# Abstract

 

## Background Cigarette smoke contains free radicals and an have adverse effect to the immune system. Supplementation of palm oil vitamin E (palmvitee), is known has antioxidant properties is thought to be beneficial for system immune protection against free radicals activity. The objective of the study was to determine the effect of palmvitee supplementation on immune response in smokers. 

## Methods This study involved a group of smokers and nonsmokers who received 200 mg/day palmvitee and placebo for the control group. Blood samples were taken at 0, 12 and 24 weeks of supplementation. Plasma tocopherol and tocotrienol were determined by HPLC, lymphocyte proliferation by lymphocyte transformation test (LTT) and enumeration of lymphocytes T and B cells by flow cytometry. Statistical analysis was performed by Mann--Whitney *U*-test for non-parametric data distribution and correlation among the variables was examined by Spearman. 

## Results Plasma tocopherol and tocotrienol were increased in vitamin E supplemented group as compared to placebo group. Urine cotinine levels and serum α~1~-antitrypsin were significantly higher in smokers compared to nonsmokers. Lymphocyte proliferation induced by PHA showed an increasing trend with palmvitee supplementation in both smokers and nonsmokers. Natural killer cells were decreased; CD4^+^ cells and B cells were increased in smokers compared to nonsmokers but were unaffected with vitamin E supplementation except in the percentage of B cells which were increased in nonsmokers supplemented palmvitee compared to placebo. CD4^+^/CD8^+^ ratio was increased in smokers compared to nonsmokers. The high TWBC count observed in smokers correlated with the increased CD4^+^ and B cells. 

## Conclusions Smoking caused alterations in certain immune parameters and palmvitee supplementation tended to cause an increase in lymphocytes transformation test but had no effect on CD3^+^, CD4^+^, CD8^+^, NK cells and B cells except B cells percentage in nonsmokers. # Results 

## Plasma total vitamin E level There was no difference in total plasma vitamin E, tocopherol and tocotrienol levels in smokers and nonsmokers at the beginning of the study (Figure  [1](#) & Table  [1](#)). After palmvitee supplementation, both plasma tocopherol and tocotrienol concentration were increased in smokers and nonsmokers (p \< 0.05) starting from 12 weeks until the end of the experiment. Tocopherol concentration was higher in plasma as compared to tocotrienol. Urinary cotinine and serum α~1~-antitrypsin of smokers were significantly higher (p \< 0.05) compared to nonsmokers (Table  [1](#)) but no changes were observed with supplementation. Urinary cotinine levels were increased accordingly to the number of cigarettes per day (Table  [2](#)). <figure> <p><img src="" /></p> <figcaption>Total plasma vitamin E concentration in smokers and nonsmokers.</figcaption> </figure> ::::table-wrap

::: caption
**The effects of palmvitee supplementation on plasma tocopherol and
tocotrienol,** α~**1**~**-antitrypsin in serum and urine cotinine levels
in smokers and nonsmokers**
:::

**Weeks** **Tocopherol (μg/ml plasma)**

______________________________________________________________________

**Nonsmokers** **Smokers**\
Placebo, n = 27 Palmvitee, n = 29 Placebo, n = 28 Palmvitee,n = 30
0 11.67 ± 3.19 11.75 ± 2.27 11.58 ± 3.19 11.39 ± 3.06
12 10.42 ± 2.17 14.22 ± 3.61\* 10.27 ± 3.33 13.92 ± 4.17\*\*
24 11.52 ± 2.94 14.56 ± 3.06\* 11.53 ± 2.78 14.11 ± 3.89\*\*
**Tocotrienol (ng/ml)**  \
0 62.49 ± 6.69 63.60 ± 5.67 60.05 ± 6.39 59.49 ± 4.21
12 63.50 ± 7.87 656.25 ± 142.50\* 61.27 ± 3.62 570.22 ± 98.64\*\*
24 60.65 ± 7.13 675.16 ± 150.00\* 60.47 ± 3.00 587.68 ± 87.50\*\*
**CCR, (μg/mg creatinine)**  \
n = 16 n = 19 n = 21 n = 20
0 0.052 ± 0.017 0.046 ± 0.017 0.927 ± 0.734# 0.941 ± 0.700#
24 0.058 ± 0.030 0.047 ± 0.017 0.866 ± 0.754# 1.179 ± 0.738#
**α**~**1**~**-antitripsin(g/L serum)**  \
n = 16 n = 19 n = 28 n = 30
0 1.48 ± 0.31 1.49 ± 0.34 1.99 ± 0.43# 1.94 ± 0.31#
24 1.47 ± 0.20 1.45 ± 0.24 1.69 ± 0.37 1.75 ± 0.39

Results are shown as mean ± SD. * indicates significant difference as
compared to nonsmokers taking placebo (p \< 0.01), \*\* indicates
significant difference as compared to smokers taking placebo
(p \< 0.01), # indicates significant difference as compared to
nonsmokers (p \< 0.0001), CCR represents cotinine:creatinine ratio.:::: ::::table-wrap
::: caption
Baseline levels of lymphocytes in nonsmokers and smokers according to
the number of cigarettes per day
:::

**Nonsmokers n = 56** **S1 (n = 7)** **S2 (n = 34)** **S3 (n = 15)**

______________________________________________________________________

Age (y) 34 ± 8 35 ± 5 35 ± 6 36 ± 6
Body mass index (kg/m^2^) 23.84 ± 3.99 24.11 ± 2.22 23.27 ± 3.26 23.55 ± 3.9822
Vitamin E μg/ml 11.76 ± 3.17 11.66 ± 3.24 11.66 ± 3.24 11.66 ± 3.98
α~1~-antitrypsin g/L serum 1.49 ± 0.32 1.98 ± 0.27\* 1.96 ± 0.39\* 1.94 ± 0.38\*
CCR μg/mg creatinine 0.049 ± 0.017 0.268 ± 0.244\* 0.974 ± 0.759\* 1.13 ± 0.56\*
PHA SI 70.54 ± 43.35 47.50 ± 34.63 89.05 ± 61.07 80.93 ± 55.85
Con A SI 54.73 ± 38.35 33.33 ± 17.07 66.94 ± 41.32 57.13 ± 23.01
White cell count (a.n. x 10^9^/L) 7.99 ± 1.74 9.83 ± 2.60\* 8.71 ± 2.16 9.55 ± 1.96\*
Lymphocytes (a.n. x 10^9^/L) 3.25 ± 0.74 3.77 ± 1.45 3.50 ± 1.00 3.58 ± 1.01
(%) 41.19 ± 7.76 37.68 ± 7.36 40.61 ± 6.74 39.83 ± 6.77\
T cell (a.n. x 10^9^/L) 2.05 ± 0.55 2.39 ± 0.10 2.36 ± 0.68 2.31 ± 0.71
(%) 62.76 ± 7.54 63.14 ± 11.31 64.64 ± 7.32 64.67 ± 7.83\
B cell (a.n. x 10^9^/L) 0.38 ± 0.20 0.67 ± 0.51\* 0.52 ± 0.29\* 0.54 ± 0.17\*
(%) 11.13 ± 4.09 16.86 ± 6.20\* 14.36 ± 4.20\* 14.80 ± 4.28\*\
NK cells (a.n. x 10^9^/L) 0.83 ± 0.35 0.67 ± 0.35 0.66 ± 0.31\* 0.67 ± 0.33
(%) 25.31 ± 903 19.00 ± 8.25\* 19.34 ± 6.67\* 18.20 ± 7.16\*\
CD4^+^ (a.n. x 10^9^/L) 0.92 ± 0.28 0.99 ± 0.30 1.18 ± 0.43\* 1.14± 0.26\*
(%) 28.27 ± 6.43 27.29 ± 4.42 33.44 ± 5.46 31.47 ± 5.82
CD8^+^ (a.n. x 10^9^/L) 1.05 ± 0.39 1.17 ± 0.60 1.00 ± 0.34 1.049± 0.42
(%) 31.53 ± 7.52 30.00 ± 7.81 28.89 ± 6.92 29.33 ± 7.16
CD4^+^/CD8^+^ (%) 0.97 ± 0.44 0.987 ± 0.383 1.22 ± 0.37\* 1.16 ± 0.42

S1 refers to \< 9 cigarettes/d, S2; 10-19 cigarettes/d and S3; >20
cigarettes/d. * indicates significant difference as compared to
nonsmokers (p \< 0.05).:::: 

## Lymphocyte proliferation There was no difference in the lymphocyte proliferation after induction with mitogens PHA and Con A between the different groups (smoking vs nonsmoking) (Table  [3](#)). When the group of smokers divided into the number of cigarettes smoked per day, it seems to reduce in S1 group and increased in S2 and S3 group (Table  [2](#)). But it remained unaffected by palmvitee supplementation (Table  [3](#)). But there seemed to be a trend for the lymphocyte proliferation to increase at 24 weeks in the palmvitee-supplemented group induced with PHA (Table  [3](#) &[4](#)). ::::table-wrap
::: caption
The effects of palmvitee supplementation on lymphocyte proliferative
presented as stimulation index (S.I.) after induction with mitogen, PHA
and Con A in nonsmokers and smokers
:::

**Weeks** **Con A (S.I)**

______________________________________________________________________

**Non-smokers** **Smokers**\
Placebo, n = 27 Palmvitee, n = 29 Placebo, n = 28 Palmvitee, n = 30
0 49.77 ± 36.42 59.17 ± 40.10 69.21 ± 41.11 52.72 ± 30.00
12 48.19 ± 28.62 51.03 ± 28.78 47.70 ± 29.73 59.50 ± 34.18
24 46.92 ± 23.64 55.10 ± 27.23 42.21 ± 26.59 42.93 ± 31.75
**PHA (S.I)**\
0 66.72 ± 48.82 73.97 ± 38.35 91.96 ± 62.15 73.45 ± 53.44
12 70.59 ± 39.43 72.14 ± 28.88 65.74 ± 34.69 75.27 ± 35.85
24 69.15 ± 28.32 86.41 ± 41.53 69.39 ± 38.07 77.10 ± 59.44

Results are shown as mean ± SD.:::: ::::table-wrap
::: caption
The effects of palmvitee supplementation on lymphocyte proliferative
presented as stimulation index (S.I.) after induction with mitogen, PHA
and Con A in nonsmokers and smokers according to the number of
cigarettes per day
:::

**Weeks** **S1** **S2** **S3**

______________________________________________________________________

**Placebo, n = 4** **Palmvitee, n = 3** **Placebo, n = 17** **Palmvitee,n = 19** **Placebo, n = 7** **Palmvitee,n = 8**
**Con A (SI)**\
0 37.75 ± 15.73 24.50 ± 21.92 80.00 ± 48.12 55.26 ± 30.92 61.00 ± 15.41 53.75 ± 39.75
12 39.25 ± 15.09 55.00 ± 35.59 53.50 ± 35.02 59.32 ± 40.01 39.29 ± 20.77 61.63 ± 28.52\*
24 36.00 ± 32.65 22.00 ± 24.56 46.24 ± 28.03 49.26 ± 35.55 36.00 ± 20.94 35.75 ± 26.99
**PHA (SI)**\
0 58.00 ± 39.30 26.50 ± 6.36 73.17 ± 40.22 70.21 ± 44.99 67.29 ± 29.84 44.60 ± 28.53
12 60.50 ± 24.96 51.67 ± 30.62 69.44 ± 40.09 76.58 ± 35.20 60.29 ± 28.19 81.00 ± 38.53
24 59.25 ± 24.07 72.67 ± 69.82 74.71 ± 44.45 78.47 ± 68.50 62.29 ± 27.49 75.50 ± 32.96

Results are shown as means ± SD. * indicates significant difference as
compared to nonsmokers.:::: 

## T-cell subsets and B cell percentage Table  [5](#) shows the baseline levels of white blood cells counts, lymphocytes and T-cell subsets in smokers and nonsmokers. Table  [2](#) shows the baseline according the number of cigarettes smoked per day. When comparing immune parameters between smokers and nonsmokers, WBC counts was higher in smokers (n = 58) compared to nonsmokers (n = 56, p \< 0.01) and unaffected by palmvitee supplementation (Table  [6](#)). No difference in the total number and percentage of lymphocytes and CD3^+^ cells was observed before and after palmvitee supplementation (Table  [6](#)). In separated group; S3, total CD3^+^ cells (T cell) increased significantly (p \< 0.05) compared to nonsmokers before supplementation (Table  [2](#)) and remained unchanged in palmvitee-supplemented group (Table  [7](#)). The percentage of B cells in smokers was higher compared to nonsmokers (p \< 0.0001) and also in S1 (p \< 0.005), S2 (p \< 0.0005) and S3 groups (p \< 0.005) (Table  [2](#)). After palmvitee supplementation, B cells percentage was significantly increased in nonsmokers at 24 weeks (p \< 0.05) but not in the smokers (Table  [6](#)). In S1 and S3 palmvitee-supplemented groups there was a trend of increment in B cells percentage. The increment was correlated at 12 (r = 0.98, 0.87) and 24 weeks (r = 0.98, 0.96) respectively. ::::table-wrap
::: caption
Baseline levels of lymphocytes in smokers and nonsmokers
:::

**Nonsmokers n = 56** **Smokers n = 58**

______________________________________________________________________

Age (y) 34 ± 8 36 ± 6 NS
Body mass index (kg/m2) (kg/m^2^) 23.84 ± 3.99 23.52 ± 3.42 NS
Vitamin E μg/ml 11.76 ± 3.17 11.66 ± 3.24 NS
α~1~-antitrypsin g/L serum 1.49 ± 0.32 1.96 ± 0.37 P \< 0.0000001
CCR μg/mg creatinine 0.049 ± 0.017 0.93 ± 0.31 P \< 0.0000001
PHA SI 70.54 ± 43.35 77.50 ± 55.34 NS
Con A SI 54.73 ± 38.35 59.98 ± 36.29 NS
White cell count (a.n. x 10^9^/L) 7.99 ± 1.74 9.06 ± 2.18 P \< 0.01
Lymphocytes (a.n. x 10^9^/L) 3.25 ± 0.74 3.58 ± 1.01 NS
(%) 41.19 ± 7.76 39.83 ± 6.77
. NS
T cell (a.n. x 10^9^/L) 2.05 ± 0.55 2.31 ± 0.71 NS
(%) 62.76 ± 7.54 64.67 ± 7.83 NS
B cell (a.n. x 10^9^/L) 0.38 ± 0.20 0.54 ± 0.29 P \< 0.00001
(%) 11.13 ± 4.09 14.78 ± 4.48 P \< 0.00001
NK cells (a.n. x 10^9^/L) 0.83 ± 0.35 0.66 ± 0.31 P \< 0.001
(%) 25.31 ± 903 19.00 ± 6.88 P \< 0.0001
CD4^+^ (a.n. x 10^9^/L) 0.92 ± 0.28 1.15± 0.38 P \< 0.0001
(%) 28.27 ± 6.43 32.19 ± 5.73 P \< 0.0001
CD8^+^ (a.n. x 10^9^/L) 1.05 ± 0.39 1.04 ± 0.39 NS
(%) 31.53 ± 7.52 29.14 ± 6.97 NS
CD4^+^/CD8^+^ (%) 0.97 ± 0.44 1.18 ± 0.38 P \< 0.0001

a.n. represents actual number.:::: ::::table-wrap
::: caption
**The effects of palmvitee supplementation on total white blood cells,
lymphocytes, CD3**^**+**^**cells, B lymphocyte, in whole blood of
nonsmokers and smokers**
:::

**Weeks** **WBC** **Lymphocytes** **CD3+ cells** **B lymphocyte**

______________________________________________________________________

**a.n. x 10**^**9**^**/L** **%** **a.n. x 10**^**9**^**/L** **%** **a.n. x 10**^**9**^**/L** **%** **a.n. x 10**^**9**^**/L**
**Non-smokers (placebo, n = 27)**\
0 7.62 ± 1.89 41.96 ± 9.34 3.12 ± 0.77 63.54 ± 7.03 1.99 ± 0.60 10.19 ± 4.06 0.35 ± 0.26
12 7.29 ± 2.10 42.39 ± 6.12 3.04 ± 0.84 65.96 ± 6.70 2.00 ± 0.56 10.89 ± 3.56 0.33 ± 0.14
24 7.14 ± 1.53 41.85 ± 8.85 2.94 ± 0.72 64.27 ± 7.12 1.88 ± 0.50 11.00 ± 3.88 0.32 ± 0.12
**Non-smokers (palmvitee, n = 29)**\
0 8.32 ± 1.55 40.52 ± 6.17 3.36 ± 0.70 62.07 ± 8.02 2.09 ± 0.52 11.97 ± 4.00 0.40 ± 0.13+
12 8.27 ± 2.08 40.39 ± 6.09 3.33 ± 0.74 62.72 ± 7.60 2.10 ± 0.55 12.45 ± 4.67 0.41 ± 0.16+
24 8.02 ± 1.50 38.05 ± 5.80+ 3.05 ± 0.77 61.66 ± 14.23 1.96 ± 0.55 13.72 ± 4.53+ 0.41 ± 0.15+
**Smokers (placebo, n = 28)**\
0 9.74 ± 2.31 40.44 ± 7.39 3.91 ± 1.14 63.50 ± 7.87 2.47 ± 0.71 15.54 ± 5.06 0.64 ± 0.38
12 9.38 ± 2.45 40.53 ± 8.20 3.76 ± 1.16 63.82 ± 8.34 2.40 ± 0.65 16.14 ± 4.70 0.59 ± 0.33
24 8.81 ± 2.30 38.00 ± 8.68 3.30 ± 1.04 66.11 ± 10.78 2.14 ± 0.63 15.25 ± 4.40 0.52 ± 0.29
**Smokers (palmvitee, n = 30)**\
0 8.43 ± 1.86 39.27 ± 6.20 3.27 ± 0.75 65.77 ± 7.75 2.16 ± 0.69 14.06 ± 3.81 0.45 ± 0.15
12 8.13 ± 2.29 40.07 ± 7.04 3.19 ± 0.72 66.27 ± 6.59 2.13 ± 0.58 14.30 ± 4.09 0.45 ± 0.16
24 8.06 ± 2.10 37.68 ± 7.29 3.01 ± 0.88 66.83 ± 7.28 2.02 ± 0.69 14.40 ± 4.03 0.43 ± 0.16

Results are shown as mean ± SD. + indicates significant difference as
compared to non-smokers taking placebo (p \< 0.05), a.n. represents
actual number.:::: ::::table-wrap
::: caption
**The effects of palmvitee supplementation on total white blood cells,
lymphocytes, CD3**^**+**^**cells, B lymphocyte, in whole blood of
nonsmokers and smokers according to the number of cigarettes per day**
:::

**Weeks** **WBC** **Lymphocytes** **CD3**^**+**^**cells** **B lymphocyte**

______________________________________________________________________

**a.n. x 10**^**9**^**/L** **%** **a.n. x 10**^**9**^**/L** **%** **a.n. x 10**^**9**^**/L** **%** **a.n. x 10**^**9**^**/L**
**S1; 1-9 cigarettes/d (placebo, n = 4)**\
0 9.80 ± 2.07 37.25 ± 8.59 3.79 ± 1.65 59.75 ± 13.25 2.21 ± 0.81 17.75 ± 8.42 0.76 ± 0.70
12 10.05 ± 3.10 38.63 ± 10.59 3.94 ± 2.08 58.00 ± 11.58 2.36 ± 0.93 18.00 ± 6.38 0.81 ± 0.73
24 9.43 ± 2.67 36.05 ± 10.39 3.56 ± 1.92 54.75 ± 21.93 1.91 ± 0.96 15.25 ± 6.90 0.63 ± 0.52
**S1; 1-9 cigarettes/d (palmvitee, n = 3)**\
0 9.87 ± 3.72 38.27 ± 7.15 3.75 ± 1.52 67.67 ± 8.14 2.62 ± 1.40 15.67 ± 2.31 0.56 ± 0.13
12 6.73 ± 1.10 39.57 ± 5.75 2.64 ± 0.35 69.00 ± 6.08 1.80 ± 0.11 17.67 ± 4.04 0.48 ± 0.16
24 6.53 ± 0.76 35.20 ± 11.15 2.29 ± 0.70 65.33 ± 5.77 1.47 ± 0.35 19.00 ± 2.65 0.45 ± 0.18
**S2; 10-19 cigarettes/d (placebo, n = 17)**\
0 9.31 ± 2.53 42.85 ± 6.63 3.96 ± 1.20 64.24 ± 6.28 2.54 ± 0.76 14.82 ± 4.25 0.63 ± 0.37
12 8.92 ± 2.61 41.19 ± 7.34 3.62 ± 0.99 65.71 ± 7.08 2.38 ± 0.62 15.47 ± 3.68 0.52 ± 0.28
24 8.22 ± 2.38 40.26 ± 8.70 3.23 ± 0.95 68.12± 7.03 2.18 ± 0.63 14.82 ± 3.59 0.50 ± 0.2
**S2; 10-19 cigarettes/d (palmvitee, n = 19)**\
0 8.17 ± 1.65 38.59 ± 6.35 3.10 ± 0.54 65.00 ± 8.30 2.00 ± 0.51 13.95 ± 4.24 0.42 ± 0.14
12 8.11 ± 2.36 38.51 ± 7.34 3.02 ± 0.60 65.74 ± 7.34 2.01 ± 0.52 13.63 ± 3.52 0.41 ± 0.14
24 7.64 ± 2.02 37.22 ± 8.70 2.81 ± 0.74 67.26 ± 7.01 1.89 ± 0.55 13.37 ± 3.99 0.38 ± 0.15
**S3; >20 cigarettes/d (placebo, n = 7)**\
0 10.76 ± 1.75 36.42 ± 7.01 3.87 ± 0.81 63.86 ± 8.65 2.46 ± 0.58 16.00 ± 5.16 0.60 ± 0.17
12 10.11 ± 1.58 39.99 ± 9.93 3.99 ± 1.10 62.57 ± 8.83 2.49 ± 0.68 16.71 ± 6.26 0.64 ± 0.23
24 9.90 ± 1.58 33.66 ± 6.62 3.31 ± 0.77 67.71± 7.09 2.18 ± 0.44 16.29 ± 5.22 0.50 ± 0.18
**S3; >20 cigarettes/d (palmvitee, n = 8)**\
0 8.50 ± 1.54 41.23 ± 6.12 3.50 ± 0.70 66.88 ± 7.95 2.37 ± 0.51 13.75 ± 3.97 0.48 ± 0.13
12 8.70 ± 2.06 43.96 ± 6.03 3.79 ± 0.74 66.50 ± 7.53 2.55 ± 0.55 14.63 ± 4.63 0.55 ± 0.16
24 9.64 ± 1.49 39.71 ± 5.75 3.75 ± 0.76 66.38 ± 14.10 2.54 ± 0.54 15.13 ± 4.49 0.56 ± 0.15

Results are shown as mean ± SD.:::: The percentage of NK cells were lower in smokers compared to nonsmokers (p \< 0.0001) at baseline (S1: p \< 0.05, S2: p \< 0.005, S3: p \< 0.01) and after palmvitee supplementation (Table  [8](#) and Table  [9](#)). CD4^+^ in nonsmokers were lower than smokers at the beginning of the study (p \< 0.0001). Total CD4^+^ in smokers were increased according to number of cigarettes per day. CD8^+^ percentage did not show any significant difference among the groups. CD4^+^/CD8^+^ ratio was found to be significantly increased (p \< 0.001) in smokers compared to nonsmokers at baseline (Table  [5](#)) and also in S2. However, no significant changes were observed after palmvitee supplementation for both smokers and nonsmokers (Table  [9](#)). There were also no changes observed in these parameters at the different times of supplementation. ::::table-wrap
::: caption
**The effects of vitamin E supplementation on NK cells,
CD4**^**+**^**cells, CD8**^**+**^**cells and
CD4**^**+**^**/CD8**^**+**^**in whole blood of nonsmokers and smokers**
:::

**Weeks** **NK cells** **CD4**^**+**^ **CD8**^**+**^ **CD4**^**+**^**/CD8**^**+**^

______________________________________________________________________

**%** **a.n. x 10**^**9**^ **%** **a.n. x 10**^**9**^ **%** **a.n. x 10**^**9**^ **%**
**Non-smokers (placebo, n = 27)**\
0 25.00 ± 9.44 0.73 ± 0.34 29.07 ± 6.16 0.91 ± 0.28 31.69 ± 7.41 0.99 ± 0.41 0.99 ± 0.40
12 22.85 ± 7.74 0.70 ± 0.35 30.81 ± 6.75 0.94 ± 0.35 31.19 ± 8.03 1.00 ± 0.36 1.02 ± 0.46
24 24.72 ± 7.94 0.75 ± 0.32 31.27 ± 6.87 0.92 ± 0.32 32.07 ± 8.30 0.94 ± 0.36 1.07 ± 0.48
**Non-smokers (palmvitee, n = 29)**\
0 25.59 ± 8.80 0.86 ± 0.36 27.55 ± 6.68 0.92 ± 0.29 31.38 ± 7.75 1.07 ± 0.30 0.96 ± 0.48
12 24.59 ± 8.45 0.81 ± 0.33 27.62 ± 7.72 0.92 ± 0.33 30.28 ± 6.75 1.03 ± 0.35 0.97 ± 0.47
24 24.72 ± 8.78 0.72 ± 0.42 30.17 ± 9.35 0.90 ± 0.30 31.00 ± 6.72 0.96 ± 0.37 1.03 ± 0.53
**Smokers (placebo, n = 28)**\
0 19.54 ± 7.50+ 0.76 ± 0.39 32.18 ± 5.98+ 1.24 ± 0.44+ 27.82 ± 5.36+ 1.08 ± 0.34 1.22 ± 0.38+
12 19.46 ± 8.55+ 0.75 ± 0.45 33.07 ± 7.32+ 1.23 ± 0.39+ 27.46 ± 6.01+ 1.05 ± 0.36 1.24 ± 0.40+
24 19.18 ± 10.93+ 0.64 ± 0.46 35.14 ± 7.93+ 1.14 ± 0.44+ 28.04 ± 7.34 0.93 ± 0.27 1.25 ± 0.36+
**Smokers (palmvitee, n = 30)**\
0 18.48 ± 6.32++ 0.58 ± 0.17++ 32.20 ± 5.54++ 1.03 ± 0.29 30.37 ± 8.09 1.00 ± 0.45 1.13 ± 0.39
12 17.87 ± 6.50++ 0.55 ± 0.21++ 33.20 ± 5.56++ 1.07 ± 0.29++ 30.17 ± 8.21 0.98 ± 0.42 1.21 ± 0.44++
24 19.03 ± 7.82 0.57 ± 0.27 33.90 ± 5.56++ 1.03 ± 0.37 30.67 ± 7.61 0.92 ± 0.38 1.19 ± 0.43++

Results are shown as mean ± SD. a.n. represents actual number. +
indicates significant difference as compared to nonsmokers taking
placebo (p \< 0.05). ++ indicates significant difference as compared to
nonsmokers taking palmvitee (p \< 0.05).:::: ::::table-wrap
::: caption
**The effects of vitamin E supplementation on NK cells,
CD4**^**+**^**cells, CD8**^**+**^**cells and
CD4**^**+**^**/CD8**^**+**^**in whole blood of nonsmokers and smokers
according to the number of cigarettes per day**
:::

**Weeks** **NK cells** **CD4**^**+**^ **CD8**^**+**^ **CD4**^**+**^**/CD8**^**+**^

______________________________________________________________________

**%** **a.n. x 10**^**9**^ **%** **a.n. x 10**^**9**^ **%** **a.n. x 10**^**9**^ **%**
**S1; 1-9 cigarettes/d (placebo, n = 4)**\
0 21.50 ± 9.57 0.78 ± 0.45 26.25 ± 5.44 0.93 ± 0.22 28.25 ± 9.11 1.09 ± 0.51 1.05 ± 0.50
12 22.00 ± 12.73 0.87 ± 0.65 25.25 ± 3.20 0.96 ± 0.41 27.25 ± 9.25 1.07 ± 0.55 1.02 ± 0.43
24 31.75 ± 22.78 1.09 ± 0.97 23.75 ± 7.27 0.83 ± 0.40 25.50 ± 11.85 0.87 ± 0.42 0.99 ± 0.22
**S1; 1-9 cigarettes/d (palmvitee, n = 3)**\
0 15.67 ± 6.11 0.53 ± 0.04 28.67 ± 3.06 1.07 ± 0.43 32.33 ± 6.66 1.28 ± 0.79 0.90 ± 0.20
12 13.67 ± 1.53 0.37 ± 0.09 34.67 ± 2.08 0.92 ± 0.16 30.67 ± 4.73 0.79 ± 0.46 1.16 ± 0.23
24 18.33 ± 6.43 0.45 ± 0.25 29.67 ± 3.79 0.69 ± 0.26 31.00 ± 6.24 0.68 ± 0.10 0.99 ± 0.29
**S2; 10-19 cigarettes/d (placebo, n = 17)**\
0 19.59 ± 6.75 0.76 ± 0.38 34.18 ± 6.15 1.37 ± 0.52 27.76 ± 4.83 1.07 ± 0.32 1.30 ± 0.39
12 20.12 ± 8.37 0.71 ± 0.39 35.65 ± 7.55 1.29 ± 0.41 27.24 ± 5.84 1.01 ± 0.29 1.33 ± 0.40
24 17.47 ± 6.69 0.55 ± 0.25 37.06 ± 7.04 1.21 ± 0.49 28.94 ± 7.26 0.93 ± 0.25 1.31 ± 0.38
**S2; 10-19 cigarettes/d (palmvitee, n = 19)**\
0 19.11 ± 6.78 0.57 ± 0.20 32.79 ± 4.85 1.02 ± 0.26 29.89 ± 8.38 0.93 ± 0.35 1.15 ± 0.34
12 18.42 ± 7.27 0.53 ± 0.20 33.05 ±5.78 1.01 ± 0.26 30.05 ± 7.71 0.93 ± 0.33 1.20 ± 0.45
24 18.58 ± 7.88 0.52 ± 0.26 34.95 ± 5.02 0.99 ± 0.34 30.74 ± 7.26 0.85 ± 0.28 1.23 ± 0.47
**S3; >20 cigarettes/d (placebo, n = 7)**\
0 18.29 ± 9.01 0.74 ± 0.47 30.71 ± 2.81 1.18 ± 0.21 27.71 ± 5.06 1.08 ± 0.35 1.15 ± 0.30
12 18.43 ± 7.46 0.76 ± 0.52 31.29 ± 4.57 1.21 ± 0.27 28.14 ± 5.30 1.13 ± 0.42 1.14 ± 0.38
24 16.14 ± 5.84 0.61 ± 0.39 37.00 ± 4.86 1.15 ± 0.25 27.29 ± 4.96 1.01 ± 0.45 1.24 ± 0.37
**S3; >20 cigarettes/d (palmvitee, n = 8)**\
0 18.13 ± 8.72 0.60 ± 0.36 32.13 ± 6.63 1.09 ± 0.29 30.75 ± 7.68 1.09 ± 0.37 1.17 ± 0.48
12 18.13 ± 8.37 0.67 ± 0.32 33.00 ± 7.65 1.24 ± 0.33 30.25 ± 6.69 1.17 ± 0.35 1.25 ± 0.46
24 20.38 ± 8.70 0.73 ± 0.42 33.00 ± 9.27 1.24 ± 0.29 30.38 ± 6.66 1.17 ± 0.37 1.19 ± 0.15

Results are shown as mean ± SD.:::: A significant positive correlation was observed in smokers when comparing total white blood cells count and total B cells count at 0 week (r = 0.57, p \< 0.05) (Table  [2](#)). Also a significant positive correlation was found between TWBC count and total CD4^+^ cell count before starting supplementation (r = 0.59, p \< 0.05).


Extracted trial result:
Outcome: Urine cotinine levels (continuous)
Intervention: 200 mg/day palmvitee smokers
Comparator: 200 mg/day palmvitee nonsmokers
Intervention group size: 30
Intervention mean: 1.179
Intervention standard deviation: 0.738
Comparator group size: 19
Comparator mean: 0.047
Comparator standard deviation: 0.017
Notes: Values are for week 24 (only timepoint after intervention).