ABSTRACT
The complexity of sepsis pathophysiology hinders patient management and therapeutic decisions. In this proof-of-concept study we characterised the underlying host immune response alterations using a standardised immune functional assay (IFA) in order to stratify a sepsis population. In septic shock patients, ex vivo LPS and SEB stimulations modulated, respectively, 5.3% (1/19) and 57.1% (12/21) of the pathways modulated in healthy volunteers (HV), highlighting deeper alterations induced by LPS than by SEB. SEB-based clustering, identified 3 severity-based groups of septic patients significantly different regarding mHLA-DR expression and TNFα level post-LPS, as well as 28-day mortality, and nosocomial infections. Combining the results from two independent cohorts gathering 20 HV and 60 patients, 1 cluster grouped all HV with 12% of patients. The second cluster grouped 42% of patients and contained all non-survivors. The third cluster grouped 46% of patients, including 78% of those with nosocomial infections. The molecular features of these clusters indicated a distinctive contribution of previously described genes defining a "healthy-immune response" and a "sepsis-related host response". The third cluster was characterised by potential immune recovery that underlines the possible added value of SEB-based IFA to capture the sepsis immune response and contribute to personalised management.
Subject(s)
Shock, Septic/classification , Shock, Septic/pathology , Aged , Biomarkers/blood , Cross Infection , Enterotoxins/immunology , Female , Gene Expression , Gene Expression Profiling/methods , HLA-DR Antigens/metabolism , Humans , Lipopolysaccharides/pharmacology , Lipopolysaccharides/standards , Male , Middle Aged , Monocytes/metabolism , Proof of Concept Study , Sepsis/metabolism , Shock, Septic/mortality , Tumor Necrosis Factor-alpha/metabolismSubject(s)
Adenosine , Drug Contamination , Escherichia coli/chemistry , Glutamic Acid , Lipopolysaccharides/standards , Animals , Dialysis , Lipopolysaccharides/pharmacology , Neuroimmunomodulation/drug effects , Norepinephrine/pharmacokinetics , Parietal Lobe/drug effects , Parietal Lobe/immunology , Rats , Sympathomimetics/pharmacokinetics , TritiumABSTRACT
A procedure for standardizing Brucella abortus smooth lipopolysaccharide used in diagnostic tests for brucellosis is proposed. The procedure is based on the reactivity of antigen preparations with a panel of sera with or without antibody to B. abortus using a set of parameters established with 13 antigen preparations. For each serum dilution, a mean and two standard deviations were calculated in the indirect and competitive enzyme immunoassays. If data obtained with an antigen preparation, using the same serum dilutions, falls within the range established using two standard deviations, the antigen would be considered acceptable for diagnostic use.
Subject(s)
Brucella abortus/chemistry , Brucella abortus/immunology , Lipopolysaccharides/standards , Animals , Antibodies, Bacterial/blood , Brucellosis/diagnosis , Cattle , Data Interpretation, Statistical , Enzyme-Linked Immunosorbent Assay/methods , Female , Lipopolysaccharides/isolation & purification , Reference StandardsSubject(s)
Endotoxins/standards , Animals , Body Temperature , Drug Stability , Endotoxins/toxicity , Humans , Limulus Test , Lipopolysaccharides/standards , Methods , Pyrogens , Rabbits , Reference Standards , SalmonellaABSTRACT
The properties of a reference bacterial endotoxin prepared from Escherichia coli and its phthalylated derivative were studied in normal human volunteers infected intravenously with the compounds. The minimal pyrogenic dose of the reference endotoxin is about 0.1-0.5 ng/kg. The increase in white blood cell count, absolute granulocyte count, absolute immature granulocyte count, and concentrations of serum amyloid A, cortisol, and growth hormone was directly related to the concentration of reference endotoxin administered. Phthalylated reference endotoxin up to 1,000 ng/kg (at least 500 ng of the patent compound/kg) was administered to normal human volunteers without significant changes in temperature, white blood cell count, absolute granulocyte count, and concentrations of serum amyloid A, cortisol, and growth hormone. Thus, this study defines biologic properties of the new reference bacterial endotoxin in humans and demonstrates effective detoxification by phthalylation of the present compound.
