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Association of Intact dupA (dupA1) rather than dupA1 cluster with duodenal ulcer in Indian population.

Alam J, Ghosh P, Ganguly M, Sarkar A, De R, Mukhopadhyay AK - Gut Pathog (2015)

Bottom Line: The distribution of long and short type dupA1 has not been significantly associated with the disease outcome.The intact dupA1 was significantly associated with DU than NUD (P = 0.029) but the dupA1 cluster has no role in the disease manifestation at India (P = 0.79).Thus, dupA1 can be considered as a biomarker for DU patients in India.

View Article: PubMed Central - PubMed

Affiliation: Division of Bacteriology, National Institute of Cholera and Enteric Diseases, P 33, CIT Road, Scheme, XM Beliaghata.

ABSTRACT

Background: The duodenal ulcer promoting gene (dupA) and dupA cluster in Helicobacter pylori have been described as a risk factor for duodenal ulcer development in some populations. Polymorphic gene dupA can be divided into two groups, intact dupA1 (long or short type based on the presence or absence of 615-bp extra sequences at the 5' region) having complete reading frame and other truncated dupA2 having frame-shift mutation. This study was aimed to elucidate the role of dupA of H. pylori and their clusters in the disease manifestation of Indian population.

Methods: A total of 170 H. pylori strains were screened for the presence of dupA, dupA alleles and dupA cluster by PCR and sequencing. Pro-inflammatory cytokine (IL-8) with different dupA variant H. pylori stimulated gastric epithelial cells (AGS cells) was measured by ELISA.

Results: A total of 50 strains (29.4%) were positive for dupA among the tested 170 strains. The prevalence of dupA1 in duodenal ulcer (DU) and non-ulcer dyspepsia (NUD) populations was found to be 25.5% (25/98) and 11.1% (8/72), respectively and 16.4% (28/170) of the tested strains had dupA1, cagA and vacAs1m1 positive. The distribution of long and short type dupA1 has not been significantly associated with the disease outcome. The dupA cluster analysis showed that 10.2% (10/98) and 8.3% (6/72) strains were positive among DU and NUD, respectively. IL-8 production was significantly higher in dupA1(+) , cagA (+), vacA (+) (902.5 ± 79.01 pg/mL) than dupA2 (+) , cagA (+) , vacA (+) (536.0 ± 100.4 pg/mL, P = 0.008) and dupA (-), cagA (+), vacA (+) (549.7 ± 104.1 pg/mL, P = 0.009). Phylogenetic analysis of dupA indicated that the Indian H. pylori strains clustered with East Asian strains but distinct from Western strains. This is the first known genetic element of Indian H. pylori that is genetically closer to the East Asian strains but differed from the Western strains.

Conclusions: The intact dupA1 was significantly associated with DU than NUD (P = 0.029) but the dupA1 cluster has no role in the disease manifestation at India (P = 0.79). Thus, dupA1 can be considered as a biomarker for DU patients in India.

No MeSH data available.


Related in: MedlinePlus

In vitro IL-8 production from AGS cells co-cultured with 6 (dupA1+,cagA+,vacA+) strains, 6 (dupA2+,cagA+,vacA+) and 6 (dupA−,cagA+,vacA+) strains. IL-8 production was significantly higher in dupA1+ (cagA+, vacA+) compared to dupA2+ (cagA+, vacA+) (536.0 ± 100.4 pg/mL, P = 0.008) and dupA-negative strains (cagA+, vacA+) (549.7 ± 104.1 pg/mL, P = 0.009). MOI of H. pylori strain was 100 for 8 hours. Experiments were repeated 3 times for the 18 strains. IL-8 in the supernatant was assayed by an ELISA in duplicate. Data are expressed as mean ± standard error.
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Fig2: In vitro IL-8 production from AGS cells co-cultured with 6 (dupA1+,cagA+,vacA+) strains, 6 (dupA2+,cagA+,vacA+) and 6 (dupA−,cagA+,vacA+) strains. IL-8 production was significantly higher in dupA1+ (cagA+, vacA+) compared to dupA2+ (cagA+, vacA+) (536.0 ± 100.4 pg/mL, P = 0.008) and dupA-negative strains (cagA+, vacA+) (549.7 ± 104.1 pg/mL, P = 0.009). MOI of H. pylori strain was 100 for 8 hours. Experiments were repeated 3 times for the 18 strains. IL-8 in the supernatant was assayed by an ELISA in duplicate. Data are expressed as mean ± standard error.

Mentions: In general, there was no difference in IL-8 production between dupA-positive but truncated (dupA2) and dupA-negative strains when tested in AGS cells co-cultured with H. pylori. However, IL-8 production was significantly higher (902.5 ± 79.01 pg/mL) in the strains with an intact dupA1 compared with a truncated dupA2 (536.0 ± 100.4 pg/mL, P = 0.008) or with dupA-negative strains (549.7 ± 104.1 pg/mL, P = 0.009) [Figure 2]. All the strains taken for IL-8 assay were positive for cagA and vacA which helped us to focus specifically the role of different dupA groups.Figure 2


Association of Intact dupA (dupA1) rather than dupA1 cluster with duodenal ulcer in Indian population.

Alam J, Ghosh P, Ganguly M, Sarkar A, De R, Mukhopadhyay AK - Gut Pathog (2015)

In vitro IL-8 production from AGS cells co-cultured with 6 (dupA1+,cagA+,vacA+) strains, 6 (dupA2+,cagA+,vacA+) and 6 (dupA−,cagA+,vacA+) strains. IL-8 production was significantly higher in dupA1+ (cagA+, vacA+) compared to dupA2+ (cagA+, vacA+) (536.0 ± 100.4 pg/mL, P = 0.008) and dupA-negative strains (cagA+, vacA+) (549.7 ± 104.1 pg/mL, P = 0.009). MOI of H. pylori strain was 100 for 8 hours. Experiments were repeated 3 times for the 18 strains. IL-8 in the supernatant was assayed by an ELISA in duplicate. Data are expressed as mean ± standard error.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4379697&req=5

Fig2: In vitro IL-8 production from AGS cells co-cultured with 6 (dupA1+,cagA+,vacA+) strains, 6 (dupA2+,cagA+,vacA+) and 6 (dupA−,cagA+,vacA+) strains. IL-8 production was significantly higher in dupA1+ (cagA+, vacA+) compared to dupA2+ (cagA+, vacA+) (536.0 ± 100.4 pg/mL, P = 0.008) and dupA-negative strains (cagA+, vacA+) (549.7 ± 104.1 pg/mL, P = 0.009). MOI of H. pylori strain was 100 for 8 hours. Experiments were repeated 3 times for the 18 strains. IL-8 in the supernatant was assayed by an ELISA in duplicate. Data are expressed as mean ± standard error.
Mentions: In general, there was no difference in IL-8 production between dupA-positive but truncated (dupA2) and dupA-negative strains when tested in AGS cells co-cultured with H. pylori. However, IL-8 production was significantly higher (902.5 ± 79.01 pg/mL) in the strains with an intact dupA1 compared with a truncated dupA2 (536.0 ± 100.4 pg/mL, P = 0.008) or with dupA-negative strains (549.7 ± 104.1 pg/mL, P = 0.009) [Figure 2]. All the strains taken for IL-8 assay were positive for cagA and vacA which helped us to focus specifically the role of different dupA groups.Figure 2

Bottom Line: The distribution of long and short type dupA1 has not been significantly associated with the disease outcome.The intact dupA1 was significantly associated with DU than NUD (P = 0.029) but the dupA1 cluster has no role in the disease manifestation at India (P = 0.79).Thus, dupA1 can be considered as a biomarker for DU patients in India.

View Article: PubMed Central - PubMed

Affiliation: Division of Bacteriology, National Institute of Cholera and Enteric Diseases, P 33, CIT Road, Scheme, XM Beliaghata.

ABSTRACT

Background: The duodenal ulcer promoting gene (dupA) and dupA cluster in Helicobacter pylori have been described as a risk factor for duodenal ulcer development in some populations. Polymorphic gene dupA can be divided into two groups, intact dupA1 (long or short type based on the presence or absence of 615-bp extra sequences at the 5' region) having complete reading frame and other truncated dupA2 having frame-shift mutation. This study was aimed to elucidate the role of dupA of H. pylori and their clusters in the disease manifestation of Indian population.

Methods: A total of 170 H. pylori strains were screened for the presence of dupA, dupA alleles and dupA cluster by PCR and sequencing. Pro-inflammatory cytokine (IL-8) with different dupA variant H. pylori stimulated gastric epithelial cells (AGS cells) was measured by ELISA.

Results: A total of 50 strains (29.4%) were positive for dupA among the tested 170 strains. The prevalence of dupA1 in duodenal ulcer (DU) and non-ulcer dyspepsia (NUD) populations was found to be 25.5% (25/98) and 11.1% (8/72), respectively and 16.4% (28/170) of the tested strains had dupA1, cagA and vacAs1m1 positive. The distribution of long and short type dupA1 has not been significantly associated with the disease outcome. The dupA cluster analysis showed that 10.2% (10/98) and 8.3% (6/72) strains were positive among DU and NUD, respectively. IL-8 production was significantly higher in dupA1(+) , cagA (+), vacA (+) (902.5 ± 79.01 pg/mL) than dupA2 (+) , cagA (+) , vacA (+) (536.0 ± 100.4 pg/mL, P = 0.008) and dupA (-), cagA (+), vacA (+) (549.7 ± 104.1 pg/mL, P = 0.009). Phylogenetic analysis of dupA indicated that the Indian H. pylori strains clustered with East Asian strains but distinct from Western strains. This is the first known genetic element of Indian H. pylori that is genetically closer to the East Asian strains but differed from the Western strains.

Conclusions: The intact dupA1 was significantly associated with DU than NUD (P = 0.029) but the dupA1 cluster has no role in the disease manifestation at India (P = 0.79). Thus, dupA1 can be considered as a biomarker for DU patients in India.

No MeSH data available.


Related in: MedlinePlus