Circadian Rhythm Disruption and Sepsis in Severe Trauma Patients.

BACKGROUND: Circadian rhythms are important regulators of immune functions. Admission to an intensive care unit may impact molecular clock activity and host response. Our objective was to assess and compare the immune circadian rhythms in trauma patients who develop and in those who do not develop sepsis. METHODS: Blood samples were collected from severe trauma patients within 4 days after admission, with collections taking place every 4 h over a 24-h period. Cortisol and cytokines were measured with immunoassays. Whole-blood expression of 3 clock genes (Bmal1, Per2, and Per3) was studied by reverse transcription quantitative polymerase chain reaction. Neutrophils, monocytes, and lymphocytes were analyzed by flow cytometry. Patients with and without sepsis were compared with the cosinor mixed model to estimate mesors, amplitudes, and acrophases. RESULTS: Thirty-eight patients were enrolled in the study, and 13 developed at least 1 septic episode. The septic patients had higher levels of cortisol than the nonseptic patients (mesor at 489 nmol/L vs. 405 nmol/L, P < 0.05) and delayed acrophases (22 h vs. 15 h, P < 0.05). They also had lower lymphocyte counts (mesor at 785 vs. 1,012 cells/µL, P < 0.05), higher neutrophil counts (mesor at 7,648 vs. 7,001 cells/µL, P < 0.05), and monocyte counts (mesor at 579 vs. 473 cells/µL, P < 0.05) than the nonseptic patients. Although no amplitude difference was identified, the acrophases were significantly different between the 2 groups for lymphocytes, interleukin 10 and tumor necrosis factor. CONCLUSION: We demonstrated that all trauma patients had impaired circadian rhythms of cortisol, cytokines, leukocytes, and clock genes. Early circadian disruption was associated with the occurrence of sepsis and might be a marker of sepsis severity.

Sepsis is associated with lack of monocyte HLA-DR expression recovery without modulating T-cell reconstitution after lung transplantation.

BACKGROUND: Immunosuppressive strategy targets mainly adaptive immunity after solid organ transplantation. We assessed the influence of early post-operative sepsis on T cell and monocyte reconstitution in anti-thymocyte globulin (ATG)-treated lung transplant recipients. METHODS: We retrospectively included recipients who underwent a first lung transplant at our Lung Transplant Center (Marseille, France) between July 2011 and February 2013. Peripheral blood T-lymphocyte subset counts and monocyte HLA-DR (mHLA-DR) expression routinely performed by flow cytometry within 60 days post-transplant were analyzed. We compared the immune kinetics of patients who did or did not develop sepsis during the post-operative intensive care unit stay. RESULTS: Among the 37 recipients included, 19 patients (51%) developed at least one episode of sepsis. At the ICU admission, septic recipients had higher SOFA score (9 [7.5-9] versus 6 [4-7]), p = .01), higher primary graft dysfunction score (1.4 ±â€¯1.4 versus 0.3 ±â€¯0.7, p = .008) and more frequent use of ECMO (47% versus 0%, p = .003). Whereas both groups had similar T-lymphocytes reconstitution in the post-operative period, mHLA-DR reconstitution was dramatically affected in septic patients after day 14, median mHLA-DR expression at 2.3 MFI [1.3-3.5] in the septic versus 8.0 MFI [5.1-10.5] in the non-septic group, p = .02. CONCLUSION: We found that sepsis is negatively correlated with the mHLA-DR expression but not adaptive T cell immune reconstitution. This finding highlights the importance of immunomonitoring after lung transplantation and questions the strategy of a lower immunosuppression therapy in context of sepsis.

Effect of Immunosuppression on Target Blood Immune Cells Within 1 Year After Lung Transplantation: Influence of Age on T Lymphocytes.

BACKGROUND Lymphocytes are targeted by immunosuppressive therapy in solid organ transplantation and they influence allograft outcome. MATERIAL AND METHODS Peripheral blood lymphocyte subsets (PBLS) determined by flow cytometry during the first year post-transplant from patients who underwent a first lung transplantation in a French University Hospital between December 2011 and July 2013 were retrospectively analyzed according to recipient characteristics and allograft outcome. RESULTS Fifty-seven recipients were enrolled and 890 PBLS were collected. T lymphocytes and NK cells were rapidly decreased, below normal range, from the first postoperative days. B cells decreased more gradually, remaining within normal range, with the lowest level reached after day 100. In multivariate analysis, greater T lymphopenia was found in older recipients (-414 [-709 to -119] cells/µL, p=0.007). According to the outcome, multivariate analysis evidenced lower levels of lymphocytes when bacterial and viral infection occurred (-177 [-310 to -44] cells/µL, p=0.009 and (-601 [-984 to -218] cells/µL, p=0.002, respectively), higher CD8+ T lymphocytes with BOS (+324 [+94 to +553] cells/µL, p=0.006), and higher leukocytes with restrictive allograft syndrome (+3770 [+418 to +7122] cells/µL, p=0.028). CONCLUSIONS Aging is associated in our cohort with more severe T lymphopenia after induction therapy for lung transplantation. The analysis of leukocytes and PBLS is associated with specific profile according to the allograft outcome.

HIV-1 infection and first line ART induced differential responses in mitochondria from blood lymphocytes and monocytes: the ANRS EP45 "Aging" study.

BACKGROUND: The ANRS EP45 "Aging" study investigates the cellular mechanisms involved in the accelerated aging of HIV-1 infected and treated patients. The data reported focus on mitochondria, organelles known to be involved in cell senescence. METHODS: 49 HIV-1 infected patients untreated with antiretroviral therapy, together with 49 seronegative age- and sex-matched control subjects and 81 HIV-1 infected and treated patients, were recruited by 3 AIDS centres (Marseille, Montpellier, Nice; France; http://clinicaltrials.gov/, NCT01038999). In more than 88% of treated patients, the viral load was <40 copies/ml and the CD4+ cell count was >500/mm(3). ROS (reactive oxygen species) production and ΔΨm (inner membrane potential) were measured by flow cytometry in blood lymphocytes and monocytes (functional parameters). Three mitochondrial network quantitative morphological parameters were computed using confocal microscopy and image analysis. Three PBMC mitochondrial proteins (porin and subunits 2 and 4 of cytochrome C oxidase encoded by mtDNA or nuclear DNA, respectively) were analysed by western blotting. RESULTS: Quantitative changes in PBMC mitochondrial proteins were not induced by either HIV-1 infection or ART. Discriminant analysis integrating functional (ROS production and ΔΨm) or morphological (network volume density, fragmentation and branching) parameters revealed HIV-1 infection and ART differential effects according to cell type. First line ART tended to rescue lymphocyte mitochondrial parameters altered by viral infection, but induced slight changes in monocytes. No statistical difference was found between the effects of three ART regimens on mitochondrial parameters. Correlations between functional parameters and viral load confirmed the damaging effects of HIV-1 in lymphocyte mitochondria. CONCLUSIONS: In patients considered to be clinically stable, mitochondria exhibited functional and morphological modifications in PBMCs resulting from either direct or indirect effects of HIV-1 infection (lymphocytes), or from first line ART (monocytes). Together with other tissue impairments, these changes may contribute to global aging.