Preview

Medical Immunology (Russia)

Advanced search

MULTI-COLOUR CYTOMETRIC ANALYSIS. IDENTIFICATION OF T LYMPHOCYTES AND THEIR SUBSETS

https://doi.org/10.15789/1563-0625-2008-2-3-115-124

Abstract

Abstract. T-lymphocytes play an important role in elimination of tumor cells, in reactions of a transplant against graft and graft versus host disease, in slow-type hypersensitivity, and other reactions directed for maintenance of homeostasis. Along with CD3, an antigen-specific T-cellular receptor (TCR) is another common marker of T-cells. There are two types of TcR – αβ-TcR and γδ-TcR that differ in ontogenetic and functional properties. γδ-T-cells play a significant role in protection of organism against various types of infections, and determination of their amounts should be an integral part of the analysis of patients’ immune status. To these purposes, a multi-colour analysis shuld be used, applying the following combinations of monoclonal antibodies: CD3/CD4/CD8/CD45 and αβ-TcR/γδ-TcR/CD3/CD45. Multi-colour staining and multi-step gating allow of carrying out multiparametric analysis of peripheral blood with high accuracy and reliability. The proposed approach considerably facilitates interpretation of results obtained, and it allows of judging about immune system functioning in various pathological conditions.

About the Authors

S. V. Khaidukov
Институт биоорганической химии им. акад. М.М. Шемякина и Ю.А. Овчинникова РАН, Москва
Russian Federation


A. V. Zurochka
ГОУ ВПО Челябинская государственная медицинская академия Росздрава, г. Челябинск
Russian Federation


V. A. Chereshnev
Институт иммунологии и физиологии УРО РАН, г. Екатеринбург
Russian Federation


References

1. Робсон А., Ройт А., Делвз П. Основы медицинской иммунологии // М.: Мир, 2006. – 320 c.

2. 1994 revised guidelines for the performance of CD4+ T-cell determinations in persons with human immunodeficiency virus (HIV) infections // MMWR Recomm. Rep. – 1994. – Vol. 43, N RR-3. – P. 1-21.

3. Autran B., Triebel F., Katlama C., Rozenbaum W., Hercend T., Debre P. T cell receptor gamma/delta+ lymphocyte subsets during HIV infection // Clin. Exp. Immunol. – 1989. – Vol. 75, N 2. – P. 206-210.

4. Bagriacik E.U., Okabe M., Klein J.R. Origins of intestinal intraepithelial lymphocytes: direct evidence for a thymus-derived gamma delta T cell component // Immunol. Lett. – 2000. – Vol. 75. – P. 77-83.

5. Battistini L., Borsellino G., Sawicki G., Poccia F., Salvetti M., Ristori G., Brosnan C.F. Phenotypic and cytokine analysis of human peripheral blood gamma delta T cells expressing NK cell receptors // J. Immunol. – 1997. – Vol. 159, N 8. – P. 3723-3730.

6. Beagley K.W., Husband A.J. Intraepithelial lymphocytes: origins, distribution, and function // Crit. Rev. Immunol. – 1998. – Vol. 18. – P. 237-254.

7. Born W.K., Lahn M., Takeda K., Kanehiro A., O'Brien R.L., Gelfand E.W. Role of gammadelta T cells in protecting normal airway function // Respir. Res. – 2000. – Vol. 1. – P. 151-158.

8. Caccamo N., Dieli F., Wesch D., Jomaa H., Eberl M. Sex-specific phenotypical and functional differences in peripheral human Vgamma9/Vdelta2 T cells // J. Leukoc. Biol. – 2006. – Vol. 79, N 4. – P. 663-666.

9. Chen J., Niu H., He W., Ba D. Antitumor activity of expanded human tumor-infiltrating gammadelta T lymphocytes // Int. Arch. Allergy. Immunol. – 2001. – Vol. 125. – P. 256-263.

10. Chen Z.W., Letvin N.L. Adaptive immune response of Vgamma2Vdelta2 T cells: a new paradigm // Trends. Immunol. – 2003. – Vol. 24. – P. 213-219.

11. Davis M.M. T cell receptor gene diversity and selection // Annu. Rev. Biochem. – 1990. – Vol. 59. – P. 475-496.

12. Davis M.M., Bjorkman P.J. T-cell antigen receptor genes and T-cell recognition // Nature. – 1988. – Vol. 334, N 6181. – P. 395-402.

13. Davis M.M., Boniface J.J., Reich Z., Lyons D., Hampl J., Arden B., Chien Y. Ligand recognition by αβ T cell receptors // Annu. Rev. Immunol. – 1998. – Vol. 16. – P. 523-544.

14. Dieli F., Troye-Blomberg M., Ivanyi J., Fourniй J.J., Krensky A.M., Bonneville M., Peyrat M.A., Caccamo N., Sireci G., Salerno A. Granulysindependent killing of intracellular and extracellular Mycobacterium tuberculosis by gamma9/Vdelta2 T lymphocytes // J. Infect. Dis. – 2001. – Vol. 184. – P. 1082-1085.

15. Ferrarini M., Ferrero E., Dagna L., Poggi A., Zocch M.R. Human gammadelta T cells: a nonredundant system in the immune-surveillance against cancer // Trends. Immunol. – 2002. – Vol. 23. – P. 14-18.

16. Ferrick D.A., King D.P., Jackson K.A., Braun R.K., Tam S., Hyde D.M., Beaman B.L. Intraepithelial gamma delta T lymphocytes: sentinel cells at mucosal barriers // Springer. Semin. Immunopathol. – 2000. – Vol. 22. – P. 283-296.

17. Fujimiya Y., Suzuki Y., Katakura R., Miyagi T., Yamaguchi T., Yoshimoto T., Ebina T. In vitro interleukin 12 activation of peripheral blood CD3(+)CD56(+) and CD3(+)CD56(–) gammadelta T cells from glioblastoma patients // Clin. Cancer. Res. – 1997. – Vol. 3, N 4. – P. 633-643.

18. Girardi M., Oppenheim D.E., Steele C.R., Lewis J.M., Glusac E., Filler R., Hobby P., Sutton B., Tigelaar R.E., Hayday A.C. Regulation of cutaneous malignancy by gammadelta T cells // Science. – 2001. – Vol. 294. – P. 605-609.

19. Gudelj L., Deniz G., Rukavina D., Johnson P.M., Christmas S.E. Expression of functional molecules by human CD3– decidual granular leucocyte clones // Immunology. – 1996. – Vol. 87, N 4. – P. 609-615.

20. Gui delines for the performance of CD4+ T-cell determinations in persons with human immunodeficiency virus infection // MMWR Recomm. Rep. – 1992. – Vol. 41 (RR-8). – P. 1-17.

21. Hayball J.D., Lake R.A. Altered superantigenic ligands demonstrate the quantitative nature of T-cell activation // Immunol. Cell Biol. – 2000. – Vol. 78, N 6. – P. 623-632.

22. Hayball J.D., Lake R.A. Partial T cell activation with an altered superantigenic ligand // Immunol. Cell Biol. – 2000. – Vol. 78, N 1. – P. 13-19.

23. Haynes B.F., Heinly C.S. Early human T cell development: analysis of the human thymus at the time of initial entry of hematopoietic stem cells into the fetal thymic microenvironment // J. Exp. Med. – 1995. – Vol. 181, N 4. – P. 1445-1458.

24. Helgeland L., Brandtzaeg P., Rolstad B., Vaage J.T. Sequential development of intraepithelial gamma delta and alpha beta T lymphocytes expressing CD8 alpha beta in neonatal rat intestine: requirement for the thymus // Immunology. – 1997. – Vol. 92. – P. 447-455.

25. Hummel S., Wilms D., Vitacolonna M., Zöller M. Donor T cell and host NK depletion improve the therapeutic efficacy of allogeneic bone marrow cell reconstitution in the nonmyeloablatively conditioned tumor-bearing host // Leukoc. Biol. – 2002. – Vol. 72, N 5. – P. 898-912.

26. Ichikawa Y., Shimizu H., Yoshida M., Takaya M., Arimori S. T cells bearing gamma/delta T cell receptor and their expression of activation antigen in peripheral blood from patients with Sjogren’s syndrome // Clin. Exp. Rheumatol. – 1991. – Vol. 9. – P. 603-609.

27. Jarry A., Cerf-Bensussan N., Brousse N., Selz F., Guy-Grand D. Subsets of CD3+ (T cell receptor alpha/beta or gamma/delta) and CD3– lymphocytes isolated from normal human gut epithelium display phenotypical features different from their counterparts in peripheral blood // Eur. J. Immunol. – 1990. – Vol. 20, N 5. – P. 1097-1103.

28. Kabelitz D., Wesch D. Role of gamma delta T-lymphocytes in HIV infection // Eur. J. Med. Res. – 2001. – Vol. 6. – P. 169-174.

29. Kato Y., Tanaka Y., Miyagawa F., Yamashita S., Minato N. Targeting of tumor cells for human gammadelta T cells by nonpeptide antigens // J. Immunol. – 2001. – Vol. 167, N 9. – P. 5092-5098.

30. Kawashima T., Norose Y, Watanabe Y, Enomoto Y., Narazaki H., Watari E., Tanaka S., Takahashi H., Yano I., Brenner M.B., Sugita M. Major CD8 T cell response to live bacillus Calmette-Gueґrin is mediated by CD1 molecules // J. Immunol. – 2003. – Vol. 170. – P. 5345-5348.

31. Kelsen J., Agnholt J., Hoffmann H.J., Kaltoft K., Dahlerup J.F. Increased expression of TCR Vbeta5.1 and 8 in mucosal T-cell lines cultured from patients with Crohn disease // Scand. J. Gastroenterol. – 2004. – Vol. 39, N 3. – P. 238-245.

32. Kobayashi H., Tanaka Y., Yagi J., Toma H., Uchyama T. Gamma/delta T cells provide innate immunity against renal cell carcinoma // Cancer. Immunol. Immunother. – 2001. – Vol. 50. – P. 115-124.

33. Komano H., Fujiurat Y., Kawaguch M., Matsumoto S., Hashimoto Y., Obana S., Mombaerts P., Tonegawa S., Yamamoto H., Itohara S., Nanno M., Ishikawa H. Homeostatic regulation of intestinal epithelia by intraepithelial gamma delta T cells // Proc. Natl. Acad. Sci. USA. – 1995. – Vol. 92. – P. 6147-6151.

34. Kretowski A., Mysliwiec J., Szelachowska M., Turowski D., Wysocka J., Kowalska I., Kinalska I. Gammadelta T-cells alterations in the peripheral blood of high risk diabetes type 1 subjects with subclinical pancreatic B-cells impairment // Immunol. Lett. – 1999. – Vol. 68. – P. 289-293.

35. Lafarge X., Merville P., Cazin M.C., Bergй F., Potaux L., Moreau J.F., Déchanet-Merville J. Cytomegalovirus infection in transplant recipients resolves when circulating gammadelta T lymphocytes expand, suggesting a protective antiviral role // J. Infect. Dis. – 2001. – Vol. 184. – P. 533-541.

36. Lahn M., Kanehiro A., Takeda K., Joetham A., Schwarze J., Köhler G., O'Brien R., Gelfand E.W., Born W. Negative regulation of airway responsiveness that is dependent on gammadelta T cells and independent of alphabeta T cells // Nat. Med. – 1999. – Vol. 5. – P. 1150-1156.

37. Lambert C., Genin C. CD3 bright lymphocyte population reveal gammadelta T cells // Cytometry B Clin. Cytom. – 2004. – Vol. 61, N 1. – P. 45-53.

38. Lehner T., Mitchell E., Bergmeier L., Singh M., Spallek R., Cranage M., Hall G., Dennis M., Villinger F., Wang Y. The role of gammadelta T cells in generating antiviral factors and beta-chemokines in protection against mucosal simian immunodeficiency virus infection // Eur. J. Immunol. – 2000. – Vol. 30. – P. 2245-2255.

39. Lesterhuis W.J., de Vries I.J., Schuurhuis D.H., Boullart A.C., Jacobs J.F., de Boer A.J., Scharenborg N.M., Brouwer H.M., van de Rakt M.W., Fig dor C.G., Ruers T.J., Adema G.J., Punt C.J. Vaccination of colorectal cancer patients with CEA-loaded dendritic cells: antigen-specific T cell responses in DTH skin tests // Ann. Oncol. – 2006. – Vol. 17, N 6. – P. 974-980.

40. Malissen M., Gillet A., Rocha B., Trucy J., Vivier E., Boyer C., Köntgen F., Brun N., Mazza G., Spanopoulou E., Guy-Grand D., Malissen B. T cell development in mice lacking the CD3-zeta/eta gene // EMBO J. – 1993. – Vol. 12, N 11. – P. 4347-4355.

41. Mandy F.F., Nicholson J.K., McDougal J.S. Guidelines for performing single-platform absolute CD4+ T-cell determinations with CD45 gating for persons infected with human immunodeficiency virus // MMWR Recomm. Rep. – 2003. – Vol. 52, N RR-2. – P. 1-13.

42. McClanahan J., Fukushima P.I., Stetler-Stevenson M. Increased peripheral blood gamma delta T-cells in patients with lymphoid neoplasia: a diagnostic dilemma in flow cytometry // Cytometry. – 1999. – Vol. 38. – P. 280-285.

43. McMahan R.H., Slansky J.E. Mobilizing the low-avidity T cell repertoire to kill tumors // Semin. Cancer. Biol. – 2007. – Vol. 17, N 4. – P. 317-329.

44. Meth M.J., Sperber K.E. Phenotypic diversity in delayed drug hypersensitivity: an immunologic explanation // Mt. Sinai. J. Med. – 2006. – Vol. 73, N 5. – P. 769-776.

45. Milasiene V., Stratilatovas E., Norkiene V. The importance of T-lymphocyte subsets on overall survival of colorectal and gastric cancer patients // Medicina (Kaunas). – 2007. – Vol. 43, N 7. – P. 548-554.

46. Mogues T., Goodrich M.E, Ryan L., LaCourse R., North R.J. The relative importance of T cell subsets in immunity and immunopathology of airborne Mycobacterium tuberculosis infection in mice // J. Exp. Med. – 2001. – Vol. 193. – P. 271-280.

47. Moore T.A., Moore B.B., Newstead M.W., Standiford T.J. Gamma delta-T cells are critical for survival and early proinflammatory cytokine gene expression during murine Klebsiella pneumonia // J. Immunol. – 2000. – Vol. 165. – P. 2643-2650.

48. Morita C.T., Beckman E.M., Bukowski J.F., Tanaka Y., Band H., Bloom B.R., Golan D.E., Brenner M.B. Direct presentation of nonpeptide prenyl pyrophosphate antigens to human γδ-T cells // Immunity. – 1995. – Vol. 3. – P. 495-507.

49. Morita C.T., Lee H.K., Leslie D.S., Tanaka Y., Bukowski J.F., Märker-Hermann E. Recognition of nonpeptide prenyl pyrophosphate antigens by human γδ-T cells // Microbes. Infect. – 1999. – Vol. 1. – P. 175-186.

50. Morita C.T., Mariuzza R.A., Brenner M.B. Antigen recognition by human γδ-T cells: pattern recognition by the adaptive immune system // Springer. Semin. Immunopathol. – 2000. – Vol. 22. – P. 191-217.

51. Morita C.T., Tanaka Y., Bloom B.R., Brenner M.B. Direct presentation of non-peptide prenyl pyrophosphate antigens to human γδ-T cells // Res. Immunol. – 1996. – Vol. 147. – P. 347-353.

52. Paz Morante M., Briones J., Canto E., Sabzevari H., Martino R., Sierra J., Rodriguez-Sanchez J.L., Vidal S. Activation-associated phenotype of CD3 T cells in acute graft-versus-host disease // Clin. Exp. Immunol. – 2006. – Vol. 145, N 1. – P. 36-43.

53. Pellegrin J.L., Taupin J.L., Dupon M., Ragnaud J.M., Maugein J., Bonneville M., Moreau J.F. Gammadelta T cells increase with Mycobacterium avium complex infection but not with tuberculosis in AIDS patients // Int. Immunol. – 1999. – Vol. 11. – P. 1475-1478.

54. Ramos A., López-Hoyos M., Labrador M., González M., Rodríguez-Sánchez J.L., Merino J. Host H-2 haplotype modulates the induction of host-versus-graft disease after the induction of neonatal tolerance to H-2 alloantigens // Int. J. Mol. Med. – 1998. – Vol. 1, N 2. – P. 431-437.

55. Reimann K.A., O’Gorman M.R., Spritzler J., Wilkening C.L., Sabath D.E., Helm K., Campbell D.E. Multisite comparison of CD4 and CD8 T-lymphocyte counting by single-versus multiple-platform methodologies: evaluation of Beckman Coulter flow-count fluorospheres and the tetraONE system. The NIAID DAIDS New Technologies Evaluation Group // Clin. Diagn. Lab. Immunol. – 2000. – Vol. 7. – P. 344-351.

56. Robak E., Blonski J.Z., Bartkowiak J., Niewiadomska H., Sysa-Jedrzejowska A., Robak T. Circulating TCR gammadelta cells in the patients with systemic lupus erythematosus // Mediators. Inflamm. – 1999. – Vol. 8. – P. 305-312.

57. Schramm C.M., Puddington L., Yiamouyiannis C.A., Lingenheld E.G., Whiteley H.E., Woly niec W.W., Noonan T.C., Thrall R.S. Proinflammatory roles of T-cell receptor (TCR) gammadelta and TCRalphabeta lymphocytes in a murine model of asthma // Am. J. Respir. Cell. Mol. Biol. – 2000. – Vol. 22. – P. 218-225.

58. Selin L.K., Santolucito P.A., Pinto A.K., Szomolanyi-Tsuda E., Welsh R.M. Innate immunity to viruses: control of vaccinia virus infection by gamma delta T cells // J. Immunol. – 2001. – Vol. 166. – P. 6784-6794.

59. Sindhu S.T., Ahmad R., Morisset R., Ahmad A., Menezes J. Peripheral blood cytotoxic gammadelta T lymphocytes from patients with human immunodeficiency virus type 1 infection and AIDS lyse uninfected CD4+ T cells, and their cytocidal potential correlates with viral load // J. Virol. – 2003. – Vol. 77. – P. 1848-1855.

60. Sun C.F., Hsieh Y.Y., Ngan K.W., Wang W.T. Search for immunomodulatory effects of blood transfusion in gastric cancer patients: flow cytometry of Th1/Th2 cells in peripheral blood // Ann. Clin. Lab. Sci. – 2001. – Vol. 31, N 2. – P. 171-178.

61. Trejdosiewicz L.K., Smart C.J., Oakes D.J., Howdle P.D., Malizia G., Campana D., Boylston A.W. Expression of T-cell receptors TcR1 (gamma/delta) and TcR2 (alpha/beta) in the human intestinal mucosa // Immunology. – 1989. – Vol. 68, N 1. – P. 7-12.

62. Van den Beemd R., Boor P.P., van Lochem E.G., Hop W.C., Langerak A.W., Wolvers-Tettero I.L., Hooijkaas H., van Dongen J.J. Flow cytometric analysis of the Vbeta repertoire in healthy controls // Cytometry. – 2000. – Vol. 40, N 4. – P. 336-345.

63. Wada T., Kurokawa T., Toma T., Shibata F., Tone Y., Hashida Y., Kaya H., Yoshida T., Yachie A. Immunophenotypic analysis of Epstein–Barr virus (EBV)-infected CD8(+) T cells in a patient with EBV-associated hemophagocytic lymphohistiocytosis // Eur. J. Haematol. – 2007. – Vol. 79, N 1. – P. 72-75.

64. Yamashita N., Kaneoka H., Kaneko S., Takeno M., Oneda K., Koizumi H., Kogure M., Inaba G., Sakane T. Role of gammadelta T lymphocytes in the development of Beh et's disease // Clin. Exp. Immunol. – 1997. – Vol. 107, N 2. – P. 241-247.

65. Yin Z., Craft J. Gamma delta T cells in autoimmunity // Springer Semin. Immunopathol. – 2000. – Vol. 22. – P. 311-320.


Review

For citations:


Khaidukov S.V., Zurochka A.V., Chereshnev V.A. MULTI-COLOUR CYTOMETRIC ANALYSIS. IDENTIFICATION OF T LYMPHOCYTES AND THEIR SUBSETS. Medical Immunology (Russia). 2008;10(2-3):115-124. (In Russ.) https://doi.org/10.15789/1563-0625-2008-2-3-115-124

Views: 1014


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


ISSN 1563-0625 (Print)
ISSN 2313-741X (Online)