…”

Section: Production Of Igg Monoclonal Antibodiesmentioning

confidence: 93%

“…After 10-14 days, ascites were collected from these mice. The growth rate of hybridomas and the volume of ascites from four clones is shown elsewhere 11,12) . Relative Luc Activity…”

Section: Establishment Of Hybridoma Cell Linesmentioning

confidence: 99%

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CD81 Mediated Regulation of Synoviolin Expression in Synovial Sarcoma Cells

Fujimoto

Matsushita

Nakajima

et al. 2016

J. Hard Tissue Biology.

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Tetraspanin CD81, which is up-regulated in rheumatoid arthritis (RA) synoviocytes, is a molecule containing four transmembrane domains. We recently showed that small interfering RNA targeting CD81 (CD81 siRNA) ameliorated arthritis in rats. In that study, the expression of synoviolin was decreased in RA joints by CD81 siRNA. We also showed that CD81 siRNA decreased the expression of synoviolin and TNF- in SW982 synovial sarcoma cells, suggesting that overexpression of CD81 in synovial cells induces the expression of both synoviolin and TNF- in the same cells. Here we raised monoclonal antibodies (MAbs) against CD81 by immunization of the outer membrane region of CD81 (LEL) that was overexpressed as a recombinant protein.One of these MAbs stimulated CD81 on the membrane of SW982 and up-regulated the expression of synoviolin. We also defined the region of the synoviolin gene promoter that is essential for induction of synoviolin gene expression by using reporter gene system. This system might be useful for the screening of anti-RA medicine.

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Section: Discussionmentioning

confidence: 95%

“…This eluted fraction was concentrated and the titer of IgG1 MAb was analyzed by ELISA. These IgG1 MAbs showed a higher titer to the antigen compared with the titer of antiserum 11,12) .…”

Section: Production Of Igg Monoclonal Antibodiesmentioning

confidence: 93%

See 1 more Smart Citation

CD81 Mediated Regulation of Synoviolin Expression in Synovial Sarcoma Cells

Fujimoto

Matsushita

Nakajima

et al. 2016

J. Hard Tissue Biology.

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show abstract

“…Given this, diagnosis using CD81 or its derivatives might open new avenues for RA treatment. We have already established a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) system for quantitating CD81 by using two MAbs against CD81 that recognize different epitopes 13) . Further improvement of this system and the identifi cation of possible molecules for RA diagnosis are in progress.…”

Section: Discussionmentioning

confidence: 99%