Use of a monoclonal rat anti-mouse Ig light chain (RAMOL-1) antibody reduces background binding in immunohistochemical and fluorescent antibody analysis.

Brodin, N. Thomas; Jansson, Bo; Hedlund, Gunnar; Sjögren, Hans

doi:10.1177/37.7.2499618

Cited by 14 publications

(7 citation statements)

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“…We considered other possibilities that might explain our observed staining. A well-known phenomenon is the specific staining of tissues by secondary antibodies due to the presence of endogenous immunoglobulins (Brodin et al, 1989;Tuson et al, 1990). For example, the use of any secondary antibody raised against mIgG will give rise to strong background staining of mouse tissue depending on the amount of plasma, lymph, and lymphoid cells present.…”

Section: Ventral Nerve Cord Of M Sextamentioning

confidence: 99%

Novel mouse IgG‐like immunoreactivity expressed by neurons in the moth Manduca sexta: Developmental regulation and colocalization with crustacean cardioactive peptide

Klukas

Brelje

Mesce

1996

Microsc. Res. Tech.

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Immunoglobulin-related molecules have been shown to play important roles in cell-cell recognition events during the development of both vertebrate and invertebrate nervous systems. In the moth, Manduca sexta, we report the presence of novel, mouse, immunoglobulin G (mIgG)-like immunoreactivity in a discrete population of identified neurosecretory neurons (the NS-Ls also known as the cell 27s) and interneurons (the IN-704s). A number of polyclonal anti-mIgG antibodies were used to immunostain these cells in wholemount. The mIgG-like-immunoreactive (IR) neurons were present during embryogenesis through the developing adult stages, but disappeared in the postemerged adult. Biochemical analysis of M. sexta ventral nerve cords revealed that the mIgG-like antigen is a membrane-associated 27-kDa protein which is likely responsible for the mIgG-like immunostaining observed. Unambiguous identification of the mIgG-like-IR neurons was based on neuronal morphology and our ability to demonstrate conclusively that these neurons expressed immunoreactivity to an antiserum against crustacean cardioactive peptide (CCAP). The NS-Ls and IN-704s were both shown to colocalize the CCAP and mIgG-like immunoreactivities. The mIgG-like and CCAP-IR neurons were identical to a subset of CCAP-IR neurons recently described by Davis et al. [(1993) J. Comp. Neurol., 338:612-627] in pupae. We found these CCAP-IR neurons, however, also to be present in larvae. The mIgG-like- and CCAP-IR neurons included the NS-L pair of the subesophageal maxillary neuromere, which projected anteriorly to the corpora cardiaca, and the NS-L of the labial neuromere whose axons projected out the dorsal nerve of the next posterior ganglion. The mIgG-like and CCAP-IR NS-Ls were also observed throughout the three thoracic ganglia, and all shared strikingly similar structural features. These cells exited out the dorsal nerve of the next posterior ganglion and eventually projected to the neurohemal release sites of the perivisceral organs. These neurons appear to be the homologues of the abdominal CCAP-IR NS-Ls, neurons that in the adult switch their neurotransmitter and release the neuropeptide bursicon. Our description of the distribution and developmental expression of this novel mIgG-like immunoreactivity may provide new insights into the regulation of neurotransmitter plasticity and/or recognition-signaling events involved in the embryonic and postembryonic assembly of the nervous system.

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Section: Ventral Nerve Cord Of M Sextamentioning

confidence: 99%

Novel mouse IgG‐like immunoreactivity expressed by neurons in the moth Manduca sexta: Developmental regulation and colocalization with crustacean cardioactive peptide

Klukas

Brelje

Mesce

1996

Microsc. Res. Tech.

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“…Tumor tissues from colon cancer patients were snap-frozen in liquid nitrogen-cooled isopentane and 4-pm frozen sections were prepared. After drying and fixation in acetone, the sections were rehydrated and stained with FabC215-SEA and SEA followed by antimouse immunoglobulin-biotinylated antibody or rabbit anti-SEA-biotinylated antibody and avidin-biotinylated peroxidase complex (17). The slides were developed in diaminobenzidine (Sigma) and mounted in DPX medium (Sigma).…”

Section: Methodsmentioning

confidence: 99%

Monoclonal antibody-superantigen fusion proteins: tumor-specific agents for T-cell-based tumor therapy.

Dohlsten

Abrahmsén

Björk

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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107

108

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The bacterial superantigen staphylococcal enterotoxin A (SEA) is an extremely potent activator of T lymphocytes when presented on major histocompatibility complex (MHC) The therapeutic use of naked monoclonal antibodies (mAbs) in human epithelial cancer has met with limited success (1). The amount of mAb found to accumulate in the tumor is generally low, tumor cells display extensive heterogeneity in antigen expression, and the effector mechanisms responsible for tumor-cell destruction are insufficient (1, 2). Recent progress in the molecular understanding of T-cell recognition has provided insight into the role of T cells in the response against allogeneic and neoplastic tissues. The vigorous and destructive response against an allograft involves a high frequency of T lymphocytes that, via the T-cell receptor (TCR), recognizes peptide antigens presented in the context of major histocompatibility complex (MHC) molecules. In contrast, the frequency of T cells responding to a tumor is generally low and insufficient to interfere with tumor growth. This suggests that an attractive approach for immunotherapy would be to target a high frequency of T cells to the tumor area. We have therefore developed an antibody-based therapy that provides tumor cells with superantigenicity. Superantigens (SAgs) are a family of bacterial and viral proteins that activate a high frequency of T cells to cytokine release and cell-mediated cytotoxicity (3)(4)(5). Bacterial SAgs bind with high affinity to MHC class II molecules and subsequently interact with T cells expressing particular sequences in the variable (V) region of the TCR , / chain (TCR V(3) (3)(4)(5) MATERIALS AND METHODSProtein Reagents. Recombinant SEA was expressed in E. coli and purified to homogeneity. The hybridomas secreting the C215 (IgG2a) and C242 (IgGl) mAbs, reacting with human colon cancer, were produced at Pharmacia (8, 9). E. coli HB101 was used as host during the DNA construction work (10).

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“…Animals were sacrificed at various times after C215Fab-SEA treatment and pieces of lung tissue containing B16-C215 tumors were snap-frozen in isopentane cooled in liquid nitrogen. Frozen sections (10 ,Im) were prepared, and after drying and fixation in ice-cold acetone, the sections were sequentially stained with the indicated mAbs, followed by biotinylated goat anti-rat IgG and avidin-biotinylated alkaline phosphatase complex (8). The slides were developed in the alkaline phosphatase substrate S-5100 (Vector Laboratories) and mounted in DPX medium (Kebo, Stockholm).…”

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confidence: 99%

Antibody-targeted superantigens are potent inducers of tumor-infiltrating T lymphocytes in vivo.

Dohlsten¹,

Hansson²,

Ohlsson³

et al. 1995

Proc. Natl. Acad. Sci. U.S.A.

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Recruitment of antigen-specific tumor-infiltrating lymphocytes (TILs) is a major goal for immunotherapy of malignant tumours. We now describe that T-cell-activating superantigens targeted to a tumor by monoclonal antibodies induced large numbers of pseudospecific TILs and eradication of micrometastases. As a model for tumor micrometastases, syngeneic B16 melanoma cells transfected with the human colon carcinoma antigen C215 were injected intravenously into C57BL/6 mice and therapy with an anti-C215 Fab fragment-staphylococcal enterotoxin A (C215Fab-SEA) fusion protein reacting with the C215 antigen was initiated when visible lung metastases were established. More than 90% reduction of the number of lung metastases was observed when mice carrying 5-day-old established lung metastases were treated with C215Fab-SEA. The antitumor effect of C215Fab-SEA was shown to be T-cell-dependent since no therapeutic effect was seen in T-cell-deficient nude mice. Depletion of T-cell subsets by injection of monoclonal antibody demonstrated that CD8+ cells were the most prominent effector cells although some contribution from CD4+ cells was also noted. C215Fab-SEA treatment induced massive tumor infiltration of CD4+ and CD8+ T cells, while only scattered T cells were observed in untreated tumors. SEA treatment alone induced a slight general inflammatory response in the lung parenchyme, but no specific accumulation of T cells was seen in the tumor. TILs induced by C215Fab-SEA were mainly CD8+ but a substantial number of CD4+ cells were also present. Immunohistochemical analysis showed strong production of the tumoricidal cytokines tumor necrosis factor alpha and interferon gamma in the tumor. Thus, the C215Fab-SEA fusion protein targets effector T lymphocytes to established tumors in vivo and provokes a strong local antitumor immune response.

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Use of a monoclonal rat anti-mouse Ig light chain (RAMOL-1) antibody reduces background binding in immunohistochemical and fluorescent antibody analysis.

Cited by 14 publications

References 0 publications

Novel mouse IgG‐like immunoreactivity expressed by neurons in the moth Manduca sexta: Developmental regulation and colocalization with crustacean cardioactive peptide

Novel mouse IgG‐like immunoreactivity expressed by neurons in the moth Manduca sexta: Developmental regulation and colocalization with crustacean cardioactive peptide

Monoclonal antibody-superantigen fusion proteins: tumor-specific agents for T-cell-based tumor therapy.

Antibody-targeted superantigens are potent inducers of tumor-infiltrating T lymphocytes in vivo.

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