Rodent Immunohistochemistry

Ward, Jerrold M.; Rehg, Jerold E.

doi:10.1177/0300985813503571

Cited by 52 publications

(27 citation statements)

References 20 publications

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“…There are several tissue handling (preanalytical) factors that can significantly affect the quality and specificity of immunostaining and its analysis. These have been discussed earlier sections of the paper and in several reviews [20,52,[60][61][62][63]. Similarly, there are many factors during the staining procedure that itself can also influence the results.…”

Section: Immunostainingmentioning

confidence: 90%

Histopathologic Evaluation and Scoring of Viral Lung Infection

Meyerholz

Beck

2019

Methods in Molecular Biology

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Emergent coronaviruses such as MERS-CoV and SARS-CoV can cause significant morbidity and mortality in infected individuals. Lung infection is a common clinical feature and contributes to disease severity as well as viral transmission. Animal models are often required to study viral infections and therapies, especially during an initial outbreak. Histopathology studies allow for identification of lesions and affected cell types to better understand viral pathogenesis and clarify effective therapies. Use of immunostaining allows detection of presumed viral receptors and viral tropism for cells can be evaluated to correlate with lesions. In the lung, lesions and immunostaining can be qualitatively described to define the cell types, microanatomic location, and type of changes seen. These features are important and necessary, but this approach can have limitations when comparing treatment groups. Semiquantitative and quantitative tissue scores are more rigorous as these provide the ability to statistically compare groups and increase the reproducibility and rigor of the study. This review describes principles, approaches, and resources that can be useful to evaluate coronavirus lung infection, focusing on MER-CoV infection as the principal example.

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Section: Immunostainingmentioning

confidence: 90%

Histopathologic Evaluation and Scoring of Viral Lung Infection

Meyerholz

Beck

2019

Methods in Molecular Biology

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“…Histiocytic sarcoma is relatively common in older C57BL/6 mice and has a similar histologic appearance to the lesions in the hu TLR7/8 mice, with infiltrates of round to oval large, monomorphic cells with dark central nuclei and abundant eosinophilic cytoplasm that are Mac-2 and variably F4/80 positive [44], [45], [46], [47]. Less differentiated neoplastic cells may be less likely to stain with F4/80 [48]. The presence of multinucleated giant cells also can be consistent with histiocytic sarcoma [44], [45].…”

Section: Discussionmentioning

confidence: 99%

Humanized TLR7/8 Expression Drives Proliferative Multisystemic Histiocytosis in C57BL/6 Mice

et al. 2014

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A humanized TLR7/TLR8 transgenic mouse line was engineered for studies using TLR7/8 ligands as vaccine adjuvants. The mice developed a spontaneous immune-mediated phenotype prior to six months of age characterized by runting, lethargy, blepharitis, and corneal ulceration. Histological examination revealed a marked, multisystemic histiocytic infiltrate that effaced normal architecture. The histological changes were distinct from those previously reported in mouse models of systemic lupus erythematosus. When the mice were crossed with MyD88−/− mice, which prevented toll-like receptor signaling, the inflammatory phenotype resolved. Illness may be caused by constitutive activation of human TLR7 or TLR8 in the bacterial artificial chromosome positive mice as increased TLR7 and TLR8 expression or activation has previously been implicated in autoimmune disease.

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“…Owing to omission of proper controls, authors often report positive labeling (often with a brown chromogen) of cells and tissues which appear to represent nonspecific background staining 46 . A good example was reported in mouse prostate epithelial cells and connective tissue 47 using an anti-human antibody that was never reported (even by the company selling the antibody) to work in mouse tissues.…”

Section: Some Examples Of Errors Seen In Reported Histopathological Dmentioning

confidence: 99%

Reproducibility of histopathological findings in experimental pathology of the mouse: a sorry tail

2017

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Reproducibility of in vivo research using the mouse as a model organism depends on many factors including experimental design, strain or stock, experimental protocols, and methods of data evaluation. Gross and histopathology are often the endpoints of such research and there is increasing concern about the accuracy and reproducibility of diagnoses in the literature. In order to reproduce histopathological results, the pathology protocol, including necropsy methods and slide preparation, should be followed by interpretation of the slides by a pathologist familiar with reading mouse slides and familiar with the consensus medical nomenclature used in mouse pathology. Likewise, it is important that pathologists are consulted as reviewers of manuscripts where histopathology is a key part of the investigation. The absence of pathology expertise in planning, executing, and reviewing in vivo research using mice leads to questionable pathology-based findings and conclusions from studies, even in high impact journals. We discuss the various aspects of this problem, give some examples from the literature, and suggest solutions.

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Rodent Immunohistochemistry

Cited by 52 publications

References 20 publications

Histopathologic Evaluation and Scoring of Viral Lung Infection

Histopathologic Evaluation and Scoring of Viral Lung Infection

Humanized TLR7/8 Expression Drives Proliferative Multisystemic Histiocytosis in C57BL/6 Mice

Reproducibility of histopathological findings in experimental pathology of the mouse: a sorry tail

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