3 Materials and Methods .1 Materials
4.6 Influence of gender on lymphocyte distribution
Breed distribution of the four gender groups are shown in table 19. The mean age of female, male, castrated male and spayed female dogs was 3.5 years, 3.8 years, 6.3 years and 6.5 years respectively.
Table 19
Breed distribution of the four gender groups in the study. f: female, m: male, mk: castrated male, fs: spayed female.
Breed Gender Total
f m mk fs
Bernese mountain dog 35 35 4 5 79
Beagle 4 10 7 1 22
Mixed breed dog 21 33 11 17 82
Weimaraner 44 28 2 1 75
Flat-coated Retriever 26 15 1 6 48
Rottweiler 10 17 4 5 36
Total 140 138 29 35 342
No significant differences of relative and absolute counts of lymphocyte and lymphocytes subsets were found (tables 20 and 21).
Results
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Table 20
Influence of gender on relative counts of lymphocyte subsets in the peripheral blood of healthy dogs. f: female, m: male, mk: castrated male, fs: spayed female.
Parameter f m mk fs
(n = 140) (n = 138) (n= 29) (n= 35) Lymphocytes (%)
median 27 25 25 24
(Q1-Q3) (22-33) (20-32) (21-31) (20-31) CD4+ T cells (%)
median 39 37 38 39
(Q1-Q3) (33-45) (30-44) (38-45) (35-43) CD8+ T cells (%)
median 20 20 21 22
(Q1-Q3) (16-24) (16-24) (17-29) (15-33) CD5+ T cells (%)
median 72 73 74 80
(Q1-Q3) (63-79) (61-79) (76-77) (75-82) CD21+B cells (%)
median 16 16 15 10
(Q1-Q3) (%) (10-21) (10-25) (11-19) (6-14) CD3+ T cells
median 73 74 77 78
(Q1-Q3) (65-80) (63-80) (71-82) (72-86) CD4+/CD8+
median 2 1.9 1.9 1.8
(Q1-Q3) (1.5-2.7) (1.4-2.6) (1.3-2.6) (1.2-2.5)
Results
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Table 21
Influence of gender on absolute counts of lymphocyte subsets in the peripheral blood of healthy dogs. f: female, m: male, mk: castrated male, fs: spayed female.
Parameter f m mk fs
(n = 140) (n = 138) (n= 29) (n= 35) Lymphocytes (x103/µL)
median 2.6 2.8 2.4 2.3
(Q1-Q3) (2.2-3.5) (2.1-3.7) (2.1-2.8) (2.3-2.9) CD4+ T cells (x103/µL)
median 1 1 0.9 0.9
(Q1-Q3) (0.8-1.3) (1.4-2.6) (1.3-2.6) (1.2-2.5) CD8+ T cells (x103/µL)
median 0.5 0.6 0.6 0.5
(Q1-Q3) (0.4-0.8) (0.4-0.8) (0.7-1.2) (0.7-1.1) CD5+ T cells (x103/µL)
median 1.9 2 1.8 1.7
(Q1-Q3) (1.5-2.4) (1.5-2.6) (1.5-2.1) (1.5-2.1) CD21+ B cells (x103/µL)
Median 0.4 0.4 0.3 0.3
(Q1-Q3) (0.3-0.7) (0.4-0.8) (0.2-0.5) (0.1-0.4) CD3+ T cells (x103/µL)
median 1.9 2 1.9 1.9
(Q1-Q3) (1.5-2.6) (1.5-2.6) (1.5-2.1) (1.5-2.1)
Results
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4.7 Effect of methylprednisolone on peripheral blood lymphocyte subsets.
4.7.1 Experiment 1
Blood samples were obtained on day 1, 2 and 3 to establish baseline values. On day 3, 4, 5 and 6, a single 1 mg/kg i.v. methylprednisolone dose was administered immediately after blood sampling. Methylprednisolone was discontinued on day 7, while blood sampling was continued until day 10. The effect of repeated daily methylprednisolone administration on the absolute counts of leucocytes, lymphocytes and subsets of lymphocytes as well as on the relative counts of lymphocytes and their subsets are shown in tables 22 and 23, respectively.
Table 22
Effect of methylprednisolone on absolute counts (x1000 cells/µl) of leucocytes, lymphocytes and subsets of lymphocytes. On days 3, 4, 5 and 6 methyprednisolone was given. STD:
Standard Deviation.
* Significant difference versus baseline values.
Results
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Table 23
Effect of methylprednisolone on the relative counts of lymphocytes and their subsets. On days 3, 4, 5 and 6 methyprednisolone was given. STD: Standard Deviation.
Parameter Baseline Day 4 Day 5 Day 6 Day 7 Day 9 Day 10
% Lymphocyte
Mean 26 30* 26 24* 27 25 27
STD 2.9 2.5 2.8 2.5 4.8 1.4 1.7
% CD4+ T cell
Mean 30 29 30 30 30 31 29 STD 6.8 5.8 7.4 6 6.2 5.8 6.3
% CD8+ T cell
Mean 30 32 32 31 28 30 29 STD 4.1 5.5 3.8 5 7 5.2 5.8
% CD3+ T cell
Mean 76 78 77 78 74 77 77 STD 3 7.5 4 5.8 8.8 9.3 8.6
% CD5+ T cell
Mean 73 74 73 73 71 72 72
STD 3 5.2 5.6 4.1 5.4 5.2 3.5
% CD21+ B cell
Mean 13 12 12.5 13 14 15 14 STD 2.3 3.4 3.7 3.2 6.8 4.3 3.2
CD4/CD8
Mean 1 0.9 0.9 1 1.1 1.1 1 STD 0.4 0.3 0.3 0.4 0.3 0.4 0.4
* Significant difference versus baseline values.
There was no significant difference between the relative and absolute counts of leucocytes, lymphocytes and subsets of lymphocytes, measured from day 1 to day 3.
The absolute counts of leucocytes, CD3+, CD5+ and CD4+ T cells were significantly (p <
0.05) lower than baseline values on the fourth day (day 10) after methylprednisolone
Results
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The relative counts of lymphocytes were significantly (p < 0.05) higher than baseline the day after the first dose of methylprednisolone, but returned to baseline the next day.
Interestingly, on day 6, i.e. three days after the first methylprednisolone dose, lymphocyte relative counts decreased significantly (p < 0.05). This decrease turned out to be short lived as relative count returned to baseline the next day.
No significant changes of the relative counts of CD4+, CD3+ and CD5+ T cells , CD21+ B cells or in the CD4/CD8 ratio were observed.
The effects of chronic methylprednisolone administration on serum alkaline phosphatase (ALP), alanine aminotransferase (ALT) and glutamate-dehydrogenase (GLDH) are shown in table 24.
Table 24
Effect of methylprednisolone on serum alkaline phosphatase (ALP), alanine aminotransferase (ALT) and glutamate-dehydrogenase (GLDH). On days 3, 4, 5 and 6 methylprednisolone was given. STD: Standard Deviation.
Parameter Baseline Day 4 Day 5 Day 6 Day 7 Day 9 Day 10
ALT
Mean 33 26* 24* 23* 21* 21* 22*
STD 7.1 4 3.8 4 4 3,6 4.2
ALP
Mean 115 207* 136* 150* 178* 207* 208* STD 51.9 113.4 65.9 73.8 76 71.7 80.1
GLDH
Mean 1.5 1.4 1.4 1.6 2.1 1.9 2*
STD 0.7 1.6 1.6 1.1 0.9 0.7 0.6
* Significant difference versus baseline values.
Serum ALT activity significantly (p < 0.05) decreased one day after initial treatment (day 4) and the values were still significantly (p < 0.05) lower than baseline 4 days after methylprednisolone discontinuation.
Results
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In contrast, serum ALP activity significantly (p < 0.05) increased one day after initial treatment, and the values were still significantly (p < 0.05) higher than baseline 4 days after discontinuing methylprednisolone.
Serum GLDH activity significantly (p < 0.05) increased 4 days (day 7) after initial treatment, returning to baseline on day 8. However, the values significantly (p < 0.05) increased again 4 days after methylprednisolone discontinuation.
4.7.2 Experiment 2
Blood samples were obtained 2 times per day on days 1 and 2 (at 8 a.m. and 4 p.m.), and at 8 a.m on day 3, to establish baseline values. There was no significant difference between relative and absolute counts of leucocytes, lymphocytes and subsets of lymphocytes measured at 8 a.m. and at 4 p.m. Two mg/kg methylprednisolone was administered i.m., at 8 a.m. on days 3, 4, 5, 6 and 7. On day 8, the dose of methyprednisolone was reduced to 1 mg/kg, and on day 11, to 0.5 mg/kg. Administration of methylprednisolone was discontinued on day 14.
The effect of repeated daily methylprednisolone administration on the absolute counts of leucocytes, neutrophils, lymphocytes, monocytes, eosinophils, and lymphocyte subsets are shown in tables 25 and 26. Similarly, the effects on relative counts are shown in tables 27 and 28.
Results
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Table 25
Effect of methylprednisolone on the absolute counts (x 1000 cells/µl) of leucocytes, neutrophils, monocytes, eosinophils and lymphocytes. STD: Standard Deviation.
Leucocyte Neutrophil Lymphocyte Monocyte Eosinophil Day Treatment mean
(STD) a Significant difference versus baseline values.
b Significant difference versus day 5 at 2 mg/kg dose (4 p.m.).
c Significant difference versus day 10 at 1 mg/kg dose (4 p.m.) d Significant difference versus day 12 at 0.5 mg/kg dose (4 p.m.).
Results
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Table 26
Effect of methylprednisolone on lymphocyte subset absolute counts (x 1000 cells/µl). STD:
Standard Deviation.
CD4+ T cell CD8+ T cell CD3+ T cell CD5+ T cell CD21+ B cell Day Treatment mean
STD
Results
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Table 27
Effect of methylprednisolone on the relative counts of leucocytes, neutrophils, monocytes, eosinophils and lymphocytes. STD: Standard Deviation.
%Neutrophil %Lymphocyte %Monocyte %Eosinophil Day Treatment a Significant difference versus baseline values.
b Significant difference versus day 5 at 2 mg/kg dose (4 p.m.).
c Significant difference versus day 10 at 1 mg/kg dose (4 p.m.) d Significant difference versus day 12 at 0.5 mg/kg dose (4 p.m.).
Results
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Table 28
Effect of methylprednisolone on lymphocyte subset relative counts. STD: Standard Deviation.
CD4+ T cell CD8+ T cell CD3+ T cell CD5+ T cell CD21+ B cell CD4/CD8 Day Treatment mean
STD a Significant difference versus baseline values.
Results
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When 2mg/kg methylprednisolone was given, the absolute counts of leucocytes (figure 35), neutrophils (figure 36) and monocytes (figure 38), and the relative counts of neutrophils (figure 37) were significantly (p < 0.05) increased 8 hours after treatment. Twenty-four hours after treatment, both relative and absolute counts of neutrophils fell, but were still significantly (p < 0.05) higher than baseline. The absolute counts of leucocytes and monocytes returned to baseline 24 hours after treatment. Interestingly, no significant change in the relative counts of monocytes was found for the time methylprednisolone was administered, but the counts significantly increased the day after stopping methylprednisolone, remaining significantly higher than baseline for three days (figure 39).
After reducing methylprednisolone to 1 mg/kg, a similar increase occurred, but the changes of the absolute counts of leucocytes and both absolute and relative counts of neutrophils were significantly less marked (p < 0.05). The decreases of the absolute counts of monocytes at a dose of 1 mg/kg were non-significant when compared to the values at a dose of 2 mg/kg (p > 0.05).
When 0.5 mg/kg methylprednisolone was given, a similar increase occurred. The decrease of the absolute and relative counts of neutrophils, and the absolute counts of monocytes were non-significant, compared to a dose of 1 mg/kg. The decrease of absolute counts of leucocytes was significantly less compared to the days when 1mg/kg was given. When the drug was discontinued (day 14), the absolute counts of leucocytes and neutrophils returned to baseline after one day (day 15), and the relative counts of neutrophils and the absolute counts of monocytes returned to baseline after three days (day 17).
Results
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Figure 35
Effect of methylprednisolone on peripheral blood leucocyte absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 10 20 30
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
Leucocytes absolute counts
* *
* *
*
* Significant difference versus baseline values.
Results
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Figure 36
Effect of methylprednisolone on peripheral blood neutrophil absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 5 10 15 20
*
*
*
*
*
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
Neutrophil absolute counts
* Significant difference versus baseline values.
Results
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Figure 37
Effect of methylprednisolone on peripheral blood neutrophil relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
40 60 80 100
Neutrophils %
*
*
*
*
*
*
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 38
Effect of methylprednisolone on peripheral blood monocyte absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0,0 0,4 0,8 1,2 1,6
Monocyte absolute counts
* *
*
*
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 39
Effect of methylprednisolone on peripheral blood monocyte relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 4 8 12
Monocyte %
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
In contrast to the above results, the absolute eosinophil counts (figure 42), and both relative and absolute lymphocyte counts (figure 40 and 41) were significantly (p < 0.05) decreased 8 hours after treatment at a dose of 2 mg/kg. The relative counts of eosinophils (figure 43) also decreased, but the differences were not statistically significant (p > 0.05).
Results
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When the three different doses are compared, no significant differences of both relative and absolute counts of lymphocytes and eosinophils at a dose of 1 mg/kg or 0.5 mg/kg were found when compared to a dose of 2 mg/kg (p > 0.05).
After the drug was discontinued, the absolute counts of eosinophils returned to baseline after one day (day 15) Both absolute and relative counts of lymphocytes were still significantly lower than baseline values after 3 days.
Figure 40
Effect of methylprednisolone on peripheral blood lymphocyte absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5
Lymphocytes absolute counts
* Significant difference versus baseline values.
Results
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Figure 41
Effect of methylprednisolone on peripheral blood lymphocyte relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 10 20 30 40
LyLmphocytes %
*
*
*
*
*
*
*
* *
Baseline D3 (4p.m.) D4 (8a.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
D4 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 42
Effect of methylprednisolone on peripheral blood eosinophil absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0,00 0,05 0,10 0,15 0,20
Eosinophils absolute counts
* *
*
*
* *
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 43
Effect of methylprednisolone on peripheral blood eosinophil relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 1 2 3 4
Eosinophils % Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
The absolute counts of CD3+ (figure 46), CD5+ (figure 47) and CD8+ T cells (figure 45), and CD21+ B cells (figure 48) were significantly decreased 8 hours after administration of 2 mg/kg methylprednisolone. However, 24 hours after treatment, all the counts with the exception of the absolute counts of CD8+ T cells returned to values that were
non-Results
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during second and third 2 mg/kg methyprednisolone doses. However, they decreased significantly on day 10, with counts returning to baseline 3 days after methylprednisolone discontinuation.
The relative counts of CD4+ T cells (figure 49) were significantly increased (p > 0.05) 8h after treatment, and the mean values returned to baseline 24 hours after treatment. In contrast, the relative counts of CD8+ T cells (figure 50) and CD21+ B cells (figure 53) were significantly decreased 8 hours after treatment. Twenty-four hours after treatment, the relative counts of CD8+ T cells reversed, but the values were still significantly lower than baseline.
The relative counts of CD21+ B cells returned to baseline 24 hours after treatment.
The relative counts of CD3+ T cells (figure 51) and CD5+ T cells (figure 52) did not change during treatment. The CD4/CD8 ratio was significantly increased 8 hours after administration and returned to baseline 24 hours after treatment (figure 54).
After reducing the dose from 2 mg/kg to 1 mg/kg, and from 1 mg/kg to 0.5 mg/kg, a similar decrease in the absolute counts of CD5+, CD3+ T cells and CD4+ T cells occurred. The decrease of the absolute and relative counts of CD21+ B cells was significantly less marked at a dose of 1 mg/kg compared to a 2 mg/kg dose and at a dose of 0.5 mg/kg compared to a 1 mg/kg dose (p < 0.05). The decrease of the absolute and relative counts of CD8+ T cells was significantly less marked at a dose of 0.5 mg/kg compared to a 2 mg/kg dose.
When the drug was discontinued the relative counts of CD4+ T cells, CD8+ T cells and CD21+ B cells returned to baseline after one day. The absolute counts of CD4+ T cells were non significantly lower than baseline after 3 days. The absolute counts of CD21+ B cells, CD3+,CD5+ T cells and CD8+ T cells were still significantly lower than baseline after 3 days.
Results
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Figure 44
Effect of methylprednisolone on peripheral blood CD4+ T cell absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0,0 0,4 0,8 1,2
CD4+ T cells absolute counts
* *
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 45
Effect of methylprednisolone on peripheral blood CD8+ T cell absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0,0 0,2 0,4 0,6 0,8 1,0
CD8+ T cells absolute counts
* *
* *
*
*
*
* *
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 46
Effect of methylprednisolone on peripheral blood CD3+ T cell absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0,4 0,8 1,2 1,6 2,0
CD3+ T cells absolute counts Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* *
*
* * * *
* Significant difference versus baseline values.
Results
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Figure 47
Effect of methylprednisolone on peripheral blood CD5+ T cell absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0,4 0,8 1,2 1,6 2,0
CD5+ T cells absolute counts Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* *
*
*
* *
*
* Significant difference versus baseline values.
Results
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Figure 48
Effect of methylprednisolone on peripheral blood CD21+ B cell absolute counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0,0 0,1 0,2 0,3 0,4
CD21+ B cells absolute counts
*
*
*
*
*
*
*
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 49
Effect of methylprednisolone on peripheral blood CD4+ T cell relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
15 25 35 45 55
CD4+ T cells %
*
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 50
Effect of methylprednisolone on peripheral blood CD8+ T cell relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 10 20 30 40 50
CD8+ T cells %
*
* *
*
*
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 51
Effect of methylprednisolone on peripheral blood CD3+ T cell relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
50 60 70 80 90
CD3+ T cells % Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 52
Effect of methylprednisolone on peripheral blood CD5+ T cell relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
50 60 70 80 90
CD5+ T cells % Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 53
Effect of methylprednisolone on peripheral blood CD21+ B cell relative counts. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 5 10 15 20
CD21+ B cells %
*
*
*
*
Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* Significant difference versus baseline values.
Results
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Figure 54
Effect of methylprednisolone on peripheral blood CD4/CD8 ratio. Days 3, 4, 5, 6 and 7 (2mg/kg i.m., 8 a.m.); Days 8, 9 and 10 (1 mg/kg i.m., 8 a.m.); Days 11, 12 and 13 (0.5 mg/kg i.m., 8 a.m.); Day 14, 15, 16, and 17 (no treatment). D: Day, •: Mean, I: Standard Deviation.
0 1 2 3 4
CD4/CD8 ratio Baseline D3 (4p.m.) D4 (8a.m.) D4 (4p.m.) D5 (8a.m.) D5 (4p.m.) D10 (4p.m.) D12 (4p.m.) D15 (4p.m.) D17 (4p.m.)
* * *
*
* Significant difference versus baseline values.
The effect of repeated, daily methylprednisolone administration on serum ALP, ALT and GLDH is shown in Table 29.
Serum ALP activity significantly increased with methylprednisolone administration on day 5 (figure 56), and the values remained significantly higher than baseline for 3 days, after therapy was discontinued.
Results
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Table 29
Effect of methylprednisolone on serum alkaline phosphatase (ALP), alanine aminotransferase (ALT) and Glutamat-Dehydrogenase (GLDH). STD: Standard Deviation.
ALP ALT GLDH
* Significant difference versus baseline values.