Tracking Anti-Staphylococcus aureusAntibodies ProducedIn VivoandEx Vivoduring Foot Salvage Therapy for Diabetic Foot Infections Reveals Prognostic Insights and Evidence of Diversified Humoral Immunity

ABSTRACTManagement of foot salvage therapy (FST) for diabetic foot infections (DFI) is challenging due to the absence of reliable diagnostics to identify the etiologic agent and prognostics to justify aggressive treatments. AsStaphylococcus aureusis the most common pathogen associated with DFI, we aimed to develop a multiplex immunoassay of IgG in serum and medium enriched for newly synthesized anti-S. aureusantibodies (MENSA) generated from cultured peripheral blood mononuclear cells of DFI patients undergoing FST. Wound samples were collected from 26 DFI patients to identify the infecting bacterial species via 16S rRNA sequencing. Blood was obtained over 12 weeks of FST to assess anti-S. aureusIgG levels in sera and MENSA. The results showed that 17 out of 26 infections were polymicrobial and 12 were positive forS. aureus. While antibody titers in serum and MENSA displayed similar diagnostic potentials to detectS. aureusinfection, MENSA showed a 2-fold-greater signal-to-background ratio. Multivariate analyses revealed increases in predictive power of diagnosingS. aureusinfections (area under the receiver operating characteristic curve [AUC] > 0.85) only when combining titers against different classes of antigens, suggesting cross-functional antigenic diversity. Anti-S. aureusIgG levels in MENSA decreased with successful FST and rose with reinfection. In contrast, IgG levels in serum remained unchanged throughout the 12-week FST. Collectively, these results demonstrate the applicability of serum and MENSA for diagnosis ofS. aureusDFI with increased power by combining functionally distinct titers. We also found that tracking MENSA has prognostic potential to guide clinical decisions during FST.

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Contribution of Surface β-Glucan Polysaccharide to Physicochemical and Immunomodulatory Properties of Propionibacterium freudenreichii

Applied and Environmental Microbiology ◽

10.1128/aem.07027-11 ◽

2012 ◽

Vol 78 (6) ◽

pp. 1765-1775 ◽

Cited By ~ 27

Author(s):

Stéphanie-Marie Deutsch ◽

Sandrine Parayre ◽

Antoine Bouchoux ◽

Fanny Guyomarc'h ◽

Joëlle Dewulf ◽

...

Keyword(s):

Mononuclear Cells ◽

Human Peripheral Blood ◽

Single Gene ◽

Bacterial Species ◽

Interleukin 10 ◽

Propionibacterium Freudenreichii ◽

Peripheral Blood Mononuclear ◽

Content Type ◽

Significant Difference ◽

Anti Inflammatory

ABSTRACTPropionibacterium freudenreichiiis a bacterial species found in Swiss-type cheeses and is also considered for its health properties. The main claimed effect is the bifidogenic property. Some strains were shown recently to display other interesting probiotic potentialities such as anti-inflammatory properties. About 30% of strains were shown to produce a surface exopolysaccharide (EPS) composed of (1→3,1→2)-β-d-glucan due to a single gene namedgtfF. We hypothesized that functional properties ofP. freudenreichiistrains, including their anti-inflammatory properties, could be linked to the presence of β-glucan. To evaluate this hypothesis,gtfFgenes of three β-glucan-producing strains were disrupted. These knockout (KO) mutants were complemented with a plasmid harboringgtfF(KO-C mutants). The absence of β-glucan in KO mutants was verified by immunological detection and transmission electron microscopy. We observed by atomic force microscopy that the absence of β-glucan in the KO mutant dramatically changed the cell's topography. The capacity to adhere to polystyrene surface was increased for the KO mutants compared to wild-type (WT) strains. Anti-inflammatory properties of WT strains and mutants were analyzed by stimulation of human peripheral blood mononuclear cells (PBMCs). A significant increase of the anti-inflammatory interleukin-10 cytokine production by PBMCs was measured in the KO mutants compared to WT strains. For one strain, the role of β-glucan in mice gut persistence was assessed, and no significant difference was observed between the WT strain and its KO mutant. Thus, β-glucan appears to partly hide the anti-inflammatory properties ofP. freudenreichii; which is an important result for the selection of probiotic strains.

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Autologous Peripheral Blood Mononuclear Cells for Limb Salvage in Diabetic Foot Patients with No-Option Critical Limb Ischemia

Journal of Clinical Medicine ◽

10.3390/jcm10102213 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2213

Author(s):

Alessia Scatena ◽

Pasquale Petruzzi ◽

Filippo Maioli ◽

Francesca Lucaroni ◽

Cristina Ambrosone ◽

...

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Critical Limb Ischemia ◽

Diabetic Foot ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Limb Ischemia ◽

Control Group ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Peripheral blood mononuclear cells (PBMNCs) are reported to prevent major amputation and healing in no-option critical limb ischemia (NO-CLI). The aim of this study is to evaluate PBMNC treatment in comparison to standard treatment in NO-CLI patients with diabetic foot ulcers (DFUs). The study included 76 NO-CLI patients admitted to our centers because of CLI with DFUs. All patients were treated with the same standard care (control group), but 38 patients were also treated with autologous PBMNC implants. Major amputations, overall mortality, and number of healed patients were evaluated as the primary endpoint. Only 4 out 38 amputations (10.5%) were observed in the PBMNC group, while 15 out of 38 amputations (39.5%) were recorded in the control group (p = 0.0037). The Kaplan–Meier curves and the log-rank test results showed a significantly lower amputation rate in the PBMNCs group vs. the control group (p = 0.000). At two years follow-up, nearly 80% of the PBMNCs group was still alive vs. only 20% of the control group (p = 0.000). In the PBMNC group, 33 patients healed (86.6%) while only one patient healed in the control group (p = 0.000). PBMNCs showed a positive clinical outcome at two years follow-up in patients with DFUs and NO-CLI, significantly reducing the amputation rate and improving survival and wound healing. According to our study results, intramuscular and peri-lesional injection of autologous PBMNCs could prevent amputations in NO-CLI diabetic patients.

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Cytokine/Chemokine Secretion and Proteomic Identification of Upregulated Annexin A1 from Peripheral Blood Mononuclear Cells Cocultured with the Liver Fluke Opisthorchis viverrini

Infection and Immunity ◽

10.1128/iai.00901-13 ◽

2014 ◽

Vol 82 (5) ◽

pp. 2135-2147 ◽

Cited By ~ 10

Author(s):

Nuttanan Hongsrichan ◽

Kitti Intuyod ◽

Porntip Pinlaor ◽

Jarinya Khoontawad ◽

Puangrat Yongvanit ◽

...

Keyword(s):

Bile Duct ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Opisthorchis Viverrini ◽

Annexin A1 ◽

Peripheral Blood Mononuclear ◽

Content Type ◽

Blood Mononuclear Cells ◽

Liver Flukes

ABSTRACTWe investigated the cytokine/chemokine secretions and alteration of protein expression from peripheral blood mononuclear cells (PBMCs) cocultured with adult liver flukes (Opisthorchis viverrini) for 6 to 24 h. PBMC-derived proteins were identified by two-dimensional electrophoresis and mass spectrometry, and the cytokines/chemokines in the supernatant were assessed using a cytokine array. Exposure toO. viverriniinduced increases in secretion of proinflammatory cytokines, costimulating protein, adhesion molecules, and chemotactic chemokines relative to untreated controls. In contrast, secretion of the CD40 ligand, interleukin 16, and macrophage inflammatory protein 1β decreased. Proteomic analysis revealed that expression of 48 proteins was significantly altered in PBMCs stimulated withO. viverrini. Annexin A1 (ANXA1) was selected for further study, and immunoblotting showed upregulation of ANXA1 expression in PBMCs after 12 and 24 h coculture with liver flukes. In anin vivostudy, transcription and translation of ANXA1 significantly increased in livers of hamsters infected withO. viverriniat 21 days and from 3 months onwards compared to normal controls. Interestingly, immunohistochemistry revealed that ANXA1 was present not only in the cytoplasm of inflammatory cells but also in the cytoplasm of cholangiocytes, which are in close contact with the parasite and its excretory/secretory products in the biliary system. Expression of ANXA1 increased with time concomitant with bile duct enlargement, bile duct formation, and epithelial cell proliferation. In conclusion, several cytokines/chemokines secreted by PBMCs and upregulation of ANXA1 in PBMCs and biliary epithelial cells might have a role in host defense againstO. viverriniinfection and tissue resolution of inflammation.

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In VitroSpectrum of Pexiganan Activity When Tested against Pathogens from Diabetic Foot Infections and with Selected Resistance Mechanisms

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.04773-14 ◽

2015 ◽

Vol 59 (3) ◽

pp. 1751-1754 ◽

Cited By ~ 30

Author(s):

Robert K. Flamm ◽

Paul R. Rhomberg ◽

Katie M. Simpson ◽

David J. Farrell ◽

Helio S. Sader ◽

...

Keyword(s):

Diabetic Foot ◽

Antimicrobial Agents ◽

Resistance Mechanisms ◽

Surveillance Program ◽

Content Type ◽

Diabetic Foot Infections ◽

Antimicrobial Surveillance ◽

Mueller Hinton ◽

The Family ◽

Infection Types

ABSTRACTPexiganan, a 22-amino-acid synthetic cationic peptide, is currently in phase 3 clinical trials as a topical antimicrobial agent for the treatment of mild infections associated with diabetic foot ulcers. Bacterial isolates from the 2013 SENTRY Antimicrobial Surveillance Program designated as pathogens from diabetic foot infections (DFI) and Gram-negative and -positive pathogens from various infection types that harbored selected resistance mechanisms/phenotypes were tested against pexiganan in reference cation-adjusted Mueller-Hinton broth. The MIC50and MIC90against all organisms tested from DFI were 16 and 32 μg/ml, respectively.Escherichia coli,Klebsiella pneumoniae,Citrobacter koseri,Enterobacter cloacae,Acinetobacterspecies, andPseudomonas aeruginosaMIC values ranged from 8 to 16 μg/ml. Pexiganan MIC values amongStaphylococcus aureus(methicillin-resistantS. aureus[MRSA] and methicillin-susceptibleS. aureus[MSSA]), beta-hemolytic streptococci, andEnterococcus faeciumranged from 8 to 32 μg/ml. Pexiganan activity was not adversely affected for members of the familyEnterobacteriaceaeorP. aeruginosathat produced β-lactamases or resistance mechanisms to other commonly used antimicrobial agents. Decreased susceptibility to vancomycin did not affect pexiganan activity againstS. aureusorE. faecium.Enterococcus faecalisappears to be intrinsically less susceptible to pexiganan (MIC, 32 to 256 μg/ml). The “all organism” MIC90of 32 μg/ml for pexiganan in this study was >250-fold below the pexiganan concentration in the cream/delivery vehicle being developed for topical use.

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Evaluation of Immunostimulatory Activities of Synthetic Mannose-Containing Structures Mimicking the β-(1→2)-Linked Cell Wall Mannans of Candida albicans

Clinical and Vaccine Immunology ◽

10.1128/cvi.00298-12 ◽

2012 ◽

Vol 19 (11) ◽

pp. 1889-1893 ◽

Cited By ~ 14

Author(s):

Kaarina Ranta ◽

Kaisa Nieminen ◽

Filip S. Ekholm ◽

Moniká Poláková ◽

Mattias U. Roslund ◽

...

Keyword(s):

Candida Albicans ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Peripheral Blood Mononuclear ◽

Content Type ◽

Mouse Macrophages ◽

Ifn Γ ◽

Low Levels ◽

Necrosis Factor

ABSTRACTImmunostimulatory properties of synthetic structures mimicking the β-(1→2)-linked mannans ofCandida albicanswere evaluatedin vitro. Contrary to earlier observations, tumor necrosis factor (TNF) production was not detected after stimulation with mannotetraose in mouse macrophages. Divalent disaccharide 1,4-bis(α-d-mannopyranosyloxy)butane induced TNF and some molecules induced low levels of gamma interferon (IFN-γ) in human peripheral blood mononuclear cells (PBMC).

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Adjusted T SPOT.TB Criteria Can Increase the Specificity of Diagnosis When Differentiating Spinal Tuberculosis From Other Spinal Infections

10.21203/rs.3.rs-142854/v1 ◽

2021 ◽

Author(s):

Zhongjing Jiang ◽

Qile Gao ◽

Mingxing Tang ◽

Hongqi Zhang ◽

Yanbing Li ◽

...

Keyword(s):

Diagnostic Criteria ◽

Sensitivity And Specificity ◽

Mononuclear Cells ◽

Characteristic Curve ◽

Spinal Tuberculosis ◽

Youden Index ◽

Mycobacterium Tuberculosis Infection ◽

Peripheral Blood Mononuclear ◽

Relative Operating Characteristic ◽

Spinal Infections

Abstract Background: The ability of T-SPOT.TB to differentiate Mycobacterium tuberculosis infection of the spine from other infections is little known. This study quantified the efficiency, sensitivity, and specificity of the T-SPOT.TB assay to distinguish between spinal tuberculosis (STB) caused by M. tuberculosis and other infections of the spine and evaluated whether diagnostic performance was improved by adjusting the T-SPOT.TB assay criteria. Methods: From January 2010 to May 2020, 147 patients with spinal infections were recruited. Peripheral blood mononuclear cells were collected, and the number of spot-forming cells was observed. Patients’ white blood cell (WBC) counts, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), procalcitonin (PCT), and TB antibodies were recorded. Specimen/tissue bacteriological culture was the reference standard for sensitivity and specificity. Results: There were 77 (52.4%) participants with confirmed TB and 70 (47.6%) with other infections. The groups were comparable in T-SPOT.TB assay results, age, sex, lesions in the segments, WBC count, CRP, procalcitonin, ESR, and TB antibodies. The sensitivity and specificity of the T-SPOT.TB assay for identifying STB was 88.3% and 40.0%, respectively. On the basis of Relative operating characteristic curve (ROC) analysis and the Youden index, when we adjusted the T-SPOT.TB assay’s diagnostic criteria, ESAT-6>12 or CFP-10>19,the sensitivity and specificity of the T-SPOT.TB assay for identifying STB was 83.1% and 64.3%, respectively. Conclusion: The T-SPOT.TB assay has great sensitivity to distinguish STB from other spinal infections; however, the specificity is extremely low. Specificity can be significantly improved while sensitivity is guaranteed by adjusting the diagnostic criteria.

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Mycoplasma bovis-Induced Inhibition of Bovine Peripheral Blood Mononuclear Cell Proliferation Is Ameliorated after Blocking the Immune-Inhibitory Programmed Death 1 Receptor

Infection and Immunity ◽

10.1128/iai.00921-17 ◽

2018 ◽

Vol 86 (3) ◽

Cited By ~ 4

Author(s):

Muhammad Suleman ◽

Farhan S. Cyprian ◽

Steve Jimbo ◽

Teresia Maina ◽

Tracy Prysliak ◽

...

Keyword(s):

T Cell ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Lung Lavage ◽

Mycoplasma Bovis ◽

Peripheral Blood Mononuclear ◽

Content Type ◽

Programmed Death ◽

Systemic Spread ◽

Programmed Death 1

ABSTRACTMycoplasma bovis-induced immune suppression is a major obstacle faced by the host for controlling infections.M. bovisimpairment of antigen-specific T-cell responses is achieved through inhibiting the proliferation of peripheral blood mononuclear cells (PBMCs). This impairment may contribute to the persistence ofM. bovisinfection in various sites, including lungs, and its systemic spread to various organs such as joints, with the underlying mechanisms remaining elusive. Here, we elucidated the role of the immune-inhibitory receptor programmed death 1 (PD-1) and its ligand (PD-L1) inM. bovisinfection. Flow cytometry (FCM) analyses revealed an upregulation of PD-L1 expression on tracheal and lung epithelial cell lines afterM. bovisinfection. In addition, we found increased PD-L1 expression on purified lung lavage macrophages followingM. bovisinfection by FCM and determined its localization by immunofluorescence analysis comparing infected and control lung tissue sections. Moreover,M. bovisinfection increased the expression of the PD-1 receptor on total PBMCs and in gated CD4+and CD8+T-cell subpopulations. We demonstrated thatM. bovisinfection induced a significant decrease in CD4+PD-1INTand CD8+PD-1INTsubsets with intermediate PD-1 expression, which functioned as progenitor pools giving rise to CD4+PD-1HIGHand CD8+PD-1HIGHsubsets with high PD-1 expression levels. We blocked PD-1 receptors on PBMCs using anti-PD-1 antibody at the beginning of infection, leading to a significant restoration of the proliferation of PBMCs. Taken together, our data indicate a significant involvement of the PD-1/PD-L1 inhibitory pathway duringM. bovisinfection and its associated immune exhaustion, culminating in impaired host immune responses.

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Human herpesvirus 6: infection and disease following autologous and allogeneic bone marrow transplantation

Blood ◽

10.1182/blood.v87.12.5341.bloodjournal87125341 ◽

1996 ◽

Vol 87 (12) ◽

pp. 5341-5354 ◽

Cited By ~ 100

Author(s):

MP Kadakia ◽

WB Rybka ◽

JA Stewart ◽

JL Patton ◽

FR Stamey ◽

...

Keyword(s):

Marrow Transplantation ◽

Mononuclear Cells ◽

Allogeneic Bone Marrow Transplantation ◽

Human Herpesvirus ◽

Clinical Event ◽

Allogeneic Bone ◽

Human Herpesvirus 6 ◽

Peripheral Blood Mononuclear ◽

Antibody Titers ◽

Herpesvirus 6

Human herpesvirus 6 activity (HHV-6) was studied in 15 allogeneic and 11 autologous marrow transplantation patients. After transplantation, HHV-6 was isolated from the peripheral blood mononuclear cells of 12 of 26 patients (6 allogeneic and 6 autologous). All isolates were variant B. Eleven of 26 and 12 of 19 patients showed salivary shedding of HHV-6 DNA before and after transplantation, respectively. The antibody titer increased in 7 of 26 patients. Thus, 23 of 26 patients showed evidence of active HHV-6 infection either by virus isolation, salivary shedding, or increases in antibody titers. The fraction of saliva specimens positive in 19 patients was negatively associated with their antibody titers (P= .005). The proportion of cultures positive increased after transplantation (P = .007). Sinusitis was associated with HHV-6 isolation in autologous recipients (P= .002). In allogeneic patients, active human cytomegalovirus infection was associated with HHV-6 isolation (P = .04). No association was observed between HHV-6 infection and GVHD, pneumonia, delay in engraftment, or marrow suppression. Of the 120 clinical events analyzed in 26 patients, HHV-6 was defined as a probable cause of 16 events in 9 patients based on the propinquity of HHV-6 activity and the clinical event plus the absence of other identified causes of the event.

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Assessing Diabetic Foot Infections With the ThermoScale: A Comparative Thermometry Device Designed as a Patient Self-Screening Tool

The International Journal of Lower Extremity Wounds ◽

10.1177/1534734620948304 ◽

2020 ◽

pp. 153473462094830

Author(s):

Laure Arts ◽

Johan De Neve ◽

Samira Baharlou ◽

Nathalie Denecker ◽

Laura Kerselaers ◽

...

Keyword(s):

Temperature Measurement ◽

Diabetic Foot ◽

Characteristic Curve ◽

Screening Method ◽

Area Under The Curve ◽

Control Group ◽

Measurement Technology ◽

Diabetic Patients ◽

Home Setting ◽

Diabetic Foot Infections

Diabetic foot infection (DFI) is an important risk factor for amputation, and late diagnosis or referral is often incriminated for poor outcome. To enable an earlier diagnosis of DFI, comparative foot thermometry has been suggested as a self-screening method for patients in a home setting. We validated the efficacy of the ThermoScale, a weighing scale outfitted with temperature sensors that allows accurate temperature measurement in both feet. Temperature differentials in DFI patients (n = 52) were compared with a control group of similar diabetic patients (n = 45) without any foot wounds. Based on these findings, we drafted a receiver operating characteristic curve to determine an area-under-the-curve of 0.8455. This value suggests that the ThermoScale, as a diagnostic test, is reasonably accurate. A cutoff value of 2.15 °C temperature difference corresponded with a sensitivity of 88.9% and a specificity of 61.5%. As wearables, portable health electronics, and telemedicine become increasingly popular, we think that comparative temperature measurement technology is valuable in improving early diagnosis of DFIs.

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Differential In Vitro Cytokine Induction by the Species of Cryptococcus gattii Complex

Infection and Immunity ◽

10.1128/iai.00958-17 ◽

2018 ◽

Vol 86 (4) ◽

Cited By ~ 4

Author(s):

Patricia F. Herkert ◽

Jessica C. dos Santos ◽

Ferry Hagen ◽

Fatima Ribeiro-Dias ◽

Flávio Queiroz-Telles ◽

...

Keyword(s):

Mononuclear Cells ◽

Human Peripheral Blood ◽

Sensu Stricto ◽

Cryptococcus Gattii ◽

The Other ◽

Human Pbmcs ◽

Peripheral Blood Mononuclear ◽

Necrosis Factor Alpha ◽

Content Type ◽

Tnf Α

ABSTRACT Cryptococcal species vary in capsule and cell size, thermotolerance, geographic distribution, and affected populations. Cryptococcus gattii sensu stricto and C. deuterogattii affect mainly immunocompetent hosts; however, C. bacillisporus , C. decagattii , and C. tetragattii cause infections mainly in immunocompromised hosts. This study aimed to compare the capacities of different species of the C. gattii species complex to induce cytokines and antimicrobial molecules in human peripheral blood mononuclear cells (PBMCs). Cryptococcus bacillisporus and C. deuterogattii induced the lowest levels of tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), and IL-6 among the five species of the C. gattii complex. Cryptococcus deuterogattii induced higher levels of IL-22 than those induced by C. tetragattii and the environmental species C. flavescens . In addition, C. bacillisporus and C. gattii sensu stricto proliferated inside human monocyte-derived macrophages after 24 h of infection. All Cryptococcus species were able to generate reactive oxygen species (ROS) in human PBMCs, with C. bacillisporus and C. deuterogattii being more efficient than the other species. In conclusion, C. bacillisporus and C. deuterogattii induce lower levels of the proinflammatory cytokines TNF-α, IL-1β, and IL-6 and higher ROS levels than those induced by the other species. Species of the Cryptococcus gattii complex have different abilities to induce cytokine and ROS production by human PBMCs.

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