Antibody Secreting Cell and Cyotokine Profiles in Neonates on ECMO
Condition: Persistent Fetal Circulation Syndrome; Diaphragmatic Hernia; Meconium Aspiration; Sepsis
MeSH terms: Meconium Aspiration Syndrome; Persistent Fetal Circulation Syndrome; Hernias, Diaphragmatic, Congenital; Hernia, Diaphragmatic
Study type: Observational

Summary: 
      Infants are placed on ECMO for correction of reversible respiratory failure. Often, because a
      few of the reasons for respiratory failure show us similar things in the baby, it is
      difficult to determine exactly which is causing the biggest problem. We are now capable of
      measuring certain cells and proteins in these infants that may help us more accurately
      diagnose the exact problem. We hypothesize that infants placed on ECMO will show unique
      antibody-secreting cells responses and patterns of cytokine and chemokine (protein) response
      to illness and to the ECMO circuit. If we find unique patterns to these cells or proteins,
      they may be able to predict outcomes or guide treatment of these infants.
    

Detailed description: 
      Specific Aims Primary Objective

      1. Determine the rise, peak, and fall of immunoglobulin isotype-specific ASC's, and
      immunoactive proteins (cytokines and chemokines) from sequential samples of peripheral blood
      from infants on ECMO.

      Secondary Objectives:

        1. Determine the most appropriate time to sample blood from infants with suspected sepsis
           for ASC diagnostic assay.

        2. Characterize the incidence of culture-negative sepsis that leads to ECMO.

        3. Determine immunoglobulin isotype-specific levels of ASC in infants with and without
           infection.

        4. Establish an archive of mononuclear cells and plasma to use in development of pathogen
           specific ASC assays.

      Hypothesis Infants on ECMO will have a high ASC response and unique cytokine/chemokine
      patterns due to possible underlying infection and exposure to many foreign antigens (blood
      products, ECMO circuit). A significant portion of these will have ASC's with specificity for
      common causes of neonatal sepsis that is not detected by routine blood culture.

      Procedures:

      Residual samples will be collected from those used in routine procedures for infants on ECMO.
      The approximate volume/sample will be 0.5-0.8ml. Specimens will be processed using methods
      well established in our laboratory. Briefly, PBMC's will be isolated via Ficoll gradient and
      archived in liquid nitrogen at -80C. Batch analysis of ASC levels and lymphocyte
      proliferation activity will be performed when sufficient number of specimens are accumulated.
      A detailed profile and quantification of immune cells will be determined by Fluorescent
      Activated Cell Sorter (FACS) staining for CD3, CD4, CD8, CD27, CD38, CD45, and HLA-DR. A bead
      micro-array will be used to detect levels of immunoactive molecules, also done on the FACS.
      The proteins detected will include, but may not be limited to, the following: IL1β, IL2, IL4,
      IL5, IL6, IL7, IL8, IL10, IL12p70, IL13, IL17, GCSF, GMCSF, IFN-γ, MCP-1, MIP-1β, TNFα. The
      ASC procedure be that established by Van de Verg modified to use membrane surface
      microculture plates in place of agar with outcomes read by CTL analyzer in place of manual
      count. LPA assays will use long established techniques.
    

Observational model: Cohort
Time perspective: Prospective
Sampling method: Probability Sample
Study population: 
        Infants requiring ECMO
      
Biospecimen description: 
      Plasma and whole blood
    
Keywords: ECMO; extracorporeal membrane oxygenation; cytokines; chemokines