Development of a new method for the determination of immune responses in the human stomach

Bergquist, Charlotta; Mattsson-Rydberg, A.; Lönroth, Hans; Svennerholm, Ann‐Mari

doi:10.1016/s0022-1759(99)00213-6

Cited by 20 publications

(17 citation statements)

References 22 publications

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“…Proteins were extracted from biopsies as previously described [49]. CXCL11, CCL17, and CCL22 concentrations were determined by Duoset ELISA (R&D systems, Abingdon, United Kingdom).…”

Section: Methodsmentioning

confidence: 99%

Accumulation of CCR4+ CTLA-4hi FOXP3+CD25hi Regulatory T Cells in Colon Adenocarcinomas Correlate to Reduced Activation of Conventional T Cells

et al. 2012

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BackgroundColorectal cancer usually gives rise to a specific anti-tumor immune response, but for unknown reasons the resulting immunity is not able to clear the tumor. Recruitment of activated effector lymphocytes to the tumor is important for efficient anti-tumor responses, while the presence of regulatory T cells (Treg) down-modulate tumor-specific immunity. We therefore aimed to determine homing mechanisms and activation stage of Treg and effector T cell infiltrating colon tumors compared to cells from the unaffected mucosa in patients suffering from colon adenocarcinoma.Methodology/Principal FindingsLymphocytes were isolated from unaffected and tumor mucosa from patients with colon adenocarcinoma, and flow cytometry, immunohistochemistry, and quantitative PCR was used to investigate the homing mechanisms and activation stage of infiltrating Treg and conventional lymphocytes. We detected significantly higher frequencies of CD25highFOXP3+CD127low putative Treg in tumors than unaffected mucosa, which had a complete demethylation in the FOXP3 promotor. Tumor-associated Treg had a high expression of CTLA-4, and some appeared to be antigen experienced effector/memory cells based on their expression of αEβ7 (CD103). There were also significantly fewer activated T cells and more CTLA-4+ conventional T cells susceptible to immune regulation in the tumor-associated mucosa. In contrast, CD8+granzyme B+ putative cytotoxic cells were efficiently recruited to the tumors. The frequencies of cells expressing α4β7 and the Th1 associated chemokine receptor CXCR3 were significantly decreased among CD4+ T cells in the tumor, while frequencies of CD4+CCR4+ lymphocytes were significantly increased.Conclusions/SignificanceThis study shows that CCR4+CTLA4hi Treg accumulate in colon tumors, while the frequencies of activated conventional Th1 type T cells are decreased. The altered lymphocyte composition in colon tumors will probably diminish the ability of the immune system to effectively attack tumor cells, and reducing the Treg activity is an important challenge for future immunotherapy protocols.

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“…Proteins were extracted from biopsies as previously described [49]. CXCL11, CCL17, and CCL22 concentrations were determined by Duoset ELISA (R&D systems, Abingdon, United Kingdom).…”

Section: Methodsmentioning

confidence: 99%

Accumulation of CCR4+ CTLA-4hi FOXP3+CD25hi Regulatory T Cells in Colon Adenocarcinomas Correlate to Reduced Activation of Conventional T Cells

et al. 2012

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“…Resuspended LPLs and PBMCs were used immediately for detecting antigen-specific IgG and IgA ASCs by ELISPOT assay. Duodenal extracts were prepared as described previously (1). In brief, a single biopsy specimen was snap-frozen at Ϫ70°C at each time point.…”

Section: Methodsmentioning

confidence: 99%

Mucosal Immunologic Responses in Cholera Patients in Bangladesh

Uddin

Harris

Bhuiyan

et al. 2011

Clin Vaccine Immunol

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Vibrio cholerae O1 causes dehydrating diarrhea with a high mortality rate if untreated. The infection also elicits long-term protective immunity. Since V. cholerae is noninvasive, mucosal immunity is likely important for protection. In this study, we compared humoral immune responses in the duodenal mucosa and blood of cholera patients at different time points after the onset of disease and compared them with those of healthy controls (HCs). Immune responses to lipopolysaccharide (LPS) and the recombinant cholera toxin B subunit (rCTB) were assessed by enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunospot (ELISPOT) assay. Significant increases in V. cholerae LPS-specific IgA and IgG antibody levels were seen in duodenal extracts on day 30, but the levels decreased to baseline by day 180; plasma V. cholerae LPS-specific IgA levels remained elevated longer. Levels of mucosal CTB antibodies also peaked on day 30, but the increase reached statistical significance only for IgG. A significant correlation was found between the CTB antibodysecreting cell (ASC) response in the circulatory system on day 7 and subsequent CTB-specific IgA levels in duodenal extracts on day 30 and the numbers of CTB-specific IgA ASCs in duodenal tissues on day 180. The proportion (0.07%) of mucosal V. cholerae LPS IgA ASCs peaked on day 30 and remained elevated through day 180 compared to that of HCs (P ‫؍‬ 0.03). These results suggest that protective immunity against V. cholerae is not likely mediated by the constitutive secretion of antibodies at the mucosal surface; our results are consistent with those of other studies that suggest instead that anamnestic immune responses of mucosal lymphocytes may play a major role in protection against cholera.

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“…To determine the types of Th responses induced following exposure to a low-dose Helicobacter inoculum, cytokine assays were performed on saponized gastric extracts from the animals using a modification of the technique of Bergquist et al (2). Gamma interferon (IFN-␥) and interleukin-4 (IL-4) levels in gastric tissue samples were assayed by a double sandwich enzyme-linked immunosorbent assay technique (Pharmingen Inc., San Diego, Calif.).…”

Section: Figmentioning

confidence: 99%

Effect of Low-Dose Antigen Exposure on Development of Immunity to Helicobacter pylori Infection in Mice

Radcliff

Ferrero

2001

Infect Immun

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Development of a new method for the determination of immune responses in the human stomach

Cited by 20 publications

References 22 publications

Accumulation of CCR4+ CTLA-4hi FOXP3+CD25hi Regulatory T Cells in Colon Adenocarcinomas Correlate to Reduced Activation of Conventional T Cells

Accumulation of CCR4+ CTLA-4hi FOXP3+CD25hi Regulatory T Cells in Colon Adenocarcinomas Correlate to Reduced Activation of Conventional T Cells

Mucosal Immunologic Responses in Cholera Patients in Bangladesh

Effect of Low-Dose Antigen Exposure on Development of Immunity to Helicobacter pylori Infection in Mice

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