Alternative Spliced CD1D Transcripts in Human Bronchial Epithelial Cells

Abstract

CD1d is a MHC I like molecule which presents glycolipid to natural killer T (NKT) cells, a group of cells with diverse but critical immune regulatory functions in the immune system. These cells are required for optimal defence against bacterial, viral, protozoan, and fungal infections, and control of immune-pathology and autoimmune diseases. CD1d is expressed on antigen presenting cells but also found on some non-haematopoietic cells. However, it has not been observed on bronchial epithelium, a site of active host defence in the lungs. Here, we identify for the first time, CD1D mRNA variants and CD1d protein expression on human bronchial epithelial cells, describe six alternatively spliced transcripts of this gene in these cells; and show that these variants are specific to epithelial cells. These findings provide the basis for investigations into a role for CD1d in lung mucosal immunity.

Matched-Comparative Modeling of Normal and Diseased Human Airway Responses Using a Microengineered Breathing Lung Chip

“…For each reaction, 2 ml cDNA, 10 ml 2 3 Universal SYBR Green Supermix (Bio-Rad) and 3 ml of forward and reverse primers (300 nM final concentration), and 2 ml molecular biology-grade water were thoroughly mixed. PCR was performed over an initial cycle at 95 C for 5 min, followed by 45 cycles of 95 C for 5 s and 60 C for 30 s. Cycle of threshold (Ct) values were extracted, and results were analyzed comparatively using 2 ÀDDCt method by normalizing against housekeeping gene hypoxanthine phosphorribosyltransferase (HPRT) as previously described (Benam et al, 2011). In details, first DCt for each gene (e.g., DC HMOX1 = Ct HMOX1 -Ct HPRT ) was calculated, and then DDCt was established by subtracting DCt of non-smoking from DCt of smoking condition.…”

Generation and characterization of CD1d‐specific single‐domain antibodies with distinct functional features

“…Similarly, iNKT cells can provide cognate (via CD1d) and non‐cognate (via DC) help to B cells and induce and/or enhance humoral immune responses to various antigens . As CD1d is also expressed on certain epithelial cells, biologically relevant interactions between iNKT and epithelial cells have been proposed . Hence iNKT cells have been recognized for their ability to orchestrate microbial immunity as well as auto‐ and antitumour immunity …”

“…1,10 As CD1d is also expressed on certain epithelial cells, biologically relevant interactions between iNKT and epithelial cells have been proposed. 11,12 Hence iNKT cells have been recognized for their ability to orchestrate microbial immunity as well as auto-and antitumour immunity. 1,10,13 Abbreviations: APC, allophycocyanin; ATRA, all-trans retinoic acid; CFU, colony-forming unit; DC, dendritic cells; FITC, fluorescein isothiocyanate; IFN-c, interferon-c; IL-2, interleukin-2; iNKT, invariant natural killer T; LPS, lipopolysaccharide; mAb, monoclonal antibody; MM, multiple myeloma; moDC, monocyte derived DC; PE, phycoerythrin; PI, propidium iodide; TCR, T-cell receptor; TNF-a, tumour necrosis factor-a; VHH, variable domain of heavy-chain-only antibodies; WT, wild-type; a-GalCer, a-galactosylceramide…”

The role of respiratory epithelium in host defence against influenza virus infection

“…iNKT cells have immunoregulatory roles [176] and have been shown to be ‘held’ within the intravascular space of the lungs, next to airway epithelium [177] . In this position iNKT cells can be activated rapidly and move into the lung interstitium and airway lumen where they may be involved in enhancing the immune response and regulating the size of myeloid responses [177] , [178] , [179] . There is evidence that human airway epithelium can express CD1d [179] , [180] , but it is unknown whether epithelial CD1d is involved in activating iNKT cells; and the cognate glycolipid which activates iNKT cells during IAV infection is also unknown However, IL-12 produced by intra-epithelial CD103 DC may activate iNKT cells in an indirect pathway [181] , [182] , [183] In the mouse, presence, and exogenous activation, of lung iNKT cells by α-GalCer, protects against lethal H1N1 and H3N2 influenza in prophylactic settings [158] , [178] [224] .…”