immunological characteristics of patients infected with common intestinal helminths : result of a...
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Immunological characteristics of patients infected with common intestinal
helminths : result of a study based on reverse-transcriptase PCR
S.Lertanekawattana,T.Wichatrong,K.Chaisari, R.Uchikawa and N.Arizono
Nongkhai Hospital and Department of Medical Zoology,
KyotoPrefectural University of Medicine
Source:Nongkhai Provincial office
Data of intestinal helminths in Nongkhai
year Positive of common parasite
by stool exam.
All age School age
2000 22.0% 19.3%
2002 33.3% 28.2%
Taenia,Hook worm,Opisthochis,Ascaris,Strongyloid
Long-term infection intestinal parasites :
-Malnutrition
-Retardation of the intelectual
-Impair physical growth of children
Oberhelman et al.,1998:
Helminth infection :
-Impair immune response to virus,bacteria protozoa and even oral vaccine
( Cooper et al, 2001)
Ascaris and Hook worm induce expansion of the Th2 lymphocyte subset and regulate IgE antibody production, eosinophilia and mastocytosis
(Pritchard et al,1995)
Th2 cytokines have antagonistic effects on the expansion of the Th1 lymphocyte subset
(Cooper et al,2001)
Anisakiasis
Clinical : Urticaria,Angioedema,
Anaphylaxis
Immunological response :
IgE , mast cell, basophil
Th 2 interleukin 4,predominant
• Nippostrongylus brasiliensis
Villus pathology induced by the rodent
nematode Nippostrongylus brasiliensis
E-cadherin expression decreases at villus tips after infection
days after infection
281410701
2
3
4
Day 0 Day 10
E-cadherin immunohistochemistry
E-cadherin score
Pathological changes develop in the intestinal mucosa,including partial villus atrophy and crypt hyperplasia,decreasing of sucrase and alkaline phosphatase activity,deterioration of epithelial permeability and barrier function
(Hyoh et al,2002 ; Sekikawa et al;2003)
**
glucose
glucose
glucose
Na
Na
Na
K
SGLT1 (Na-driven glucose cotransporter)
Na-K ATPase
Lumen
Lamina propria
Absorptivecell
Glucose transport in the small intestine
Infection with intestinal nematodes in rodents causes induction Th2 cytokines such as IL-4.IL-5and IL-13 and induce IgE antibody responses, eosinophilia and intestinal mastocytosis
(Matsuda et al, 1995 ; Garside et al,2000)
Aim of study :
To determine whether common helminth infections could modify the intestinal immunopathological status of the host,by the expression in the human duodenal mucosa of cytokines,eosinophil,mast cell specific molecule,monosaccharide transporters of the glucose-transporter (GLUT)
Subjects :
31 patients with gastrointestinal problems and indicated for gastroduodenal fiberscopy as diagnotic measurement and did not have dysenteric disease and gave their written,informed consent for the study
Density analysis of PCR products
Electrophoresis on agar each band was evaluated using NIH Image software, the band densities were normalized relative to those of ß-actin
Table1 The primer pairs used for the PCR amplifications Size of expected product(bp)
IFN- 5'-GGGTTCTCTTGGCTGTTACTG-3‘5'-GACAGTTCAGCCATCACTTGGA-3'
384
IL-5 5'-GAAATTCCCACAAGTGCATTGG-3'5'-CTTTCTATTATCCACTCGGTGTTC-3'
335
IL-13 5'-AGGAGCTGGTCAACATCACC-3'5'-GTTGAACCGTCCCTCGCGAA-3'
296
FceRI1 5'-TCAGTGACTGGCTGCTCCTT-3'5'-GGGGTTTGGCTTAGGATGTG-3'
437
McTr2 5'-TCAGCAGGATCATCGTGCAC-3'5'-TGGGGACATAGTGGTGGATC-3'
507
CLC3 5'-TACCCGTGCCATACACAGAG-3'5'-CTCTCCACACTTGCACCATC-3'
375
GLUT-1
5'-ATCGTCAACACGGCCTTCAC-3'5'-AAGCCGGAAGCGATCTCATC-3'
458
GLUT-5 5'-GGTACAACGTGGCTGCTGTC-3'5'-CATGGGGACCACGTTGGAAG-3'
347
-Actin 5'-TCAGAAGGATTCCTATGTGGGC-3'5'-CCATCACGATGCCAGTGGTA-3'
317
Table 2 The baseline demographic and laboratory data for the 31 subjects
Total
N=31
Helminth
infected
9
Uninfected
22
Ulcer
6
Non-ulcer
25
Male/female 22/9 9/0 13/9 5/1 17/8
Age(years) 45.8 ( 13.5)(31)
43.3 (10.8)(9)
46.8(14.5)(22)
47.8(8.9)(6) 45.3 (14.6)(25)
Albumin(g/dl) 3.9(0.7)(23) 4.1(0.6)(8) 3.8(0.7)(15) 3.5(1.0)(5) 4.0(0.6)(18)
Haemoglobin (g/dl)
13.3(2.8)(29) 13.5(2.2)(8) 13.0(3.0)(21) 12.3(4.1)(6) 13.4(2.4)(23)
Eosinophils(%0f leucocyte)
4.8(4.6)(31) 5.3(4.6)(9) 4.6(4.7)(22) 3.1(3.2)(6) 5.2(4.9)(25)
Figure 1 Some of PCR results,showing the expression of IL-13,IL-5,IFN-,andfor reference,-actin in the duodenal mucosa of the 31 patients
Figure 2 (a) The frequencies of expression of IL-5( )and IL-13 ( )
In the duodenal mucosa of the 10 patients with eosinophilia and the 21 without
Figure 3 (b) In the 9 patients with helminth infection(on presentation or in the previous 3 months ) and the 22 without
P=0.041
Table 3 Expression levels of IL-5, IFN-, CLC, FcRI, McTr, GLUT1 and GLUT5 in the duodenal mucosa.
Non-eosinophilics Eosinophilics
Total
(N=21)
Uninfected
(N=16)
Infected
(N=5)
Total
(N=10)
Uninfected
(N=6)
Infected
(N=4)
IL-5 0.21(0.09) 0.14(0.08) 0.45(0.33) 0.36(0.10) 0.33(0.11) 0.39(0.24)
IFN- 0.65(0.11) 0.66(0.14) 0.62(0.17) 1.20(0.22)* 1.32(0.28)* 1.02(0.42)
IL-5/ IFN- 0.27(0.09) 0.15(0.06) 0.65(0.29)† 0.41(0.17) 0.39(0.24) 0.45(0.25)
CLC 0.69(0.10) 0.66(0.11) 0.78(0.31) 0.74(0.19) 0.92(0.27) 0.48(0.21)
FcRI 0.52(0.11) 0.42(0.09) 0.82(0.32) 0.58(0.09) 0.56(0.12) 0.60(0.14)
McTr 2.16(0.19) 1.98(0.18) 2.76(0.52) 2.79(0.44) 3.41(0.57) 1.87(0.13)
FcRI/McTr 0.21(0.04) 0.20(0.04) 0.26(0.06) 0.23(0.05) 0.16(0.03) 0.33(0.09)‡
GLUT1 1.17(0.15) 1.13(0.20) 1.23(0.17) 0.90(0.23) 1.01(0.32) 0.74(0.32)
GLUT5 1.17(0.09) 1.12(0.10) 1.32(0.25) 1.25(0.13) 1.41(0.17) 1.00(0.12)
Table 4 The results of the principal-components analysis
Parameter Value
Eigenvalue(and percentage of variation explained) 2.13966(42.8)
Unrotated facter loadings
Eosinophils
Interleukin-5
Interferon- IgE receptor type (FcRI)
Mast cell tryptase (McTr)
-0.55202
-0.43733
-0.72827
-0.61319
-0.85866
Mean (S.E.) component scores
Others (uninfected patients without eosinophilia;N=16)
Helminth-infected (N=9)
Eosinophilia (uninfected patients with eosinophilia; N=6)
0.761(0.283)
-0.327(0.375)
-1.538(0.733)
With intestinal helminth infection on presentation or within previous 3 months.
Significantly different to other component scores (p<0.05)
Significantly different to other component scores (p<0.01)
Summary:
The present findings indicate that infections with common intestinal helminths, including Taeniasis are likely to modify mucosal immunology so that it shifts to a Th2-predominant status.
SGUT-1 and GLUT-5 no significant decrease in patients with helminth infection
Acknowledgment:
This work was supported by Grants-in-Aid for Research from the Japan Society for the Promotion of Science and the Japanese Ministry of Health,Labour and Welfare