5 research outputs found
Modulation of invariant NKT cell activity by cytokines and receptors in human disease
Invariant natural killer T (NKT) cells are innate-like lymphocytes with
both immunoregulatory and cytotoxic functions that play a role as
activators and regulators of the immune response in many diseases. NKT
cells are believed to bridge the innate and adaptive immune systems by
rapidly producing large quantities of cytokines after recognition of
CD1d-presented lipid antigens. NKT cells can be divided into two
phenotypically and functionally distinct subsets based on the expression
of CD4. How the NKT cells and their subsets are regulated, and how they
integrate signals from their environment to modulate immune responses is
still not fully understood.
In this thesis I have investigated how the activity of NKT cells can be
modulated by factors other than the TCR, such as cell surface receptors
and cytokines. These investigations have been based on blood samples from
healthy controls, and from patients suffering from HIV-1 infection or
atopic eczema (AE). We have found that CD4- NKT cells are able to
degranulate and kill target cells in an NKG2D-dependent but
TCR-independent manner in response to NKG2D stimulus. Moreover, we have
shown that NKG2D+ NKT cells frequently express perforin that polarizes
toward NKG2D-ligand expressing tumor cells. These data demonstrate that
the CD4- subset of human NKT cells can mediate direct lysis of
CD1d-negative target cells upon NKG2D engagement. We have further
characterized the phenotype and function of NKT cells in patients with
chronic diseases. In patients with chronic HIV- 1 infection, the CD4- NKT
cell subset showed increased expression of the inhibitory programmed
death-1 (PD-1) receptor, and displayed severe functional defects.
However, the functional impairment was not caused by PD-1 expression per
se because the defect could not be reversed by PD-1 blockade. In
addition, we have studied the effect of interleukin-2 (IL-2) on NKT cells
and natural killer (NK) cells in patients with chronic HIV-1 infection.
Material for this study was obtained from a longitudinally study, where
administration of IL-2 was added to the antiretroviral treatment (ART)
for one year. We found that NKT cells and NK cells responded with
different kinetics and in different ways to the IL-2 administration. The
NKT cells responded with a gradual numerical increase, but with no
significant functional changes. NK cells responded rapidly with an
expansion of the cytotoxic CD56dim NK cell subset and increased IFN-γ
production. However, the effects of IL-2 on these cells were generally
not sustained post treatment. NKT cells were also studied in the chronic
inflammatory skin disease AE where the patients have elevated levels of
plasma IL-18. Our data provide evidence that IL-18 is a potent activator
of human NKT cells promoting an acute pro-inflammatory CD1d-dependent
response, even in the absence of exogenous lipid antigens. Interestingly,
chronic exposure of NKT cells to IL-18 is inhibitory and skews the NKT
cell pool by selectively suppressing the proliferation of CD4+ NKT cells.
Importantly, our in vitro data are reflected in AE patients where reduced
numbers of CD4+ NKT cells are associated with elevated levels of IL-18
and disease severity.
In conclusion, the work presented here contributes to our understanding
of the function and role of NKT cells in human diseases including
infections and allergie
Evolution of DC-SIGN use revealed by fitness studies of R5 HIV-1 variants emerging during AIDS progression
<p>Abstract</p> <p>Background</p> <p>At early stages of infection CCR5 is the predominant HIV-1 coreceptor, but in approximately 50% of those infected CXCR4-using viruses emerge with disease progression. This coreceptor switch is correlated with an accelerated progression. However, those that maintain virus exclusively restricted to CCR5 (R5) also develop AIDS. We have previously reported that R5 variants in these "non-switch virus" patients evolve during disease progression towards a more replicative phenotype exhibiting altered CCR5 coreceptor interactions. DC-SIGN is a C-type lectin expressed by dendritic cells that HIV-1 may bind and utilize for enhanced infection of T cells in <it>trans</it>. To further explore the evolution of the R5 phenotype we analyzed sequential R5 isolates obtained before and after AIDS onset, i.e. at the chronic stage and during end-stage disease, with regard to efficiency of DC-SIGN use in <it>trans</it>-infections.</p> <p>Results</p> <p>Results from binding and <it>trans</it>-infection assays showed that R5 viruses emerging during end-stage AIDS disease displayed reduced ability to use DC-SIGN. To better understand viral determinants underlying altered DC-SIGN usage by R5 viruses, we cloned and sequenced the HIV-1 <it>env </it>gene. We found that end-stage R5 viruses lacked potential N-linked glycosylation sites (PNGS) in the gp120 V2 and V4 regions, which were present in the majority of the chronic stage R5 variants. One of these sites, amino acid position 160 (aa160) in the V2 region, also correlated with efficient use of DC-SIGN for binding and <it>trans</it>-infections. In fitness assays, where head-to-head competitions between chronic stage and AIDS R5 viruses were setup in parallel direct and DC-SIGN-mediated infections, results were further supported. Competitions revealed that R5 viruses obtained before AIDS onset, containing the V2 PNGS at aa160, were selected for in the <it>trans</it>-infection. Whereas, in agreement with our previous studies, the opposite was seen in direct target cell infections where end-stage viruses out-competed the chronic stage viruses.</p> <p>Conclusion</p> <p>Results of our study suggest R5 virus variants with diverse fitness for direct and DC-SIGN-mediated <it>trans</it>-infections evolve within infected individuals at end-stage disease. In addition, our results point to the importance of a glycosylation site within the gp120 V2 region for efficient DC-SIGN use of HIV-1 R5 viruses.</p
Lower cytokine secretion ex vivo by natural killer T cells in HIV-infected individuals is associated with higher CD161 expression
Objective: Natural killer T (NKT) cells are efficiently targeted by HIV and severely reduced in numbers in the circulation of infected individuals. the functional capacity of the remaining NKT cells in HIV-infected individuals is poorly characterized. This study measured NKT cell cytokine production directly ex vivo and compared these responses with both the disease status and NKT subset distribution of individual patients.Methods: NKT cell frequencies, subsets, and ex-vivo effector functions were measured in the peripheral blood mononuclear cells of HIV-infected patients and healthy controls by flow cytometry. We measured cytokines from NKT cells after stimulation with either a-galactosyl ceramide-loaded CD1d dimers (DimerX-alpha GalCer) or phorbol myristate acetate and ionomycin.Results: the frequencies of NKT cells secreting interferon-gamma and tumor necrosis factor-alpha were significantly lower in HIV-infected patients than healthy controls after DimerX-alpha GalCer treatment, but responses were similar after treatment with phorbol myristate acetate and ionomycin. the magnitude of the interferon-gamma response to DimerX-alpha GalCer correlated inversely with the number of years of infection. Both interferon-gamma and tumor necrosis factor-alpha production in response to DimerX-alpha GalCer correlated inversely with CD161 expression.Conclusion: the ex-vivo Th1 responses of circulating NKT cells to CD1d-glycolipid complexes are impaired in HIV-infected patients. NKT cell functions may be progressively lost over time in HIV infection, and CD161 is implicated in the regulation of NKT cell responsiveness. (C) 2009 Wolters Kluwer Health vertical bar Lippincott Williams & WilkinsNIAIDBrazilian Program for STD and AIDSMinistry of HealthSão Paulo City Health DepartmenFundacao de Arnparo a Pesquisa do Estado de São PauloJohn E. Fogarty International CenterAIDS Research Institute of the AIDS Biology ProgramCoordenação de Aperfeiçoamento de Pessoal de NÃvel Superior (CAPES)Brazilian Ministry of EducationUniv Calif San Francisco, Div Expt Med, Dept Med, San Francisco, CA 94110 USAUniversidade Federal de São Paulo, São Paulo, BrazilUniv São Paulo, São Paulo, BrazilKarolinska Univ Hosp, CIM, Dept Med, Karolinska Inst, Stockholm, SwedenSan Francisco Gen Hosp, Posit Hlth Program, San Francisco, CA 94110 USAAlbert Einstein Coll Med, Jacobi Med Ctr, Bronx, NY 10467 USAUniversidade Federal de São Paulo, São Paulo, BrazilNIAID: R37-A152731Ministry of Health: 914/BRA/3014 - UNESCO/KallasSão Paulo City Health Departmen: 20040.168.922-7/KallasFundacao de Arnparo a Pesquisa do Estado de São Paulo: 04/15856-9/KallasJohn E. Fogarty International Center: D43 TW00003Web of Scienc