Applications in diagnosis and research & literature
Literature References: EliSpot Applications in Research and Diagnostics
Elispot is now widely used as a method to determine cytokine secretion, especially in vaccine development. The antigen specific secretion of IFN-gamma also is used for diagnostic procedures. We include some references without any claim to completeness. Use medline (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi) and type "elispot and HIV", " elispot and HCV" etc. to get recent publications online.
Replacing the Skin Prick: TB Diagnostics
Arend SM, van Meijgaarden KE, de Boer K, de Palou EC, van Soolingen D, Ottenhoff TH, van Dissel JT. (2002): Tuberculin skin testing and in vitro T cell responses to ESAT-6 and culture filtrate protein 10 after infection with Mycobacterium
marinum or M. kansasii. J Infect Dis. 186(12):1797-807
Ewer K, Deeks J, Alvarez L, Bryant G, Waller S, Andersen P, Monk P, Lalvani A. (2003): Comparison of T-cell-based assay with tuberculin skin test for diagnosis of Mycobacterium tuberculosis infection in a school tuberculosis outbreak.
Chapman AL, Munkanta M, Wilkinson KA, Pathan AA, Ewer K, Ayles H, Reece WH, Mwinga A, Godfrey-Faussett P, Lalvani A. (2002): Rapid detection of active and latent tuberculosis infection in HIV-positive individuals by enumeration of Mycobacterium tuberculosis-specific T cells.
Lalvani A, Pathan AA, Durkan H, Wilkinson KA, Whelan A, Deeks JJ, Reece WH, Latif M, Pasvol G, Hill AV. (2001): Enhanced contact tracing and spatial tracking of Mycobacterium tuberculosis infection by enumeration of antigen-specific T cells.
Lalvani A, Pathan AA, McShane H, Wilkinson RJ, Latif M, Conlon CP, Pasvol G, Hill AV. (2001): Rapid detection of Mycobacterium tuberculosis infection by enumeration of antigen-specific T cells.
Am J Respir Crit Care Med. 163(4):824-8.
Pathan AA, Wilkinson KA, Klenerman P, McShane H, Davidson RN, Pasvol G, Hill AV, Lalvani A. (2001): Direct ex vivo analysis of antigen-specific IFN-gamma-secreting CD4 T cells in Mycobacterium tuberculosis-infected individuals: associations with clinical disease state and effect of treatment.
J Immunol. 1;167(9):5217-25.
Lalvani A, Nagvenkar P, Udwadia Z, Pathan AA, Wilkinson KA, Shastri JS, Ewer K, Hill AV, Mehta A, Rodrigues C. (2001): Enumeration of T cells specific for RD1-encoded antigens suggests a high prevalence of latent Mycobacterium tuberculosis infection in healthy urban Indians.
J Infect Dis. 1;183(3):469-77. Epub 2001 Jan 09.
Lalvani A. (2003): Spotting latent infection: the path to better tuberculosis control.
Almeida D, Rodrigues C, Udwadia ZF, Lalvani A, Gothi GD, Mehta P, Mehta A. (2003) Incidence of multidrug-resistant tuberculosis in urban and rural India and implications for prevention.
Clin Infect Dis. 15;36(12):e152-4. Epub 2003 Jun 03.
Cockle PJ, Gordon SV, Lalvani A, Buddle BM, Hewinson RG, Vordermeier HM. (2002): Identification of novel Mycobacterium tuberculosis antigens with potential as diagnostic reagents or subunit vaccine candidates by comparative genomics.
Infect Immun. 70(12):6996-7003.
Lalvani A. (2002): CD8 cytotoxic T cells and the development of new tuberculosis vaccines.
Am J Respir Crit Care Med. 15;166(6):789-90.
Pathan AA, Wilkinson KA, Wilkinson RJ, Latif M, McShane H, Pasvol G, Hill AV, Lalvani A. (2000) High frequencies of circulating IFN-gamma-secreting CD8 cytotoxic T cells specific for a novel MHC class I-restricted Mycobacterium tuberculosis epitope in M. tuberculosis-infected subjects without disease.
Eur J Immunol. 30(9):2713-21.
Lalvani A, Hill AV. (1998) Cytotoxic T-lymphocytes against malaria and tuberculosis: from natural immunity to vaccine design.
Clin Sci (Lond). 95(5):531-8.
The Lyme-Disease can trigger an autoimmune reaction. The surface antigen OSP (Outer Surface Protein) A is highly similar to the human protein LFA-1, which can lead to misdirection of the immunological reaction against the host.
Gross DM, Forsthuber T, Tary-Lehmann M et al. 1998: Identification of LFA-1 as a candidate autoantigen in treatment-resistant Lyme-Arthritis. Science 281: 703-706
With the EliSpot method the production of anti-chlamydial antibodies can be detected at the cellular level.
Daugharty H, Messmer TO, Fields BS 1997: Elispot assay for Chlamydia-specific, antibody-producing cells correlated with conventional complement fixation and microimmunofluorescence. J Clin Lab Anal 11: 45-52.
Helicobacter pylori Infection
A Helicobacter-Infection induces a Th1-response with an increased production of IFN-gamma.
Karttunen R, Karttunen T, Ekre HP & MacDonald TT 1995: IFN gamma and interleukin 4 secreting cells in the gastric antrum in Helicobacter pylori positive and negative gastritis. Gut 36(3): 341-345.
The loss of CD4+-cells leads to a reduced function of the whole immune system. The quotient CD4+/CD8+ ist shifted in favour of the CD8+-cells. The response of the T-lymphocytes to stimulating antigens or mitogens is significantly reduced.
Design and validation of an ELISPOT assay are described for use in clinical trials of candidate human immunodeficiency virus (HIV) vaccines, using a particular immunogen termed HIVA M.
Mwau, et al. Design and validation of an enzyme-linked immunosorbent spot assay for use in clinical trials of candidate HIV vaccines. AIDS Research and Human Retroviruses 18 (9): 611-618 JUN 10 2002
Keir ME, Rosenberg MG, Sandberg JK, Jordan KA, Wiznia A, Nixon DF, Stoddart CA, McCune JM. Generation of CD3+CD8low thymocytes in the HIV type 1-infected thymus. J Immunol. 2002 Sep 1;169(5):2788-96.
Jennes W, Kestens L, Nixon D, Shacklett B. Enhanced ELISPOT detection of antigen-specific T cell responses from cryopreserved specimens with addition of both IL-7 and IL-15-the Amplispot assay. J Immunol Methods. 2002 Dec 1;270(1):99.
Shacklett BL, Shaw KE, Adamson LA, Wilkens DT, Cox CA, Montefiori DC, Gardner MB, Sonigo P, Luciw PA. Live, Attenuated Simian Immunodeficiency Virus SIVmac-M4, with Point Mutations in the Env Transmembrane Protein Intracytoplasmic Domain, Provides Partial Protection from Mucosal Challenge with Pathogenic SIVmac251. J Virol. 2002 Nov 15;76(22):11365-11378.
Shacklett BL, Means RE, Larsson M, Wilkens DT, Beadle TJ, Merritt MJ, Bhardwaj N, Palumbo PE, Skurnick JH, Louria DB, Nixon DF. Dendritic cell amplification of HIV type 1-specific CD8+ T cell responses in exposed, seronegative heterosexual women. AIDS Res Hum Retroviruses. 2002 Jul 20;18(11):805-15.
Skurnick JH, Palumbo P, DeVico A, Shacklett BL, Valentine FT, Merges M, Kamin-Lewis R, Mestecky J, Denny T, Lewis GK, Lloyd J, Praschunus R, Baker A, Nixon DF, Stranford S, Gallo R, Vermund SH, Louria DB. Correlates of nontransmission in US women at high risk of human immunodeficiency virus type 1 infection through sexual exposure. J Infect Dis. 2002 Feb 15;185(4):428-38.
A Th1-response against (INF-, IL-2) Myeline Basic Proteine seems to be causally involved in the genesis of Multiple Sklerosis.
McCutcheon M, Wehner N, Wensky A et al. 1997: A sensitive Elispot assay to detect low-frequency human T-lymphocytes. J Immunol Methods 210(2): 149-166.
Interleukin-4, produced by Th2 cells, induces class switch in plasma cells from IgM and IgG to IgE. IFN-gamma, produced by Th1-cells, counters this effect. Allergen-specific IL-4 producing Th2-cells can be detected in vitro with the EliSpot method. They correlate with the IgE-titers in sera of persons suffering from atopies.
Gabrielsson S, Paulie S, Rak S et al. 1997: Specific induction of interleukin-4-producing cells in response to in vitro allergen stimulation in atopic individuals. Clin Exp Allergy 27: 808-815.
In recipients of allogenetic renal transplants a quotient of IFN-gamma to Interleukin-5 higher than 15 correlates with an allograft rejection.
Tary-Lehmann M, Hricik D, Justice AC et al. 1998: Enzyme-linked immunosorbent assay spot detection of IFN-gamma ind interleukin 5-producing cells as a predictive marker for renal allograft failure. Transplantation 66: 219-224.