Generation of procoagulant activity by mononuclear phagocytes: Optimisation of an in vitro assay

Morrice, Lora M.; McIntosh, L. C.; Webster, Laura M.; Thomson, Angus W.

doi:10.1016/0022-1759(85)90151-6

Cited by 7 publications

(5 citation statements)

References 29 publications

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“…In pig artery patch recipients, pro-inflammatory CD14 + CD11c + monocytes were significantly increased with IS, together with significantly higher percentages of activated monocytes and DC and significant upregulation of TF expression (Figure 3). The pro-coagulant properties of innate immune cells are well characterized (51, 52). Activated monocytes and dendritic cells are known to upregulate TF expression, and to mediate inflammation and coagulation (53-55).…”

Section: Discussionmentioning

confidence: 99%

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Systemic inflammation in xenograft recipients precedes activation of coagulation

Ezzelarab

Ekser

Azimzadeh

et al. 2014

Xenotransplantation

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114

173

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Background Dysregulation of coagulation is considered a major barrier against successful pig organ xenotransplantation in nonhuman primates. Inflammation is known to promote activation of coagulation. The role of pro-inflammatory factors as well as the relationship between inflammation and activation of coagulation in xenograft recipients is poorly understood. Methods Baboons received kidney (n=3), heart (n=4) or artery patch (n=8) xenografts from α1,3-galactosyltransferase gene-knockout (GTKO) pigs or GTKO pigs additionally transgenic for human complement regulatory protein CD46 (GTKO/CD46). Immunosuppression (IS) was based on either CTLA4-Ig or anti-CD154 costimulation blockade. Three artery patch recipients did not receive IS. Pro-inflammatory cytokines, chemokines and coagulation parameters were evaluated in the circulation after transplantation. In artery patch recipients, monocytes and dendritic cells (DC) were monitored in peripheral blood. Expression of tissue factor (TF) and CD40 on monocytes and DC were assessed by flow cytometry. C-reactive protein (C-RP) levels in the blood and C-RP deposition in xenografts as well as native organs were evaluated. Baboon and pig C-RP mRNA in heart and kidney xenografts were evaluated. Results In heart and kidney xenograft recipients, the levels of INFγ, TNF-α, IL-12 and IL-8 were not significantly higher after transplantation. However, MCP-1 and IL-6 levels were significantly higher after transplantation, particularly in kidney recipients. Elevated C-RP levels preceded activation of coagulation in heart and kidney recipients, where high levels of C-RP were maintained until the time of euthanasia in both heart and kidney recipients. In artery patch recipients, INFγ, TNF-α, IL-12, IL-8 and MCP-1 were elevated with no IS, while IL-6 was not. With IS, INFγ, TNF-α, IL-12, IL-8 and MCP-1 were reduced, but IL-6 was elevated. Elevated IL-6 levels were observed as early as 2 weeks in artery patch recipients. While IS was associated with reduced thrombin activation, fibrinogen and C-RP levels were increased when IS was given. There was a significant positive-correlation between C-RP, IL-6, and fibrinogen levels. Additionally, absolute numbers of monocytes were significantly increased when IS was given, but not without IS. This was associated with increased CD40 and TF expression on CD14+ monocytes and lineageneg CD11c+ DC, with increased differentiation of the pro-inflammatory CD14+ CD11c+ monocyte population. At the time of euthanasia, C-RP deposition in kidney and heart xenografts, C-RP positive cells in artery patch xenograft and native lungs were detected. Finally, high levels of both pig and baboon C-RP mRNA were detected in heart and kidney xenografts. Conclusions Inflammatory responses precede activation of coagulation after organ xenotransplantation. Early upregulation of C-RP and IL-6 levels may amplify activation of coagulation through upregulation of TF on innate immune cells. Prevention of systemic inflammation in xenograft recipients (SIXR) may be required ...

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Section: Discussionmentioning

confidence: 99%

“…). The procoagulant properties of innate immune cells are well characterized . Activated monocytes and dendritic cells are known to upregulate TF expression and to mediate inflammation and coagulation .…”

Section: Discussionmentioning

confidence: 99%

Systemic inflammation in xenograft recipients precedes activation of coagulation

Ezzelarab

Ekser

Azimzadeh

et al. 2014

Xenotransplantation

Self Cite

114

173

View full text Add to dashboard Cite

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“…The procoagulant ac tivity of the cells was then measured in an automatic coagulometer (Schnitger and Gross, Amelung GmbH, Lemgo, FRG) by mixing 0.2 ml of cell suspension, 0.1 ml of platelet-poor, citrated human plasma and 0.1 ml CaCL (0.025 mol/l). The plasma was prepared and used following the precautions outlined by Geczy and Meyer [6] and by Morrice et al [10], Each test was conducted in triplicate. A standard thromboplastin calibration curve was constructed for each batch of plasma, using serial dilutions of standardised throm boplastin (Manchester comparative reagent, National UK Refer ence Laboratory, Withington Hospital, Manchester).…”

Section: Leucocyte Procoagulant Activitymentioning

confidence: 99%

“…A standard thromboplastin calibration curve was constructed for each batch of plasma, using serial dilutions of standardised throm boplastin (Manchester comparative reagent, National UK Refer ence Laboratory, Withington Hospital, Manchester). The clotting times were expressed in standard thromboplastin units [10],…”

Section: Leucocyte Procoagulant Activitymentioning

confidence: 99%

Factors Influencing the Enhancement of Delayed-Type Hypersensitivity to Ovalbumin by Cyclosporin A in the Guinea Pig: Possible Role of Suppressor Cells

Aldridge¹,

Thomson²

1986

Int Arch Allergy Immunol

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Enhancement of delayed-type hypersensitivity (DTH) reactions to ovalbumin (OVA) was demonstrated in guinea pigs given a single, high dose of cyclosporin A (CsA) intraperitoneally, 2 days before immunization. Courses of oral CsA, commencing at the time of immunization and lasting until day 4 or 13 also resulted in augmented DTH responses at days 14 and 28, respectively. However, the enhancing protocol (CsA; day 0–4) did not significantly affect circulating anti-OVA antibody titres. The capacity to express increased DTH could be adoptively transferred to naive recipients by systemic injection of pooled spleen and peritoneal exudate cells. Moreover, the expression of augmented responses was inhibited by transfer of cells from normally immunized donors. Although augmentation of DTH was accompanied by increased lymphocyte proliferative responses to OVA in vitro, there was no similar effect on T cell responses to polyclonal mitogens. The data support the view that augmentation of DTH by CsA is attributable to suppressor cell dysfunction, but that this is unlikely to be a non-specific suppressor cell impairment.

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“…It was stored in 2-ml aliquots at -70 °C for more than 2 months. Im- mediately prior to use, the plasma was thawed at 37 °C for 30 min, followed by 30 min at 4°C to allow stabili zation [12].…”

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confidence: 99%

Stimulation of Human Blood Mononuclear Cell Procoagulant Activity by Cyclosporin A

Webster¹,

Thomson²

1986

Int Arch Allergy Immunol

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Using a one-stage assay of plasma recalcifícation time, we have examined the procoagulant activity (PA) of human blood mononuclear cells following incubation for 20 h in the presence of cyclosporin A (CsA; 50–5,000 ng/ml). CsA alone caused a significant, dose-related increase in PA above basal values at 1,000 and 5,000 ng/ml. At these concentrations, it also enhanced the increases in PA produced by concanavalin A, phytohaemagglutinin and bacterial endotoxin. These effects of CsA were totally inhibited by actinomycin D or cycloheximide.

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Generation of procoagulant activity by mononuclear phagocytes: Optimisation of an in vitro assay

Cited by 7 publications

References 29 publications

Systemic inflammation in xenograft recipients precedes activation of coagulation

Systemic inflammation in xenograft recipients precedes activation of coagulation

Factors Influencing the Enhancement of Delayed-Type Hypersensitivity to Ovalbumin by Cyclosporin A in the Guinea Pig: Possible Role of Suppressor Cells

Stimulation of Human Blood Mononuclear Cell Procoagulant Activity by Cyclosporin A

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