Detection of a ∼100 kD protein with strong immunoreactivity to antibodies specific for inducible nitric oxide synthase (iNOS) but without NOS activity in neutrophils of patients suffering from sepsis: Results of a preliminary study

Richter, Nadja; Petersson, Ann-Sofi; Wennmalm, Åke; Schimke, Ingolf

doi:10.1007/s000110050517

Cited by 6 publications

(5 citation statements)

References 21 publications

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“…Our study also found a protein band at approximately 100 kDa in several cases. This band has also been reported by other workers who studied NOS2 expression in neutrophils from patients with septic shock (22). It was concluded that further studies were required to elucidate the nature of this protein.…”

Section: Discussionsupporting

confidence: 82%

Type II nitric oxide synthase (NOS2) expression correlates with lymph node status in oral squamous cell carcinoma

Brennan

Palacios-Callender²,

Zaki

et al. 2001

J Oral Pathology Medicine

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In tumour biology, nitric oxide (NO) has a complex array of concentration-dependent actions, including both inhibitory and promoting effects. It is thought that the levels of NO found in many human cancers lead to enhanced angiogenesis and tumour dissemination. In the current study, we assessed the immunohistochemical expression of the enzyme type II nitric oxide synthase (NOS2) in 41 cases of oral squamous cell carcinoma and correlated the findings with lymph node status. A significant relationship was found between NOS2 expression and lymph node metastasis (P<0.0002). Furthermore, lymph node metastasis correlated with the degree and intensity of staining seen (P<0.001). No correlation was found between the size of the primary tumour, degree of tumour differentiation or smoking status and NOS2 staining. Western blotting confirmed NOS2 protein expression in select cases. As with many other human tumours, NOS2 is not a ubiquitous finding in oral cancer. Its expression may be of value in assessing lymph node status prior to surgery, and it represents a target for possible therapeutic manipulation.

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Section: Discussionsupporting

confidence: 82%

Type II nitric oxide synthase (NOS2) expression correlates with lymph node status in oral squamous cell carcinoma

Brennan

Palacios-Callender²,

Zaki

et al. 2001

J Oral Pathology Medicine

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“…Inactivated resting rodent neutrophils and macrophages have very low or nondetectable expression of iNOS2, and expression is greatly enhanced upon activation (Nair et al 2003;McCafferty et al 1999;Richter et al 1999).…”

Section: Eosinophil Antigens: Major Basic Proteinmentioning

confidence: 99%

“…Inducible NO synthase 2 (iNOS2) is a cytoplasmic biomarker of activated neutrophils and macrophages activated by CD4Th1 proinflammatory cytokines IFN-gamma and TNF-alpha (Figure 34). Inactivated resting rodent neutrophils and macrophages have very low or nondetectable expression of iNOS2, and expression is greatly enhanced upon activation (Nair et al 2003; McCafferty et al 1999; Richter et al 1999).…”

Section: Tissue Histiocyte (Macrophage) Antigens: Cd68 Cd163 F4/80 Iba1 Lysozyme Mac2 Mac3 S100p Arginase 1 Inos2 Ym1mentioning

confidence: 99%

“…A major differentiating factor between these two host reactions is whether the neutrophil is in an activated state. Unlike resting neutrophils, activated neutrophils show strong cytoplasmic expression of iNOS2 (Richter et al 1999; McCaffery et al 1999; Morton et al 2008). Figures 80 and 81, correlate resting neutrophils with weak low levels of iNOS2 and activated neutrophils with strong high levels of iNOS2 (Figures 82 and 83).…”

Section: Examples Of the Use Of Ihc In Ffpe Sections To Evaluate The Mouse Hematopoietic And Lymphoid Systemsmentioning

confidence: 99%

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The Utility of Immunohistochemistry for the Identification of Hematopoietic and Lymphoid Cells in Normal Tissues and Interpretation of Proliferative and Inflammatory Lesions of Mice and Rats

2012

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Expression of antigens in cells and tissues can be readily studied immunohistochemically with the use of antibodies. A panel of antibodies to cellspecific markers can be used to diagnose lesions, including tumors, in the hematopoietic and lymphoid systems. This review discusses the use of readily available antibodies and procedures to identify antigens expressed in normal tissues and in proliferative and inflammatory lesions in formalin-fixed, paraffin-embedded (FFPE) murine specimens.

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“…This corroborates the findings of Wheeler et al, who reported that deoxycholate was superior to other detergents for the extraction of neutrophil NOS II, which they found to be tightly membrane bound to >90% (13). Thus, failure to demonstrate a 130 kD protein corresponding to NOS II in whole cell lysates of neutrophils (7, 8, 20, 21) may be explained both by insufficient proteinase inhibition and by insufficient release of the membrane‐bound enzyme.…”

Section: Discussionmentioning

confidence: 99%

Inducible nitric oxide synthase (NOS II) is constitutive in human neutrophils

Cedergren

Follin

Forslund

et al. 2003

APMIS

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The objective was to study the expression of inducible nitric oxide synthase (NOS II) in and NO production by human blood neutrophils and in in vivo exudated neutrophils. Cellular expression of NOS II was evaluated by flow cytometry in whole blood, in isolated blood neutrophils, and in neutrophils obtained by exudation in vivo into skin chambers. Neutrophil NOS II was also demonstrated by Western blotting. Uptake of 3H-labelled L-arginine was studied in vitro and NOS activity measured in a whole cell assay by the conversion of 3H-arginine to 3H-citrulline. In contrast to unseparated blood cells, NOS II was demonstrable both in isolated blood neutrophils and exudated cells. The failure to detect NOS II by flow cytometry in whole blood cells thus proved to be due to the quenching effect of hemoglobin. Western blotting revealed a 130 kD band corresponding to NOS II in isolated blood neutrophils, but detection was dependent on diisopropylfluorophosphate for proteinase inhibition. L-arginine was taken up by neutrophils, but enzymatic activity could not be demonstrated. We conclude that human neutrophils constitutively express NOS II, but that its demonstration by FITC-labelling is inhibited by hemoglobin-mediated quenching in whole blood samples.

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Detection of a ∼100 kD protein with strong immunoreactivity to antibodies specific for inducible nitric oxide synthase (iNOS) but without NOS activity in neutrophils of patients suffering from sepsis: Results of a preliminary study

Abstract: The detection of an approximately 100 kD protein with immunoreactivity to antibodies recognizing human iNOS but without NOS activity in neutrophils of patients with sepsis should initiate studies to elucidate the origin and function of this protein.

Cited by 6 publications

References 21 publications

Type II nitric oxide synthase (NOS2) expression correlates with lymph node status in oral squamous cell carcinoma

Type II nitric oxide synthase (NOS2) expression correlates with lymph node status in oral squamous cell carcinoma

The Utility of Immunohistochemistry for the Identification of Hematopoietic and Lymphoid Cells in Normal Tissues and Interpretation of Proliferative and Inflammatory Lesions of Mice and Rats

Inducible nitric oxide synthase (NOS II) is constitutive in human neutrophils

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