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Evaluation of Immunostimulatory Potential of Branded and US-Generic Enoxaparins in an In Vitro Human Immune System Model.

Luna E, Agrawal P, Mehta R, Vernhes C, Viskov C, Amiral J, Warren WL, Drake DR - Clin. Appl. Thromb. Hemost. (2014)

Bottom Line: Low-molecular-weight heparins (LMWHs) have several positive therapeutic effects and can also form immunostimulatory complexes with plasma proteins, such as platelet factor 4 (PF4).Production of tissue factor pathway inhibitor (TFPI), a physiologic heparin-induced inhibitor of tissue factor-induced coagulation that was used as a functional readout of biological activity of enoxaparins in these assays, was heightened in the presence of branded enoxaparin complexes, but its levels were variable in cultures treated with complexes containing US-generic enoxaparins.Analytical analyses suggest that the heightened immunostimulatory potential of some of the US-generic enoxaparin product lots could be tied to their capacity to form ultra-large and/or more stable complexes with PF4 than the other LMWHs included in this study.

View Article: PubMed Central - PubMed

Affiliation: Sanofi Pasteur, VaxDesign Campus, Orlando, FL, USA.

No MeSH data available.


Related in: MedlinePlus

Branded and generic enoxaparins have different capacities to activate APCs in the MIMIC® PTE construct. MIMIC® PTE assays were treated for 48 hours with the indicated concentrations of PF4 complexes prepared with (A) UFH, (B) Sanofi enoxaparin, (C) Sandoz enoxaparin, or (D) Amphastar enoxaparin. Thereafter, the cells were harvested and examined for CD83 expression on the CD14+HLA-DR+ DC population by multiparameter flow cytometry. The data plotted show the means ± SD for 20 donors examined in 5 distinct-blinded experiments. Ag indicates antigen; APC, antigen-presenting cell; DC, dendritic cell; MIMIC® PTE, Modular IMmune In vitro Construct system-peripheral tissue equivalent; PF4, platelet factor 4; SD, standard deviation; UFH, unfractionated heparin.
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fig3-1076029614562037: Branded and generic enoxaparins have different capacities to activate APCs in the MIMIC® PTE construct. MIMIC® PTE assays were treated for 48 hours with the indicated concentrations of PF4 complexes prepared with (A) UFH, (B) Sanofi enoxaparin, (C) Sandoz enoxaparin, or (D) Amphastar enoxaparin. Thereafter, the cells were harvested and examined for CD83 expression on the CD14+HLA-DR+ DC population by multiparameter flow cytometry. The data plotted show the means ± SD for 20 donors examined in 5 distinct-blinded experiments. Ag indicates antigen; APC, antigen-presenting cell; DC, dendritic cell; MIMIC® PTE, Modular IMmune In vitro Construct system-peripheral tissue equivalent; PF4, platelet factor 4; SD, standard deviation; UFH, unfractionated heparin.

Mentions: Consistent with the pilot study results (Figure 2), PF4–UFH complexes prepared for this phase of work triggered a significant dose-dependent reduction in cell numbers (data not shown). Likewise, PF4–UFH complexes induced high levels of APC activation in a dose-dependent manner; Figure 3A shows the peak response was 40.2% ± 9.5%; ANOVA, P ≤ .0001 at the highest treatment dose. Regarding the evaluation of variant enoxaparins, no reduction in cell viability was observed when MIMIC® PTE cultures were treated with PF4 complexes prepared with either the branded or the generic enoxaparins (data not shown). However, although the APC activation profile for PF4–branded enoxaparin was consistently low across the 2 test lots, innate response profiles triggered by PF4 complexes prepared with generic enoxaparins (Sandoz and Amphastar) were significantly variable between the 2 lots of each product (Figure 3B, C, and D; ANOVA, P ≤ .0001 at the highest treatment dose). Also, statistical analyses revealed that the PF4–enoxaparin complexes prepared with Lot 1 Sandoz and Amphastar enoxaparins generated significantly different innate responses than complexes prepared with either lot of the branded product (ANOVA, P ≤ .0001 at the highest treatment dose). See Table 3 for all statistical comparisons generated for this data set.


Evaluation of Immunostimulatory Potential of Branded and US-Generic Enoxaparins in an In Vitro Human Immune System Model.

Luna E, Agrawal P, Mehta R, Vernhes C, Viskov C, Amiral J, Warren WL, Drake DR - Clin. Appl. Thromb. Hemost. (2014)

Branded and generic enoxaparins have different capacities to activate APCs in the MIMIC® PTE construct. MIMIC® PTE assays were treated for 48 hours with the indicated concentrations of PF4 complexes prepared with (A) UFH, (B) Sanofi enoxaparin, (C) Sandoz enoxaparin, or (D) Amphastar enoxaparin. Thereafter, the cells were harvested and examined for CD83 expression on the CD14+HLA-DR+ DC population by multiparameter flow cytometry. The data plotted show the means ± SD for 20 donors examined in 5 distinct-blinded experiments. Ag indicates antigen; APC, antigen-presenting cell; DC, dendritic cell; MIMIC® PTE, Modular IMmune In vitro Construct system-peripheral tissue equivalent; PF4, platelet factor 4; SD, standard deviation; UFH, unfractionated heparin.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4401814&req=5

fig3-1076029614562037: Branded and generic enoxaparins have different capacities to activate APCs in the MIMIC® PTE construct. MIMIC® PTE assays were treated for 48 hours with the indicated concentrations of PF4 complexes prepared with (A) UFH, (B) Sanofi enoxaparin, (C) Sandoz enoxaparin, or (D) Amphastar enoxaparin. Thereafter, the cells were harvested and examined for CD83 expression on the CD14+HLA-DR+ DC population by multiparameter flow cytometry. The data plotted show the means ± SD for 20 donors examined in 5 distinct-blinded experiments. Ag indicates antigen; APC, antigen-presenting cell; DC, dendritic cell; MIMIC® PTE, Modular IMmune In vitro Construct system-peripheral tissue equivalent; PF4, platelet factor 4; SD, standard deviation; UFH, unfractionated heparin.
Mentions: Consistent with the pilot study results (Figure 2), PF4–UFH complexes prepared for this phase of work triggered a significant dose-dependent reduction in cell numbers (data not shown). Likewise, PF4–UFH complexes induced high levels of APC activation in a dose-dependent manner; Figure 3A shows the peak response was 40.2% ± 9.5%; ANOVA, P ≤ .0001 at the highest treatment dose. Regarding the evaluation of variant enoxaparins, no reduction in cell viability was observed when MIMIC® PTE cultures were treated with PF4 complexes prepared with either the branded or the generic enoxaparins (data not shown). However, although the APC activation profile for PF4–branded enoxaparin was consistently low across the 2 test lots, innate response profiles triggered by PF4 complexes prepared with generic enoxaparins (Sandoz and Amphastar) were significantly variable between the 2 lots of each product (Figure 3B, C, and D; ANOVA, P ≤ .0001 at the highest treatment dose). Also, statistical analyses revealed that the PF4–enoxaparin complexes prepared with Lot 1 Sandoz and Amphastar enoxaparins generated significantly different innate responses than complexes prepared with either lot of the branded product (ANOVA, P ≤ .0001 at the highest treatment dose). See Table 3 for all statistical comparisons generated for this data set.

Bottom Line: Low-molecular-weight heparins (LMWHs) have several positive therapeutic effects and can also form immunostimulatory complexes with plasma proteins, such as platelet factor 4 (PF4).Production of tissue factor pathway inhibitor (TFPI), a physiologic heparin-induced inhibitor of tissue factor-induced coagulation that was used as a functional readout of biological activity of enoxaparins in these assays, was heightened in the presence of branded enoxaparin complexes, but its levels were variable in cultures treated with complexes containing US-generic enoxaparins.Analytical analyses suggest that the heightened immunostimulatory potential of some of the US-generic enoxaparin product lots could be tied to their capacity to form ultra-large and/or more stable complexes with PF4 than the other LMWHs included in this study.

View Article: PubMed Central - PubMed

Affiliation: Sanofi Pasteur, VaxDesign Campus, Orlando, FL, USA.

No MeSH data available.


Related in: MedlinePlus