Scientific journal
European Journal of Natural History
ISSN 2073-4972

IMMUNE SYSTEM STATE IN CHRONIC CHOLECYSTITIS AND METABOLIC SYNDROME PATIENTS

Yurenko A.V., Antonyuk M.V., Khodosova K.K., Demyanenko N.B.
On the ground of 127 chronic cholecystitis patients’ examination results the immunological aspects of metabolic syndrome and chronic cholecystitis comorbid course were elucidated. The research results showed that in the patients with chronic cholecystitis the immune status changes take place together with the development of metabolic disorders (dyslipodemia, insulin-resistance) due to suppressing the cellular component of the immune system, nonspecific resistance factors’ stimulation, cytokine level increase.

The development of chronic cholecystitis is conditioned, as a rule, by the bacterial flora, the penetration of which into the bile cyst happens by the enterogenous, hematogenous or lymphogenous ways [1]. The chronic inflammation at the chronic cholecystitis long course results in the metabolic and immune status disorders. In spite of the fact that cytokines are not specific factors of inflammation the determination of their concentration in blood gives the information about functional activity of various types of immunocompetent cells, the inflammatory process severity and disease prognostication [6].

The connection between the immune state and metabolic syndrome (MS), which often takes course against the background of chronic cholecystitis, is widely discussed in literature. At that, the data got by different investigators are rather contradictory [10].

Up to the present day the immunological aspects at the MS and chronic cholecystitis (CC) comorbid course remain one of the poorly studied problems.

The purpose of the research is to evaluate the immune system state in the patients with chronic cholecystitis at the MS formation.

127 persons aged from 20 to 55 years old, 39 men and 88 women, took part in the investigation. According to the research tasks three study groups were formed. The first group (control) was represented by 32 patients without chronic cholecystitis and MS; the second group - were 49 patients with chronic cholecystitis without MS manifestations and the third group - 49 patients with chronic cholecystitis and MS. Diabetes was diagnosed in 15%, the overweight - in 30%, adiposis - in 65% of the third group patients, in 82% of the patients all these changes were combined with hypertension.

The chronic cholecystitis diagnosis was made on the basis of clinical, laboratorial and functional research methods. The chronic non-calculous cholecystitis was detected in 75 patients, the chronic calculous cholecystitis - in 20 patients. The metabolic syndrome was diagnosed according to the USA National Committee criteria on cholesterol (ATR, 2001) [5].

In blood serum the level of total cholesterol, high-density lipoprotein cholesterol, triglycerides («OLVEX DIAGNOSTICUM» sets), apo-A1 and apo-B («DiaSys» sets) were determined. The apo-protein atherogenicity coefficient was calculated on the quotient of apo-proteins - apo-B to apo-A1 [7]. The carbohydrate metabolism investigation included the detection of glucose content in blood serum on the empty stomach, insulin - by the enzyme multiplied immunoassay method («DRG Diagnostics» sets). To determine the insulin resistance the NOMA index was used [9]. For the purpose of inflammation markers detecting in blood serum the tumor necrosis factor level - alpha (TNF- α) and receptor to TNF- α (TNF RI), were determined by the enzyme multiplied immunoassay method (the chemical reagents of the "BD Bioscience" firm). The peripheral blood immunocompetent cells phenotyping was carried out using monoclonal antibodies to the cells CD3, CD4, CD8, CD 16, CD 22, CD 25, HLA-DR (Vitebsk) [3]. To determine the nonspecific resistance of the body the immune system monocytic macrophageal element cells´ functional abilities were investigated. The neutrophils´ metabolic activity study was carried out by means of the Nitro Blue Tetrazolium Reduction Test (NTR-test), the Nitro Blue Tetrazolium Reduction Test reserve (NTRR), the neutrophils´ activation index (NAI) and the neutrophils´ activation index reserve (NAIR) on the method of Park B.H. in the modification of Shmelev Ye.V. [12]. The A, M, G immunoglobulines concentration was determined in blood serum by the enzyme multiplied immunoassay ("Vector-Best" sets). The detection of the circulating immune complexes (CIC) of large (C3) and small (C4) sizes was carried out by the method of M. Digeon, M. H. Jover, J. Rizo in the modification of Struchkov P.V. [9]. As the pathogenicity factor their quotient (К = С4/С3) was used. At the static processing of the data the Student´s t-test was determined; the differences were considered authentic at p<0,05.

In the patients with CC and MS the level of arterial blood tension, body-weight index, waist and thighs circuit, and also the studied factors of lipid and carbohydrate metabolism differed from the analogous factors in the first and second groups (p<0,05).

The immunological status analysis revealed a tendency to the immune system imbalance formation in chronic cholecystitis patients at metabolic disorders augmentation. As it is seen from table 1, in the second group patients the T suppressors´ quantity reduction, tellingly for the chronic inflammatory process, takes place against the background of the T helpers´ quantity reduction. In the third group patients there was a tendency to the "adult" T lymphocytes´ (CD3), T helpers´ (CD4) quantity reduction and cytotoxic T lymphocytes´ (CD8) quantity increase compared to the control group marked. The CD4/CD8 index fell in the third group patients compared to the 1st and 2nd groups.

In the third group patients there was a modest rise of the B "adult" lymphocytes´ level against Ig G level increase (р<0,05) registered compared to the 1st and 2nd groups.

The given changes testify to the immune resistance suppression in CC and MS patients. On the one hand, the humoral immunity activation happens in this group; on the other hand - the immune system cellular component´s functional abilities decrease takes place.

In the patients of the 2nd and 3rd groups there was an increase of the level of the cells containing receptors to the interleukin-2 (IL-2) (р<0,05) compared to the control group registered, that reflects the production stimulation of IL-2 promoting, in its turn, the activation and maturation of different subpopulations of T-cells, B-cells, natural killers and macrophages. The decrease of the "adult" T lymphocytes´, T helpers´ CD4/CD8 index´, natural killers´ (CD 16) level in the third group patients testifies to a negative activation of lymphocytes at the MS formation against the background of chronic cholecystitis [4].

The results of antigens´ state study are of concern. The HLA-DR increase in the second and third groups compared to the control was registered (p<0,05). There are no analogous investigations of the HLA antigens´ state in the CC and MS patients described in the works of other authors. The findings, in the course of which the associative relations between the HLA antigens and various diseases were set, it letting high risk groups be emphasized, the preconditions for various pathological forms combination be found out and the course be prognosticated; are submitted in literature [4].

Table 1. Immunity factors in chronic cholecystitis patients

Factors

1 group, control, n=32

2 group,

n=49

3group,

 n=46

Leukocytes, g/l

5,55±0,27

5,60±0,28

6,02±0,34

Lymphocytes, %

27,70±1,38

27,85±1,3

26,2±1,48

Т - lymphocytes "adult" (CD3), %

33,39±2,19

30,24±1,55

29,97±1,55

Т -helpers (CD4), %

33,30±1,85

29,42±1,66

27,83±1,57

р 3-1< 0,05

Т -suppressors (CD8), %

21,35±0,91

19,42±0,93

23,53±1,47

р 2-3 < 0,05

CD4/ CD8 index

1,50±0,06

1,55±0,08

1,24±0,07

р 3-1, р 2-3 < 0,05

B - lymphocytes "adult" (CD22), %

24,26±1,9

24,18±1,33

26,03±1,79

Natural killers (CD16), %

19,74±1,43

20,33±1,13

19,8±1,34

IL-2-receptor bearing Т- and В-lymphocytes (early stage of activation) (CD 25), %

11,26±0,86

18,09±1,39

р 2-1 < 0,05

18,47±1,52

р 3-1 < 0,05

Activated Т- and В-lymphocytes (late activation stage), bearing receptors HLA-DR , %

13,61±0,81

18,55±1,44

р 2-1 < 0,05

16,69±1,07

р 3-1< 0,05

Oxidative metabolism of neutrophils (NTR-test), %

10,14±0,8

12,94±1,55

16,55±1,48

р 3-1< 0,05

NTR reserve,c.u.

1,30±0,09

1,12±0,08

1,09±0,1

Neutrophils´ activation index (NAI), %

0,17±0,05

0,24±0,06

0,19±0,02

NAI reserve, c.u.

1,32±0,09

1,13±0,08

1,07±0, 1

Ig A, mg/ml

1,63±0,1

1,88±0,07

р 2-1 < 0,05

1,64±0,1

р 3-2< 0,05

Ig M, mg/ml

1,25±0,1

1,22±0,08

1,36±0,08

Ig G, mg/ml

9,70±0,2

9,84±0,18

10,41±0,2

р 3-1, р 2-3 < 0,05

CIC: С3, c.u.

0,44±0,02

0,52±0,04

0,49±0,03

CIC: С4, c.u.

0,51±0,02

0,55±0,03

0,56±0,03

К, c.u.

1,17±0,04

1,12±0,03

1,15±0,04

TNF-α, pg/ml

4,88±0,17

5,68±0,38

6,62±0,35

р 3-1 < 0,05

TNF RI pg/ml

1039,1±209,11

1926,35±132,7

р 2-1 < 0,05

2520,2±123,36

р 3-1, р 3-2 < 0,05

Note: р 3-1, р 2-1 -reliability of distinctions between factors in comparison with the control; р 2-3 - reliability of distinctions between the 2nd and 3rd group.

The state of non-specific resistance factors was characterized by the neutrophils´ oxidative metabolism activity (NTR) increase with the decrease of neutrophilic granulocytes´ reserve opportunities and increase of C3 levels by 11% and C4 CIC by 8% in the patients of the 2nd and 3rd groups compared to the control (p<0,05). An authentic increase of the TNF- α factors and TNF- α receptors is registered in the CC and MS patients compared to the control (p<0,05). The anti-inflammatory cytokines´ level increase and neutrophils´ oxidative metabolism activation is considered by many authors as the factor of vessels´ endothelium injury through the intensification of oxidation processes resulting in nitrogen oxide inactivation, the release of the enzymes, which catalyze the formation of kinins, which promote vascular permeability and change the vascular wall structure [11, 2].

Thus, the research results testified that in CC patients together with the development of metabolic disorders (dyslipodemia, insulin-resistance) the immunological status changes take place due to the immunity due to suppressing the immune system cellular component, nonspecific resistance factors´ stimulation, cytokine level increase. The revealed changes reflect systemic reactions of the body at chronic inflammation and are associated with metabolic disorders resulting in the development of MS clinical implications - hypertension, diabetes, in chronic cholecystitis patients.

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