Localization of Antigen in Tissue Cells

Coons, Albert H.; Kaplan, Melvin H.

doi:10.1084/jem.91.1.1

Cited by 1,689 publications

(258 citation statements)

References 11 publications

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“…in cold acetone and stained by the indirect fluorescent antibody technique (Coons & Kaplan, 1950), using hyperimmune rabbit anti-herpes simplex type 1 serum (Watson et al 1966) and fluorescein isothiocyanate conjugated to anti-rabbit IgG (Wellcome Reagents Ltd). The sections were examined on a Zeiss Ultraphot II fluorescent microscope using incident illumination.…”

Section: Examination Of Tissue Sections By Immunofluorescent Microscopymentioning

confidence: 99%

The pathogenicity of thymidine kinase-deficient mutants of herpes simplex virus in mice

Field¹,

Wildy²

1978

J. Hyg.

326

148

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SUMMARYThe pathogenicity for mice of two mutants of herpes simplex virus (type 1 and type 2), which fail to induce thymidine kinase, were compared with their respective parent strains. The mutants were much less virulent than the parents following either intracerebral or peripheral inoculation. The replication of the virus at the site ofinoculation and its progression into the nervous system were studied. Following a very large inoculum in the ear, the type 1 mutant was found to establish a latent infection in the cervical dorsal root ganglia. Mice inoculated intracerebrally with small doses of the mutant viruses were solidly immune to challenge with lethal doses of the parent strain.

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Section: Examination Of Tissue Sections By Immunofluorescent Microscopymentioning

confidence: 99%

The pathogenicity of thymidine kinase-deficient mutants of herpes simplex virus in mice

Field¹,

Wildy²

1978

J. Hyg.

326

148

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“…Immunofluorescent Method.--The fluorescent antibody technique as described by Coons and Kaplan (14) was employed with minor modifications (6). One important modification was an initial washing of the tissue sections 30 seconds in buffered saline before fixation.…”

Section: Sacrifice and Morphological Study Ofmentioning

confidence: 99%

Pathogenic Factors in Vascular Lesions of Experimental Serum Sickness

Kniker

Cochrane

1965

The Journal of Experimental Medicine

150

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The present studies suggest that polymorphonuclear leukocytes (PMN's) are essential for the development of cardiovascular lesions in serum sickness. In the absence of PMN's, necrotic vascular lesions were never seen and endothelial proliferation in arteries was inhibited. Zones of fibrinoid deposits did not occur. By contrast, at least two-thirds of the control animals exhibited endothelial proliferation, and half had necrosis of arterial walls, usually with fibrinoid deposits. In arterial lesions that involved the intima and media, the internal elastic lamina was disrupted. This was associated with accumulations of PMN's and was prevented when PMN's were depleted. The observations suggested that the elastic lamina acts as a barrier to the outward spread of inflammation in arteries and that it is an important substrate of PMN action. Although glomerulitis and proteinuria developed in PMN-depleted animals, no conclusions could be drawn concerning the pathogenic role of PMN's in renal lesions, since PMN depletion could not be effected before the onset of immune elimination without influencing the immune response itself. Host complement (ß1C-globulin) was localized along with the antigen and rabbit gamma globulin in glomeruli and arteries showing lesions. In the glomeruli these deposits formed a granular lining along the area of the basement membrane. In arteries the fluorescent amorphous particles were in the intima and media of inflamed vessels. The immune response to BSA and the incidence and severity of cardiovascular and renal lesions were enhanced by the intravenous administration of pooled rabbit antiserum to BSA given 18 hours before BSA antigen and by injecting endotoxin along with the BSA. These additions to the usual procedure of inducing serum sickness did not appear to change the quality of the disease. In normal rabbits, the peak incidence of cardiovascular lesions was early in immune elimination of antigen, at a time when levels of circulating complexes was maximal. Conversely, the severest renal injury was noted several days later, at the completion of immune elimination.

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“…Albert H. Coons showed in the early 1940s that it was possible to use fluorescently tagged antibodies to localize antigens in tissue sections (Coons and Kaplan, 1950;Coons, 1971). Since then a multitude of different immunochemical methods have been developed.…”

Section: Introductionmentioning

confidence: 99%

Strategies for immunohistochemical protein localization using antibodies: What did we learn from neurotransmitter transporters in glial cells and neurons

Danbolt

Zhou

Furness

et al. 2016

Glia

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Immunocytochemistry and Western blotting are still major methods for protein localization, but they rely on the specificity of the antibodies. Validation of antibody specificity remains challenging mostly because ideal negative controls are often unavailable. Further, immunochemical labeling patterns are also influenced by a number of other factors such as postmortem changes, fixation procedures and blocking agents as well as the general assay conditions (e.g., buffers, temperature, etc.). Western blotting similarly depends on tissue collection and sample preparation as well as the electrophoretic separation, transfer to blotting membranes and the immunochemical probing of immobilized molecules. Publication of inaccurate information on protein distribution has downstream consequences for other researchers because the interpretation of physiological and pharmacological observations depends on information on where ion channels, receptors, enzymes or transporters are located. Despite numerous reports, some of which are strongly worded, erroneous localization data are being published. Here we describe the extent of the problem and illustrate the nature of the pitfalls with examples from studies of neurotransmitter transporters. We explain the importance of supplementing immunochemical observations with other measurements (e.g., mRNA levels and distribution, protein activity, mass spectrometry, electrophysiological recordings, etc.) and why quantitative considerations are integral parts of the quality control. Further, we propose a practical strategy for researchers who plan to embark on a localization study. We also share our thoughts about guidelines for quality control. GLIA 2016;64:2045–2064

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Localization of Antigen in Tissue Cells

Cited by 1,689 publications

References 11 publications

The pathogenicity of thymidine kinase-deficient mutants of herpes simplex virus in mice

The pathogenicity of thymidine kinase-deficient mutants of herpes simplex virus in mice

Pathogenic Factors in Vascular Lesions of Experimental Serum Sickness

Strategies for immunohistochemical protein localization using antibodies: What did we learn from neurotransmitter transporters in glial cells and neurons

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