Rheumatoid Arthritis
Conditions
Brief summary
This open-label, single arm study will assess the effect of RoActemra/Actemra (tocilizumab) on neutrophils and monitor safety and benefit-risk of RoActemra/Actemra treatment in patients with active rheumatoid arthritis who have an inadequate response to current biologic or non-biologic disease-modifying antirheumatic drugs (DMARDs). Patients will receive RoActemra/Actemra at a dose of 8 mg/kg intravenously every 4 weeks, either as monotherapy or in combination with their current non-biologic DMARD. Anticipated time on study treatment is 52 weeks.
Interventions
DRUGtocilizumab [RoActemra/Actemra]
8 mg/kg iv every 4 weeks, 52 weeks
Sponsors
Hoffmann-La Roche
Study design
Allocation
NON_RANDOMIZED
Intervention model
SINGLE_GROUP
Primary purpose
TREATMENT
Masking
NONE
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
No
Inclusion criteria
* Adult patients, \>/= 18 years of age * Moderate to severe active rheumatoid arthritis of \>/= 6 months duration * DAS28 \>/= 3.2 at screening and baseline * Inadequate response to biologic or non-biologic DMARDs * Biologic DMARDs must be withdrawn (approximately 5 half-lives for the agent) before first dose of study drug * If continuing on a non-biologic DMARD, dose should be stable for at least 8 weeks * Oral corticosteroids must have been at stable dose for at least 25 out of 28 days prior to baseline
Exclusion criteria
* Major surgery (including joint surgery) within 8 weeks prior to screening or not recovered from prior surgery * Rheumatic autoimmune disease other then RA * Functional class IV as defined by the American College of Rheumatology (ACR) classification * Prior history of or current inflammatory joint disease other than RA * Previous treatment with any cell-depleting therapies * Intraarticular or parenteral corticosteroids within 4 weeks prior to baseline * Active infection or history of recurrent infection * Positive for HIV or hepatitis B or C * History of or current primary or secondary immunodeficiency
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Percentage of Neutrophils Positive for Dihydrorhodamine-123 (DHR) Oxidation | Visit 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24) | Phagocytosis can be measured by incubating neutrophils with PI-labeled heat killed S. aureus following incubation for 30 minutes. Neutrophils are co-incubated with DHR, which becomes oxidized by the products of the respiratory burst generated during phagocytosis. Fluorescence can then be measured by flow cytometry. |
| Mean Chemiluminescence (AUC) of Neutrophil Reactive Species Production Using Phorbol 12-Myristate 13-Acetate (PMA) Stimulation | Visit 2, 3, 5, and 8 (Baseline and predose at Weeks 4, 12 and 24) | Using luminol as a substrate for reactive oxidants, a chemical reaction is produced resulting in photon emission (chemiluminescence). PMA is a receptor-independent stimulator of the respiratory burst and the PMA response measures the total capacity of neutrophils to generate reactive oxidants. Measurements of reactive oxygen species are calculated as total chemiluminescence or the AUC. |
| Percentage of Neutrophils Positive for Propidium Iodide (PI)-Labeled Staphylococcus Aureus (S. Aureus) Uptake | Visit 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24) | S. aureus were heat killed then labeled with PI and opsonized with AB serum (SAPI). S. aureus was then incubated with the neutrophils for 30 minutes at 37 degrees Celsius. The neutrophils were washed, then the percentage of cells positive for the labeled S. aureus (that is, phagocytosed) was calculated via flow cytometry. A higher percentage represented more active phagocytosis. |
| Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visits 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24) | Aging 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 fluorescein isothiocyanate (FITC)-labeled annexin V to bind exposed phosphatidylserine. Cells that stain positive to Annexin V binding are apoptotic. At 4 hours (hrs) and 20 hrs stimulated and control samples were analyzed for levels of apoptosis. |
| Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visits 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24) | Aging 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 FITC-labeled annexin V to bind exposed phosphatidylserine. Cells that stain positive to Annexin V binding are apoptotic. At 4 hrs and 20 hrs stimulated and control samples were analyzed for levels of apoptosis. GM-CSF is an agent that delays apoptosis. Percentage of cells that stained positive for Annexin V binding in the presence or absence of GM-CSF (GM-CSF delayed or constitutive) were determined by flow cytometry. |
| Mean Fluorescence Intensity of CD11b on Neutrophil Surface | Visits 2, 3, and 5 (Baseline and Weeks 4 and 12) | Neutrophils were incubated with labeled antibodies against CD11b. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater adhesion, migration, and ingestion of complement-opsonized particles. |
| Mean Fluorescence Intensity of CD18 on Neutrophil Surface | Visits 2, 3, and 5 (Baseline and Weeks 4 and 12) | Neutrophils were incubated with labeled antibodies against CD18. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater adhesion, migration, and ingestion of complement-opsonized particles. |
| Mean Fluorescence Intensity of CD62L (L Selectin) on Neutrophil Surface | Visits 2, 3 and 5 (Baseline and Weeks 4 and 12) | Neutrophils were incubated with labeled antibody against CD62L (L selectin). Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater adhesion of neutrophils to vessel walls. |
| Mean Fluorescence Intensity of CD63 on Neutrophil Surface | Visits 2, 3, and 5 (Baseline and Weeks 4 and 12) | Neutrophils were incubated with labeled antibody against CD63b. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater azurophilic degranulation, an indicator of greater microbe killing. |
| Mean Fluorescence Intensity of Interleukin-6 Receptor (Il-6R) on Neutrophil Surface | Visits 2, 3, and 5 (Baseline and Weeks 4 and 12) | Neutrophils were incubated with labeled antibody against IL-6R. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater density of membrane bound IL-6 receptor. |
| Mean Fluorescence Intensity of Membrane Bound Tumor Necrosis Factor Alpha (mTNFα) on Neutrophil Surface | Visits 2, 3, and 5 (Baseline and Weeks 4 and 12) | Neutrophils were incubated with labeled antibody against mTNF. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates to a greater density of membrane bound TNF. |
| Mean Chemiluminescence (Area Under the Concentration-time Curve [AUC]) of Neutrophil Reactive Species Production Using Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) Stimulation | Visit 2, 3, 5, and 8 (Baseline and predose at Weeks 4, 12 and 24) | Using luminol as a substrate for reactive oxidants, a chemical reaction is produced resulting in photon emission (chemiluminescence). fMLP stimulation is mediated through the fMLP receptor on the cell surface. The fMLP response is only observed in primed neutrophils and response is a measure of in vivo priming. Measurements of reactive oxygen species are calculated as total chemiluminescence or the AUC. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments | Weeks 12, 24, and 36 | Benefit:Risk was defined at the participant level. It was considered acceptable if the DAS28 improvement represented at least a moderate European League Against Rheumatism (EULAR) response. The risks were based on the known adverse event (AE) profile of tocilizumab rather than on the actual AEs experienced by each participant |
| Disease Activity Score Based on 28-Joint Count (DAS28) | Screening, Baseline, and Weeks 4, 8, 12, 16, 20, 24, 36, 48, and 52 | The DAS28 is a combined index for measuring disease activity in rheumatoid arthritis. The index includes swollen and tender joint counts, acute phase response (erythrocyte sedimentation rate \[ESR\] or C-reactive protein \[CRP\]), and general health status. The DAS28, which uses a 28 joint count, is derived from the original DAS, which includes a 44 swollen joint count. The DAS28 scale ranges from 0 to 10, where higher scores represent higher disease activity. |
Countries
United Kingdom
Participant flow
Participants by arm
| Arm | Count |
|---|---|
| Tocilizumab 8 mg/kg Participants received tocilizumab 8 mg/kg (maximum dose 800 mg) IV, once every 4 weeks up to 52 weeks (total of 13 infusions). | 21 |
| Total | 21 |
Withdrawals & dropouts
| Period | Reason | FG000 |
|---|---|---|
| Overall Study | Adverse Event | 1 |
| Overall Study | Lack of Efficacy | 3 |
Baseline characteristics
| Characteristic | Tocilizumab 8 mg/kg |
|---|---|
| Age, Continuous | 49.2 Years STANDARD_DEVIATION 11.2 |
| Sex: Female, Male Female | 18 Participants |
| Sex: Female, Male Male | 3 Participants |
Adverse events
| Event type | EG000 affected / at risk |
|---|---|
| deaths Total, all-cause mortality | — / — |
| other Total, other adverse events | 21 / 21 |
| serious Total, serious adverse events | 4 / 21 |
Outcome results
Primary
Mean Chemiluminescence (Area Under the Concentration-time Curve [AUC]) of Neutrophil Reactive Species Production Using Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) Stimulation
Using luminol as a substrate for reactive oxidants, a chemical reaction is produced resulting in photon emission (chemiluminescence). fMLP stimulation is mediated through the fMLP receptor on the cell surface. The fMLP response is only observed in primed neutrophils and response is a measure of in vivo priming. Measurements of reactive oxygen species are calculated as total chemiluminescence or the AUC.
Time frame: Visit 2, 3, 5, and 8 (Baseline and predose at Weeks 4, 12 and 24)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (Area Under the Concentration-time Curve [AUC]) of Neutrophil Reactive Species Production Using Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) Stimulation | Visit 2 (n=19) | 8671 chemiluminescence units*hours | Standard Deviation 4597.18 |
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (Area Under the Concentration-time Curve [AUC]) of Neutrophil Reactive Species Production Using Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) Stimulation | Visit 3 (n=18) | 9971 chemiluminescence units*hours | Standard Deviation 7139.36 |
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (Area Under the Concentration-time Curve [AUC]) of Neutrophil Reactive Species Production Using Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) Stimulation | Visit 5 (n=17) | 32361 chemiluminescence units*hours | Standard Deviation 72947.87 |
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (Area Under the Concentration-time Curve [AUC]) of Neutrophil Reactive Species Production Using Formyl-Methionyl-Leucyl-Phenylalanine (fMLP) Stimulation | Visit 8 (n=14) | 10573 chemiluminescence units*hours | Standard Deviation 5537.06 |
Comparison: Visit 3 versus Visit 2p-value: 0.313ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.083ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.092ANOVA
Comparison: Visit 8 versus Visit 2p-value: 0.145ANOVA
Comparison: Visit 8 versus Visit 3p-value: 0.398ANOVA
Comparison: Visit 8 versus Visit 5p-value: 0.138ANOVA
Primary
Mean Chemiluminescence (AUC) of Neutrophil Reactive Species Production Using Phorbol 12-Myristate 13-Acetate (PMA) Stimulation
Using luminol as a substrate for reactive oxidants, a chemical reaction is produced resulting in photon emission (chemiluminescence). PMA is a receptor-independent stimulator of the respiratory burst and the PMA response measures the total capacity of neutrophils to generate reactive oxidants. Measurements of reactive oxygen species are calculated as total chemiluminescence or the AUC.
Time frame: Visit 2, 3, 5, and 8 (Baseline and predose at Weeks 4, 12 and 24)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (AUC) of Neutrophil Reactive Species Production Using Phorbol 12-Myristate 13-Acetate (PMA) Stimulation | Visit 2 (n=19) | 141039 chemiluminescence units * hours | Standard Deviation 72621.95 |
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (AUC) of Neutrophil Reactive Species Production Using Phorbol 12-Myristate 13-Acetate (PMA) Stimulation | Visit 3 (n=18) | 145304 chemiluminescence units * hours | Standard Deviation 48836.93 |
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (AUC) of Neutrophil Reactive Species Production Using Phorbol 12-Myristate 13-Acetate (PMA) Stimulation | Visit 5 (n=17) | 157028 chemiluminescence units * hours | Standard Deviation 73054.72 |
| Tocilizumab 8 mg/kg | Mean Chemiluminescence (AUC) of Neutrophil Reactive Species Production Using Phorbol 12-Myristate 13-Acetate (PMA) Stimulation | Visit 8 (n=14) | 116968 chemiluminescence units * hours | Standard Deviation 51291.31 |
Comparison: Visit 3 versus Visit 2p-value: 0.467ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.25ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.06ANOVA
Comparison: Visit 8 versus Visit 2p-value: 0.149ANOVA
Comparison: Visit 8 versus Visit 3p-value: 0.061ANOVA
Comparison: Visit 8 versus Visit 5p-value: 0.047ANOVA
Primary
Mean Fluorescence Intensity of CD11b on Neutrophil Surface
Neutrophils were incubated with labeled antibodies against CD11b. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater adhesion, migration, and ingestion of complement-opsonized particles.
Time frame: Visits 2, 3, and 5 (Baseline and Weeks 4 and 12)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD11b on Neutrophil Surface | Visit 2 (n=19) | 286.40 fluorescence intensity unit | Standard Deviation 105.86 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD11b on Neutrophil Surface | Visit 3 (n=17) | 275.33 fluorescence intensity unit | Standard Deviation 127.37 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD11b on Neutrophil Surface | Visit 5 (n=16) | 240.93 fluorescence intensity unit | Standard Deviation 77.5 |
Comparison: Visit 3 versus Visit 2p-value: 0.39ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.08ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.18ANOVA
Primary
Mean Fluorescence Intensity of CD18 on Neutrophil Surface
Neutrophils were incubated with labeled antibodies against CD18. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater adhesion, migration, and ingestion of complement-opsonized particles.
Time frame: Visits 2, 3, and 5 (Baseline and Weeks 4 and 12)
Population: ITT Population; n=number of participants analyzed at each visit
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD18 on Neutrophil Surface | Visit 2 (n=19) | 20.33 fluorescence intensity unit | Standard Deviation 8.3 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD18 on Neutrophil Surface | Visit 3 (n=17) | 20.47 fluorescence intensity unit | Standard Deviation 12.31 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD18 on Neutrophil Surface | Visit 5 (n=16) | 22.47 fluorescence intensity unit | Standard Deviation 15.42 |
Comparison: Visit 3 versus Visit 2p-value: 0.48ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.3ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.34ANOVA
Primary
Mean Fluorescence Intensity of CD62L (L Selectin) on Neutrophil Surface
Neutrophils were incubated with labeled antibody against CD62L (L selectin). Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater adhesion of neutrophils to vessel walls.
Time frame: Visits 2, 3 and 5 (Baseline and Weeks 4 and 12)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD62L (L Selectin) on Neutrophil Surface | Visit 2 (n=19) | 20.56 fluorescence intensity unit | Standard Deviation 10.05 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD62L (L Selectin) on Neutrophil Surface | Visit 3 (n=17) | 23.40 fluorescence intensity unit | Standard Deviation 11.55 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD62L (L Selectin) on Neutrophil Surface | Visit 5 (n=16) | 19.93 fluorescence intensity unit | Standard Deviation 15.32 |
Comparison: Visit 3 versus Visit 2p-value: 0.22ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.44ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.23ANOVA
Primary
Mean Fluorescence Intensity of CD63 on Neutrophil Surface
Neutrophils were incubated with labeled antibody against CD63b. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater azurophilic degranulation, an indicator of greater microbe killing.
Time frame: Visits 2, 3, and 5 (Baseline and Weeks 4 and 12)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD63 on Neutrophil Surface | Visit 5 (n=16) | 30.28 fluorescence intensity unit | Standard Deviation 16.53 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD63 on Neutrophil Surface | Visit 2 (n=19) | 31.00 fluorescence intensity unit | Standard Deviation 20.68 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of CD63 on Neutrophil Surface | Visit 3 (n=17) | 26.34 fluorescence intensity unit | Standard Deviation 14.55 |
Comparison: Visit 3 versus Visit 2p-value: 0.22ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.46ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.24ANOVA
Primary
Mean Fluorescence Intensity of Interleukin-6 Receptor (Il-6R) on Neutrophil Surface
Neutrophils were incubated with labeled antibody against IL-6R. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates with greater density of membrane bound IL-6 receptor.
Time frame: Visits 2, 3, and 5 (Baseline and Weeks 4 and 12)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of Interleukin-6 Receptor (Il-6R) on Neutrophil Surface | Visit 3 (n=17) | 3.69 fluorescence intensity unit | Standard Deviation 1.97 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of Interleukin-6 Receptor (Il-6R) on Neutrophil Surface | Visit 2 (n=19) | 2.72 fluorescence intensity unit | Standard Deviation 1.5 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of Interleukin-6 Receptor (Il-6R) on Neutrophil Surface | Visit 5 (n=16) | 4.42 fluorescence intensity unit | Standard Deviation 2.55 |
Comparison: Visit 3 versus Visit 2p-value: 0.05ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.01ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.18ANOVA
Primary
Mean Fluorescence Intensity of Membrane Bound Tumor Necrosis Factor Alpha (mTNFα) on Neutrophil Surface
Neutrophils were incubated with labeled antibody against mTNF. Flow cytometry was used to determine the mean fluorescence intensity. Greater fluorescence correlates to a greater density of membrane bound TNF.
Time frame: Visits 2, 3, and 5 (Baseline and Weeks 4 and 12)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of Membrane Bound Tumor Necrosis Factor Alpha (mTNFα) on Neutrophil Surface | Visit 2 (n=19) | 3.49 fluorescence intensity unit | Standard Deviation 3.32 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of Membrane Bound Tumor Necrosis Factor Alpha (mTNFα) on Neutrophil Surface | Visit 3 (n=17) | 3.55 fluorescence intensity unit | Standard Deviation 3.29 |
| Tocilizumab 8 mg/kg | Mean Fluorescence Intensity of Membrane Bound Tumor Necrosis Factor Alpha (mTNFα) on Neutrophil Surface | Visit 5 (n=16) | 3.75 fluorescence intensity unit | Standard Deviation 4.85 |
Comparison: Visit 3 versus Visit 2p-value: 0.48ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.43ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.44ANOVA
Primary
Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis
Aging 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 fluorescein isothiocyanate (FITC)-labeled annexin V to bind exposed phosphatidylserine. Cells that stain positive to Annexin V binding are apoptotic. At 4 hours (hrs) and 20 hrs stimulated and control samples were analyzed for levels of apoptosis.
Time frame: Visits 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24)
Population: Intent-to-treat (ITT) Population: all participants in the Safety Population who provided follow-up data for neutrophils or at least 1 efficacy variable. n (number) equals (=) number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 2, 4 hrs (n=19) | 8.95 percentage of cells staining positive | Standard Deviation 5.46 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 3, 4 hrs (n=19) | 7.36 percentage of cells staining positive | Standard Deviation 4.18 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 5, 4 hrs (n=19) | 8.45 percentage of cells staining positive | Standard Deviation 4.64 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 8, 4 hrs (n=15) | 12.43 percentage of cells staining positive | Standard Deviation 6.34 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 2, 20 hrs (n=19) | 53.97 percentage of cells staining positive | Standard Deviation 8.97 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 3, 20 hrs (n=19) | 50.32 percentage of cells staining positive | Standard Deviation 10.64 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 5, 20 hrs (n=19) | 50.50 percentage of cells staining positive | Standard Deviation 15.51 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding in Apoptosis | Visit 8, 20 hrs (n=15) | 48.24 percentage of cells staining positive | Standard Deviation 10.37 |
Comparison: 4 hrs: Visit 3 versus Visit 2p-value: 0.308t-test, 2 sided
Comparison: 4 hrs: Visit 5 versus Visit 2p-value: 0.415t-test, 2 sided
Comparison: 4 hrs: Visit 5 versus Visit 3p-value: 0.335t-test, 2 sided
Comparison: 4 hrs: Visit 8 versus Visit 2p-value: 0.12t-test, 2 sided
Comparison: 4 hrs: Visit 8 versus Visit 3p-value: 0.061t-test, 2 sided
Comparison: 4 hrs: Visit 8 versus Visit 5p-value: 0.031t-test, 2 sided
Comparison: 20 hrs: Visit 3 versus Visit 2p-value: 0.131t-test, 2 sided
Comparison: 20 hrs: Visit 5 versus Visit 2p-value: 0.202t-test, 2 sided
Comparison: 20 hrs: Visit 5 versus Visit 3p-value: 0.484t-test, 2 sided
Comparison: 20 hrs: Visit 8 versus Visit 2p-value: 0.047t-test, 2 sided
Comparison: 20 hrs: Visit 8 versus Visit 3p-value: 0.285t-test, 2 sided
Comparison: 20 hrs: Visit 8 versus Visit 5p-value: 0.315t-test, 2 sided
Primary
Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF)
Aging 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 FITC-labeled annexin V to bind exposed phosphatidylserine. Cells that stain positive to Annexin V binding are apoptotic. At 4 hrs and 20 hrs stimulated and control samples were analyzed for levels of apoptosis. GM-CSF is an agent that delays apoptosis. Percentage of cells that stained positive for Annexin V binding in the presence or absence of GM-CSF (GM-CSF delayed or constitutive) were determined by flow cytometry.
Time frame: Visits 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 8, 4 hrs (n=15) | 6.34 percentage of cells staining positive | Standard Deviation 2.58 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 2, 4 hrs (n=19) | 5.46 percentage of cells staining positive | Standard Deviation 6.73 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 3, 4 hrs (n=19) | 4.18 percentage of cells staining positive | Standard Deviation 7.13 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 5, 4 hrs (n=19) | 4.64 percentage of cells staining positive | Standard Deviation 3.18 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 2, 20 hrs (n=19) | 27.00 percentage of cells staining positive | Standard Deviation 11.16 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 3, 20 hrs (n=19) | 28.35 percentage of cells staining positive | Standard Deviation 10.95 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 5, 20 hrs (n=19) | 29.68 percentage of cells staining positive | Standard Deviation 14.78 |
| Tocilizumab 8 mg/kg | Mean Percentage of Cells Staining Positive for Annexin V Binding With Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) | Visit 8, 20 hrs (n=15) | 27.79 percentage of cells staining positive | Standard Deviation 12.01 |
Comparison: 4 hrs: Visit 8 versus Visit 5p-value: 0.052t-test, 2 sided
Comparison: 4 hrs: Visit 3 versus Visit 2p-value: 0.288t-test, 2 sided
Comparison: 4 hrs: Visit 5 versus Visit 2p-value: 0.318t-test, 2 sided
Comparison: 4 hrs: Visit 5 versus Visit 3p-value: 0.4t-test, 2 sided
Comparison: 4 hrs: Visit 8 versus Visit 2p-value: 0.317t-test, 2 sided
Comparison: 4 hrs: Visit 8 versus Visit 3p-value: 0.137t-test, 2 sided
Comparison: 20 hrs: Visit 3 versus Visit 2p-value: 0.354t-test, 2 sided
Comparison: 20 hrs: Visit 5 versus Visit 2p-value: 0.266t-test, 2 sided
Comparison: 20 hrs: Visit 5 versus Visit 3p-value: 0.378t-test, 2 sided
Comparison: 20 hrs: Visit 8 versus Visit 2p-value: 0.422t-test, 2 sided
Comparison: 20 hrs: Visit 8 versus Visit 3p-value: 0.444t-test, 2 sided
Comparison: 20 hrs: Visit 8 versus Visit 5p-value: 0.345t-test, 2 sided
Primary
Percentage of Neutrophils Positive for Dihydrorhodamine-123 (DHR) Oxidation
Phagocytosis can be measured by incubating neutrophils with PI-labeled heat killed S. aureus following incubation for 30 minutes. Neutrophils are co-incubated with DHR, which becomes oxidized by the products of the respiratory burst generated during phagocytosis. Fluorescence can then be measured by flow cytometry.
Time frame: Visit 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Dihydrorhodamine-123 (DHR) Oxidation | Visit 3 (n=19) | 99.20 percentage of positive neutrophils | Standard Deviation 1.93 |
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Dihydrorhodamine-123 (DHR) Oxidation | Visit 2 (n=19) | 99.21 percentage of positive neutrophils | Standard Deviation 2.43 |
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Dihydrorhodamine-123 (DHR) Oxidation | Visit 5 (n=17) | 99.91 percentage of positive neutrophils | Standard Deviation 0.18 |
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Dihydrorhodamine-123 (DHR) Oxidation | Visit 8 (n=15) | 99.92 percentage of positive neutrophils | Standard Deviation 0.22 |
Comparison: Visit 3 versus Visit 2p-value: 0.496ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.122ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.07ANOVA
Comparison: Visit 8 versus Visit 2p-value: 0.135ANOVA
Comparison: Visit 8 versus Visit 3p-value: 0.082ANOVA
Comparison: Visit 8 versus Visit 5p-value: 0.461ANOVA
Primary
Percentage of Neutrophils Positive for Propidium Iodide (PI)-Labeled Staphylococcus Aureus (S. Aureus) Uptake
S. aureus were heat killed then labeled with PI and opsonized with AB serum (SAPI). S. aureus was then incubated with the neutrophils for 30 minutes at 37 degrees Celsius. The neutrophils were washed, then the percentage of cells positive for the labeled S. aureus (that is, phagocytosed) was calculated via flow cytometry. A higher percentage represented more active phagocytosis.
Time frame: Visit 2, 3, 5, and 8 (Baseline and Weeks 4, 12 and 24)
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Propidium Iodide (PI)-Labeled Staphylococcus Aureus (S. Aureus) Uptake | Visit 2 (n=19) | 96.59 percentage of positive neutrophils | Standard Deviation 3.01 |
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Propidium Iodide (PI)-Labeled Staphylococcus Aureus (S. Aureus) Uptake | Visit 3 (n=19) | 97.47 percentage of positive neutrophils | Standard Deviation 2.56 |
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Propidium Iodide (PI)-Labeled Staphylococcus Aureus (S. Aureus) Uptake | Visit 5 (n=17) | 97.53 percentage of positive neutrophils | Standard Deviation 2.68 |
| Tocilizumab 8 mg/kg | Percentage of Neutrophils Positive for Propidium Iodide (PI)-Labeled Staphylococcus Aureus (S. Aureus) Uptake | Visit 8 (n=15) | 97.25 percentage of positive neutrophils | Standard Deviation 2.27 |
Comparison: Visit 3 versus Visit 2p-value: 0.169ANOVA
Comparison: Visit 5 versus Visit 2p-value: 0.165ANOVA
Comparison: Visit 5 versus Visit 3p-value: 0.471ANOVA
Comparison: Visit 8 versus Visit 2p-value: 0.243ANOVA
Comparison: Visit 8 versus Visit 3p-value: 0.396ANOVA
Comparison: Visit 8 versus Visit 5p-value: 0.375ANOVA
Secondary
Disease Activity Score Based on 28-Joint Count (DAS28)
The DAS28 is a combined index for measuring disease activity in rheumatoid arthritis. The index includes swollen and tender joint counts, acute phase response (erythrocyte sedimentation rate \[ESR\] or C-reactive protein \[CRP\]), and general health status. The DAS28, which uses a 28 joint count, is derived from the original DAS, which includes a 44 swollen joint count. The DAS28 scale ranges from 0 to 10, where higher scores represent higher disease activity.
Time frame: Screening, Baseline, and Weeks 4, 8, 12, 16, 20, 24, 36, 48, and 52
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Screening (n=21) | 5.67 score on a scale | Standard Deviation 1.22 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Baseline (n=21) | 6.15 score on a scale | Standard Deviation 1.26 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 4 (n=21) | 4.90 score on a scale | Standard Deviation 1.4 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 8 (n=20) | 4.31 score on a scale | Standard Deviation 1.64 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 12 (n=20) | 3.83 score on a scale | Standard Deviation 1.45 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 16 (n=20) | 3.81 score on a scale | Standard Deviation 1.69 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 20 (n=20) | 3.92 score on a scale | Standard Deviation 1.45 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 24 (n=16) | 3.26 score on a scale | Standard Deviation 1.49 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 36 (n=12) | 3.27 score on a scale | Standard Deviation 1.33 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 48 (n=11) | 3.59 score on a scale | Standard Deviation 1.62 |
| Tocilizumab 8 mg/kg | Disease Activity Score Based on 28-Joint Count (DAS28) | Week 52 (n=18) | 3.58 score on a scale | Standard Deviation 1.51 |
Secondary
Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments
Benefit:Risk was defined at the participant level. It was considered acceptable if the DAS28 improvement represented at least a moderate European League Against Rheumatism (EULAR) response. The risks were based on the known adverse event (AE) profile of tocilizumab rather than on the actual AEs experienced by each participant
Time frame: Weeks 12, 24, and 36
Population: ITT Population; n=number of participants analyzed for the specified parameter at a given visit.
| Arm | Measure | Group | Value (NUMBER) |
|---|---|---|---|
| Tocilizumab 8 mg/kg | Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments | Acceptable, Week 12 | 100 percentage of participants |
| Tocilizumab 8 mg/kg | Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments | Not Acceptable, Week 12 | 0 percentage of participants |
| Tocilizumab 8 mg/kg | Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments | Acceptable, Week 24 | 94 percentage of participants |
| Tocilizumab 8 mg/kg | Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments | Not Acceptable, Week 24 | 6 percentage of participants |
| Tocilizumab 8 mg/kg | Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments | Acceptable, Week 36 | 92 percentage of participants |
| Tocilizumab 8 mg/kg | Percentage of Participants With Acceptable and Not Acceptable Benefit-Risk Assessments | Not Acceptable, Week 36 | 8 percentage of participants |