Human Immunology - Dr. Daniel Douek

Research Laboratories
Human Immunology Section
Daniel Douek, M.D., MRCP, Ph.D.

Telephone: 301-594-8484

One of the main goals of the lab is the analysis of T cell immune responses in humans. HIV disease - its pathogenesis, treatment and prevention - provides us with an ideal system in which to study this, and at the same time allows us to contribute directly to the design and analysis of therapies that are potentially of enormous clinical benefit. Determining the precise role of HIV-specific T cells in the control of HIV infection is a key step in the effort to understand the composition of protective, or at least effective, immune responses after infection or immunization with vaccines. The most practical way to achieve this in humans is to characterize, precisely and quantitatively, correlations between the fundamental aspects of the HIV-specific T cell response, and clinical and virologic parameters of HIV disease. This involves the analysis of the frequency, function and specificity of HIV-specific T cells in response to HIV peptide antigens. We perform this primarily using intracellular cytokine analysis, and a novel technique we have developed which combines in vitro antigen stimulation and real-time quantitative clonotypic PCR. The subjects studied include those with HIV infection, those undergoing therapeutic vaccination, and those administered candidate prophylactic vaccines. This will afford us with a valuable framework in which to develop strategies to amplify, broaden or focus T cell immunity to HIV infection.

In addition to analyzing T cell responses in HIV disease and its vaccination, the lab is also interested in understanding and augmenting immune reconstitution - especially via the thymic pathway. Our previous work in this field showed that, contrary to the accepted dogma, the human thymus maintains substantial output and plays a role in immune homeostasis throughout the lifetime of an individual. The implications of this are far reaching, notably in the fields of HIV infection, cancer chemotherapy and stem cell transplantation, where reconstitution of a complete T cell repertoire would require the generation of new naïve T cells from the thymus. We have shown that thymic output is suppressed by HIV infection and that in the majority of patients treated with antiretroviral therapy, the thymus indeed contributes to immune reconstitution. Furthermore, the adult thymus can contribute to immune reconstitution following myeloablative chemotherapy and SCT in adults. Through a number of collaborations we are currently assessing the role and relevance of the thymus in immune reconstitution after different transplantation protocols. Using human thymus organ culture and the NOD-scid-hu thymic explant system we are analyzing the effect of cytokines such as IL-7 on augmenting thymopoietc capacity.

Selected Publications:

Llewelyn M, Sriskandan S, Peakman M, Ambrozak DR, Douek DC, Kwok WW, Cohen J, Altmann DM. HLA class II polymorphisms determine responses to bacterial superantigens. J Immunol. 2004 Feb 1;172(3):1719-26.

Brenchley JM, Hill BJ, Ambrozak DR, Price DA, Guenaga FJ, Casazza JP, Kuruppu J, Yazdani J, Migueles SA, Connors M, Roederer M, Douek DC, Koup RA. T-cell subsets that harbor human immunodeficiency virus (HIV) in vivo: implications for HIV pathogenesis. J Virol. 2004 Feb;78(3):1160-8.

Fujiwara H, El Ouriaghli F, Grube M, Price DA, Rezvani K, Gostick E, Sconocchia G, Melenhorst J, Hensel N, Douek DC, Barrett AJ. Identification and in vitro expansion of CD4+ and CD8+ T cells specific for human neutrophil elastase. Blood. 2003 Dec 4.

Douek DC. Disrupting T-cell homeostasis: how HIV-1 infection causes disease. AIDS Rev. 2003 Jul-Sep;5(3):172-7.

Betts MR, Brenchley JM, Price DA, De Rosa SC, Douek DC, Roederer M, Koup RA. Sensitive and viable identification of antigen-specific CD8+ T cells by a flow cytometric assay for degranulation. J Immunol Methods. 2003 Oct 1;281(1-2):65-78.

Storek J, Douek DC, Keesey JC, Boehmer L, Storer B, Maloney DG. Low T cell receptor excision circle levels in patients thymectomized 25-54 years ago. Immunol Lett. 2003 Oct 31;89(2-3):91-2.

Mascola JR, Lewis MG, VanCott TC, Stiegler G, Katinger H, Seaman M, Beaudry K, Barouch DH, Korioth-Schmitz B, Krivulka G, Sambor A, Welcher B, Douek DC, Montefiori DC, Shiver JW, Poignard P, Burton DR, Letvin NL. Cellular immunity elicited by human immunodeficiency virus type 1/ simian immunodeficiency virus DNA vaccination does not augment the sterile protection afforded by passive infusion of neutralizing antibodies. J Virol. 2003 Oct;77(19):10348-56.

Bahceci E, Epperson D, Douek DC, Melenhorst JJ, Childs RC, Barrett AJ. Early reconstitution of the T-cell repertoire after non-myeloablative peripheral blood stem cell transplantation is from post-thymic T-cell expansion and is unaffected by graft-versus-host disease or mixed chimaerism. Br J Haematol. 2003 Sep;122(6):934-43.

Permar SR, Moss WJ, Ryon JJ, Douek DC, Monze M, Griffin DE. Increased thymic output during acute measles virus infection. J Virol. 2003 Jul;77(14):7872-9.

Rezvani K, Grube M, Brenchley JM, Sconocchia G, Fujiwara H, Price DA, Gostick E, Yamada K, Melenhorst J, Childs R, Hensel N, Douek DC, Barrett AJ. Functional leukemia-associated antigen-specific memory CD8+ T cells exist in healthy individuals and in patients with chronic myelogenous leukemia before and after stem cell transplantation. Blood. 2003 Oct 15;102(8):2892-900. Epub 2003 Jun 26.

Lederman MM, Douek DC. Sometimes help may not be enough. AIDS. 2003 May 23;17(8):1249-51.

Diaz M, Douek DC, Valdez H, Hill BJ, Peterson D, Sanne I, Piliero PJ, Koup RA, Green SB, Schnittman S, Lederman MM. T cells containing T cell receptor excision circles are inversely related to HIV replication and are selectively and rapidly released into circulation with antiretroviral treatment. AIDS. 2003 May 23;17(8):1145-9.

Aquino VM, Douek DC, Berryman B, Johnson M, Jain VK, Collins RH. Evaluation of thymic output by measurement of T-cell-receptor gene rearrangement excisional circles (TREC) in patients who have received fludarabine. Leuk Lymphoma. 2003 Feb;44(2):343-8.

Michalek J, Collins RH, Hill BJ, Brenchley JM, Douek DC. Identification and monitoring of graft-versus-host specific T-cell clone in stem cell transplantation. Lancet. 2003 Apr 5;361(9364):1183-5.

Michalek J, Collins RH, Durrani HP, Vaclavkova P, Ruff LE, Douek DC, Vitetta ES. Definitive separation of graft-versus-leukemia- and graft-versus-host-specific CD4+ T cells by virtue of their receptor beta loci sequences. Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):1180-4. Epub 2003 Jan 16.

Douek DC, Picker LJ, Koup RA. T cell dynamics in HIV-1 infection. Annu Rev Immunol. 2003;21:265-304. Epub 2001 Dec 19. Review.

Douek DC. The contribution of the thymus to immune reconstitution after hematopoietic stem-cell transplantation. Cytotherapy. 2002;4(5):425-6. Review.

K. Weinberg, B.R. Blazar, J. Wagner, B. J. Hill, M. Smogorzewska, R.A. Koup, Michael R. Betts, R.H. Collins, and DC Douek. Factors affecting thymic function after allogeneic hematopoietic stem cell transplantation. Blood 2000.

Douek DC, Vescio RA, Betts MR, Brenchley JM, Hill BJ, Zhang L, Berenson JR, Collins R, Koup RA. Assessment of Thymic Output in Adults after Haematopoietic Stem Cell Transplant and Prediction of T Cell Reconstitution. Lancet 2000; 355:1875-1881.

McFarland RD, Douek DC, Koup RA, Picker LJ. Identification of a Human Recent Thymic Emigrant Phenotype. Proc Natl Acad Sci U S A 2000;97(8):4215-4220.

DL Sodora, DC Douek, G Silvestri, L Montgomery, M Rosenzweig, T Igarashi, B Bernacky, RP Johnson, M Feinberg, M A Martin, and RA Koup. Quantification of Thymic Function by Measuring T cell Receptor Excision Circles within Peripheral Blood and Lymphoid Tissues in Monkeys. Eur J Immunol. 2000;30:1145-1153.

Douek DC, K Luzzuriaga, J Sullivan, and RA Koup. Effects of pediatric HIV infection and its treatment on the thymus. Journal of Infectious Diseases 2000;181(4):1479-1482.

Douek DC and Altmann DM. T cell apoptosis and differential human leucocyte antigen class II expression in human thymus. Immunology 2000;99:249-256.

Nixon DF, Douek DC, Kuebler PJ, Jin X, Vesanen M, Bonhoeffer S, Cao Y, Koup RA, Ho DD, Markowitz M. Molecular tracking of an Human Immunodeficiency Virus nef specific cytotoxic T-cell clone shows persistence of clone-specific T-cell receptor DNA but not mRNA following early combination antiretroviral therapy. Immunol Lett 1999;66:219-28.

Jamieson BD, Douek DC, Killian S, Hultin LE, Scripture-Adams DD, Giorgi JV, Marelli D, Koup RA, Zack JA. Generation of functional thymocytes in the human adult. Immunity 1999;10(5):569-75.

Markert ML, Boeck A, Hale LP, Kloster AL, McLaughlin TM, Batchvarova MN, Douek DC, Koup RA, Kostyu DD, Ward FE, Rice HE, Mahaffey SM, Schiff SE, Buckley RH, Haynes BF. Transplantation of thymus tissue in complete DiGeorge syndrome. N Engl J Med 1999;341(16):1180-9.

Douek DC, McFarland RD, Keiser PH, Gage EA, Massey JM, Haynes BF, Polis MA, Haase AT, Feinberg MB, Sullivan JL, Jamieson BD, Zack JA, Picker LJ, Koup RA. Changes in thymic function with age and during the treatment of HIV infection. Nature 1998;396(6712):690-5.

Douek DC, Altmann DM. HLA-DO is an intracellular class II molecule with distinctive thymic expression. Int Immunol 1997;9(3):355-64.

Douek DC, Corley KT, Zal T, Mellor A, Dyson PJ, Altmann DM. Negative selection by endogenous antigen and superantigen occurs at multiple thymic sites. Int Immunol 1996;8(9):1413-20.

Altmann DM, Lider O, Douek DC, Cohen IR. Activation of specific T cell lines by the antigens avidin and myelin basic protein in the absence of antigen-presenting cells. Eur J Immunol 1987;17(11):1635-40.

If you are interested in a Research Fellowship, please send your CV to:

Dr. Daniel Douek
NIH/Vaccine Research Center
40 Convent Drive
Bldg. 40, Room 4500
Bethesda, MD 20892-3005