Development of a Quantitative Cell-Based Intracellular ELISA for the Screening of B Cell Hybridoma Supernatants: A Novel Rapid Assay to Detect Positive Clones

Renukaradhya, Gourapura J.; Sriram, Venkataraman; Poláková, K; Russ, G; Brutkiewicz, Randy R.

doi:10.1089/hyb.2004.23.373

Cited by 7 publications

(9 citation statements)

References 15 publications

(18 reference statements)

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“…57 We were able to detect significant differences in vivo regarding tumor growth and mouse survival (Fig. 2), although the overall level of CD1d on the cell surface of RMA/S-CD1 cells was about 62% greater (as measured by the 8F3 antibody 24 ) than the RMA/ S-V line (Fig. 1).…”

Section: Discussionmentioning

confidence: 92%

“…Thus, how does this difference manifest itself in the observations reported here? When we stained these two lines with a panel of 10 mouse CD1d-specific mAb (which have distinct epitope specificities for the murine CD1d1 molecule, 24 and data not shown), it was apparent that the staining patterns were different when compared to that observed with CD1d-transfected murine L cells (data not shown). This suggested that there were both quantitative and especially ''qualitative'' differences between the two tumor-cell lines; namely, the RMA/S lipids that are presented by CD1d to NKT cells may be inhibitory in nature.…”

Section: Discussionmentioning

confidence: 99%

“…To address this question, CD1d was overexpressed in the murine T-cell lymphoma RMA/S. 21 As measured by staining with the 8F3 mouse CD1d-specific mAb, 24 there was approximately 62% more CD1d on the surface of RMA/S cells upon transfection with the cd1d1 cDNA ( Fig. 1).…”

Section: T-cell Lymphomamentioning

confidence: 99%

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Inhibition of antitumor immunity by invariant natural killer T cells in a T‐cell lymphoma model in vivo

Renukaradhya

Sriram

et al. 2006

Intl Journal of Cancer

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We have investigated the role of the host's CD1d-dependent innate antitumor immune response in a murine T-cell lymphoma model in vivo. We found that C57BL/6 wildtype (WT) mice inoculated with RMA/S cells transfected with murine CD1d1 died at the same rate as mice inoculated with vector-transfected cells. In contrast, natural killer T (NKT) cell-deficient CD1d or Ja18 knockout mice inoculated with CD1d-transfected RMA/S cells survived significantly longer than mice inoculated with vector-transfected RMA/S cells, implicating the involvement of invariant NKT (iNKT) cells in inhibiting antitumor activity in vivo. In contrast to the mutant mice, which produced more of the proinflammatory cytokines IFN-c and GM-CSF, WT mice produced significantly elevated amounts of IL-13. Antitumor activity in the knockout mice was not due to the development of CD1d-specific cytotoxic T lymphocytes or circulating antibodies. However, iNKT cell numbers were elevated in tumor-bearing mice. Thus, iNKT cells may be playing a negative role in the host's antitumor immune response against T-cell lymphomas in a CD1d-dependent manner. ' 2006 Wiley-Liss, Inc.Key words: T-cell lymphoma; RMA/S; invariant NKT cells; CD1d molecules; cytokines Despite apparent immunocompetence in the host, lymphomas can arise nonetheless, evading the body's attempts at tumor immunosurveillance. Following transformation, the host's innate immune response is essential in controlling the dissemination and growth of metastatic disease. Immunosurveillance against spontaneously occurring lymphomas is not well understood. Understanding the molecular mechanisms that regulate divergent arms of the immune response is a key to develop effective immunotherapies for many diseases, including cancer.1 Recent studies have suggested an immunosurveillance function for a unique subpopulation of T cells that play an important role in innate immunity called natural killer T (NKT) cells.2 NKT cells are defined as T cells expressing markers on the surface shared with NK cells, and that rapidly secrete both Th1 and Th2 cytokines upon T-cell receptor (TCR) ligation. 3,4 Rather than recognizing peptides presented by MHC class I or class II molecules, NKT cells recognize lipid antigens presented by the MHC class I-like CD1d molecule.5 NKT cells are key players in the regulation of antitumor immunity, particularly in experimental models of tumor immunotherapy, including IL-12 or a-galactosylceramide (a-GalCer) administration.6 In a mouse model in which tumors spontaneously regress after initial growth and then recur, the negative regulation of tumor immunosurveillance by IL-13, possibly produced by CD41 NKT cells, has been reported. 7,8 In a more recent study, the same group has shown that a reduction in pulmonary metastases in the CT26 nonregressor colon carcinoma model occurs following the elimination of CD4 1 NKT cells, further illustrating an inhibitory role for NKT cells in antitumor immunity. 9 To date, the role of invariant (Va14Ja18 1 ) NKT (iNKT) cells in the evasion of hematopoieti...

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Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

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Inhibition of antitumor immunity by invariant natural killer T cells in a T‐cell lymphoma model in vivo

Renukaradhya

Sriram

et al. 2006

Intl Journal of Cancer

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“…Normal rat serum was purchased from Sigma-Aldrich (St. Louis, MO). The anti-mouse CD1d1 mAbs: 1H6 [25], 1E2, 8F3, 9E4, and 1A8 generated by us have been described previously [29]. The anti-MHC class I (K b ) exon-8 specific rabbit antiserum and human TAP1-expressing recombinant vaccinia virus (VV) [30] were kindly provided by Drs.…”

Section: Methodsmentioning

confidence: 99%

Forming a Complex with MHC Class I Molecules Interferes with Mouse CD1d Functional Expression

Renukaradhya¹,

Khan²,

Gallo³

et al. 2013

PLoS ONE

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CD1d molecules are structurally similar to MHC class I, but present lipid antigens as opposed to peptides. Here, we show that MHC class I molecules physically associate with (and regulate the functional expression of) mouse CD1d on the surface of cells. Low pH (3.0) acid stripping of MHC class I molecules resulted in increased surface expression of murine CD1d on antigen presenting cells as well as augmented CD1d-mediated antigen presentation to NKT cells. Consistent with the above results, TAP1-/- mice were found to have a higher percentage of type I NKT cells as compared to wild type mice. Moreover, bone marrow-derived dendritic cells from TAP1-/- mice showed increased antigen presentation by CD1d compared to wild type mice. Together, these results suggest that MHC class I molecules can regulate NKT cell function, in part, by masking CD1d.

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“…8 ␣-and ␤-galactosylceramides (GalCer) were purchased from Enzo Life Sciences. Anti-CD1d Abs were: 1B1, 20 1H6, 21 1A8, 22 1E2, 22 6F7, 22 and 9G1 22 and isotype-matched controls were generated as described previously.…”

Section: Reagentsmentioning

confidence: 99%

CD1d expression on and regulation of murine hematopoietic stem and progenitor cells

Broxmeyer

Christopherson

Hangoc

et al. 2012

Blood

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Development of a Quantitative Cell-Based Intracellular ELISA for the Screening of B Cell Hybridoma Supernatants: A Novel Rapid Assay to Detect Positive Clones

Cited by 7 publications

References 15 publications

Inhibition of antitumor immunity by invariant natural killer T cells in a T‐cell lymphoma model in vivo

Inhibition of antitumor immunity by invariant natural killer T cells in a T‐cell lymphoma model in vivo

Forming a Complex with MHC Class I Molecules Interferes with Mouse CD1d Functional Expression

CD1d expression on and regulation of murine hematopoietic stem and progenitor cells

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