Trial Outcomes & Findings for A Pharmacodynamic Study to Evaluate Neutrophil Distribution Kinetics and Function Following Single-Dose RoActemra/Actemra (Tocilizumab) in Healthy Volunteers (NCT NCT01991990)
NCT ID: NCT01991990
Last Updated: 2015-11-11
Results Overview
On Day 4 participants had neutrophils isolated from 100 milliliters (mL) of acid-citrate dextrose (ACD)-anti-coagulated autologous venous blood and labeled in autologous plasma with up to 2.5 megaBecquerel (MBq) 111 Indium (111In)-tropolonate before being reinjected. Participants rested for 45 minutes (min) post-injection to allow for neutrophil equilibrium between the circulating and marginating neutrophil pools. Whole-body profiling was performed in a heavily shielded dedicated whole-body counter with 2 highly sensitive scintillation detectors with the recorded counts corrected for the physical decay of 111In to allow measurement of the effect of TCZ on the normal redistribution pattern of neutrophils and assessment of margination of neutrophils in the presence of TCZ. Distribution of radiolabelled neutrophils on Day 4 (45 min post re-injection) in the blood, liver/spleen and pelvic bone marrow, expressed as percentages of total body counts (TBCs).
COMPLETED
PHASE4
18 participants
Day 4
2015-11-11
Participant Flow
The screening visit was up to 3 weeks before randomization to the first dose of study medication. Out of 23 screened participants; 5 participants discontinued (4=met exclusion criteria; 1=withdrew), 18 participants were included.
Participant milestones
| Measure |
Placebo
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab
Participants received a single dose of intravenous (IV) tocilizumab (TCZ) at a dose of 8 milligrams (mg) per kilogram (kg) body weight infusion over 1 hour on Day 0.
|
|---|---|---|
|
Overall Study
STARTED
|
6
|
12
|
|
Overall Study
COMPLETED
|
6
|
12
|
|
Overall Study
NOT COMPLETED
|
0
|
0
|
Reasons for withdrawal
Withdrawal data not reported
Baseline Characteristics
A Pharmacodynamic Study to Evaluate Neutrophil Distribution Kinetics and Function Following Single-Dose RoActemra/Actemra (Tocilizumab) in Healthy Volunteers
Baseline characteristics by cohort
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab
n=12 Participants
Participants received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0.
|
Total
n=18 Participants
Total of all reporting groups
|
|---|---|---|---|
|
Age, Continuous
|
29.5 years
STANDARD_DEVIATION 11.0 • n=93 Participants
|
34.5 years
STANDARD_DEVIATION 12.6 • n=4 Participants
|
32.7 years
STANDARD_DEVIATION 12.01 • n=27 Participants
|
|
Sex: Female, Male
Female
|
0 Participants
n=93 Participants
|
0 Participants
n=4 Participants
|
0 Participants
n=27 Participants
|
|
Sex: Female, Male
Male
|
6 Participants
n=93 Participants
|
12 Participants
n=4 Participants
|
18 Participants
n=27 Participants
|
PRIMARY outcome
Timeframe: Day 4Population: Safety analysis population: Includes all the participants who received the single dose of randomized study medication. One participant in the polymorphonuclear leukocyte (PMN)-high group was excluded due to external contamination affecting profiling data.
On Day 4 participants had neutrophils isolated from 100 milliliters (mL) of acid-citrate dextrose (ACD)-anti-coagulated autologous venous blood and labeled in autologous plasma with up to 2.5 megaBecquerel (MBq) 111 Indium (111In)-tropolonate before being reinjected. Participants rested for 45 minutes (min) post-injection to allow for neutrophil equilibrium between the circulating and marginating neutrophil pools. Whole-body profiling was performed in a heavily shielded dedicated whole-body counter with 2 highly sensitive scintillation detectors with the recorded counts corrected for the physical decay of 111In to allow measurement of the effect of TCZ on the normal redistribution pattern of neutrophils and assessment of margination of neutrophils in the presence of TCZ. Distribution of radiolabelled neutrophils on Day 4 (45 min post re-injection) in the blood, liver/spleen and pelvic bone marrow, expressed as percentages of total body counts (TBCs).
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=4 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Redistribution Analysis on Day 4 (Neutrophil Nadir)
Blood
|
26.7 percentage of total body count
Standard Error 4.7
|
26.5 percentage of total body count
Standard Error 1.9
|
30.8 percentage of total body count
Standard Error 5
|
|
Neutrophil Redistribution Analysis on Day 4 (Neutrophil Nadir)
Liver/Spleen
|
50.4 percentage of total body count
Standard Error 1.6
|
55 percentage of total body count
Standard Error 3.1
|
52.4 percentage of total body count
Standard Error 1.1
|
|
Neutrophil Redistribution Analysis on Day 4 (Neutrophil Nadir)
Pelvic marrow
|
12.6 percentage of total body count
Standard Error 0.9
|
11.7 percentage of total body count
Standard Error 1
|
10.8 percentage of total body count
Standard Error 1
|
PRIMARY outcome
Timeframe: Day 5Population: Safety analysis population. One participant in the PMN-high group was excluded due to external contamination affecting profiling data.
On Day 4 participants had neutrophils isolated from 100 mL of ACD-anti-coagulated autologous venous blood and labeled with up to 2.5 MBq 111In-tropolonate before being reinjected. Participants rested for 45 min post-injection to allow for neutrophil equilibrium between the circulating and marginating neutrophil pools. Whole-body profiling was performed in a heavily shielded dedicated whole-body counter with 2 highly sensitive scintillation detectors with the recorded counts corrected for the physical decay of 111In to allow measurement of the effect of TCZ on the normal redistribution pattern of neutrophils and assessment of margination of neutrophils in the presence of TCZ. Distribution of radiolabelled neutrophils and peak counts, on Day 5 (24-hours post re-injection) in liver/spleen and pelvic bone marrow were decay corrected and expressed as percentages of Day 4 (45 minutes post re-injection).
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=4 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Redistribution Analysis on Day 5
Liver/spleen peak counts (Day 5)
|
91.0 percentage of Day 4 counts
Standard Error 5.1
|
90.3 percentage of Day 4 counts
Standard Error 5.0
|
105.2 percentage of Day 4 counts
Standard Error 3.3
|
|
Neutrophil Redistribution Analysis on Day 5
Pelvic peak counts (Day 5)
|
187.8 percentage of Day 4 counts
Standard Error 14.1
|
178.4 percentage of Day 4 counts
Standard Error 18.9
|
129.1 percentage of Day 4 counts
Standard Error 7.0
|
PRIMARY outcome
Timeframe: Day 10Population: Safety analysis population. One participant in the PMN-high group was excluded due to external contamination affecting profiling data.
On Day 4 participants had neutrophils isolated from 100 mL of ACD-anti-coagulated autologous venous blood and labeled with up to 2.5 MBq 111In-tropolonate before being reinjected. Participants rested for 45 min post-injection to allow for neutrophil equilibrium between the circulating and marginating neutrophil pools. Whole-body profiling was performed in a heavily shielded dedicated whole-body counter with 2 highly sensitive scintillation detectors with the recorded counts corrected for the physical decay of 111In to allow measurement of the effect of TCZ on the normal redistribution pattern of neutrophils and assessment of margination of neutrophils in the presence of TCZ. Distribution of radiolabelled neutrophils and peak counts, on Day 10 (6 days post re-injection) in liver/spleen and pelvic bone marrow were decay corrected and expressed as percentages of Day 4 (45 minutes post re-injection).
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=4 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Redistribution Analysis on Day 10
Liver/spleen peak counts (Day 10)
|
84.1 percentage of Day 4 counts
Standard Error 6.2
|
76.6 percentage of Day 4 counts
Standard Error 3.3
|
96.2 percentage of Day 4 counts
Standard Error 2.9
|
|
Neutrophil Redistribution Analysis on Day 10
Pelvic peak counts (Day 10)
|
180.3 percentage of Day 4 counts
Standard Error 12.9
|
175.6 percentage of Day 4 counts
Standard Error 14.4
|
132.6 percentage of Day 4 counts
Standard Error 5.0
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population.
Neutrophil phagocytosis was assessed by flow cytometry using heat-killed Staphylococcal pneumonia (S.pneumonia) bacteria labeled with eFluor670. Phagocytosis was quantified by measuring the eFluor670 fluorescence from neutrophils containing phagocytosed bacteria. Experiments were performed using neutrophils (PMN) only, PMN plus S. pneumonia at 4 degrees(˚) centigrade (C) (to control for non-specific bacterial adherence to PMN cell surface), and PMN plus S. pneumonia at 37˚C. Change from baseline in the percentage of eFluor670+ neutrophils was calculated on Day 4.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Phagocytosis: Change From Baseline to Nadir (Day 4) in the Percentage of eFluor670-Positive (eFluoro670+) Neutrophils
PMN only
|
0 percentage of eFlouro+ neutrophils
Standard Error 0
|
0 percentage of eFlouro+ neutrophils
Standard Error 0
|
0 percentage of eFlouro+ neutrophils
Standard Error 0
|
|
Neutrophil Phagocytosis: Change From Baseline to Nadir (Day 4) in the Percentage of eFluor670-Positive (eFluoro670+) Neutrophils
PMN and S.pneumonia at 4˚C
|
-1 percentage of eFlouro+ neutrophils
Standard Error 1
|
2 percentage of eFlouro+ neutrophils
Standard Error 1
|
5 percentage of eFlouro+ neutrophils
Standard Error 2
|
|
Neutrophil Phagocytosis: Change From Baseline to Nadir (Day 4) in the Percentage of eFluor670-Positive (eFluoro670+) Neutrophils
PMN and S.pneumonia at 37˚C
|
4 percentage of eFlouro+ neutrophils
Standard Error 2
|
7.5 percentage of eFlouro+ neutrophils
Standard Error 2
|
9 percentage of eFlouro+ neutrophils
Standard Error 2
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population
Neutrophil phagocytosis was assessed by flow cytometry using heat-killed Staphylococcal pneumonia bacteria labeled with eFluor670. Phagocytosis was quantified by measuring the eFluor670 fluorescence from neutrophils containing phagocytosed bacteria. Experiments were performed using neutrophils (PMN) only, PMN plus S. pneumonia at 4˚C (to control for non-specific bacterial adherence to PMN cell surface), and PMN plus S. pneumonia at 37˚C. Change from baseline in the eFluor670+ MFI was calculated on Day 4.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Phagocytosis: Change From Baseline to Nadir (Day 4) in Median Fluorescence Intensity (MFI) of eFluor670+ Neutrophils
PMN and S.pneumonia at 37˚C
|
685 median fluoresence intensity
Standard Error 443
|
979 median fluoresence intensity
Standard Error 350
|
810 median fluoresence intensity
Standard Error 217
|
|
Neutrophil Phagocytosis: Change From Baseline to Nadir (Day 4) in Median Fluorescence Intensity (MFI) of eFluor670+ Neutrophils
PMN only
|
-1 median fluoresence intensity
Standard Error 3
|
2 median fluoresence intensity
Standard Error 7
|
2 median fluoresence intensity
Standard Error 2
|
|
Neutrophil Phagocytosis: Change From Baseline to Nadir (Day 4) in Median Fluorescence Intensity (MFI) of eFluor670+ Neutrophils
PMN and S.pneumonia at 4˚C
|
-2 median fluoresence intensity
Standard Error 3
|
39 median fluoresence intensity
Standard Error 18
|
32 median fluoresence intensity
Standard Error 7
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population
Neutrophils generate a respiratory burst using reactive oxygen species (ROS) to kill invading pathogens. When luminol is used as a substrate for ROS, a chemical reaction is produced resulting in photon emission (chemiluminescence) in primed and unprimed neutrophils following formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation which is quantifiable. fMLP stimulation of the respiratory burst is mediated through activation of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in primed neutrophils. The maximal fMLP response is observed in primed neutrophils and is an ex vivo measure of the capacity of neutrophils to respond to pathogenic stimuli. In the current experiments, neutrophils were primed with tumor necrosis factor alpha (TNFα). Light emission was recorded on a luminometer. Absolute change from baseline in the production of ROS on Day 4 was reported.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Respiratory Burst: Change From Baseline to Nadir (Day 4) in the Production of Reactive Oxygen Species as Measured by Chemiluminescence (Relative Light Units - Absolute)
Unprimed neutrophils
|
-16710 relative light units
Standard Error 12346
|
-5553 relative light units
Standard Error 6409
|
-2766 relative light units
Standard Error 3291
|
|
Neutrophil Respiratory Burst: Change From Baseline to Nadir (Day 4) in the Production of Reactive Oxygen Species as Measured by Chemiluminescence (Relative Light Units - Absolute)
TNF α primed neutrophils
|
82465 relative light units
Standard Error 16727
|
-45257 relative light units
Standard Error 17540
|
64072 relative light units
Standard Error 16130
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population
Neutrophil apoptosis was measured using microscopy method with slides stained with Diff-Quik (modified Wright Giemsa stain) and morphology examined under oil immersion light microscopy with 100 times magnification. Neutrophils constitutively undergo apoptosis when cultured ex vivo, and this can be delayed by the addition of agents such as granulocyte-macrophage colony-stimulating factor (GM-CSF) or TNFα. Apoptotic neutrophils were characterized with dark and pyknotic nuclei compared to the viable neutrophils. Change from baseline in the percentage of apoptotic neutrophils on Day 4 measured by microscopy is reported.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Survival: Change From Baseline to the Nadir (Day 4) in the Percentage of Apoptotic Neutrophils as Measured by Microscopic Morphology
Untreated neutrophils
|
-6.0 percentage of apoptotic neutrophils
Standard Error 3.0
|
-1.8 percentage of apoptotic neutrophils
Standard Error 4.0
|
-6.0 percentage of apoptotic neutrophils
Standard Error 5.0
|
|
Neutrophil Survival: Change From Baseline to the Nadir (Day 4) in the Percentage of Apoptotic Neutrophils as Measured by Microscopic Morphology
GM-CSF neutrophils
|
-1.0 percentage of apoptotic neutrophils
Standard Error 1.0
|
-7.0 percentage of apoptotic neutrophils
Standard Error 5.0
|
-12.0 percentage of apoptotic neutrophils
Standard Error 6.0
|
|
Neutrophil Survival: Change From Baseline to the Nadir (Day 4) in the Percentage of Apoptotic Neutrophils as Measured by Microscopic Morphology
TNFα neutrophils
|
-2.0 percentage of apoptotic neutrophils
Standard Error 4.0
|
3.0 percentage of apoptotic neutrophils
Standard Error 4.0
|
-4.0 percentage of apoptotic neutrophils
Standard Error 5.0
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population
Ageing neutrophils translocate phosphatidylserine from the inner leaflet of the plasma membrane to the outer leaflet during the early stages of apoptosis. This translocation can be measured due to the affinity of Annexin V (AV) to bind exposed phosphatidylserine. Propidium Iodide (PI) is normally membrane-impermeable but enters cells in late apoptosis when their plasma membrane becomes leaky. Neutrophils constitutively undergo apoptosis when cultured ex vivo, and this can be delayed by the addition of agents such as granulocyte-macrophage colony-stimulating factor (GM-CSF) or TNFα. Apoptosis was assessed by flow cytometry with fluorescein isocyanate-labeled recombinant human AV (AV-FITC) and PI staining and the change from baseline in the percentage of apoptotic neutrophils on Day 4 measured by flow cytometry is reported.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Survival: Change From Baseline to the Nadir in the Percentage of Apoptotic Neutrophils as Measured by Flow Cytometry
Untreated neutrophils
|
-5.0 percentage of apoptotic neutrophils
Standard Error 2.0
|
-8.0 percentage of apoptotic neutrophils
Standard Error 4.0
|
-7.0 percentage of apoptotic neutrophils
Standard Error 5.0
|
|
Neutrophil Survival: Change From Baseline to the Nadir in the Percentage of Apoptotic Neutrophils as Measured by Flow Cytometry
GM-CSF neutrophils
|
-4.0 percentage of apoptotic neutrophils
Standard Error 2.0
|
-13.0 percentage of apoptotic neutrophils
Standard Error 4.0
|
-9.0 percentage of apoptotic neutrophils
Standard Error 8.0
|
|
Neutrophil Survival: Change From Baseline to the Nadir in the Percentage of Apoptotic Neutrophils as Measured by Flow Cytometry
TNFα neutrophils
|
-3.0 percentage of apoptotic neutrophils
Standard Error 4.0
|
3.0 percentage of apoptotic neutrophils
Standard Error 2.0
|
3.0 percentage of apoptotic neutrophils
Standard Error 5.0
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population
Neutrophil shape change is an indicator of the chemotactic ability of neutrophils to respond to and migrate to sites of inflammation. For determination of neutrophil shape change, fresh (0 min control), phosphate-buffered saline (PBS) control (30 min control) and formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated (30 min fMLP) PMNs (at 5 × 10\^6 PMNs/ milliliter \[mL\]) were fixed with CellFIX (organic solvent used as fixative for adherent cells), 90 microliters (μL) transferred to each sample tube, and cold PBS added to stop further reaction. Shape change was assessed by measuring forward scatter (FSC) on flow cytometry. Change from baseline in the number of neutrophils with shape change on Day 4 was reported.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Morphology: Change From Baseline to Nadir in the Number of Neutrophils With Shape Change Measured Using Flow Cytometry
0 min control
|
1613 neutrophils with shape change
Standard Error 842
|
-359 neutrophils with shape change
Standard Error 2613
|
4790 neutrophils with shape change
Standard Error 1341
|
|
Neutrophil Morphology: Change From Baseline to Nadir in the Number of Neutrophils With Shape Change Measured Using Flow Cytometry
30 min control
|
4882 neutrophils with shape change
Standard Error 3069
|
-2309 neutrophils with shape change
Standard Error 4439
|
8814 neutrophils with shape change
Standard Error 3016
|
|
Neutrophil Morphology: Change From Baseline to Nadir in the Number of Neutrophils With Shape Change Measured Using Flow Cytometry
30 min fMLP
|
-1889 neutrophils with shape change
Standard Error 2800
|
-1667 neutrophils with shape change
Standard Error 4093
|
1524 neutrophils with shape change
Standard Error 3515
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population
Neutrophil shape change is an indicator of the chemotactic ability of neutrophils to respond to and migrate to sites of inflammation. For determination of neutrophil shape change, fresh (0 min control), PBS control (30 min control) and fMLP-stimulated (30 min fMLP) PMNs (at 5 × 10\^6 PMNs/ mL) were fixed with CellFIX, 90 μL transferred to each sample tube, and cold PBS added to stop further reaction. Shape change was assessed by measuring FSC on flow cytometry. Change from baseline in the percentage of neutrophils with shape change on Day 4 was reported.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Morphology: Change From Baseline to the Nadir (Day 4) in the Percentage of Neutrophils With Shape Change Measured by Flow Cytometry (FSC-High Cells)
0 min control
|
14.0 percentage of shape changed neutrophils
Standard Error 11.0
|
0.0 percentage of shape changed neutrophils
Standard Error 3.0
|
2.0 percentage of shape changed neutrophils
Standard Error 2.0
|
|
Neutrophil Morphology: Change From Baseline to the Nadir (Day 4) in the Percentage of Neutrophils With Shape Change Measured by Flow Cytometry (FSC-High Cells)
30 min control
|
17.0 percentage of shape changed neutrophils
Standard Error 8.0
|
1.0 percentage of shape changed neutrophils
Standard Error 4.0
|
6.0 percentage of shape changed neutrophils
Standard Error 3.0
|
|
Neutrophil Morphology: Change From Baseline to the Nadir (Day 4) in the Percentage of Neutrophils With Shape Change Measured by Flow Cytometry (FSC-High Cells)
30 min fMLP
|
1.0 percentage of shape changed neutrophils
Standard Error 3.0
|
0.0 percentage of shape changed neutrophils
Standard Error 2.0
|
-1.0 percentage of shape changed neutrophils
Standard Error 3.0
|
PRIMARY outcome
Timeframe: Baseline, Day 4Population: Safety analysis population
Neutrophil shape change is an indicator of the chemotactic ability of neutrophils to respond to and migrate to sites of inflammation. For determination of neutrophil shape change, fresh (0 min control), PBS control (30 min control) and fMLP-stimulated (30 min fMLP) PMNs (at 5 × 10\^6 PMNs/ mL) were fixed with CellFIX, 90 μL transferred to each sample tube, and cold PBS added to stop further reaction. Shape change was assessed by microscopy with neutrophils classified as shape-changed if they contained \> 1 cell surface bleb or irregularity and change from baseline in percentage of neutrophil with shape change on Day 4 was reported.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Neutrophil Morphology: Change From Baseline to the Nadir (Day 4) in the Percentage of Neutrophils With Shape Change Measured by Microscopic Morphology
0 min control
|
12.0 percentage of shape changed neutrophils
Standard Error 10.0
|
-3.0 percentage of shape changed neutrophils
Standard Error 4.0
|
0.0 percentage of shape changed neutrophils
Standard Error 2.0
|
|
Neutrophil Morphology: Change From Baseline to the Nadir (Day 4) in the Percentage of Neutrophils With Shape Change Measured by Microscopic Morphology
30 min control
|
13.0 percentage of shape changed neutrophils
Standard Error 11.0
|
-2.0 percentage of shape changed neutrophils
Standard Error 4.0
|
-1.0 percentage of shape changed neutrophils
Standard Error 1.0
|
|
Neutrophil Morphology: Change From Baseline to the Nadir (Day 4) in the Percentage of Neutrophils With Shape Change Measured by Microscopic Morphology
30 min fMLP
|
2.0 percentage of shape changed neutrophils
Standard Error 6.0
|
4.0 percentage of shape changed neutrophils
Standard Error 5.0
|
-1.0 percentage of shape changed neutrophils
Standard Error 3.0
|
PRIMARY outcome
Timeframe: Day 4Population: Safety analysis population
Neutrophil surface receptor expression may be used to characterize the activation status of neutrophils. Fresh (0 min), PBS control (30 min) and fMLP-stimulated (30 min) PMNs (5 × 10\^6 PMNs/mL) were fixed with CellFIX, and 90 μL transferred to each tube containing antibody mixture (2 μL cluster of differentiation \[CD\] 11b-brilliant violet (BV) 421, 2 μL CD16-FITC, 5 μL CD62L-allophycocyanin (APC) and 5 μL CD162-phycoerythrin \[PE\]) or isotype control mixture of equivalent volumes. After 30 minutes of incubation on ice and in the dark, cold PBS was added to stop further reaction. Surface marker expressions were quantified by flow cytometry.
Outcome measures
| Measure |
Placebo
n=6 Participants
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab (Polymorphonuclear Leukocyte (PMN)]-High Group)
n=5 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with less than or equal to (≤) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
Tocilizumab (PMN-Low Group)
n=7 Participants
Participants who received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0 and with greater than (\>) 50% neutrophil count decrease at Day 4 relative to baseline were included in this group.
|
|---|---|---|---|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD11b - 0 min control
|
12633 median fluoresence intensity
Standard Error 2062
|
15751 median fluoresence intensity
Standard Error 2043
|
17764 median fluoresence intensity
Standard Error 1910
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD11b - 30 min control
|
13144 median fluoresence intensity
Standard Error 2416
|
16047 median fluoresence intensity
Standard Error 2058
|
18134 median fluoresence intensity
Standard Error 1836
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD11b - 30 min fMLP
|
39629 median fluoresence intensity
Standard Error 2699
|
44481 median fluoresence intensity
Standard Error 5049
|
41339 median fluoresence intensity
Standard Error 2723
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD16 - 0 min control
|
15822 median fluoresence intensity
Standard Error 741
|
16872 median fluoresence intensity
Standard Error 1874
|
15901 median fluoresence intensity
Standard Error 1476
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD16 - 30 min control
|
13771 median fluoresence intensity
Standard Error 706
|
14885 median fluoresence intensity
Standard Error 1744
|
13475 median fluoresence intensity
Standard Error 1269
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD16 - 30 min fMLP
|
17145 median fluoresence intensity
Standard Error 884
|
18229 median fluoresence intensity
Standard Error 2613
|
14707 median fluoresence intensity
Standard Error 2205
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD62L - 0 min control
|
563 median fluoresence intensity
Standard Error 37
|
650 median fluoresence intensity
Standard Error 201
|
767 median fluoresence intensity
Standard Error 65
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD62L - 30 min control
|
493 median fluoresence intensity
Standard Error 38
|
588 median fluoresence intensity
Standard Error 181
|
615 median fluoresence intensity
Standard Error 37
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD62L - 30 min fMLP
|
6 median fluoresence intensity
Standard Error 3
|
5 median fluoresence intensity
Standard Error 9
|
9 median fluoresence intensity
Standard Error 12
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD162 - 0 min control
|
3393 median fluoresence intensity
Standard Error 454
|
3253 median fluoresence intensity
Standard Error 895
|
3849 median fluoresence intensity
Standard Error 314
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD162 - 30 min control
|
3088 median fluoresence intensity
Standard Error 433
|
2972 median fluoresence intensity
Standard Error 820
|
3355 median fluoresence intensity
Standard Error 347
|
|
Absolute Median Fluorescence Intensities of Neutrophil Adhesion Molecules
CD162 - 30 min fMLP
|
1571 median fluoresence intensity
Standard Error 219
|
1568 median fluoresence intensity
Standard Error 472
|
1733 median fluoresence intensity
Standard Error 154
|
Adverse Events
Placebo
Tocilizumab
Serious adverse events
Adverse event data not reported
Other adverse events
| Measure |
Placebo
n=6 participants at risk
Participants received a single dose of placebo-matched to tocilizumab on Day 0.
|
Tocilizumab
n=12 participants at risk
Participants received a single dose of IV TCZ at a dose of 8 mg/kg body weight infusion over 1 hour on Day 0.
|
|---|---|---|
|
Investigations
Aspartate aminotransferase increased
|
0.00%
0/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
16.7%
2/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
|
Investigations
Alanine aminotransferase increased
|
0.00%
0/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
8.3%
1/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
|
Investigations
Blood bilirubin increased
|
0.00%
0/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
8.3%
1/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
|
Investigations
Eosinophil count increased
|
16.7%
1/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
0.00%
0/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
|
Nervous system disorders
Headache
|
33.3%
2/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
8.3%
1/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
|
Respiratory, thoracic and mediastinal disorders
Oropharyngeal pain
|
0.00%
0/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
16.7%
2/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
|
Gastrointestinal disorders
Nausea
|
0.00%
0/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
8.3%
1/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
|
Skin and subcutaneous tissue disorders
Rash erythematous
|
0.00%
0/6 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
8.3%
1/12 • From baseline to 8 weeks of safety follow-up (approximately 8 months)
|
Additional Information
Results disclosure agreements
- Principal investigator is a sponsor employee The Study being conducted under this Agreement is part of the Overall Study. Investigator is free to publish in reputable journals or to present at professional conferences the results of the Study, but only after the first publication or presentation that involves the Overall Study. The Sponsor may request that Confidential Information be deleted and/or the publication be postponed in order to protect the Sponsor's intellectual property rights.
- Publication restrictions are in place
Restriction type: OTHER