The identification of eosinophil colonies in soft-agar cultures by differential staining for peroxidase.

Zucker‐Franklin, Dorothea; Grusky, George

doi:10.1177/24.12.63511

Cited by 51 publications

(20 citation statements)

References 8 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Based on their characteristic morphologies, the polymorphonuclear leukocytes in brains of MRL-GKO mice appeared to represent eosinophilic granulocytes. Staining for cyanide-resistant peroxidase activity (which specifically detects eosinophilic granulocytes [72]) confirmed the identity of these cells (Fig. 6D).…”

Section: Resultsmentioning

confidence: 71%

“…Treatment of brain sections and immunohistochemistry were done as described previously (15). For the specific detection of eosinophils, we took advantage of the fact that the peroxidase activity of eosinophils is highly resistant to cyanide treatment, whereas peroxidase of other granulocytes, mast cells, and endothelial cells can be poisoned by cyanide (72). Frozen sections of mouse brains were fixed with a 1:1 (vol/vol) mixture of methanol/acetone, air dried, and incubated for 20 min with cyanide-containing diaminobenzidine substrate solution (5 l of 30% H 2 O 2 and 5 drops of 100 mM KCN for 5 ml of diaminobenzidine substrate solution) at room temperature.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

CD8 T Cells Require Gamma Interferon To Clear Borna Disease Virus from the Brain and Prevent Immune System-Mediated Neuronal Damage

Hausmann¹,

Pagenstecher

Baur³

et al. 2005

J Virol

View full text Add to dashboard Cite

Borna disease virus (BDV) frequently causes meningoencephalitis and fatal neurological disease in young but not old mice of strain MRL. Disease does not result from the virus-induced destruction of infected neurons. Rather, it is mediated by H-2 k -restricted antiviral CD8 T cells that recognize a peptide derived from the BDV nucleoprotein N. Persistent BDV infection in mice is not spontaneously cleared. We report here that N-specific vaccination can protect wild-type MRL mice but not mutant MRL mice lacking gamma interferon (IFN-␥) from persistent infection with BDV. Furthermore, we observed a significant degree of resistance of old MRL mice to persistent BDV infection that depended on the presence of CD8 T cells. We found that virus initially infected hippocampal neurons around 2 weeks after intracerebral infection but was eventually cleared in most wild-type MRL mice. Unexpectedly, young as well as old IFN-␥-deficient MRL mice were completely susceptible to infection with BDV. Moreover, neurons in the CA1 region of the hippocampus were severely damaged in most diseased IFN-␥-deficient mice but not in wild-type mice. Furthermore, large numbers of eosinophils were present in the inflamed brains of IFN-␥-deficient mice but not in those of wild-type mice, presumably because of increased intracerebral synthesis of interleukin-13 and the chemokines CCL1 and CCL11, which can attract eosinophils. These results demonstrate that IFN-␥ plays a central role in host resistance against infection of the central nervous system with BDV and in clearance of BDV from neurons. They further indicate that IFN-␥ may function as a neuroprotective factor that can limit the loss of neurons in the course of antiviral immune responses in the brain.

show abstract

Section: Resultsmentioning

confidence: 71%

Section: Methodsmentioning

confidence: 99%

CD8 T Cells Require Gamma Interferon To Clear Borna Disease Virus from the Brain and Prevent Immune System-Mediated Neuronal Damage

Hausmann¹,

Pagenstecher

Baur³

et al. 2005

J Virol

View full text Add to dashboard Cite

show abstract

“…Sections were cut on a cryostat, mounted on glass slides precoated with aminopropyltriethoxysilane (Sigma Chemical), and fixed in chloroform-acetone (Merck, Rio de Janeiro, Brazil) vol/vol for 10 min at room temperature. A histochemical method for cyanideresistant EPO activity employing diaminobenzidine (Sigma Chemical), H 2O2, and potassium cyanide (Sigma Chemical) was used to stain eosinophils, as previously described (25,44,52). Counterstaining with hematoxylin was employed to reveal cellular nuclei, and counts of positive cells in the airway walls were performed as previously described for morphometric studies (44).…”

Section: Methodsmentioning

confidence: 99%

Neurokinins and inflammatory cell iNOS expression in guinea pigs with chronic allergic airway inflammation

Prado¹,

Leick-Maldonado²,

Arata³

et al. 2005

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

In the present study we evaluated the role of neurokinins in the modulation of inducible nitric oxide synthase (iNOS) inflammatory cell expression in guinea pigs with chronic allergic airway inflammation. In addition, we studied the acute effects of nitric oxide inhibition on this response. Animals were anesthetized and pretreated with capsaicin (50 mg/kg sc) or vehicle 10 days before receiving aerosolized ovalbumin or normal saline twice weekly for 4 wk. Animals were then anesthetized, mechanically ventilated, given normal saline or N G -nitro-L-arginine methyl ester (L-NAME, 50 mg/kg ic), and challenged with ovalbumin. Prechallenge exhaled NO increased in ovalbumin-exposed guinea pigs (P Ͻ 0.05 compared with controls), and capsaicin reduced this response (P Ͻ 0.001). Compared with animals inhaled with normal saline, ovalbumin-exposed animals presented increases in respiratory system resistance and elastance and numbers of total mononuclear cells and eosinophils, including those expressing iNOS (P Ͻ 0.001). Capsaicin reduced all these responses (P Ͻ 0.05) except for iNOS expression in eosinophils. Treatment with L-NAME increased postantigen challenge elastance and restored both resistance and elastance previously attenuated by capsaicin treatment. Isolated L-NAME administration also reduced total eosinophils and mononuclear cells, as well as those cells expressing iNOS (P Ͻ 0.05 compared with ovalbumin alone). Because L-NAME treatment restored lung mechanical alterations previously attenuated by capsaicin, NO and neurokinins may interact in controlling airway tone. In this experimental model, NO and neurokinins modulate eosinophil and lymphocyte infiltration in the airways. nitric oxide; inducible nitric oxide synthase; asthma; lymphocytes; eosinophils NEUROKININS such as substance P and neurokinin A are involved in the modulation of several aspects of inflammatory responses in the lungs. These peptides induce airway smooth muscle contraction, peribronchial edema, and airway mucous secretion and are considered mediators of the excitatory nonadrenergicnoncholinergic response (43). It has been previously shown that substance P and neurokinin A play significant roles in priming eosinophils and in lymphocyte recruitment in allergic lung inflammation (33,43,44). In guinea pigs and other mammals, pretreatment with capsaicin has been used as a research tool to elucidate the physiological effects of neurokinins (18,29). It has been previously show that administration of high doses of capsaicin reduces tissue concentrations of substance P and other neurokinins (28,29).Nitric oxide (NO), a gaseous molecule that is generated during the conversion of the amino acid L-arginine to Lcitrulline by NO synthase (NOS), is considered a mediator of the inhibitory nonadrenergic-noncholinergic response (3,26,32). The role of NO in the modulation of allergic inflammation and bronchial hyperresponsiveness is currently unclear. There is some evidence that NO has both inflammatory and antiinflammatory effects in experimental models of ai...

show abstract

“…One eye was frozen for later RNA preparation or microdissection, and the other eye was used for histological analysis. Eye sections were stained by conventional H&E or by an eosinophil stain that highlights cyanide-resistant eosinophil peroxidase (19,20). Briefly, eye frozen sections were fixed in 1% Formalin in acetone for 30 s, then stained for 10 min in PBS containing 0.4 mg/ml NaCN, 3 l/ml 3% hydrogen peroxide, and 0.75 mg/ml diaminobenzidine (Sigma-Aldrich).…”

Section: Adoptive Transfer Of Th1 and Th2 Cellsmentioning

confidence: 99%

Inflammatory Mediators in Uveitis: Differential Induction of Cytokines and Chemokines in Th1- Versus Th2-Mediated Ocular Inflammation

Foxman

Zhang

Hurst³

et al. 2002

The Journal of Immunology

135

116

View full text Add to dashboard Cite

Ocular inflammation leads to vision loss through the destruction and scarring of delicate tissues along the visual axis. To identify inflammatory mediators involved in this process, we used real time RT-PCR to quantify the expression of mRNA transcripts of 34 cytokines, 26 chemokines, and 14 chemokine receptors at certain time points during T cell-mediated ocular inflammation. We induced disease by adoptive transfer of Ag-specific Th1 or Th2 cells into recipients expressing the target Ag in their eyes. We also compared the mediator expression patterns seen in adoptive transfer-induced inflammation with that seen in mouse eyes developing experimental autoimmune uveoretinitis. In addition, we used laser capture microdissection to examine chemokine mRNA production by both retinal pigment epithelium cells and infiltrating leukocytes in inflamed eyes. Major findings included the following: 1) Three patterns of expression of the inflammation-related molecules were seen in recipients of adoptively transferred Th cells: preferential expression in Th1 recipients, or in Th2 recipients, or similar expression in both recipient groups. 2) In experimental autoimmune uveoretinitis, the inflammatory mediator expression pattern largely paralleled that seen in Th1-induced disease. 3) Both retinal pigment epithelium and infiltrating leukocytes expressed chemokine transcripts in distinct, but overlapping patterns in inflamed eyes. 4) Interestingly, trancripts of multiple cytokines, chemokines, and chemokine receptors were constitutively expressed in high levels in mouse eyes. Seven of these molecules have not been previously associated with the eye. These data underscore the multiplicity of mediators that participate in the pathogenesis of eye inflammation and point to upstream cytokines as potential therapeutic targets.

show abstract

The identification of eosinophil colonies in soft-agar cultures by differential staining for peroxidase.

Cited by 51 publications

References 8 publications

CD8 T Cells Require Gamma Interferon To Clear Borna Disease Virus from the Brain and Prevent Immune System-Mediated Neuronal Damage

CD8 T Cells Require Gamma Interferon To Clear Borna Disease Virus from the Brain and Prevent Immune System-Mediated Neuronal Damage

Neurokinins and inflammatory cell iNOS expression in guinea pigs with chronic allergic airway inflammation

Inflammatory Mediators in Uveitis: Differential Induction of Cytokines and Chemokines in Th1- Versus Th2-Mediated Ocular Inflammation

Contact Info

Product

Resources

About