Distinct Adaptive Immunophenotype in duodenal mucosa but not peripheral blood of patients with functional dyspepsia

Background and aims: Functional dyspepsia is characterised by chronic symptoms of post-prandial distress or epigastric pain not associated with defined structural pathology. Increased peripheral gut-homing T cell have been previously identified in patients. To date, it is unknown if these T cells were antigen-experienced, or if a specific immunophenotype was associated with FD. This study aimed to characterise immune populations in the blood and duodenal mucosa of FD patients that may be implicated in disease pathophysiology. Methods: We identified duodenal T cell populations from 23 controls and 49 Rome III FD patients by flow cytometry. We also analysed duodenal eosinophils and T cell populations in peripheral blood from 37 controls and 49 patients and investigated if subtyping patients based on reported symptoms or co-morbidity identified specific immunophenoptypes. Results: In addition to increased duodenal mucosal CD4+ effector cells, FD patients demonstrated a shift in the T helper cell balance compared to controls. Patients had increased duodenal mucosal Th2 populations in the effector (13.03±16.11, 19.84±15.51, p=0.038), central memory (23.75±18.97, 37.52±17.51, p=0.007) and effector memory (9.80±10.50 vs 20.53±14.15, p=0.001) populations. Th17 populations were also increased in the effector (31.74±24.73 vs 45.57±23.75, p=0.03) and effector memory (11.95±8.42 vs 18.44±15.63, p=0.027) subsets. Conclusion: Our findings confirm the involvement of adaptive responses in the aetiopathogenesis of FD, specifically a Th2 and Th17 signature in the duodenal mucosa. The presence of effector and memory cells suggest that the microinflammation in FD is antigen driven.

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Significant mobilization of both conventional and regulatory T cells with AMD3100

Blood ◽

10.1182/blood-2011-06-359331 ◽

2011 ◽

Vol 118 (25) ◽

pp. 6580-6590 ◽

Cited By ~ 48

Author(s):

Leslie S. Kean ◽

Sharon Sen ◽

Olusegun Onabajo ◽

Karnail Singh ◽

Jennifer Robertson ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Effector Memory ◽

Cell Populations ◽

Cell Repertoire ◽

Hematopoietic Stem ◽

Macaque Model ◽

The Impact

AbstractIn this study, we used the rhesus macaque model to determine the impact that AMD3100 has on lymphocyte mobilization, both alone and in combination with G-CSF. Our results indicate that, unlike G-CSF, AMD3100 substantially mobilizes both B and T lymphocytes into the peripheral blood. This led to significant increases in the peripheral blood content of both effector and regulatory T-cell populations, which translated into greater accumulation of these cells in the resulting leukapheresis products. Notably, CD4+/CD25high/CD127low/FoxP3+ Tregs were efficiently mobilized with AMD3100-containing regimens, with as much as a 4.0-fold enrichment in the leukapheresis product compared with G-CSF alone. CD8+ T cells were mobilized to a greater extent than CD4+ T cells, with accumulation of 3.7 ± 0.4-fold more total CD8+ T cells and 6.2 ± 0.4-fold more CD8+ effector memory T cells in the leukapheresis product compared with G-CSF alone. Given that effector memory T-cell subpopulations may mediate less GVHD compared with other effector T-cell populations and that Tregs are protective against GVHD, our results indicate that AMD3100 may mobilize a GVHD-protective T-cell repertoire, which would be of benefit in allogeneic hematopoietic stem cell transplantation.

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A novel 120-kD surface antigen expressed by a subset of human lymphocytes. Evidence that lymphokine-activated killer cells express this molecule and use it in their effector function.

Journal of Experimental Medicine ◽

10.1084/jem.166.2.319 ◽

1987 ◽

Vol 166 (2) ◽

pp. 319-326 ◽

Cited By ~ 22

Author(s):

M R Zocchi ◽

C Bottino ◽

S Ferrini ◽

L Moretta ◽

A Moretta

Keyword(s):

T Cell ◽

Cell Lines ◽

Peripheral Blood ◽

Cell Clone ◽

Human Lymphocytes ◽

Cytolytic Activity ◽

Cell Populations ◽

Target Cells ◽

Lymphokine Activated Killer Cells ◽

Effector Cells

A human cell clone (SF-16) displaying strong cytolytic activity against fresh tumor target cells was used for production of murine mAbs against surface antigens expressed by lymphokine-activated killer (LAK) cells and their peripheral blood precursors. The preliminary screening of hybridoma supernatants was performed according to the ability to bind SF-16 cells. Selected mAbs were further analyzed for their reactivity with several T and B cell lines and with peripheral blood T and non-T cell populations. A selected mAb, termed anti-LAK-1, only reacted with some T cell lines and with 15-30% of PBMC. Approximately 10-15% E-rosetting (T) cells and 40-50% E-rosette-negative cells were LAK-1+, as determined by cytofluorometric analysis. As the fluorescence distribution of LAK-1 antigen was clearly bimodal, LAK-1+ and LAK-1- cells could be separated by FACS. Positive cells were composed of large granular lymphocytes (LGL), whereas negative cells were mostly small lymphocytes and monocytes without LGL. After culture in rIL-2, purified LAK-1+ (but not LAK-1-) cells acquired the ability to lyse NK-resistant fresh melanoma target cells. In addition, only the LAK-1+ fraction of PBMC cultured for 5 d in rIL-2 lysed fresh tumor targets, thus indicating that the LAK-1 antigen is expressed also on LAK effector cells. Unlike some other LGL/NK cell markers, LAK-1 antigen is characterized by a stable expression: thus, LAK-1+ cell populations cultured for up to 20 d in rIL-2 maintained the LAK-1 antigen expression, whereas HNK-1 and, partially, CD16 were lost. Finally the cytolytic activity of LAK effector cells generated from PBMC cultured for 3 d in rIL-2 was susceptible to inhibition by the anti-LAK-1 mAb.

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Single-Cell Analysis Reveals Characterization of Infiltrating T Cells in Moderately Differentiated Colorectal Cancer

Frontiers in Immunology ◽

10.3389/fimmu.2020.620196 ◽

2021 ◽

Vol 11 ◽

Author(s):

Xi Yang ◽

Quan Qi ◽

Yuefen Pan ◽

Qing Zhou ◽

Yinhang Wu ◽

...

Keyword(s):

Colorectal Cancer ◽

Rectal Cancer ◽

Colon Cancer ◽

T Cells ◽

T Cell ◽

Single Cell ◽

Peripheral Blood ◽

Strong Correlation ◽

Tumor Tissue ◽

Cell Populations

ObjectiveThis study aimed to characterize the tumor-infiltrating T cells in moderately differentiated colorectal cancer.MethodsUsing single-cell RNA sequencing data of isolated 1632 T cells from tumor tissue and 1252 T cells from the peripheral blood of CRC patients, unsupervised clustering analysis was performed to identify functionally distinct T cell populations, followed by correlations and ligand-receptor interactions across cell types. Finally, differential analysis of the tumor-infiltrating T cells between colon cancer and rectal cancer were carried out.ResultsA total of eight distinct T cell populations were identified from tumor tissue. Tumor-Treg showed a strong correlation with Th17 cells. CD8+TRM was positively correlated with CD8+IEL. Seven distinct T cell populations were identified from peripheral blood. There was a strong correlation between CD4+TN and CD4+blood-TCM. Colon cancer and rectal cancer showed differences in the composition of tumor-infiltrating T cell populations. Tumor-infiltrating CD8+IEL cells were found in rectal cancer but not in colon cancer, while CD8+ TN cells were found in the peripheral blood of colon cancer but not in that of rectal cancer. A larger number of tumor-infiltrating CD8+ Tex (88.94%) cells were found in the colon cancer than in the rectal cancer (11.06%). The T cells of the colon and rectal cancers showed changes in gene expression pattern.ConclusionsWe characterized the T cell populations in the CRC tumor tissue and peripheral blood.

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High-Dimensional Immunophenotyping of Murine T-cell and B-cell Subsets

10.1101/2020.05.07.082214 ◽

2020 ◽

Author(s):

Kyle T. Mincham ◽

Jacob D. Young ◽

Deborah H. Strickland

Keyword(s):

T Cell ◽

B Cell ◽

Plasma Cells ◽

Effector Memory ◽

High Dimensional ◽

Cell Populations ◽

Lymphoid Tissues ◽

Murine Models ◽

Follicular Helper ◽

B Cell Subsets

Purpose and appropriate sample typesThis 19-parameter, 18-colour flow cytometry panel was designed and optimised to enable the comprehensive and simultaneous immunophenotyping of distinct T-cell and B-cell subsets within murine lymphoid tissues (Table 1). Cellular populations identified by employing this OMIP include 4 major subsets of B-cells (memory, activated, plasma cells and plasmablasts) and 7 major subsets of CD4+ T-cells (naïve, central memory, effector memory, helper, regulatory, follicular helper and follicular regulatory). Staining was performed on freshly isolated splenocytes from 21-day-old neonatal BALB/c mice, however due to the omission of mouse strain-specific markers, this OMIP can be implemented across a range of murine models where in-depth immunophenotyping of the diverse repertoire of T-cell and B-cell populations localised within lymphoid tissues is required.

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Distinct cellular immune profiles in the airways and blood of critically ill patients with COVID-19

10.1101/2020.10.29.360586 ◽

2020 ◽

Author(s):

Anno Saris ◽

Tom D.Y. Reijnders ◽

Esther J. Nossent ◽

Alex R. Schuurman ◽

Jan Verhoeff ◽

...

Keyword(s):

Immune Response ◽

T Cells ◽

T Cell ◽

T Cell Activation ◽

Peripheral Blood ◽

Cell Activation ◽

Effector Memory ◽

Activated T Cells ◽

Immune Profile ◽

Prolonged Icu Stay

AbstractOur understanding of the coronavirus disease-19 (COVID-19) immune response is almost exclusively derived from studies that examined blood. To gain insight in the pulmonary immune response we analysed BALF samples and paired blood samples from 17 severe COVID-19 patients. Macrophages and T cells were the most abundant cells in BALF. In the lungs, both CD4 and CD8 T cells were predominantly effector memory cells and expressed higher levels of the exhaustion marker PD-1 than in peripheral blood. Prolonged ICU stay associated with a reduced proportion of activated T cells in peripheral blood and even more so in BALF. T cell activation in blood, but not in BALF, was higher in fatal COVID-19 cases. Increased levels of inflammatory mediators were more pronounced in BALF than in plasma. In conclusion, the bronchoalveolar immune response in COVID-19 has a unique local profile that strongly differs from the immune profile in peripheral blood.SummaryThe bronchoalveolar immune response in severe COVID-19 strongly differs from the peripheral blood immune profile. Fatal COVID-19 associated with T cell activation blood, but not in BALF.

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Short-Lived Effector CD8 T Cells Induced by Genetically Attenuated Malaria Parasite Vaccination Express CD11c

Infection and Immunity ◽

10.1128/iai.00871-13 ◽

2013 ◽

Vol 81 (11) ◽

pp. 4171-4181 ◽

Cited By ~ 20

Author(s):

Laura A. Cooney ◽

Megha Gupta ◽

Sunil Thomas ◽

Sebastian Mikolajczak ◽

Kimberly Y. Choi ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Inflammatory Cytokines ◽

Plasmodium Yoelii ◽

T Cell Response ◽

Parasite Development ◽

Effector Memory ◽

Cell Phenotype ◽

Effector Cells ◽

Ifn Γ

ABSTRACTVaccination with a single dose of genetically attenuated malaria parasites can induce sterile protection against sporozoite challenge in the rodentPlasmodium yoeliimodel. Protection is dependent on CD8+T cells, involves perforin and gamma interferon (IFN-γ), and is correlated with the expansion of effector memory CD8+T cells in the liver. Here, we have further characterized vaccine-induced changes in the CD8+T cell phenotype and demonstrated significant upregulation of CD11c on CD3+CD8b+T cells in the liver, spleen, and peripheral blood. CD11c+CD8+T cells are predominantly CD11ahiCD44hiCD62L−, indicative of antigen-experienced effector cells. Followingin vitrorestimulation with malaria-infected hepatocytes, CD11c+CD8+T cells expressed inflammatory cytokines and cytotoxicity markers, including IFN-γ, tumor necrosis factor alpha (TNF-α), interleukin-2 (IL-2), perforin, and CD107a. CD11c−CD8+T cells, on the other hand, expressed negligible amounts of all inflammatory cytokines and cytotoxicity markers tested, indicating that CD11c marks multifunctional effector CD8+T cells. Coculture of CD11c+, but not CD11c−, CD8+T cells with sporozoite-infected primary hepatocytes significantly inhibited liver-stage parasite development. Tetramer staining for the immunodominant circumsporozoite protein (CSP)-specific CD8+T cell epitope demonstrated that approximately two-thirds of CSP-specific cells expressed CD11c at the peak of the CD11c+CD8+T cell response, but CD11c expression was lost as the CD8+T cells entered the memory phase. Further analyses showed that CD11c+CD8+T cells are primarily KLRG1+CD127−terminal effectors, whereas all KLRG1−CD127+memory precursor effector cells are CD11c−CD8+T cells. Together, these results suggest that CD11c marks a subset of highly inflammatory, short-lived, antigen-specific effector cells, which may play an important role in eliminating infected hepatocytes.

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P09.15 Targeting the stroma to enhance effector memory T cell infiltration and anti-tumor response to anti-PD1 antibody in pancreatic ductal adenocarcinoma

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2020-itoc7.115 ◽

2020 ◽

Vol 8 (Suppl 2) ◽

pp. A59.2-A60

Author(s):

A Osipov ◽

L Zheng

Keyword(s):

T Cell ◽

Myeloid Cells ◽

Myeloid Cell ◽

Cxcr4 Expression ◽

Cell Infiltration ◽

Ductal Adenocarcinoma ◽

Effector Memory ◽

Cell Populations ◽

Memory T Cell ◽

T Cell Infiltration

BackgroundPancreatic ductal adenocarcinoma (PDAC) is resistant to immune checkpoint inhibition. One of the major resistance mechanisms is attributed to myeloid cells as an immunosuppressive element within the stroma of PDAC. It has been reported that focal adhesion kinase inhibitor (FAKi) can suppress immunosuppressive myeloid cells such as tumor associated macrophages (TAMs) and myeloid derived suppressor cells (MDSC), consequently sensitizing tumor to anti-PD1 antibody in mouse models of PDAC. Our group has previously shown in a murine model that targeting the stroma via PEGylated recombinant human hyaluronidase (PEGPH20) enhanced the anti-tumor activity of the whole cell vaccine (GVAX) by targeting CXCR4-expressing myeloid cells and led to an increase in infiltration of CCR7- effector memory T cell subsets. Here, we evaluate the hypothesis that FAK expressing myeloid cell subsets modulate T cell infiltration in human PDAC and FAKi can synergize with PEGPH20 by targeting myeloid cells in PDAC.Material and MethodsResected human PDAC tissue specimens treated with GVAX and anti-PD1 therapy was used to assess FAK expression in myeloid cell subsets and its impact on T cell infiltration. A sequential staining and stripping multiplex IHC technique that incorporates 28 myeloid and lymphoid biomarkers, as well as phosphorylated FAK (pFAK) combined with computational image processing was used to assess myeloid cell populations, T cell infiltration and FAK expression.An established murine model of metastatic PDAC treated with and without anti-PD1 therapy was used to assess the synergy and immune-modulating effect of FAKi and stromal degradation of hyaluronan via PEGPH20.ResultsIn human PDAC, FAK is widely expressed in TAMs and neutrophils. Increased FAK expression is associated with increased CXCR4 expression. Lower pFAK density in neutrophils and M2 TAMs, but not lower pFAK density in M1 TAMs, is associated with higher CD8+ T cell infiltration.FAKi and combination of FAKi with anti-PD1 extends survival in the mouse metastasis model of PDAC. Adding PEGPH20 to FAKi and anti-PD1 antibody significantly prolonged survival in this model. Comparing to the combination of FAKi and anti-PD1 antibody, adding PEGPH20 significantly decreased the number of CXCR4-expressing myeloid cells in the tumor microenvironment (TME) of PDAC and consequently led to an increase in the amount of CCR7+ central memory T cells. Additionally, the amount of G-MDSCs, inflammatory resident monocytes and PDL1 expressing myeloid cells in the TME of PDAC, was also decreased in PDAC treated with the triple combination of PEGPH20, FAKi and anti-PD1 antibody compared to FAKi and anti-PD1 antibody.ConclusionFAK is widely expressed in myeloid cell populations, directly correlated with CXCR4 expression and decreased FAK expression in a myeloid (M2 TAMs, neutrophil) inflamed stroma is associated with infiltration of effector CD8 T cells in human PDAC. Stromal degradation of hyaluronan via PEGPH20 combined with FAKi and anti-PD1 antibody further depletes immunosuppressive cells in the TME including G-MDSCs, inflammatory resident monocytes and PDL1 expressing myeloid cells and appears to target the CXCR4 pathway through PEGPH20. These findings support testing the combination of FAKi and anti-PD1 antibody with agents targeting CXCR4 directly or indirectly by PEGPH20 in human PDAC.Disclosure InformationA. Osipov: None. L. Zheng: None.

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