Kelly Summers | Western University Canada (original) (raw)

Papers by Kelly Summers

Medical Science Monitor, 2011

Background: To examine the effect of probiotics as adjunctive therapy for the treatment of rheuma... more Background: To examine the effect of probiotics as adjunctive therapy for the treatment of rheumatoid arthritis (RA). A sample size of 30 subjects was calculated to determine a moderate effect. Material/Methods: A three month double-blind, placebo-controlled study was performed using probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 capsules administered orally. Inclusion criteria required at least 4 swollen and 4 tender joints and stable medications with no steroids for at least 1 month prior to and during the study. Twenty-nine patients with RA were randomized to treatment. ACR20 responses, serum cytokine levels and safety parameters were assessed. Results: Fifteen patients were randomized to the probiotic group, and 14 to placebo. Three subjects in the probiotic (20%) and one in the placebo group (7%) achieved an ACR20 response (p= 0.33). There was no statistically significant difference between individual components of the ACR20 criteria. Changes in cytokines favored placebo over probiotic. There was a significant improvement in the Health Assessment Questionnaire (HAQ) score in the probiotic group from visit 1 to visit 3 (p=0.02) but no between-group differences. Conclusions: Due to inclusion criteria, patients selected for the study had stable RA with chronic synovitis, and thus it may have been difficult for an adjunctive therapy to demonstrate improvement within 3 months. Although probiotics did not clinically improve RA as measured by the ACR20, it is interesting that there was functional improvement seen within the probiotic group compared to placebo.

Microcirculation, 2013

ObjectiveTo determine if the DKA‐induced inflammation in juvenile mice provokes activation and dy... more ObjectiveTo determine if the DKA‐induced inflammation in juvenile mice provokes activation and dysfunction of CVECs.MethodsDKA in juvenile mice was induced with administration of STZ and ALX. Blood from DKA mice was assessed for cytokines and soluble cell adhesion proteins, and either DKA plasma or exogenous compounds were applied to immortalized bEND3.ResultsDKA increased circulating levels of IL‐6, IL‐8(KC), MCP‐1, IL‐10, sE‐selectin, sICAM‐1, and sVCAM‐1. Stimulation of bEND3 with DKA plasma caused cellular activation (increased ROS and activation of NF‐κΒ), upregulation of a proadhesive phenotype (E‐selectin, ICAM‐1, and VCAM‐1), and increased leukocyte‐bEND3 interaction (leukocyte rolling/adhesion). TEER, a measure of bEND3 monolayer integrity, was decreased by DKA plasma. Activation and dysfunction of bEND3 with DKA plasma were suppressed by plasma heat treatment (56°C, 1 hour) and replicated with the application of DKA recombinant cytomix (IL‐6, IL‐8[KC], MCP‐1, and IL‐10), i...

Microbiology and Immunology, Sep 1, 2009

ABSTRACTVulvovaginal candidiasis, a high prevailing infection worldwide, is mainly caused by Cand... more ABSTRACTVulvovaginal candidiasis, a high prevailing infection worldwide, is mainly caused by Candida albicans. Probiotic Lactobacillus reuteri RC‐14 and Lactobacillus rhamnosus GR‐1 have been previously shown to be useful as adjuvants in the treatment of women with VVC. In order to demonstrate and better understand the anti‐Candida activity of the probiotic microorganisms in an in vitro model simulating vaginal candidiasis, a human vaginal epithelial cell line (VK2/E6E7) was infected with C.albicans 3153a and then challenged with probiotic L. rhamnosus GR‐1 and/or L. reuteri RC‐14 or their respective CFS (alone or in combination). At each time point (0, 6, 12 and 24 hr), numbers of yeast, lactobacilli and viable VK2/E6E7 cells were determined and, at 0, 6 and 12 hr, the supernatants were measured for cytokine levels. We found that C. albicans induced a significant increase in IL‐1α and IL‐8 production by VK2/E6E7 cells. After lactobacilli challenge, epithelial cells did not alter IL‐6, IL‐1α, RANTES and VEGF levels. However, CFS from the probiotic microorganisms up‐regulated IL‐8 and IP‐10 levels secreted by VK2/E6E7 cells infected with C. albicans. At 24 hr of co‐incubation, L. reuteri RC‐14 alone and in combination with L. rhamnosus GR‐1 decreased the yeast population recoverable from the cells. In conclusion, L. reuteri RC‐14 alone and together with L. rhamnosus GR‐1 have the potential to inhibit the yeast growth and their CFS may up‐regulate IL‐8 and IP‐10 secretion by VK2/E6E7 cells, which could possibly have played an important role in helping to clear VVC in vivo.

Orthopaedic Proceedings, Mar 1, 2008

T cells have been implicated in the pathogenesis of osteolysis. The goal of this study was to com... more T cells have been implicated in the pathogenesis of osteolysis. The goal of this study was to compare the ratios of CD4+ T cell populations in total hip arthroplasty (THA) patients with and without osteolysis. We found no significant differences in the frequency of peripheral blood CD4+CD25+ regulatory and effector T cells, serum IL-10 and TGF-β concentrations, and immuno-suppressive ability of regulatory T cells from patients with osteoarthritis prior to THA, and THA patients with and without radiographic evidence of osteolysis. CD4+ T cells are critical in regulating immune-mediated conditions. This study compared the frequency of CD4+ T cell subpopulations in the peripheral blood of patients with and without osteolysis following total hip arthroplasty (THA). Numbers of CD4+CD25hi regulatory T cells, CD4+CD25moderate effector T cells, and CD4+CD25+ T cells in the peripheral blood of thirty patients with osteoarthritis prior to primary THA, thirty patients with asymptomatic THAs and no radiographic evidence of osteolysis, nineteen patients with asymptomatic THAs with radiographic evidence of early osteolysis (not requiring revision surgery) and nine patients scheduled for revision THA for osteolysis were determined by flow cytometry. Serum IL-10 and TGF-β levels were measured using ELISA kits. Results were compared by t-test and rank sum test. CD4+ CD25hi regulatory T cells and CD4+ CD25neg T cells were isolated from blood using a MACS cell isolation kit, co-cultured for three days, and T cell proliferation determined by [3H]-thy-midine uptake. The frequency of CD4+CD25hi regulatory T cells, CD4+CD25moderate effector T cells, and CD4+CD25+ T cells were similar in each study group. Regulatory T cells from patients with and without osteolysis had a normal functional ability to inhibit CD4+ T cell proliferation. Serum levels of the regulatory T cell-derived cytokines IL-10 and TGF-β were also comparable between groups. Our data suggests that CD4+ T cell immune responses are normal in THA regardless of the level of osteolysis, in contrast to previous studies that have implicated T cell hypersensitivity in the pathogenesis of osteolysis surrounding THA.

The Open Biomarkers Journal, Jan 29, 2010

Background: Progressive fibrosis and cirrhosis, clinically presenting as end-stage liver disease ... more Background: Progressive fibrosis and cirrhosis, clinically presenting as end-stage liver disease are common outcomes in alcoholic hepatitis as well as non-alcoholic fatty liver disease(NAFLD). In these processes, a series of changes occurs in liver tissues leading to cell death, remodeling, fibrosis and regeneration. The aim of this study is to identify potential novel biomarkers for non-invasive diagnosis of cirrhosis due to alcoholic etiology or NAFLD. Methods: Serum from patients with biopsy proven end-stage liver disease of various etiologies, namely NAFLD(n=9), alcohol(n=5), and other end-stage liver diseases(n=6), who underwent liver transplant during the first six months of 2007 were utilized for retrospective analysis. Serum samples were also collected from a group of healthy volunteers (n=7). The samples were analysed using Luminex technology or ELISA for 27 biomarkers that are known to be involved in pathologic processes such as cell death, regeneration and fibrosis. Results: Of the 27 serum markers examined, 16 were elevated in the serum in all groups with end-stage liver diseases compared with the control group. They include adipokines, apoptosis and inflammatory mediators and growth factors. Interestingly, the serum of NAFLD patients showed significantly elevated HGF levels and trend towards increase in sFAS, TGF 1, TNFR-1, TNFR-2 and leptin. The level of serum markers showed excellent correlation with each other indicating a complex interdependent pathogenetic mechanism. Conclusions: The data from this study indicate that a large number of serum markers are altered in end-stage liver diseases. A panel of such markers may potentially be useful in assessing advanced fibrosis and cirrhosis in patients with chronic end stage liver diseases.

Archivum Immunologiae Et Therapiae Experimentalis, Jul 21, 2017

Transforming growth factor (TGF)-b has been implicated in regulation of the immune system, includ... more Transforming growth factor (TGF)-b has been implicated in regulation of the immune system, including autoimmunity. We have found that TGF-b is readily produced by T cells following immunization with self-peptide epitopes that downregulate autoimmune responses in type 1 diabetes (T1D) prone nonobese diabetic (NOD) mice. These include multiple peptide epitopes derived from the islet b-cell antigens GAD65 (GAD65 p202-221, GAD65 p217-236), GAD67 (GAD67 p210-229, GAD67 p225-244), IGRP (IGRP p123-145, IGRP p195-214) and insulin B-chain (Ins. B:9-23) that protected NOD mice from T1D. Immunization of NOD mice with the self-MHC class II I-A g7 b-chain-derived peptide, I-Ab g7 p54-76 also induced large amounts of TGF-b and also protected these mice from diabetes development. These results indicate that peptides derived from disease related self-antigens and MHC class II molecules primarily induce TGF-b producing regulatory Th3 and Tr1-like cells. TGF-b produced by these cells could enhance the differentiation of induced regulatory iTreg and iTreg17 cells to prevent induction and progression of autoimmune diseases. We therefore suggest that peripheral immune tolerance could be induced and maintained by immunization with self-peptides that induce TGF-b producing T cells.

PubMed, Jul 1, 1995

Objective: To characterize chondrocytes in normal and arthritic joints and compare their phenotyp... more Objective: To characterize chondrocytes in normal and arthritic joints and compare their phenotype to that of synovial macrophages present in rheumatoid joints. Methods: Using an immunoperoxidase staining technique, we examined the presence and distribution of a number of leukocyte activation and differentiation antigens on samples of cartilage obtained from resected joints of normal controls and subjects with rheumatoid arthritis or osteoarthritis. Results: Chondrocytes in each group were CD14+, CD68+, Thy-1+, CD11a+, CD18+, MAX.3-, and MAX.24-. Staining was variable for MAX.1 and CD45. HLA-DR and CD71 were expressed only on cells located in the superficial layer of rheumatoid cartilage. We found lower levels of expression of CD14 on chondrocytes in arthritic joints, whereas CD58 was expressed at higher levels. Surface expression of CD14 was confirmed on normal chondrocytes using flow cytometry and further supported by the detection of CD14 mRNA by polymerase chain reaction. Conclusion: Our findings demonstrate that chondrocytes express several antigens that are also found on monocytes and macrophages.

Orthopaedic Proceedings, Mar 1, 2008

The pathogenesis of osteolysis in failed total hip arthroplasty is not fully understood. The purp... more The pathogenesis of osteolysis in failed total hip arthroplasty is not fully understood. The purpose of this study is to identify CD4+CD25+ Regulatory T cells in periprosthetic tissues in failed total hip replacements secondary to osteolysis. Intra-operative tissue samples and peripheral blood were collected from patients undergoing revision total hip arthroplasty surgery. Regulatory T cells were present in the tissues, and significantly increased in the peripheral blood in patients with failed total hips compared to normal controls. Further characterization of these regulatory T cells are warranted as they may play a role in osteolysis in loose total hip replacements. Osteolysis remains the most common complication following total joint arthroplasty. To date, no authors have investigated the role of CD4+CD25+ regulatory T cells (T REG ) participating in the osteolytic pathogenesis. The purpose of this study is to quantitate the presence of T REG cells in periprosthetic tissues in failed total hip replacements secondary to osteolysis. Fifteen consecutive patients booked for revision total hip arthroplasty secondary to osteolysis were included. Tissue samples were collected: peripheral blood (PB), synovial fluid (SF), synovial tissue (ST), and interface tissue (IT) between the failed component and the bone defect. Total lymphocytes were isolated and analyzed using fluorescent-tagged antibody cell sorting (FACS) for the presence of T REG cells. Frozen sections of ST and IT were analyzed with immunohistochemistry for T REG cells. T REG cells were significantly upregulated (p REG cells in the ST and IT were confirmed with immunohistochemistry. T REG cells are upregulated in the peripheral blood of patients with failed total hips secondary to osteolysis. The TREG cells are also present in the synovial tissue and interface tissue. Evidence for involvement of regulatory T cells contribute to our understanding of this complex biologic response to artificial wear particles. Functional studies of these T REG cells are warranted as they are upregulated in patients with loose total hip replacements.

Gastroenterology, May 1, 2013

Advances in Experimental Medicine and Biology, 1995

Rheumatology, May 20, 2014

Objective. Data from a small study testing imatinib to treat SSc were used to determine if cytoki... more Objective. Data from a small study testing imatinib to treat SSc were used to determine if cytokine changes were related to differences in clinical parameters to model future early phase trials pairing cytokine changes and clinical parameters. Methods. Plasma and punch skin biopsy specimens collected at baseline and 6 months were analysed for levels of 26 fibrotic and inflammatory cytokines using multiplexed immunoassays and ELISA. Seven of nine patients on active treatment had paired data. Biopsies were biopulverized and standardized to protein levels in the tissue homogenate. Plasma was frozen at À80 C and analysed using multiplexed immunoassays or ELISAs standardized to CRP. Correlations between fold changes in cytokines and differences in clinical parameters (skin score, physician and patient global assessments and HAQ) were performed. P < 0.01 was considered significant. Results. After 6 months of imatinib treatment, plasma levels of soluble vascular cell adhesion molecule 1 decreased significantly (P < 0.001), while tissue levels of soluble intercellular adhesion molecule 1 increased (P < 0.01). Some significant correlations between fold changes in certain plasma fibrotic and inflammatory cytokines and changes in clinical parameters after 6 months of treatment were found: patient global scores and IL-13 (r = 0.964, P < 0.0001); ESR and IL-12p70 (r = À0.903, P < 0.01); in tissue samples, patient global score and soluble E-selectin (r = 0.913, P < 0.01); and physician global score with sCD40L (r = À0.883, P < 0.01). Conclusion. Some serum and tissue cytokines may have a role in early phase clinical trials of SSc, correlating with changes in clinical parameters. Serum and tissue samples could be analysed in early phase trials to determine whether they support the clinical observations.

Cellular Immunology, May 1, 2019

Pathogenic lymphocytes aberrantly recognize and mount an immune response against self-antigens, l... more Pathogenic lymphocytes aberrantly recognize and mount an immune response against self-antigens, leading to the destruction of healthy cells, tissues and organs. Recent studies have shown that both B and T lymphocytes contribute to the development, prevention and modulation of various autoimmune diseases. Regulatory T and B cell subsets appear to play a prominent role in the prevention of autoimmune diseases. The recent identification of novel regulatory Th17 cells, termed as Treg17 cells, has expanded the scope of regulatory T lymphocytes (Treg cells) in the prevention of autoimmune diseases. Similarly, novel regulatory B cell subsets, termed as Breg cells, acting on their own or by inducing Treg cells have extended the role of B lymphocytes in the prevention and regulation of autoimmune diseases. We suggest that Treg17 cells and Breg cells have an important immunoregulatory role in autoimmune diseases.

Journal of Immunology, Apr 15, 2008

Arthritis Research & Therapy, Jan 26, 2001

We discuss the presence of anti-keratin antibodies (AKA) of the IgG class in patients with define... more We discuss the presence of anti-keratin antibodies (AKA) of the IgG class in patients with defined juvenile idiopathic arthritis (JIA). An indirect immunofluorescence test and rat oesophagus substrate was used for the detection and quantification of AKA antibodies in patients´ sera. Overall 33/60 patients with JIA had sera positive for AKA (55 %, P = 0,0001) ranging from 1:10 to 1:160 dilutions. Following idiopathic arthritis of childhood classification criteria AKA occurred in 2/7 patients with systemic disease (28,6 %), in 13/30 patients with RF negative polyarthritis (43,3 %, P = 0,008) and in 15/18 RF positive polyarthritis (83,3 %, P = 0,000002). AKA were also found in a small cohort of patients with oligoarthritis (1/3) and psoriatic arthritis (2/2). AKA positivity occurred in 3/26 healthy controls at a 1:20 dilution. The presence of AKA was correlated as well as with the severity of the disease. Our study revealed that AKA was present overall in 18/29 patients (62%) with severe JIA and in 12/26 patients (46,2 %) with non-severe disease, however this did not reach statistical significance (P = 0,18). We also observed that AKA remained positive regardless of disease activity. AKA were detectable in 55,6 % patients with active JIA and in 48,6 % patients in the complete or near remission. Acknowledgement: This research was supported by a European Commission (Acronym: EUROBANK, contract no: QOL-2000-14.1), web site http://www.ncl.ac.uk and by grant of 2nd Medical Faculty, Charles University in Prague, VZ no. 111300003. P2 The significance of antibodies to cyclic citrullinated peptide, antikeratin antibodies, antiperinuclear factor, rheumatoid factor isotypes and HLA shared epitope in prediction of erosive disease in early rheumatoid arthritis patients

Journal of Maternal-fetal & Neonatal Medicine, May 17, 2012

To establish gender-specific differences in maternal and fetal immune response in healthy human f... more To establish gender-specific differences in maternal and fetal immune response in healthy human fetuses at term. Methods: Forty-five women with elective caesarean sections for uncomplicated singleton pregnancies were recruited for two studies. Using a multiplex biomarker immunoassay system, unstimulated maternal and fetal plasma concentrations of interleukin (IL)-1β, IL-1ra, IL-6, IL-8, macrophage inflammatory protein (MIP)-1α, and tumor necrosis factor (TNF)-α were measured from one study population. Lipopolysaccharide (LPS)stimulated cytokine response was measured in a second study. Results: There were no significant gender differences in either maternal or fetal unstimulated plasma cytokine concentrations, but concentrations of the proinflammatory cytokines IL-1β and IL-6 were significantly greater in male fetal LPS-stimulated samples than in female fetal samples. Conclusions: Blood of male fetuses mounts a larger pro-inflammatory response to lipopolysaccharide (LPS). This heightened response could be a critical pathway in promoting premature rupture of membranes (PPROM) and may be associated with life long differential gender response to infection.

Clinical and Experimental Immunology, Jan 2, 2015

The increased risk and persistence of infections in diabetic condition is probably associated wit... more The increased risk and persistence of infections in diabetic condition is probably associated with defects in the cellular immune responses. We have previously shown a decrease in the production of interferon (IFN)-α by dendritic cells (DCs) in diabetic subjects. The basal level of IFN-α in splenic plasmacytoid DCs (pDCs) is also lower in non-obese diabetic (NOD) mice compared to prediabetic mice. The objective of this study was to analyse the ability of diabetic mice to mobilize innate and CD8 + T cell-mediated immune response to influenza A virus (IAV) with the live influenza A/Puerto Rico/8/1934 H1N1 (PR8) strain or with its immunodominant CD8 + T cell epitopes. We found that following immunization with IAV, the level of IFN-α in diabetic mice was increased to the level in prediabetic mice. Immunization of NOD mice with the immunodominant IAV PR8 peptide induced clonal expansion of IFN-γ-producing CD8 + T cells similar to the response observed in prediabetic mice. Thus, diabetic and prediabetic NOD mice have a similar capacity for IFN-α and IFN-γ production by pDCs and CD8 + T cells, respectively. Therefore, the DC-related immune defect in diabetic NOD mice does not impair their capacity to develop an effective immune response to IAV. Our results suggest that reduced IFN-α production by diabetic human and mouse DCs is not an impediment to an effective immunity to IAV in type 1 diabetic subjects vaccinated with live attenuated influenza vaccine.

Advances in Experimental Medicine and Biology, 1997

Dendritic cells (DC) are specialist antigen presenting cells (APC) derived in common with other l... more Dendritic cells (DC) are specialist antigen presenting cells (APC) derived in common with other leukocytes from bone marrow stem cells.1,2 A myeloid derived precursor3 gives rise to immature circulating blood DC which enter the tissues and after interacting with antigen migrate to the T lymphocyte dependent areas of lymph nodes, where they deliver stimulatory signals to responding T lymphocytes. Recent studies suggest the growth and differentiation of myeloid DC is heavily influenced by cytokines, notably flt-3 ligand, SCF, GM-CSF, IL-4 and TNFα4–6. A unique DC precursor in blood can he distinguished from freshly isolated blood monocytes.7,8 However, considerable evidence suggests that monocytes exposed in vitro to certain cytokine combinations (notably including IL-4, which downregulates CD14) can acquire many, if not most DC characteristics.9 The DC series also includes a lymphoid precursor derived cell10 a subset of which, in the mouse, has an inhibiting effect on responding T lymphocytes. Lymphoid precursor cells have been described in man11,12 hut their function is unknown.

Advances in Urology, 2009

The management of urinary tract infection (UTI) in individuals with spinal cord injury (SCI) cont... more The management of urinary tract infection (UTI) in individuals with spinal cord injury (SCI) continues to be of concern, due to complications that can occur. An emerging concept that is a common underlying pathophysiological process is involved, wherein pathogens causing UTI have a role in inflammatory progression. We hypothesized that members of the commensal flora, such as lactobacilli, may counter this reaction through anti-inflammatory mediation. This was assessed in a pilot two-patient study in which probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri were administered to one patient and placebo to another, both along with antibiotics to treat acute UTI. Urinary TNF-alpha was significantly downregulated (P = .015) in the patient who received the probiotic and who used intermittent catheterization compared with patient on placebo and using an indwelling catheter. The extent to which this alteration resulted in improved well-being in spinal cord injured patients remains to be determined in a larger study.

Social Science Research Network, 2020

Long-term stress disrupts the neuroimmune interface, often resulting in immunosuppression. The de... more Long-term stress disrupts the neuroimmune interface, often resulting in immunosuppression. The deleterious effects of stress on mainstream T cells are well-documented. However, innate-like invariant T cells have been overlooked. Chief among these are invariant natural killer T (iNKT) and mucosa-associated invariant T (MAIT) cells, which fulfill important functions in anticancer and antimicrobial immunity. We found stress to surprisingly abrogate both TH1- and TH2-type responses mediated by iNKT cells. This was not due to cell death since iNKT cells were unusually refractory to stress-inflicted apoptosis. Activated iNKT cells in stressed mice triggered an abnormal response characterized by a ‘split’ inflammatory signature along with a sudden and robust induction of IL-10, IL-23 and IL-27. Pharmacological inhibitors, conditional knockout systems and gene expression analyses revealed that dysregulated iNKT cell responses were driven by iNKT cell-intrinsic glucocorticoid receptor signaling. Accordingly, the inhibitory effects of chronic stress on iNKT cell functions were lost upon habituation to predictable stressors. Importantly, iNKT cells in stressed mice failed to potentiate cytotoxicity against lymphoma targets or promote immune surveillance against metastatic melanoma. Using a congenic mouse model in which MAIT cells are abundant, these cells remained resistant to stress-induced death but demonstrated hindered TH1- and TH2-type cytokine responses, reminiscent of their iNKT cell counterparts. This was corroborated in culture systems in which we interrogated human peripheral blood and hepatic iNKT and MAIT cells. Our findings uncover a previously unappreciated mechanism of stress-induced immunosuppression and implicate the stress response as a potential barrier to iNKT cell- and MAIT cell-based immunotherapies.

Medical Science Monitor, 2011

Background: To examine the effect of probiotics as adjunctive therapy for the treatment of rheuma... more Background: To examine the effect of probiotics as adjunctive therapy for the treatment of rheumatoid arthritis (RA). A sample size of 30 subjects was calculated to determine a moderate effect. Material/Methods: A three month double-blind, placebo-controlled study was performed using probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 capsules administered orally. Inclusion criteria required at least 4 swollen and 4 tender joints and stable medications with no steroids for at least 1 month prior to and during the study. Twenty-nine patients with RA were randomized to treatment. ACR20 responses, serum cytokine levels and safety parameters were assessed. Results: Fifteen patients were randomized to the probiotic group, and 14 to placebo. Three subjects in the probiotic (20%) and one in the placebo group (7%) achieved an ACR20 response (p= 0.33). There was no statistically significant difference between individual components of the ACR20 criteria. Changes in cytokines favored placebo over probiotic. There was a significant improvement in the Health Assessment Questionnaire (HAQ) score in the probiotic group from visit 1 to visit 3 (p=0.02) but no between-group differences. Conclusions: Due to inclusion criteria, patients selected for the study had stable RA with chronic synovitis, and thus it may have been difficult for an adjunctive therapy to demonstrate improvement within 3 months. Although probiotics did not clinically improve RA as measured by the ACR20, it is interesting that there was functional improvement seen within the probiotic group compared to placebo.

Microcirculation, 2013

ObjectiveTo determine if the DKA‐induced inflammation in juvenile mice provokes activation and dy... more ObjectiveTo determine if the DKA‐induced inflammation in juvenile mice provokes activation and dysfunction of CVECs.MethodsDKA in juvenile mice was induced with administration of STZ and ALX. Blood from DKA mice was assessed for cytokines and soluble cell adhesion proteins, and either DKA plasma or exogenous compounds were applied to immortalized bEND3.ResultsDKA increased circulating levels of IL‐6, IL‐8(KC), MCP‐1, IL‐10, sE‐selectin, sICAM‐1, and sVCAM‐1. Stimulation of bEND3 with DKA plasma caused cellular activation (increased ROS and activation of NF‐κΒ), upregulation of a proadhesive phenotype (E‐selectin, ICAM‐1, and VCAM‐1), and increased leukocyte‐bEND3 interaction (leukocyte rolling/adhesion). TEER, a measure of bEND3 monolayer integrity, was decreased by DKA plasma. Activation and dysfunction of bEND3 with DKA plasma were suppressed by plasma heat treatment (56°C, 1 hour) and replicated with the application of DKA recombinant cytomix (IL‐6, IL‐8[KC], MCP‐1, and IL‐10), i...

Microbiology and Immunology, Sep 1, 2009

ABSTRACTVulvovaginal candidiasis, a high prevailing infection worldwide, is mainly caused by Cand... more ABSTRACTVulvovaginal candidiasis, a high prevailing infection worldwide, is mainly caused by Candida albicans. Probiotic Lactobacillus reuteri RC‐14 and Lactobacillus rhamnosus GR‐1 have been previously shown to be useful as adjuvants in the treatment of women with VVC. In order to demonstrate and better understand the anti‐Candida activity of the probiotic microorganisms in an in vitro model simulating vaginal candidiasis, a human vaginal epithelial cell line (VK2/E6E7) was infected with C.albicans 3153a and then challenged with probiotic L. rhamnosus GR‐1 and/or L. reuteri RC‐14 or their respective CFS (alone or in combination). At each time point (0, 6, 12 and 24 hr), numbers of yeast, lactobacilli and viable VK2/E6E7 cells were determined and, at 0, 6 and 12 hr, the supernatants were measured for cytokine levels. We found that C. albicans induced a significant increase in IL‐1α and IL‐8 production by VK2/E6E7 cells. After lactobacilli challenge, epithelial cells did not alter IL‐6, IL‐1α, RANTES and VEGF levels. However, CFS from the probiotic microorganisms up‐regulated IL‐8 and IP‐10 levels secreted by VK2/E6E7 cells infected with C. albicans. At 24 hr of co‐incubation, L. reuteri RC‐14 alone and in combination with L. rhamnosus GR‐1 decreased the yeast population recoverable from the cells. In conclusion, L. reuteri RC‐14 alone and together with L. rhamnosus GR‐1 have the potential to inhibit the yeast growth and their CFS may up‐regulate IL‐8 and IP‐10 secretion by VK2/E6E7 cells, which could possibly have played an important role in helping to clear VVC in vivo.

Orthopaedic Proceedings, Mar 1, 2008

T cells have been implicated in the pathogenesis of osteolysis. The goal of this study was to com... more T cells have been implicated in the pathogenesis of osteolysis. The goal of this study was to compare the ratios of CD4+ T cell populations in total hip arthroplasty (THA) patients with and without osteolysis. We found no significant differences in the frequency of peripheral blood CD4+CD25+ regulatory and effector T cells, serum IL-10 and TGF-β concentrations, and immuno-suppressive ability of regulatory T cells from patients with osteoarthritis prior to THA, and THA patients with and without radiographic evidence of osteolysis. CD4+ T cells are critical in regulating immune-mediated conditions. This study compared the frequency of CD4+ T cell subpopulations in the peripheral blood of patients with and without osteolysis following total hip arthroplasty (THA). Numbers of CD4+CD25hi regulatory T cells, CD4+CD25moderate effector T cells, and CD4+CD25+ T cells in the peripheral blood of thirty patients with osteoarthritis prior to primary THA, thirty patients with asymptomatic THAs and no radiographic evidence of osteolysis, nineteen patients with asymptomatic THAs with radiographic evidence of early osteolysis (not requiring revision surgery) and nine patients scheduled for revision THA for osteolysis were determined by flow cytometry. Serum IL-10 and TGF-β levels were measured using ELISA kits. Results were compared by t-test and rank sum test. CD4+ CD25hi regulatory T cells and CD4+ CD25neg T cells were isolated from blood using a MACS cell isolation kit, co-cultured for three days, and T cell proliferation determined by [3H]-thy-midine uptake. The frequency of CD4+CD25hi regulatory T cells, CD4+CD25moderate effector T cells, and CD4+CD25+ T cells were similar in each study group. Regulatory T cells from patients with and without osteolysis had a normal functional ability to inhibit CD4+ T cell proliferation. Serum levels of the regulatory T cell-derived cytokines IL-10 and TGF-β were also comparable between groups. Our data suggests that CD4+ T cell immune responses are normal in THA regardless of the level of osteolysis, in contrast to previous studies that have implicated T cell hypersensitivity in the pathogenesis of osteolysis surrounding THA.

The Open Biomarkers Journal, Jan 29, 2010

Background: Progressive fibrosis and cirrhosis, clinically presenting as end-stage liver disease ... more Background: Progressive fibrosis and cirrhosis, clinically presenting as end-stage liver disease are common outcomes in alcoholic hepatitis as well as non-alcoholic fatty liver disease(NAFLD). In these processes, a series of changes occurs in liver tissues leading to cell death, remodeling, fibrosis and regeneration. The aim of this study is to identify potential novel biomarkers for non-invasive diagnosis of cirrhosis due to alcoholic etiology or NAFLD. Methods: Serum from patients with biopsy proven end-stage liver disease of various etiologies, namely NAFLD(n=9), alcohol(n=5), and other end-stage liver diseases(n=6), who underwent liver transplant during the first six months of 2007 were utilized for retrospective analysis. Serum samples were also collected from a group of healthy volunteers (n=7). The samples were analysed using Luminex technology or ELISA for 27 biomarkers that are known to be involved in pathologic processes such as cell death, regeneration and fibrosis. Results: Of the 27 serum markers examined, 16 were elevated in the serum in all groups with end-stage liver diseases compared with the control group. They include adipokines, apoptosis and inflammatory mediators and growth factors. Interestingly, the serum of NAFLD patients showed significantly elevated HGF levels and trend towards increase in sFAS, TGF 1, TNFR-1, TNFR-2 and leptin. The level of serum markers showed excellent correlation with each other indicating a complex interdependent pathogenetic mechanism. Conclusions: The data from this study indicate that a large number of serum markers are altered in end-stage liver diseases. A panel of such markers may potentially be useful in assessing advanced fibrosis and cirrhosis in patients with chronic end stage liver diseases.

Archivum Immunologiae Et Therapiae Experimentalis, Jul 21, 2017

Transforming growth factor (TGF)-b has been implicated in regulation of the immune system, includ... more Transforming growth factor (TGF)-b has been implicated in regulation of the immune system, including autoimmunity. We have found that TGF-b is readily produced by T cells following immunization with self-peptide epitopes that downregulate autoimmune responses in type 1 diabetes (T1D) prone nonobese diabetic (NOD) mice. These include multiple peptide epitopes derived from the islet b-cell antigens GAD65 (GAD65 p202-221, GAD65 p217-236), GAD67 (GAD67 p210-229, GAD67 p225-244), IGRP (IGRP p123-145, IGRP p195-214) and insulin B-chain (Ins. B:9-23) that protected NOD mice from T1D. Immunization of NOD mice with the self-MHC class II I-A g7 b-chain-derived peptide, I-Ab g7 p54-76 also induced large amounts of TGF-b and also protected these mice from diabetes development. These results indicate that peptides derived from disease related self-antigens and MHC class II molecules primarily induce TGF-b producing regulatory Th3 and Tr1-like cells. TGF-b produced by these cells could enhance the differentiation of induced regulatory iTreg and iTreg17 cells to prevent induction and progression of autoimmune diseases. We therefore suggest that peripheral immune tolerance could be induced and maintained by immunization with self-peptides that induce TGF-b producing T cells.

PubMed, Jul 1, 1995

Objective: To characterize chondrocytes in normal and arthritic joints and compare their phenotyp... more Objective: To characterize chondrocytes in normal and arthritic joints and compare their phenotype to that of synovial macrophages present in rheumatoid joints. Methods: Using an immunoperoxidase staining technique, we examined the presence and distribution of a number of leukocyte activation and differentiation antigens on samples of cartilage obtained from resected joints of normal controls and subjects with rheumatoid arthritis or osteoarthritis. Results: Chondrocytes in each group were CD14+, CD68+, Thy-1+, CD11a+, CD18+, MAX.3-, and MAX.24-. Staining was variable for MAX.1 and CD45. HLA-DR and CD71 were expressed only on cells located in the superficial layer of rheumatoid cartilage. We found lower levels of expression of CD14 on chondrocytes in arthritic joints, whereas CD58 was expressed at higher levels. Surface expression of CD14 was confirmed on normal chondrocytes using flow cytometry and further supported by the detection of CD14 mRNA by polymerase chain reaction. Conclusion: Our findings demonstrate that chondrocytes express several antigens that are also found on monocytes and macrophages.

Orthopaedic Proceedings, Mar 1, 2008

The pathogenesis of osteolysis in failed total hip arthroplasty is not fully understood. The purp... more The pathogenesis of osteolysis in failed total hip arthroplasty is not fully understood. The purpose of this study is to identify CD4+CD25+ Regulatory T cells in periprosthetic tissues in failed total hip replacements secondary to osteolysis. Intra-operative tissue samples and peripheral blood were collected from patients undergoing revision total hip arthroplasty surgery. Regulatory T cells were present in the tissues, and significantly increased in the peripheral blood in patients with failed total hips compared to normal controls. Further characterization of these regulatory T cells are warranted as they may play a role in osteolysis in loose total hip replacements. Osteolysis remains the most common complication following total joint arthroplasty. To date, no authors have investigated the role of CD4+CD25+ regulatory T cells (T REG ) participating in the osteolytic pathogenesis. The purpose of this study is to quantitate the presence of T REG cells in periprosthetic tissues in failed total hip replacements secondary to osteolysis. Fifteen consecutive patients booked for revision total hip arthroplasty secondary to osteolysis were included. Tissue samples were collected: peripheral blood (PB), synovial fluid (SF), synovial tissue (ST), and interface tissue (IT) between the failed component and the bone defect. Total lymphocytes were isolated and analyzed using fluorescent-tagged antibody cell sorting (FACS) for the presence of T REG cells. Frozen sections of ST and IT were analyzed with immunohistochemistry for T REG cells. T REG cells were significantly upregulated (p REG cells in the ST and IT were confirmed with immunohistochemistry. T REG cells are upregulated in the peripheral blood of patients with failed total hips secondary to osteolysis. The TREG cells are also present in the synovial tissue and interface tissue. Evidence for involvement of regulatory T cells contribute to our understanding of this complex biologic response to artificial wear particles. Functional studies of these T REG cells are warranted as they are upregulated in patients with loose total hip replacements.

Gastroenterology, May 1, 2013

Advances in Experimental Medicine and Biology, 1995

Rheumatology, May 20, 2014

Objective. Data from a small study testing imatinib to treat SSc were used to determine if cytoki... more Objective. Data from a small study testing imatinib to treat SSc were used to determine if cytokine changes were related to differences in clinical parameters to model future early phase trials pairing cytokine changes and clinical parameters. Methods. Plasma and punch skin biopsy specimens collected at baseline and 6 months were analysed for levels of 26 fibrotic and inflammatory cytokines using multiplexed immunoassays and ELISA. Seven of nine patients on active treatment had paired data. Biopsies were biopulverized and standardized to protein levels in the tissue homogenate. Plasma was frozen at À80 C and analysed using multiplexed immunoassays or ELISAs standardized to CRP. Correlations between fold changes in cytokines and differences in clinical parameters (skin score, physician and patient global assessments and HAQ) were performed. P < 0.01 was considered significant. Results. After 6 months of imatinib treatment, plasma levels of soluble vascular cell adhesion molecule 1 decreased significantly (P < 0.001), while tissue levels of soluble intercellular adhesion molecule 1 increased (P < 0.01). Some significant correlations between fold changes in certain plasma fibrotic and inflammatory cytokines and changes in clinical parameters after 6 months of treatment were found: patient global scores and IL-13 (r = 0.964, P < 0.0001); ESR and IL-12p70 (r = À0.903, P < 0.01); in tissue samples, patient global score and soluble E-selectin (r = 0.913, P < 0.01); and physician global score with sCD40L (r = À0.883, P < 0.01). Conclusion. Some serum and tissue cytokines may have a role in early phase clinical trials of SSc, correlating with changes in clinical parameters. Serum and tissue samples could be analysed in early phase trials to determine whether they support the clinical observations.

Cellular Immunology, May 1, 2019

Pathogenic lymphocytes aberrantly recognize and mount an immune response against self-antigens, l... more Pathogenic lymphocytes aberrantly recognize and mount an immune response against self-antigens, leading to the destruction of healthy cells, tissues and organs. Recent studies have shown that both B and T lymphocytes contribute to the development, prevention and modulation of various autoimmune diseases. Regulatory T and B cell subsets appear to play a prominent role in the prevention of autoimmune diseases. The recent identification of novel regulatory Th17 cells, termed as Treg17 cells, has expanded the scope of regulatory T lymphocytes (Treg cells) in the prevention of autoimmune diseases. Similarly, novel regulatory B cell subsets, termed as Breg cells, acting on their own or by inducing Treg cells have extended the role of B lymphocytes in the prevention and regulation of autoimmune diseases. We suggest that Treg17 cells and Breg cells have an important immunoregulatory role in autoimmune diseases.

Journal of Immunology, Apr 15, 2008

Arthritis Research & Therapy, Jan 26, 2001

We discuss the presence of anti-keratin antibodies (AKA) of the IgG class in patients with define... more We discuss the presence of anti-keratin antibodies (AKA) of the IgG class in patients with defined juvenile idiopathic arthritis (JIA). An indirect immunofluorescence test and rat oesophagus substrate was used for the detection and quantification of AKA antibodies in patients´ sera. Overall 33/60 patients with JIA had sera positive for AKA (55 %, P = 0,0001) ranging from 1:10 to 1:160 dilutions. Following idiopathic arthritis of childhood classification criteria AKA occurred in 2/7 patients with systemic disease (28,6 %), in 13/30 patients with RF negative polyarthritis (43,3 %, P = 0,008) and in 15/18 RF positive polyarthritis (83,3 %, P = 0,000002). AKA were also found in a small cohort of patients with oligoarthritis (1/3) and psoriatic arthritis (2/2). AKA positivity occurred in 3/26 healthy controls at a 1:20 dilution. The presence of AKA was correlated as well as with the severity of the disease. Our study revealed that AKA was present overall in 18/29 patients (62%) with severe JIA and in 12/26 patients (46,2 %) with non-severe disease, however this did not reach statistical significance (P = 0,18). We also observed that AKA remained positive regardless of disease activity. AKA were detectable in 55,6 % patients with active JIA and in 48,6 % patients in the complete or near remission. Acknowledgement: This research was supported by a European Commission (Acronym: EUROBANK, contract no: QOL-2000-14.1), web site http://www.ncl.ac.uk and by grant of 2nd Medical Faculty, Charles University in Prague, VZ no. 111300003. P2 The significance of antibodies to cyclic citrullinated peptide, antikeratin antibodies, antiperinuclear factor, rheumatoid factor isotypes and HLA shared epitope in prediction of erosive disease in early rheumatoid arthritis patients

Journal of Maternal-fetal & Neonatal Medicine, May 17, 2012

To establish gender-specific differences in maternal and fetal immune response in healthy human f... more To establish gender-specific differences in maternal and fetal immune response in healthy human fetuses at term. Methods: Forty-five women with elective caesarean sections for uncomplicated singleton pregnancies were recruited for two studies. Using a multiplex biomarker immunoassay system, unstimulated maternal and fetal plasma concentrations of interleukin (IL)-1β, IL-1ra, IL-6, IL-8, macrophage inflammatory protein (MIP)-1α, and tumor necrosis factor (TNF)-α were measured from one study population. Lipopolysaccharide (LPS)stimulated cytokine response was measured in a second study. Results: There were no significant gender differences in either maternal or fetal unstimulated plasma cytokine concentrations, but concentrations of the proinflammatory cytokines IL-1β and IL-6 were significantly greater in male fetal LPS-stimulated samples than in female fetal samples. Conclusions: Blood of male fetuses mounts a larger pro-inflammatory response to lipopolysaccharide (LPS). This heightened response could be a critical pathway in promoting premature rupture of membranes (PPROM) and may be associated with life long differential gender response to infection.

Clinical and Experimental Immunology, Jan 2, 2015

The increased risk and persistence of infections in diabetic condition is probably associated wit... more The increased risk and persistence of infections in diabetic condition is probably associated with defects in the cellular immune responses. We have previously shown a decrease in the production of interferon (IFN)-α by dendritic cells (DCs) in diabetic subjects. The basal level of IFN-α in splenic plasmacytoid DCs (pDCs) is also lower in non-obese diabetic (NOD) mice compared to prediabetic mice. The objective of this study was to analyse the ability of diabetic mice to mobilize innate and CD8 + T cell-mediated immune response to influenza A virus (IAV) with the live influenza A/Puerto Rico/8/1934 H1N1 (PR8) strain or with its immunodominant CD8 + T cell epitopes. We found that following immunization with IAV, the level of IFN-α in diabetic mice was increased to the level in prediabetic mice. Immunization of NOD mice with the immunodominant IAV PR8 peptide induced clonal expansion of IFN-γ-producing CD8 + T cells similar to the response observed in prediabetic mice. Thus, diabetic and prediabetic NOD mice have a similar capacity for IFN-α and IFN-γ production by pDCs and CD8 + T cells, respectively. Therefore, the DC-related immune defect in diabetic NOD mice does not impair their capacity to develop an effective immune response to IAV. Our results suggest that reduced IFN-α production by diabetic human and mouse DCs is not an impediment to an effective immunity to IAV in type 1 diabetic subjects vaccinated with live attenuated influenza vaccine.

Advances in Experimental Medicine and Biology, 1997

Dendritic cells (DC) are specialist antigen presenting cells (APC) derived in common with other l... more Dendritic cells (DC) are specialist antigen presenting cells (APC) derived in common with other leukocytes from bone marrow stem cells.1,2 A myeloid derived precursor3 gives rise to immature circulating blood DC which enter the tissues and after interacting with antigen migrate to the T lymphocyte dependent areas of lymph nodes, where they deliver stimulatory signals to responding T lymphocytes. Recent studies suggest the growth and differentiation of myeloid DC is heavily influenced by cytokines, notably flt-3 ligand, SCF, GM-CSF, IL-4 and TNFα4–6. A unique DC precursor in blood can he distinguished from freshly isolated blood monocytes.7,8 However, considerable evidence suggests that monocytes exposed in vitro to certain cytokine combinations (notably including IL-4, which downregulates CD14) can acquire many, if not most DC characteristics.9 The DC series also includes a lymphoid precursor derived cell10 a subset of which, in the mouse, has an inhibiting effect on responding T lymphocytes. Lymphoid precursor cells have been described in man11,12 hut their function is unknown.

Advances in Urology, 2009

The management of urinary tract infection (UTI) in individuals with spinal cord injury (SCI) cont... more The management of urinary tract infection (UTI) in individuals with spinal cord injury (SCI) continues to be of concern, due to complications that can occur. An emerging concept that is a common underlying pathophysiological process is involved, wherein pathogens causing UTI have a role in inflammatory progression. We hypothesized that members of the commensal flora, such as lactobacilli, may counter this reaction through anti-inflammatory mediation. This was assessed in a pilot two-patient study in which probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri were administered to one patient and placebo to another, both along with antibiotics to treat acute UTI. Urinary TNF-alpha was significantly downregulated (P = .015) in the patient who received the probiotic and who used intermittent catheterization compared with patient on placebo and using an indwelling catheter. The extent to which this alteration resulted in improved well-being in spinal cord injured patients remains to be determined in a larger study.

Social Science Research Network, 2020

Long-term stress disrupts the neuroimmune interface, often resulting in immunosuppression. The de... more Long-term stress disrupts the neuroimmune interface, often resulting in immunosuppression. The deleterious effects of stress on mainstream T cells are well-documented. However, innate-like invariant T cells have been overlooked. Chief among these are invariant natural killer T (iNKT) and mucosa-associated invariant T (MAIT) cells, which fulfill important functions in anticancer and antimicrobial immunity. We found stress to surprisingly abrogate both TH1- and TH2-type responses mediated by iNKT cells. This was not due to cell death since iNKT cells were unusually refractory to stress-inflicted apoptosis. Activated iNKT cells in stressed mice triggered an abnormal response characterized by a ‘split’ inflammatory signature along with a sudden and robust induction of IL-10, IL-23 and IL-27. Pharmacological inhibitors, conditional knockout systems and gene expression analyses revealed that dysregulated iNKT cell responses were driven by iNKT cell-intrinsic glucocorticoid receptor signaling. Accordingly, the inhibitory effects of chronic stress on iNKT cell functions were lost upon habituation to predictable stressors. Importantly, iNKT cells in stressed mice failed to potentiate cytotoxicity against lymphoma targets or promote immune surveillance against metastatic melanoma. Using a congenic mouse model in which MAIT cells are abundant, these cells remained resistant to stress-induced death but demonstrated hindered TH1- and TH2-type cytokine responses, reminiscent of their iNKT cell counterparts. This was corroborated in culture systems in which we interrogated human peripheral blood and hepatic iNKT and MAIT cells. Our findings uncover a previously unappreciated mechanism of stress-induced immunosuppression and implicate the stress response as a potential barrier to iNKT cell- and MAIT cell-based immunotherapies.