Identification of Rhodococcus equi lipids recognized by host cytotoxic T lymphocytes.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
The researchers investigated the immune response of adult horses to Rhodococcus equi, a type of bacteria, with a focus on identifying the bacterial lipids that cytotoxic T lymphocytes (immune system cells) recognize. Results showed the immune system recognition of unique lipids present within the bacterial cell wall, particularly trehalose monomycolate and cardiolipin, plays a key role in effective immune responses.
Research Objectives and Hypothesis
- The primary aim of the research was to establish whether cytotoxic T lymphocytes recognize unique cell-wall lipids in Rhodococcus equi, which relate to mycobacterial lipids, based on the fact that the immune response was not restricted in the way it typically is for protein antigens.
Methodology
- Initial experiments involved treating soluble R. equi antigens with proteases to see if the immune response would weaken. This was not the case, which suggested that proteins were not the antigens triggering the response.
- Researchers extracted the lipid from R. equi and exposed it to T lymphocytes, and revealed that the lymphocytes were able to destroy target cells (macrophages) pulsed with the lipid.
- The team used thin-layer chromatography and mass spectroscopy to further analyze the lipid, finding it contained mainly trehalose monomycolate (TMM), trehalose dimycolate (TDM) and cardiolipin (CL).
- Cells exposed to TMM and CL were recognized and destroyed by the T lymphocytes, but those exposed to TDM were not.
- The researchers also measured cytokine (a type of immune response protein) transcription in response to the R. equi lipids. The study showed increased transcription of interferon-gamma but not interleukin-4, the former being associated with bacterial clearance.
Findings and Conclusion
- The results pointed to the immune system’s recognition of these unique bacterial cell wall lipids as a significant part of the protective response against R. equi.
- The findings also identified potential lipid antigens that have not previously been recognized as key elements in the immune response to this bacterial pathogen. The two isolated lipid fractions, TMM and CL, were shown to trigger the response of cytotoxic T lymphocytes without any restriction, leading to the destruction of infected cells.
These results help broaden the overall understanding of how the immune systems of horses (and potentially other organisms) combat bacterial infections on the cellular level. This could lead to improved treatment strategies for diseases caused by similar bacteria.
Cite This Article
Publication
Researcher Affiliations
- Department of Veterinary Microbiology and Pathology, Washington State University, College of Veterinary Medicine, PO Box 647040, Pullman, WA 99164-7040, USA.
- Department of Bacteriology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-Ku, Osaka 545-8585, Japan.
- Department of Veterinary Microbiology and Pathology, Washington State University, College of Veterinary Medicine, PO Box 647040, Pullman, WA 99164-7040, USA.
- Department of Veterinary Microbiology and Pathology, Washington State University, College of Veterinary Medicine, PO Box 647040, Pullman, WA 99164-7040, USA.
- Department of Bacteriology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-Ku, Osaka 545-8585, Japan.
- Department of Bacteriology, Osaka City University, Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-Ku, Osaka 545-8585, Japan.
- Department of Veterinary Microbiology and Pathology, Washington State University, College of Veterinary Medicine, PO Box 647040, Pullman, WA 99164-7040, USA.
MeSH Terms
- Animals
- Antigens, Bacterial / immunology
- Cell Wall / immunology
- Horses / immunology
- Horses / microbiology
- Interferon-gamma / immunology
- Interleukin-4 / genetics
- Interleukin-4 / immunology
- Leukocytes, Mononuclear / immunology
- Lipids / immunology
- Rhodococcus equi / cytology
- Rhodococcus equi / immunology
- T-Lymphocytes, Cytotoxic / immunology
Grant Funding
- T32 AI007025 / NIAID NIH HHS
- T32 AI007025-24 / NIAID NIH HHS
References
- Beckman EM, Porcelli SA, Morita CT, Behar SM, Furlong ST, Brenner MB. Recognition of a lipid antigen by CD1-restricted alpha beta+ T cells.. Nature 1994 Dec 15;372(6507):691-4.
- Bhatt A, Fujiwara N, Bhatt K, Gurcha SS, Kremer L, Chen B, Chan J, Porcelli SA, Kobayashi K, Besra GS, Jacobs WR Jr. Deletion of kasB in Mycobacterium tuberculosis causes loss of acid-fastness and subclinical latent tuberculosis in immunocompetent mice.. Proc Natl Acad Sci U S A 2007 Mar 20;104(12):5157-62.
- Boom WH, Canaday DH, Fulton SA, Gehring AJ, Rojas RE, Torres M. Human immunity to M. tuberculosis: T cell subsets and antigen processing.. Tuberculosis (Edinb) 2003;83(1-3):98-106.
- Brennan PJ, Nikaido H. The envelope of mycobacteria.. Annu Rev Biochem 1995;64:29-63.
- Bricard G, Porcelli SA. Antigen presentation by CD1 molecules and the generation of lipid-specific T cell immunity.. Cell Mol Life Sci 2007 Jul;64(14):1824-40.
- Brigl M, Brenner MB. CD1: antigen presentation and T cell function.. Annu Rev Immunol 2004;22:817-90.
- Canaday DH, Wilkinson RJ, Li Q, Harding CV, Silver RF, Boom WH. CD4(+) and CD8(+) T cells kill intracellular Mycobacterium tuberculosis by a perforin and Fas/Fas ligand-independent mechanism.. J Immunol 2001 Sep 1;167(5):2734-42.
- Catucci L, Depalo N, Lattanzio VM, Agostiano A, Corcelli A. Neosynthesis of cardiolipin in Rhodobacter sphaeroides under osmotic stress.. Biochemistry 2004 Nov 30;43(47):15066-72.
- Dascher CC, Hiromatsu K, Xiong X, Morehouse C, Watts G, Liu G, McMurray DN, LeClair KP, Porcelli SA, Brenner MB. Immunization with a mycobacterial lipid vaccine improves pulmonary pathology in the guinea pig model of tuberculosis.. Int Immunol 2003 Aug;15(8):915-25.
- DITTMER JC, LESTER RL. A SIMPLE, SPECIFIC SPRAY FOR THE DETECTION OF PHOSPHOLIPIDS ON THIN-LAYER CHROMATOGRAMS.. J Lipid Res 1964 Jan;5:126-7.
- Fischer K, Chatterjee D, Torrelles J, Brennan PJ, Kaufmann SH, Schaible UE. Mycobacterial lysocardiolipin is exported from phagosomes upon cleavage of cardiolipin by a macrophage-derived lysosomal phospholipase A2.. J Immunol 2001 Aug 15;167(4):2187-92.
- FOLCH J, LEES M, SLOANE STANLEY GH. A simple method for the isolation and purification of total lipides from animal tissues.. J Biol Chem 1957 May;226(1):497-509.
- Grotzke JE, Lewinsohn DM. Role of CD8+ T lymphocytes in control of Mycobacterium tuberculosis infection.. Microbes Infect 2005 Apr;7(4):776-88.
- Hava DL, Brigl M, van den Elzen P, Zajonc DM, Wilson IA, Brenner MB. CD1 assembly and the formation of CD1-antigen complexes.. Curr Opin Immunol 2005 Feb;17(1):88-94.
- Hines MT, Paasch KM, Alperin DC, Palmer GH, Westhoff NC, Hines SA. Immunity to Rhodococcus equi: antigen-specific recall responses in the lungs of adult horses.. Vet Immunol Immunopathol 2001 May 10;79(1-2):101-14.
- Hines SA, Stone DM, Hines MT, Alperin DC, Knowles DP, Norton LK, Hamilton MJ, Davis WC, McGuire TC. Clearance of virulent but not avirulent Rhodococcus equi from the lungs of adult horses is associated with intracytoplasmic gamma interferon production by CD4+ and CD8+ T lymphocytes.. Clin Diagn Lab Immunol 2003 Mar;10(2):208-15.
- Hiromatsu K, Dascher CC, LeClair KP, Sugita M, Furlong ST, Brenner MB, Porcelli SA. Induction of CD1-restricted immune responses in guinea pigs by immunization with mycobacterial lipid antigens.. J Immunol 2002 Jul 1;169(1):330-9.
- Hondalus MK. Pathogenesis and virulence of Rhodococcus equi.. Vet Microbiol 1997 Jun 16;56(3-4):257-68.
- Hooper-McGrevy KE, Wilkie BN, Prescott JF. Virulence-associated protein-specific serum immunoglobulin G-isotype expression in young foals protected against Rhodococcus equi pneumonia by oral immunization with virulent R. equi.. Vaccine 2005 Dec 30;23(50):5760-7.
- Kanaly ST, Hines SA, Palmer GH. Failure of pulmonary clearance of Rhodococcus equi infection in CD4+ T-lymphocyte-deficient transgenic mice.. Infect Immun 1993 Nov;61(11):4929-32.
- Kanaly ST, Hines SA, Palmer GH. Cytokine modulation alters pulmonary clearance of Rhodococcus equi and development of granulomatous pneumonia.. Infect Immun 1995 Aug;63(8):3037-41.
- Kanaly ST, Hines SA, Palmer GH. Transfer of a CD4+ Th1 cell line to nude mice effects clearance of Rhodococcus equi from the lung.. Infect Immun 1996 Apr;64(4):1126-32.
- Karakousis PC, Bishai WR, Dorman SE. Mycobacterium tuberculosis cell envelope lipids and the host immune response.. Cell Microbiol 2004 Feb;6(2):105-16.
- Kurano S, Sugimoto N, Sumi Y, Sawai H, Kato Y, Kaneda K, Yano I. [Newly isolated glycolipids from Rhodococcus terrae cell wall and their granuloma forming activities].. Yakugaku Zasshi 1987 Jan;107(1):46-52.
- Li B, Rossman MD, Imir T, Oner-Eyuboglu AF, Lee CW, Biancaniello R, Carding SR. Disease-specific changes in gammadelta T cell repertoire and function in patients with pulmonary tuberculosis.. J Immunol 1996 Nov 1;157(9):4222-9.
- Linder R. Rhodococcus equi and Arcanobacterium haemolyticum: two "coryneform" bacteria increasingly recognized as agents of human infection.. Emerg Infect Dis 1997 Apr-Jun;3(2):145-53.
- Lopez AM, Hines MT, Palmer GH, Alperin DC, Hines SA. Identification of pulmonary T-lymphocyte and serum antibody isotype responses associated with protection against Rhodococcus equi.. Clin Diagn Lab Immunol 2002 Nov;9(6):1270-6.
- McGuire TC, Tumas DB, Byrne KM, Hines MT, Leib SR, Brassfield AL, O'Rourke KI, Perryman LE. Major histocompatibility complex-restricted CD8+ cytotoxic T lymphocytes from horses with equine infectious anemia virus recognize Env and Gag/PR proteins.. J Virol 1994 Mar;68(3):1459-67.
- Moody DB, Porcelli SA. Intracellular pathways of CD1 antigen presentation.. Nat Rev Immunol 2003 Jan;3(1):11-22.
- Moody DB, Reinhold BB, Reinhold VN, Besra GS, Porcelli SA. Uptake and processing of glycosylated mycolates for presentation to CD1b-restricted T cells.. Immunol Lett 1999 Jan;65(1-2):85-91.
- Moody DB, Briken V, Cheng TY, Roura-Mir C, Guy MR, Geho DH, Tykocinski ML, Besra GS, Porcelli SA. Lipid length controls antigen entry into endosomal and nonendosomal pathways for CD1b presentation.. Nat Immunol 2002 May;3(5):435-42.
- Natsuhara Y, Yoshinaga J, Shogaki T, Sumi-Nishikawa Y, Kurano S, Kato Y, Kaneda K, Oka S, Yano I. Granuloma-forming activity and antitumor activity of newly isolated mycoloyl glycolipid from Rhodococcus terrae 70012 (Rt. GM-2).. Microbiol Immunol 1990;34(1):45-53.
- Nishiuchi Y, Baba T, Yano I. Mycolic acids from Rhodococcus, Gordonia, and Dietzia.. J Microbiol Methods 2000 Mar;40(1):1-9.
- Patton KM, McGuire TC, Fraser DG, Hines SA. Rhodococcus equi-infected macrophages are recognized and killed by CD8+ T lymphocytes in a major histocompatibility complex class I-unrestricted fashion.. Infect Immun 2004 Dec;72(12):7073-83.
- Patton KM, McGuire TC, Hines MT, Mealey RH, Hines SA. Rhodococcus equi-specific cytotoxic T lymphocytes in immune horses and development in asymptomatic foals.. Infect Immun 2005 Apr;73(4):2083-93.
- Perez MG, Vassilev T, Kemmerly SA. Rhodococcus equi infection in transplant recipients: a case of mistaken identity and review of the literature.. Transpl Infect Dis 2002 Mar;4(1):52-6.
- Porcelli SA. Cutting glycolipids down to size.. Nat Immunol 2001 Mar;2(3):191-2.
- Rahman MT, Herron LL, Kapur V, Meijer WG, Byrne BA, Ren J, Nicholson VM, Prescott JF. Partial genome sequencing of Rhodococcus equi ATCC 33701.. Vet Microbiol 2003 Jul 1;94(2):143-58.
- Rao V, Fujiwara N, Porcelli SA, Glickman MS. Mycobacterium tuberculosis controls host innate immune activation through cyclopropane modification of a glycolipid effector molecule.. J Exp Med 2005 Feb 21;201(4):535-43.
- Roura-Mir C, Moody DB. Sorting out self and microbial lipid antigens for CD1.. Microbes Infect 2003 Oct;5(12):1137-48.
- Serbina NV, Liu CC, Scanga CA, Flynn JL. CD8+ CTL from lungs of Mycobacterium tuberculosis-infected mice express perforin in vivo and lyse infected macrophages.. J Immunol 2000 Jul 1;165(1):353-63.
- Siliciano RF, Keegan AD, Dintzis RZ, Dintzis HM, Shin HS. The interaction of nominal antigen with T cell antigen receptors. I. Specific binding of multivalent nominal antigen to cytolytic T cell clones.. J Immunol 1985 Aug;135(2):906-14.
- Skinner MA, Yuan S, Prestidge R, Chuk D, Watson JD, Tan PL. Immunization with heat-killed Mycobacterium vaccae stimulates CD8+ cytotoxic T cells specific for macrophages infected with Mycobacterium tuberculosis.. Infect Immun 1997 Nov;65(11):4525-30.
- Stenger S, Modlin RL. T cell mediated immunity to Mycobacterium tuberculosis.. Curr Opin Microbiol 1999 Feb;2(1):89-93.
- Szereday L, Baliko Z, Szekeres-Bartho J. Gamma/delta T cell subsets in patients with active Mycobacterium tuberculosis infection and tuberculin anergy.. Clin Exp Immunol 2003 Feb;131(2):287-91.
- Takai S. Epidemiology of Rhodococcus equi infections: a review.. Vet Microbiol 1997 Jun 16;56(3-4):167-76.
- Tonge PJ. Another brick in the wall.. Nat Struct Biol 2000 Feb;7(2):94-6.
- Torres-Tortosa M, Arrizabalaga J, Villanueva JL, Gálvez J, Leyes M, Valencia ME, Flores J, Peña JM, Pérez-Cecilia E, Quereda C. Prognosis and clinical evaluation of infection caused by Rhodococcus equi in HIV-infected patients: a multicenter study of 67 cases.. Chest 2003 Jun;123(6):1970-6.
- Ueda S, Fujiwara N, Naka T, Sakaguchi I, Ozeki Y, Yano I, Kasama T, Kobayashi K. Structure-activity relationship of mycoloyl glycolipids derived from Rhodococcus sp. 4306.. Microb Pathog 2001 Feb;30(2):91-9.
- Ulrichs T, Moody DB, Grant E, Kaufmann SH, Porcelli SA. T-cell responses to CD1-presented lipid antigens in humans with Mycobacterium tuberculosis infection.. Infect Immun 2003 Jun;71(6):3076-87.
- Willcox BE, Willcox CR, Dover LG, Besra G. Structures and functions of microbial lipid antigens presented by CD1.. Curr Top Microbiol Immunol 2007;314:73-110.
- Zhang W, Lonning SM, McGuire TC. Gag protein epitopes recognized by ELA-A-restricted cytotoxic T lymphocytes from horses with long-term equine infectious anemia virus infection.. J Virol 1998 Dec;72(12):9612-20.
- Zinner SH. Changing epidemiology of infections in patients with neutropenia and cancer: emphasis on gram-positive and resistant bacteria.. Clin Infect Dis 1999 Sep;29(3):490-4.
Citations
This article has been cited 6 times.- Giri SS, Kim HJ, Kim SG, Kim SW, Kwon J, Lee SB, Park SC. Immunomodulatory Role of Microbial Surfactants, with Special Emphasis on Fish.. Int J Mol Sci 2020 Sep 23;21(19).
- Harvey DJ. Analysis of carbohydrates and glycoconjugates by matrix-assisted laser desorption/ionization mass spectrometry: an update for 2009-2010.. Mass Spectrom Rev 2015 May-Jun;34(3):268-422.
- Dossa RG, Alperin DC, Hines MT, Hines SA. The equine CD1 gene family is the largest and most diverse yet identified.. Immunogenetics 2014 Jan;66(1):33-42.
- Bordin AI, Suchodolski JS, Markel ME, Weaver KB, Steiner JM, Dowd SE, Pillai S, Cohen ND. Effects of administration of live or inactivated virulent Rhodococccus equi and age on the fecal microbiome of neonatal foals.. PLoS One 2013;8(6):e66640.
- Sydor T, von Bargen K, Hsu FF, Huth G, Holst O, Wohlmann J, Becken U, Dykstra T, Söhl K, Lindner B, Prescott JF, Schaible UE, Utermöhlen O, Haas A. Diversion of phagosome trafficking by pathogenic Rhodococcus equi depends on mycolic acid chain length.. Cell Microbiol 2013 Mar;15(3):458-73.
- Harris SP, Hines MT, Mealey RH, Alperin DC, Hines SA. Early development of cytotoxic T lymphocytes in neonatal foals following oral inoculation with Rhodococcus equi.. Vet Immunol Immunopathol 2011 Jun 15;141(3-4):312-6.