Phenotypic alteration of neutrophils in the blood of HIV seropositive patients

Tom Cloke, Markus Munder, Philip Bergin, Shanthi Herath, Manuel Modolell, Graham Taylor, Ingrid Müller, Pascale Kropf, Tom Cloke, Markus Munder, Philip Bergin, Shanthi Herath, Manuel Modolell, Graham Taylor, Ingrid Müller, Pascale Kropf

Abstract

We have recently identified a novel population of activated low-density granulocytes (LDGs) in peripheral blood mononuclear cells of HIV seropositive patients. LDGs have a similar morphology to normal density granulocytes (NDGs), but are phenotypically different. Here we measured the expression levels of different phenotypic markers of granulocytes in the blood of HIV seropositive patients at different stages of HIV infection to determine whether the phenotype of NDGs and LDGs are affected by disease severity. Our results reveal that the phenotype of NDGs, but not that of LDGs, varies according to the severity of the disease.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1. Phenotypic analysis of NDGs.
Figure 1. Phenotypic analysis of NDGs.
NDGs were isolated from the blood of HIV+ patients with CD4+ T cell counts >350 (n = 11) or <350 cells/µL (n = 10) as described in materials and methods and the expression levels of phenotypic markers were determined by flow cytometry. Isotype controls: <1%. Statistical significance was determined by a two-tailed Mann-Whitney test. Box = interquartile range and median; whiskers = range.
Figure 2. Correlation between CD4 + T…
Figure 2. Correlation between CD4+ T cells and phenotypic markers.
NDGs were isolated from the blood of HIV+ patients (n = 21) as described in materials and methods and the expression levels of phenotypic markers were determined by flow cytometry. Correlation between CD4+ T cell counts and phenotypic markers was determined by a Spearman's rank test.
Figure 3. Correlation between viral load and…
Figure 3. Correlation between viral load and phenotypic markers.
NDGs were isolated from the blood of HIV+ patients (n = 21) as described in materials and methods and the expression levels of phenotypic markers were determined by flow cytometry. Correlation between viral load and phenotypic markers was determined by a Spearman's rank test.
Figure 4. Phenotypes of NDGs and LDGs…
Figure 4. Phenotypes of NDGs and LDGs in CD4low and CD4high HIV+ patients.
PBMCs and NDGs were isolated from the blood of HIV+ patients with CD4+ T cell counts >350 (n = 11) or <350 cells/µL (n = 10) as described in materials and methods and the expression levels of phenotypic markers were determined by flow cytometry. Isotype controls: <1%. Statistical significance was determined by a two-tailed Mann-Whitney test. Box = interquartile range and median; whiskers = range.

References

    1. Drescher B, Bai F (2012) Neutrophil in viral infections, friend or foe? Virus Res
    1. Kuritzkes DR (2000) Neutropenia, neutrophil dysfunction, and bacterial infection in patients with human immunodeficiency virus disease: the role of granulocyte colony-stimulating factor. Clin Infect Dis 30: 256–260.
    1. Saitoh T, Komano J, Saitoh Y, Misawa T, Takahama M, et al. (2012) Neutrophil extracellular traps mediate a host defense response to human immunodeficiency virus-1. Cell Host Microbe 12: 109–116.
    1. Munder M, Mollinedo F, Calafat J, Canchado J, Gil-Lamaignere C, et al. (2005) Arginase I is constitutively expressed in human granulocytes and participates in fungicidal activity. Blood 105: 2549–2556.
    1. Cederbaum SD, Yu H, Grody WW, Kern RM, Yoo P, et al. (2004) Arginases I and II: do their functions overlap? Mol Gen Metabol 81: S38–S44.
    1. Munder M (2009) Arginase: an emerging key player in the mammalian immune system. Br J Pharmacol 158: 638–651.
    1. Ochoa AC, Zea AH, Hernandez C, Rodriguez PC (2007) Arginase, prostaglandins, and myeloid-derived suppressor cells in renal cell carcinoma. Clin Cancer Res 13: 721s–726s.
    1. Popovic PJ, Zeh HJ 3rd, Ochoa JB (2007) Arginine and immunity. J Nutr 137: 1681S–1686S.
    1. Bronte V, Zanovello P (2005) Regulation of immune responses by L-arginine metabolism. Nat Rev Immunol 5: 641–654.
    1. Munder M, Choi BS, Rogers M, Kropf P (2009) L-arginine deprivation impairs Leishmania major-specific T cell responses. European Journal of Immunology 39: 2161–2172.
    1. Weinberg JB, Lopansri BK, Mwaikambo E, Granger DL (2008) Arginine, nitric oxide, carbon monoxide, and endothelial function in severe malaria. Curr Opin Infect Dis 21: 468–475.
    1. Zea AH, Culotta KS, Ali J, Mason C, Park HJ, et al. (2006) Decreased Expression of CD3 zeta and Nuclear Transcription Factor kappa B in Patients with Pulmonary Tuberculosis: Potential Mechanisms and Reversibility with Treatment. J Infect Dis 194: 1385–1393.
    1. Abebe T, Hailu A, Woldeyes M, Mekonene W, Bilch K, et al. (2012) Local increase of arginase activity in lesions of patients with cutaneous leishmaniasis in Ethiopia. PLoS NTD 6: e1684.
    1. Takele Y, Abebe T, Weldegebreal T, Hailu A, Hailu W, et al. (2012) Arginase activity in the blood of patients with visceral leishmaniasis and HIV infection. PLoS NTD 7 (1) e1977.
    1. Abebe T, Takele T, Weldegebreal T, Cloke T, Closs E, et al. (2013) Arginase: a marker of disease status in patients with visceral leishmaniasis. PLoS NTD in press.
    1. Cloke T, Garvery L, Choi BS, Abebe T, Hailu A, et al. (2010) Increased arginase activity correlates with disease severity in HIV seropositive patients. Journal of Infectious Diseases 202: 374–385.
    1. Cloke T, Munder M, Taylor GP, Müller I, Kropf P (2012) Characterization of a novel population of low-density granulocytes associated with disease severity in HIV seropositive patients. PLOS ONE 7: e48939.
    1. Kropf P, Baud D, Marshall SE, Munder M, Mosley A, et al. (2007) Arginase activity mediates reversible T cell hyporesponsiveness in human pregnancy. Eur J Immunol 37: 935–945.
    1. Phillips A, Pezzotti P (2004) Short-term risk of AIDS according to current CD4 cell count and viral load in antiretroviral drug-naive individuals and those treated in the monotherapy era. AIDS 18: 51–58.
    1. Gazzard BG (2008) British HIV Association Guidelines for the treatment of HIV-1-infected adults with antiretroviral therapy 2008. HIV Med 9: 563–608.
    1. Elbim C, Prevot MH, Bouscarat F, Franzini E, Chollet-Martin S, et al. (1994) Polymorphonuclear neutrophils from human immunodeficiency virus-infected patients show enhanced activation, diminished fMLP-induced L-selectin shedding, and an impaired oxidative burst after cytokine priming. Blood 84: 2759–2766.
    1. Rotondo R, Bertolotto M, Barisione G, Astigiano S, Mandruzzato S, et al. (2011) Exocytosis of azurophil and arginase 1-containing granules by activated polymorphonuclear neutrophils is required to inhibit T lymphocyte proliferation. J Leukoc Biol 89: 721–727.
    1. Kuijpers TW, Tool AT, van der Schoot CE, Ginsel LA, Onderwater JJ, et al. (1991) Membrane surface antigen expression on neutrophils: a reappraisal of the use of surface markers for neutrophil activation. Blood 78: 1105–1111.
    1. Martin-Martin B, Nabokina SM, Blasi J, Lazo PA, Mollinedo F (2000) Involvement of SNAP-23 and syntaxin 6 in human neutrophil exocytosis. Blood 96: 2574–2583.

Source: PubMed

Sponsors and Collaborators

Medical Conditions

Drug Interventions

3
Subscribe