title: Investigating the role of Natural Killer T-cells in Gram negative infections of patients with type 2 diabetes mellitus.
abstract: BACKGROUND
  Invariant Natural Killer T (iNKT) cells belong to innate immunity and combine T-cell receptor specificity with Natural Killer surface markers. They can produce cytokines immediately after stimulation and direct immunity to either Th1 or Th2 cytokine production. iNKT cells participate in a variety of immune responses, such as microbial infections, autoimmunity, and cancer. Type 2 Diabetes Mellitus (T2DM) has been associated with activated innate immunity and certain cytokine profile during microbial infections. This study aimed to evaluate whether iNKT cells have a role in the immune response of T2DM patients during infections with gram-negative bacteria.
  
  
  METHODS
  The T2DM group consisted of patients (n =11) who had a diagnosis of T2DM for at least six months and febrile illness for three days, while the control group consisted of patients (n =11) who had not T2DM, but were febrile for three days. All patients were infected by gram-negative bacteria. Physical examination was performed, and peripheral blood was drawn on days three and six of febrile illness. Flow cytometry was utilized for iNKT cell identification with monoclonal antibodies Phycoerythrin (PE) - Cyanin (CY) 5 anti-Human CD3, Fluorescein isothiocyanate (FITC) anti-Human CD4, PE anti-human invariant NKT T-Cell Receptor. For intracellular staining, we used Alexa Fluor anti-Human interferon-γ (IFN-γ) and Allophycocyanin (APC) anti-human interleukin-4 (IL-4). The variables processed were: CD3+IL-4+iNKT+ , CD4+IL-4+iNKT+, CD3+IFNγ+iNKT+, CD4+IFNγ+iΝΚΤ+, CD3+iNKT+, CD4+iNKT+ ,CD3+IL4+, CD4+IL-4+, CD3+IFNγ+, CD4+IFNγ+ on days three and six of febrile illness (CD3+, CD4+: T lymphocyte surface markers, iNKT+: invariant Natural Killer T- Cell Receptor, IL4+: interleukin 4, IFNγ+: interferon γ).
  
  
  RESULTS
  Comparisons between T2DM patients and controls revealed no statistically significant difference in any of the study's variable. Regarding within T2DM patients comparisons CD4+IL4+iNKT+, CD3+IL4+iNKT+, CD4+IFN+iNKT+, CD3+IFN+iNKT+, and CD3+iNKT+  decreased, whereas CD3+IL4+ was increased at day six compared to day three. Within control group CD4+IL4+iNKT+, CD3+IL4+iNKT+, and CD3+iNKT+ were decreased, whereas CD4+IFN+, CD3+IFN+ were increased at day six compared to day three.
  
  
  CONCLUSIONS
  The absence of statistical difference between T2DM patients and controls implies that the role of iNKT cells is virtually the same in both groups of patients during the course of gram-negative infections and that there is no numerical variance of this cell population between the two groups. Despite the small sample size, we notice that all iNKT parameters (both IL4/IFNγ) are suppressed in the T2DM group during the later phase, but only those concerning IL4+iNKT+ in the control group, suggesting that IFNγ production remains elevated in the controls. A compensatory anti-inflammatory type-response could provide an explanation for the prevalence of IL4 production during the later phase of infection in T2DM and the sustained production of IFNγ in controls. Hippokratia 2015; 19 (3): 231-234.
authors.affiliation: Second Internal Medicine Department, Hippokratio General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece.
authors.forename: P
authors.initials: P
authors.lastname: Karagianni
authors.affiliation: Second Internal Medicine Department, Hippokratio General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece.
authors.forename: S A
authors.initials: SA
authors.lastname: Polyzos
authors.affiliation: Hematology Department, Theagenion Hospital, Thessaloniki, Greece.
authors.forename: D
authors.initials: D
authors.lastname: Bougiouklis
authors.affiliation: Second Internal Medicine Department, Hippokratio General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece.
authors.forename: A
authors.initials: A
authors.lastname: Tsapas
authors.affiliation: Internal Medicine Department, Papageorgiou General Hospital, Aristotle University of Thessaloniki, Thessaloniki, Greece.
authors.forename: K
authors.initials: K
authors.lastname: Paletas
keywords: Type 2 diabetes mellitus; gram-negative infection; iNKT cells; invariant natural killer T cells