source:nongkhai provincial office
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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 - PowerPoint PPT PresentationTRANSCRIPT
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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
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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
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Long-term infection intestinal parasites :
-Malnutrition
-Retardation of the intelectual
-Impair physical growth of children
Oberhelman et al.,1998:
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Helminth infection :
-Impair immune response to virus,bacteria protozoa and even oral vaccine
( Cooper et al, 2001)
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Ascaris and Hook worm induce expansion of the Th2 lymphocyte subset and regulate IgE antibody production, eosinophilia and mastocytosis
(Pritchard et al,1995)
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Th2 cytokines have antagonistic effects on the expansion of the Th1 lymphocyte subset
(Cooper et al,2001)
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Anisakiasis
Clinical : Urticaria,Angioedema,
Anaphylaxis
Immunological response :
IgE , mast cell, basophil
Th 2 interleukin 4,predominant
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• Nippostrongylus brasiliensis
Villus pathology induced by the rodent
nematode Nippostrongylus brasiliensis
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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
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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)
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**
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
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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)
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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)
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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
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Materials:
Duodenal biopsy specimens
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Methods :
Semi-quantitative RT-PCR for IL-5 ,IL-13, IFN- , GLUT-1 and GLUT-5
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Laboratory study:
At central lab. of Kyoto Prefectural University of Medicine
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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
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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
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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)
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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
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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
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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
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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)
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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)
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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
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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
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